Metalaxyl Degradation by Mucorales Strains Gongronella sp. and Rhizopus oryzae.

Science.gov (United States)

Martins, Maria Rosário; Santos, Cledir; Pereira, Pablo; Cruz-Morais, Júlio; Lima, Nelson

2017-12-14

In this study, the degradation of metalaxyl was investigated in the presence of two Mucorales strains, previously isolated from soil subjected to repeated treatments with this fungicide and selected after enrichment technique. Fungal strains were characterised by a polyphasic approach using phylogenetic analysis of the Internal Transcribed Spacer (ITS) gene region, phenotypic characterisation by Matrix-Assisted Laser Desorption Ionization Time-Of-Flight Mass Spectrometry (MALDI-TOF MS) spectral analysis, and growth kinetics experiments. The strains were identified as Gongronella sp. and Rhizopus oryzae . The fungal growth kinetics in liquid cultures containing metalaxyl fits with Haldane model. Under laboratory conditions, the ability of Gongronella sp. and R. oryzae cultures to degrade metalaxyl was evaluated in liquid cultures and soil experiments. Both species were able to: (a) use metalaxyl as the main carbon and energy source; and (b) degrade metalaxyl in polluted soils, with rates around 1.0 mg kg - ¹ d - ¹. This suggests these strains could degrade metalaxyl in soils contaminated with this fungicide.

Metalaxyl Degradation by Mucorales Strains Gongronella sp. and Rhizopus oryzae

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Maria Rosário Martins

2017-12-01

Full Text Available In this study, the degradation of metalaxyl was investigated in the presence of two Mucorales strains, previously isolated from soil subjected to repeated treatments with this fungicide and selected after enrichment technique. Fungal strains were characterised by a polyphasic approach using phylogenetic analysis of the Internal Transcribed Spacer (ITS gene region, phenotypic characterisation by Matrix-Assisted Laser Desorption Ionization Time-Of-Flight Mass Spectrometry (MALDI-TOF MS spectral analysis, and growth kinetics experiments. The strains were identified as Gongronella sp. and Rhizopus oryzae. The fungal growth kinetics in liquid cultures containing metalaxyl fits with Haldane model. Under laboratory conditions, the ability of Gongronella sp. and R. oryzae cultures to degrade metalaxyl was evaluated in liquid cultures and soil experiments. Both species were able to: (a use metalaxyl as the main carbon and energy source; and (b degrade metalaxyl in polluted soils, with rates around 1.0 mg kg−1 d−1. This suggests these strains could degrade metalaxyl in soils contaminated with this fungicide.

Effect of dietary supplementation with Rhizopus oryzae or Chrysonilia crassa on growth performance, blood profile, intestinal microbial population, and carcass traits in broilers exposed to heat stress

OpenAIRE

S. Sugiharto; T. Yudiarti; I. Isroli; E. Widiastuti; F. D. Putra

2017-01-01

Dietary supplementation of additives has recently been part of strategies to deal with the detrimental effects of heat stress (HS) on the performance and carcass traits in broiler chicks. This study aimed to investigate the effect of dietary supplementation with the fungi Rhizopus oryzae or Chrysonilia crassa on growth, blood profile, intestinal microbial population and carcass traits in broiler chicks subjected to HS. R. oryzae and C. crassa are filamentous fungi isolated from...

Carbohydrate-active enzymes from the zygomycete fungus Rhizopus oryzae: a highly specialized approach to carbohydrate degradation depicted at genome level

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Henrissat Bernard

2011-01-01

Full Text Available Abstract Background Rhizopus oryzae is a zygomycete filamentous fungus, well-known as a saprobe ubiquitous in soil and as a pathogenic/spoilage fungus, causing Rhizopus rot and mucomycoses. Results Carbohydrate Active enzyme (CAZy annotation of the R. oryzae identified, in contrast to other filamentous fungi, a low number of glycoside hydrolases (GHs and a high number of glycosyl transferases (GTs and carbohydrate esterases (CEs. A detailed analysis of CAZy families, supported by growth data, demonstrates highly specialized plant and fungal cell wall degrading abilities distinct from ascomycetes and basidiomycetes. The specific genomic and growth features for degradation of easily digestible plant cell wall mono- and polysaccharides (starch, galactomannan, unbranched pectin, hexose sugars, chitin, chitosan, β-1,3-glucan and fungal cell wall fractions suggest specific adaptations of R. oryzae to its environment. Conclusions CAZy analyses of the genome of the zygomycete fungus R. oryzae and comparison to ascomycetes and basidiomycete species revealed how evolution has shaped its genetic content with respect to carbohydrate degradation, after divergence from the Ascomycota and Basidiomycota.

Rhizopus oryzae - Ancient microbial resource with importance in modern food industry.

Science.gov (United States)

Londoño-Hernández, Liliana; Ramírez-Toro, Cristina; Ruiz, Héctor A; Ascacio-Valdés, Juan A; Aguilar-Gonzalez, Miguel A; Rodríguez-Herrera, Raúl; Aguilar, Cristóbal N

2017-09-18

Filamentous fungi are microorganisms widely known for their diverse biochemical features. Fungi can efficiently invade a wide variety of substrates under operational conditions producing numerous bioproducts of interest, such as enzymes, organic acids, aromatic compounds and colorants. An additional interesting characteristic of some fungi is their safety classification for different uses, which guarantees that the bioproducts obtained from them do not contain any toxic component deleterious to humans. Rhizopus oryzae is among this group of fungi and is classified as a GRAS filamentous fungus, commonly used for production of some oriental traditional foods. It is mainly recognized as a good producer of lactic acid; however, its potential for other biotechnological processes is under study. This review analyzes and discusses the current scientific and technical contributions which may maximize the potential of R. oryzae as a producer of different compounds of industrial interest. Copyright © 2017 Elsevier B.V. All rights reserved.

Inhibition of lipases from Chromobacterium viscosum and Rhizopus oryzae by tetrahydrolipstatin.

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Potthoff, A P; Haalck, L; Spener, F

1998-01-15

Tetrahydrolipstatin is known as an inhibitor for pancreatic lipase but not for microbial lipases. In this paper we demonstrate that in the presence of water-insoluble substrates like tributyrin or olive oil, tetrahydrolipstatin inhibits the lipases of Chromobacterium viscosum and Rhizopus oryzae, although with different potency. In contrast to porcine pancreatic lipase, which forms an irreversible and covalent enzyme-inhibitor complex with tetrahydrolipstatin, the inhibition of the microbial lipases is reversible as the inhibitor can be removed from the enzyme-inhibitor complex by solvent extraction. Moreover, after inhibition of Chromobacterium viscosum lipase tetrahydrolipstatin remains chemically unchanged.

Damage to Aspergillus fumigatus and Rhizopus oryzae Hyphae by Oxidative and Nonoxidative Microbicidal Products of Human Neutrophils In Vitro

OpenAIRE

Diamond, Richard D.; Clark, Robert A.

1982-01-01

Our previous studies established that human neutrophils could damage and probably kill hyphae of Aspergillus fumigatus and Rhizopus oryzae in vitro, primarily by oxygen-dependent mechanisms active at the cell surface. These studies were extended, again quantitating hyphal damage by reduction in uptake of 14C-labeled uracil or glutamine. Neither A. fumigatus nor R. oryzae hyphae were damaged by neutrophils from patients with chronic granulomatous disease, confirming the importance of oxidative...

Enhanced L-(+)-lactic acid production by an adapted strain of Rhizopus oryzae using corncob hydrolysate

DEFF Research Database (Denmark)

Bai, Dongmei; Li, S.Z.; Liu, Z.L.

2008-01-01

-added production of a variety of bioproducts. Lactic acid can be used as a precursor for poly-lactic acid production. Although current industrial lactic acid is produced by lactic acid bacteria using enriched medium, production by Rhizopus oryzae is preferred due to its exclusive formation of the......-isomer and a simple nutrition requirement by the fungus. Production of-L-(+)-lactic acid by R. oryzae using xylose has been reported; however, its yield and conversion rate are poor compared with that of using glucose. In this study, we report an adapted R. oryzae strain HZS6 that significantly improved efficiency...... of substrate utilization and enhanced production of L-(+)-lactic acid from corncob hydrolysate. It increased L-(+)-lactic acid final concentration, yield, and volumetric productivity more than twofold compared with its parental strain. The optimized growth and fermentation conditions for Strain HZS6 were...

High-level expression and characterization of a chimeric lipase from Rhizopus oryzae for biodiesel production.

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Yu, Xiao-Wei; Sha, Chong; Guo, Yong-Liang; Xiao, Rong; Xu, Yan

2013-02-21

Production of biodiesel from non-edible oils is receiving increasing attention. Tung oil, called "China wood oil" is one kind of promising non-edible biodiesel oil in China. To our knowledge, tung oil has not been used to produce biodiesel by enzymatic method. The enzymatic production of biodiesel has been investigated extensively by using Rhizopus oryzae lipase as catalyst. However, the high cost of R. oryzae lipase remains a barrier for its industrial applications. Through different heterologous expression strategies and fermentation techniques, the highest expression level of the lipase from R. oryzae reached 1334 U/mL in Pichia pastoris, which is still not optimistic for industry applications. The prosequence of lipases from Rhizopus sp. is very important for the folding and secretion of an active lipase. A chimeric lipase from R. oryzae was constructed by replacing the prosequence with that from the R. chinensis lipase and expressed in P. pastoris. The maximum activity of the chimera reached 4050 U/mL, which was 11 fold higher than that of the parent. The properties of the chimera were studied. The immobilized chimera was used successfully for biodiesel production from tung oil, which achieved higher FAME yield compared with the free chimeric lipase, non-chimeric lipase and mature lipase. By response surface methodology, three variables, water content, methanol to tung oil molar ratio and enzyme dosage were proved to be crucial parameters for biosynthesis of FAME and the FAME yield reached 91.9±2.5% at the optimized conditions by adding 5.66 wt.% of the initial water based on oil weight, 3.88 of methanol to tung oil molar ratio and 13.24 wt.% of enzyme concentration based on oil weight at 40°C. This is the first report on improving the expression level of the lipase from R. oryzae by replacing prosequences. The immobilized chimera was used successfully for biodiesel production from tung oil. Using tung oil as non-edible raw material and a chimeric lipase

Fob1 and Fob2 Proteins Are Virulence Determinants of Rhizopus oryzae via Facilitating Iron Uptake from Ferrioxamine.

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Mingfu Liu

2015-05-01

Full Text Available Dialysis patients with chronic renal failure receiving deferoxamine for treating iron overload are uniquely predisposed for mucormycosis, which is most often caused by Rhizopus oryzae. Although the deferoxamine siderophore is not secreted by Mucorales, previous studies established that Rhizopus species utilize iron from ferrioxamine (iron-rich form of deferoxamine. Here we determined that the CBS domain proteins of Fob1 and Fob2 act as receptors on the cell surface of R. oryzae during iron uptake from ferrioxamine. Fob1 and Fob2 cell surface expression was induced in the presence of ferrioxamine and bound radiolabeled ferrioxamine. A R. oryzae strain with targeted reduced Fob1/Fob2 expression was impaired for iron uptake, germinating, and growing on medium with ferrioxamine as the sole source of iron. This strain also exhibited reduced virulence in a deferoxamine-treated, but not the diabetic ketoacidotic (DKA, mouse model of mucormycosis. The mechanism by which R. oryzae obtains iron from ferrioxamine involves the reductase/permease uptake system since the growth on ferrioxamine supplemented medium is associated with elevated reductase activity and the use of the ferrous chelator bathophenanthroline disulfonate abrogates iron uptake and growth on medium supplemented with ferrioxamine as a sole source of iron. Finally, R. oryzae mutants with reduced copies of the high affinity iron permease (FTR1 or with decreased FTR1 expression had an impaired iron uptake from ferrioxamine in vitro and reduced virulence in the deferoxamine-treated mouse model of mucormycosis. These two receptors appear to be conserved in Mucorales, and can be the subject of future novel therapy to maintain the use of deferoxamine for treating iron-overload.

Development of a system for integrative and stable transformation of the zygomycete Rhizopus oryzae by Agrobacterium-mediated DNA transfer

NARCIS (Netherlands)

Michielse, C.B.; Salim, K.; Ragas, P.; Ram, A.F.J.; Kudla, B.; Jarry, B.; Punt, P.J.; Hondel, C.A.M.J.J. van den

2004-01-01

Two transformation systems, based on the use of CaCl2/PEG and Agrobacterium tumefaciens, respectively, were developed for the zygomycete Rhizopus oryzae. Irrespective of the selection marker used, a pyr4 marker derived from R. niveus or a dominant amdS+ marker from Aspergillus nidulans, and

Influence of rice straw-derived dissolved organic matter on lactic acid fermentation by Rhizopus oryzae.

Science.gov (United States)

Chen, Xingxuan; Wang, Xiahui; Xue, Yiyun; Zhang, Tian-Ao; Li, Yuhao; Hu, Jiajun; Tsang, Yiu Fai; Zhang, Hongsheng; Gao, Min-Tian

2018-01-31

Rice straw can be used as carbon sources for lactic acid fermentation. However, only a small amount of lactic acid is produced even though Rhizopus oryzae can consume glucose in rice straw-derived hydrolysates. This study correlated the inhibitory effect of rice straw with rice straw-derived dissolved organic matter (DOM). Lactic acid fermentations with and without DOM were conducted to investigate the effect of DOM on lactic acid fermentation by R. oryzae. Fermentation using control medium with DOM showed a similar trend to fermentation with rice straw-derived hydrolysates, showing that DOM contained the major inhibitor of rice straw. DOM assay indicated that it mainly consisted of polyphenols and polysaccharides. The addition of polyphenols and polysaccharides derived from rice straw confirmed that lactic acid fermentation was promoted by polysaccharides and significantly inhibited by polyphenols. The removal of polyphenols also improved lactic acid production. However, the loss of polysaccharides during the removal of polyphenols resulted in low glucose consumption. This study is the first to investigate the effects of rice straw-derived DOM on lactic acid fermentation by R. oryzae. The results may provide a theoretical basis for identifying inhibitors and promoters associated with lactic acid fermentation and for establishing suitable pretreatment methods. Copyright © 2018 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Cosmeceutical potentials and bioactive compounds of rice bran fermented with single and mix culture of Aspergillus oryzae and Rhizopus oryzae

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Dang Lelamurni Abd Razak

2017-04-01

Full Text Available In the present study, rice bran, one of the most abundant agricultural by-products in Malaysia, was fermented with single and mixed cultures of Aspergillus oryzae and Rhizopus oryzae. The fermented rice bran extracts were tested for their functional properties and compared to the non-fermented counterparts. Antioxidant activities as well as phenolics and organic acid contents were evaluated. Skincare-related functionalities were also tested by evaluating tyrosinase and elastase inhibition activities. Tyrosinase inhibition activity, measured to determine the anti-pigmentation effect of extracts, was found to be the highest in the extract of rice bran fermented with A. oryzae (56.18% compared to other extracts. In determining the anti-aging effect of fermented rice bran extracts, the same extract showed the highest elastase inhibition activity with a value of 60.52%. Antioxidant activities were found to be highest in the mix-cultured rice bran extract. The results of phenolic and organic acid content were varied; the major phenolic acid detected was ferulic acid with a value of 43.19 μg/ml in the mix-cultured rice bran extract. On the other hand, citric acid was the major organic acid detected, with the highest content found in the same extract (214.6 mg/g. The results of this study suggest that the fermented rice bran extracts may have the potential to be further exploited as ingredients in cosmetics as well as in antioxidant-rich products.

Antifungal Activity of Colistin against Mucorales Species In Vitro and in a Murine Model of Rhizopus oryzae Pulmonary Infection▿

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Ben-Ami, Ronen; Lewis, Russell E.; Tarrand, Jeffrey; Leventakos, Konstantinos; Kontoyiannis, Dimitrios P.

2010-01-01

In immunosuppressed hosts, mucormycosis is a life-threatening infection with few treatment options. We studied the activity of colistin (polymyxin E) against Mucorales species in vitro and in a murine model of pulmonary Rhizopus oryzae infection. Colistin exhibited fungicidal activity in vitro against Mucorales spores and mycelia. At the colistin MIC, initial R. oryzae hyphal damage was followed by rapid regrowth; however, regrowth was prevented by combining colistin with a subinhibitory concentration of amphotericin B. Using electron microscopy and FM4-64 staining, we demonstrated that colistin disrupts R. oryzae cytoplasmic and vacuolar membranes, resulting in the leakage of intracellular contents. The prophylactic intranasal treatment of immunosuppressed mice with colistimethate significantly reduced the mortality rate and pulmonary fungal burden resulting from inhalational challenge with R. oryzae spores, whereas intraperitoneal colistimethate treatment had no effect. We conclude that colistin has modest in vitro and in vivo fungicidal activity against Mucorales spp. Further studies are warranted to assess the use of this drug in the prevention and treatment of mucormycosis. PMID:19858263

Antifungal activity of colistin against mucorales species in vitro and in a murine model of Rhizopus oryzae pulmonary infection.

Science.gov (United States)

Ben-Ami, Ronen; Lewis, Russell E; Tarrand, Jeffrey; Leventakos, Konstantinos; Kontoyiannis, Dimitrios P

2010-01-01

In immunosuppressed hosts, mucormycosis is a life-threatening infection with few treatment options. We studied the activity of colistin (polymyxin E) against Mucorales species in vitro and in a murine model of pulmonary Rhizopus oryzae infection. Colistin exhibited fungicidal activity in vitro against Mucorales spores and mycelia. At the colistin MIC, initial R. oryzae hyphal damage was followed by rapid regrowth; however, regrowth was prevented by combining colistin with a subinhibitory concentration of amphotericin B. Using electron microscopy and FM4-64 staining, we demonstrated that colistin disrupts R. oryzae cytoplasmic and vacuolar membranes, resulting in the leakage of intracellular contents. The prophylactic intranasal treatment of immunosuppressed mice with colistimethate significantly reduced the mortality rate and pulmonary fungal burden resulting from inhalational challenge with R. oryzae spores, whereas intraperitoneal colistimethate treatment had no effect. We conclude that colistin has modest in vitro and in vivo fungicidal activity against Mucorales spp. Further studies are warranted to assess the use of this drug in the prevention and treatment of mucormycosis.

Effect of Inoculum Dosage Aspergillus niger and Rhizopus oryzae mixture with Fermentation Time of Oil Seed Cake (Jatropha curcas L) to the content of Protein and Crude Fiber

Science.gov (United States)

Kurniati, T.; Nurlaila, L.; Iim

2017-04-01

Jatropha curcas L already widely cultivated for its seeds pressed oil used as an alternative fuel. This plant productivity per hectare obtained 2.5-5 tonnes of oil/ha / year and jatropha seed cake from 5.5 to 9.5 tonnes/ha/year, nutrient content of Jatropha curcas seed L potential to be used as feed material, However, the constraints faced was the low crude protein and high crude protein. The purpose of the research was to determine the dosage of inoculum and fermentation time of Jatropha seed cake by a mixture of Aspergillus niger and Rhizopus oryzae on crude protein and crude fibre. The study was conducted by an experimental method using a Completely Randomised Design (CRD) factorial design (3×3). The treatment consisted of a mixture of three dosage levels of Aspergillus niger and Rhizopus oryzae (= 0.2% d1, d2 and d3 = 0.3% = 0.4%) and three levels of fermentation time (w1 = 72 hours, 96 hours and w2 = w3 = 120 hours) each repeated three times. The parameters measured were crude protein and crude fibre. The results showed that dosages of 0.3% (Aspergillus niger Rhizopus oryzae 0.15% and 0.15%) and 72 hours (d2w1) is the dosage and the optimal time to generate the highest crude protein content of 21.11% and crude fibre amounted to 21.36%.

Pb(II) and Cd(II) removal from aqueous solution, shipyard wastewater, and landfill leachate by modified Rhizopus oryzae biomass

Science.gov (United States)

Naeimi, Behrouz; Foroutan, Rauf; Ahmadi, Bahram; Sadeghzadeh, Farzaneh; Ramavandi, Bahman

2018-04-01

This study was designed to remove Pb(II) and Cd(II) from aqueous solution, shipyard wastewater, and sanitary landfill leachate using an alkaline-modified Rhizopus oryzae biomass. According to the Fourier transform infrared test, different functional groups like O–H, N–H, C=O, and P–O were detected in the bioadsorbent. The x-ray fluorescence (XRF) analysis showed that CaO, P2O3, and SO3 oxides have the highest content in the bioadsorbent. The surface area of modified Rhizopus oryzae was obtained as 20.32 m2 g‑1. The effect of initial pH, temperature, contact time, and bioadsorbent dose on the metals removal was discussed. At optimal conditions, maximum Pb(II) and Cd(II) removal was obtained 95.66% and 94.55%, respectively. Freundlich model was well- accurately described the equilibrium data. Among four studied models, the pseudo-second-order was better able to describe the kinetic behavior of the bioadsorption process. The amount of enthalpy, free energy of Gibbs, and entropy parameters indicated that the bioadsorption process of studied heavy metals is negative, exothermic, and spontaneous. The amount of heavy metals in a shipyard wastewater and sanitary landfill leachate was significantly decreased by using the developed bioadsorbent.

Rhizopus oryzae hyphae are damaged by human natural killer (NK) cells, but suppress NK cell mediated immunity.

Science.gov (United States)

Schmidt, Stanislaw; Tramsen, Lars; Perkhofer, Susanne; Lass-Flörl, Cornelia; Hanisch, Mitra; Röger, Frauke; Klingebiel, Thomas; Koehl, Ulrike; Lehrnbecher, Thomas

2013-07-01

Mucormycosis has a high mortality and is increasingly diagnosed in hematopoietic stem cell transplant (HSCT) recipients. In this setting, there is a growing interest to restore host defense to combat infections by adoptively transferring donor-derived immunocompetent cells. Natural killer (NK) cells exhibit antitumor and antiinfective activity, but the interaction with Mucormycetes is unknown. Our data demonstrate that both unstimulated and IL-2 prestimulated human NK cells damage Rhizopus oryzae hyphae, but do not affect resting conidia. The damage of the fungus is mediated, at least in part, by perforin. R. oryzae hyphae decrease the secretion of immunoregulatory molecules by NK cells, such as IFN-γ and RANTES, indicating an immunosuppressive effect of the fungus. Our data indicate that NK cells exhibit activity against Mucormycetes and future research should evaluate NK cells as a potential tool for adoptive immunotherapy in HSCT. Copyright © 2012 Elsevier GmbH. All rights reserved.

Structure, function, and phylogeny of the mating locus in the Rhizopus oryzae complex.

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Andrii P Gryganskyi

2010-12-01

Full Text Available The Rhizopus oryzae species complex is a group of zygomycete fungi that are common, cosmopolitan saprotrophs. Some strains are used beneficially for production of Asian fermented foods but they can also act as opportunistic human pathogens. Although R. oryzae reportedly has a heterothallic (+/- mating system, most strains have not been observed to undergo sexual reproduction and the genetic structure of its mating locus has not been characterized. Here we report on the mating behavior and genetic structure of the mating locus for 54 isolates of the R. oryzae complex. All 54 strains have a mating locus similar in overall organization to Phycomyces blakesleeanus and Mucor circinelloides (Mucoromycotina, Zygomycota. In all of these fungi, the minus (- allele features the SexM high mobility group (HMG gene flanked by an RNA helicase gene and a TP transporter gene (TPT. Within the R. oryzae complex, the plus (+ mating allele includes an inserted region that codes for a BTB/POZ domain gene and the SexP HMG gene. Phylogenetic analyses of multiple genes, including the mating loci (HMG, TPT, RNA helicase, ITS1-5.8S-ITS2 rDNA, RPB2, and LDH genes, identified two distinct groups of strains. These correspond to previously described sibling species R. oryzae sensu stricto and R. delemar. Within each species, discordant gene phylogenies among multiple loci suggest an outcrossing population structure. The hypothesis of random-mating is also supported by a 50:50 ratio of plus and minus mating types in both cryptic species. When crossed with tester strains of the opposite mating type, most isolates of R. delemar failed to produce zygospores, while isolates of R. oryzae produced sterile zygospores. In spite of the reluctance of most strains to mate in vitro, the conserved sex locus structure and evidence for outcrossing suggest that a normal sexual cycle occurs in both species.

Role of N-terminal 28-amino-acid region of Rhizopus oryzae lipase in directing proteins to secretory pathway of Aspergillus oryzae.

Science.gov (United States)

Hama, Shinji; Tamalampudi, Sriappareddy; Shindo, Naoki; Numata, Takao; Yamaji, Hideki; Fukuda, Hideki; Kondo, Akihiko

2008-07-01

To develop a new approach for improving heterologous protein production in Aspergillus oryzae, we focused on the functional role of the N-terminal region of Rhizopus oryzae lipase (ROL). Several N-terminal deletion variants of ROL were expressed in A. oryzae. Interestingly, a segment of 28 amino acids from the C-terminal region of the propeptide (N28) was found to be critical for secretion of ROL into the culture medium. To further investigate the role of N28, the ROL secretory process was visualized in vivo using ROL-green fluorescent protein (GFP) fusion proteins. In cells producing ROL with N28, fluorescence observations showed that the fusion proteins are transported through endoplasmic reticulum (ER), Golgi, and cell wall, which is one of the typical secretory processes in a eukaryotic cell. Because the expression of the mature ROL-GFP fusion protein induced fluorescence accumulation without its translocation into the ER, N28 is considered to play a crucial role in protein transport. When N28 was inserted between the secretion signal and GFP, fluorescence observations showed that GFP, which is originally a cytoplasmic protein, was efficiently translocated into the ER of A. oryzae, resulting in an enhanced secretion of mature GFP after proteolytic cleavage of N28. These findings suggest that N28 facilitates protein translocation into ER and can be a promising candidate for improving heterologous protein production in A. oryzae.

Mitochondrial respiratory pathways inhibition in Rhizopus oryzae potentiates activity of posaconazole and itraconazole via apoptosis.

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Fazal Shirazi

Full Text Available The incidence of mucormycosis has increased drastically in immunocompromised patients. Also the array of targets whose inhibition results in Mucorales death is limited. Recently, researchers identified mitochondria as important regulators of detoxification and virulence mechanisms in fungi. In this context, targeting the mitochondrial respiratory chain may provide a new platform for antifungal development. We hypothesized that targeting respiratory pathways potentiates triazoles activity via apoptosis. We found that simultaneous administration of antimycin A (AA and benzohydroxamate (BHAM, inhibitors of classical and alternative mitochondrial pathways respectively, resulted in potent activity of posaconazole (PCZ and itraconazole (ICZ against Rhizopus oryzae. We observed cellular changes characteristic of apoptosis in R. oryzae cells treated with PCZ or ICZ in combination with AA and BHAM. The fungicidal activity of this combination against R. oryzae was correlated with intracellular reactive oxygen species accumulation (ROS, phosphatidylserine externalization, mitochondrial membrane depolarization, and increased caspase like activity. DNA fragmentation and condensation assays also revealed apoptosis of R. oryzae cells. These apoptotic features were prevented by the addition of the ROS scavenger N-acetyl-cysteine. Taken together, these findings suggest that the use of PCZ or ICZ in combination with AA and BHAM makes R. oryzae exquisitely sensitive to treatment with triazoles via apoptosis. This strategy may serve as a new model for the development of improved or novel antifungal agents.

PENGARUH RHIZOPUS ORYZAE DAN ASPERGILLUS ORYZAE TERHADAP KUALITAS KECAP

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Dewi Sabita Slamet

2012-11-01

Full Text Available Telah diteliti pengganti fermentasi mikroorganisme Aspergillus oryzae Rhyzopus oryzae dan campuran Aspergillus dan Rhyzopus oryzae, dengan perendaman dalam larutan garam 20% dalam waktu yang berbeda terhadap kualitas kecap.Lamanya perendaman dalam larutan garam 20% yang berbeda menghasilkan kadar protein kecap yang berbeda. Aspergillus oryzae lebih baik dalam menghasilkan enzima protease dari pada Rhyzopus oryzae.Uji organoleptik menunjukkan perbedaan tidak bermakna dalam hal rasa maupun aroma antar kecap yang dibuat dengan strain jamur yang berlainan serta waktu perendaman yang berbeda. Untuk membuat kecap, sebaiknya dilakukan perendaman dalam larutan garam 20% selama 14 hari.

Optimization of xylanase production by Mucor indicus, Mucor hiemalis, and Rhizopus oryzae through solid state fermentation

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Sanaz Behnam

2016-03-01

Full Text Available Introduction: Xylan is the main hemicellulosic polymer in a number of lignocelluloses which can be hydrolyzed by xylanolytic enzymes. One of the main ways for enzymes production is solid state fermentation (SSF. The ability of three fungal strains (Mucor indicus, Mucor hiemalis, and Rhizopus oryzae for xylanase production on wheat bran by SSF was investigated. Materials and methods: The effects of cultivation temperature, medium moisture content, and cultivation time on the enzyme production were investigated. Experiments were designed with an orthogonal central composite design on three variables using response surface methodology (RSM. Analysis of variance was applied and the enzyme production was expressed with a mathematical equation as a function of the three factors. The optimum operating conditions for the enzyme production was obtained. Results: For xylanase production by M. indicus, M. hiemalis and R. oryzae the optimum temperatures were 40.0, 43.4 and 43.4ºC respectively. These values were 49.8, 54.2 and 71.8% for moisture percent and 51.3, 53.2 and 53.5 h for cultivation time. The highest enzyme activities per g of dry substrate (gds were 43.1, 43.8 and 25.9 U/gds for M. indicus, M. hiemalis and R. oryzae respectively. Discussion and conclusion: All the fungi were able to produce xylanase. Maximum xylanase production was predicted by M. indicus and M. hiemalis at similar optimum conditions, while R. oryzae produced relatively lower xylanase activity even at the best condition. 

Biosorption of Cr(VI) in Aqueous Solution using Microorganisms: Comparison of the Use of Rhizopus oryzae, Bacillus firmus, and Trichoderma viride

Science.gov (United States)

Safitri, Anna; Mahardini, Putri; Prasetyawan, Sasangka; Roosdiana, Anna

2018-01-01

In this work, the study of biosorption of Cr(VI) from aqueous solution was conducted using Rhizopus oryzae, Bacillus firmus, and Trichoderma viride as microorganisms that can absorb Cr(VI). The research is focused on determination of optimum conditions including pH, the number of R. oryzae, B. firmus, and T. viride (inoculums), and initial concentrations of Cr(VI) used. Optimum pH was obtained at pH 5, 4.5 and 6, for biosorption of Cr(VI) with R. oryzae, B. firmus, and T. viride, respectively, in the capacity of 45.3%, 24.5%, and 90.3%. The highest amount of Cr(VI) adsorbed for biosorption with R. oryzae, B. firmus, and T. viride, were 55.4%, 18.5%, and 74.5%, respectively, using 6-mL inoculums. The equilibrium concentrations achieved for R. oryzae, B. firmus, and T. viride were 60 mg/mL, 40 mg/mL, and 40 mg/mL, with the amount of Cr(VI) adsorbed were 32.4%, 28.2%, and 89.3%, respectively. The adsorption capacity for R. oryzae, B. firmus, and T. viride were 45.3 mg/1×106 colonies, 36.2 mg/1×106 cells, and 77.8 mg/1×106 colonies, respectively. Overall, the biosorbents effectivity order in the biosorption process of Cr(VI) are T. viride > R. oryzae > B. firmus.

Protein concentrations of sweet soysauces from Rhizopus oryzae and R. oligosporus fermentation without moromi fermentation

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NOOR SOESANTI HANDAJANI

2007-07-01

Full Text Available Soy sauce was produce from soybean that fermented with koji/tempeh fungi and thenfermented under salt solution or moromi fermentation. The objectives of this experiment was to compare of protein (total and soluble content of sweet soy sauce that produced from soybean fermented with Rhizopus oryzae and R. oligosporus without moromi fermentation to the sweet soysauce with moromi fermentation one. The total and soluble proteins of sweet soy sauces that produce from soybean without moromi fermentation were higher that sweet soy sauces that produce with moromi fermentation. Soluble protein of sweet soy sauce that produced from soybean fermented with R. oligosporus without moromi fermentation was 8.2% and meet to the highest quality of sweet soy sweet sauce based on Indonesia Industrial Standard. Soluble protein of sweet soy sauce that produced from soybean fermented with R. oryzae without moromi fermentation was 4.1% and meet to the medium quality of sweet soy sweet sauce based on Indonesia Industrial Standard.

Enhancement of L(+)-Lactic Acid Production of Immobilized Rhizopus Oryzae Implanted by Ion Beams

International Nuclear Information System (INIS)

Fan Yonghong; Yang Yingge; Zheng Zhiming; Li Wen; Wang Peng; Yao Liming; Yu Zengliang

2008-01-01

Immobilized Rhizopus oryzae culturing may be a solution to the inhibited production of L(+)-lactic acid in submerged fermentation, which is caused by aggregated mycelia floc. In the present study, a R. oryzae mutant (RL6041) with a 90% conversion rate of glucose into L-lactic acid was obtained by N + implantation under the optimized conditions of a beam energy of 15 keV and a dose of 2.6 x 10 15 ions/cm 2 . Using polyurethane foam as the immobilization matrix, the optimal L-lactic acid production conditions were determined as 4 mm polyurethane foam, 150 r/min, 50 g/L ∼ 80 g/L of initial glucose, 38 deg. C and pH 6.0. 15-cycle repeated productions of L-lactic acid by immobilized RL6041 were performed under the optimized culturing conditions and over 80% of the glucose was converted into L-lactic acid in 30 hours on average. The results show that immobilized RL6041 is a promising candidate for continuous L-lactic acid production.

In vitro antifungal susceptibility of clinical species belonging to Aspergillus genus and Rhizopus oryzae.

Science.gov (United States)

Kachuei, R; Khodavaisy, S; Rezaie, S; Sharifynia, S

2016-03-01

Among filamentous fungal pathogens, Aspergillus spp. and zygomycetes account for highest rates of morbidity and mortality among immunocompromised patients. Recently developed antifungal drugs offer the potential to improve management and therapeutic outcomes of fungal infections. The aim of this study was to analyse the in vitro activities of voriconazole, itraconazole, amphotericin B and caspofungin against clinical isolates of Aspergillus spp. and Rhizopus oryzae. The in vitro antifungal susceptibility of 54 isolates belonging to different clinical isolates of Aspergillus spp. and R. oryzae was tested for four antifungal agents using a microdilution reference method (CLSI, M38-A2). All isolates were identified by typical colony and microscopic characteristics, and also characterized by molecular methods. Caspofungin (MEC range: 0.008-0.25 and MEC50: 0.0023μg/mL) was the most active drug in vitro against Aspergillus spp., followed by voriconazole (MIC range: 0.031-8 and MIC50: 0.5μg/mL), itraconazole (MIC range: 0.031-16 and MIC50: 0.25μg/mL), and amphotericin B (MIC range: 0.125-4 and MIC50: 0.5μg/mL), in order of decreasing activity. The caspofungin, voriconazole, and itraconazole demonstrated poor in vitro activity against R. oryzae isolates evaluated, followed by amphotericin B. This study demonstrates that caspofungin had good antifungal activity and azole agents had better activity than amphotericin B against Aspergillus species. Although, azole drugs are considered ineffective against R. oryzae. This result is just from a small scale in vitro susceptibility study and we did not take other factors into consideration. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

Stimulation with lysates of Aspergillus terreus, Candida krusei and Rhizopus oryzae maximizes cross-reactivity of anti-fungal T cells.

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Deo, Shivashni S; Virassamy, Balaji; Halliday, Catriona; Clancy, Leighton; Chen, Sharon; Meyer, Wieland; Sorrell, Tania C; Gottlieb, David J

2016-01-01

Invasive fungal diseases caused by filamentous fungi and yeasts are significant causes of morbidity and mortality in immunosuppressed hematology patients. We previously published a method to expand Aspergillus fumigatus-specific T cells for clinical cell therapy. In the present study, we investigated expansion of T cells specific for other fungal pathogens and creation of a broadly reactive panfungal T-cell product. Fungal strains selected were those frequently observed in the clinical hematology setting and included Aspergillus, Candida, Fusarium, Rhizopus and Lomentospora/Scedosporium. Four T-cell cultures specific to each fungus were established. We selected lysates of Aspergillus terreus, Candida krusei and Rhizopus oryzae to expand panfungal T cells. Allelic restriction of anti-fungal activity was determined through the use of specific major histocompatibility complex class II-blocking antibodies. Individual T-cell cultures specific to each fungus could be expanded in vitro, generating predominantly CD4(+) T cells of which 8% to 20% were fungus-specific. We successfully expanded panfungal T cells from the peripheral blood (n = 8) and granulocyte-colony-stimulating factor-primed stem cell products (n = 3) of normal donors by using a combination of lysates from Aspergillus terreus, Candida krusei and Rhizopus oryzae. Anti-fungal activity was mediated through human leukocyte antigen (HLA)-DR alleles and was maintained when antigen-presenting cells from partially HLA-DRB1-matched donors were used to stimulate T cells. We demonstrate a method to manufacture panfungal T-cell products with specificity against a range of clinical fungal pathogens by use of the blood and stem cells of healthy donors as the starting material. The safety and efficacy of these products will need to be tested clinically. Copyright © 2015 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

An optimized fed-batch culture strategy integrated with a one-step fermentation improves L-lactic acid production by Rhizopus oryzae.

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Fu, Yongqian; Sun, Xiaolong; Zhu, Huayue; Jiang, Ru; Luo, Xi; Yin, Longfei

2018-05-21

In previous work, we proposed a novel modified one-step fermentation fed-batch strategy to efficiently generate L-lactic acid (L-LA) using Rhizopus oryzae. In this study, to further enhance efficiency of L-LA production through one-step fermentation in fed-batch cultures, we systematically investigated the initial peptone- and glucose-feeding approaches, including different initial peptone and glucose concentrations and maintained residual glucose levels. Based on the results of this study, culturing R. oryzae with initial peptone and glucose concentrations of 3.0 and 50.0 g/l, respectively, using a fed-batch strategy is an effective approach of producing L-LA through one-step fermentation. Changing the residual glucose had no obvious effect on the generation of L-LA. We determined the maximum LA production and productivity to be 162 g/l and 6.23 g/(l·h), respectively, during the acid production stage. Compared to our previous work, there was almost no change in L-LA production or yield; however, the productivity of L-LA increased by 14.3%.

Omics-based approaches reveal phospholipids remodeling of Rhizopus oryzae responding to furfural stress for fumaric acid-production from xylose.

Science.gov (United States)

Pan, Xinrong; Liu, Huanhuan; Liu, Jiao; Wang, Cheng; Wen, Jianping

2016-12-01

In order to relieve the toxicity of furfural on Rhizopus oryzae fermentation, the molecular mechanism of R. oryzae responding to furfural stress for fumaric acid-production was investigated by omics-based approaches. In metabolomics analysis, 29 metabolites including amino acid, sugars, polyols and fatty acids showed significant changes for maintaining the basic cell metabolism at the cost of lowering fumaric acid production. To further uncover the survival mechanism, lipidomics was carried out, revealing that phosphatidylcholine, phosphatidylglycerol, phosphatidylinositol and polyunsaturated acyl chains might be closely correlated with R. oryzae's adapting to furfural stress. Based on the above omics analysis, lecithin, inositol and soybean oil were exogenously supplemented separately with an optimized concentration in the presence of furfural, which increased fumaric acid titer from 5.78g/L to 10.03g/L, 10.05g/L and 12.13g/L (increased by 73.5%, 73.8% and 110%, respectively). These findings provide a methodological guidance for hemicellulose-fumaric acid development. Copyright © 2016 Elsevier Ltd. All rights reserved.

Kinetics Study of Extracellular Detergent Stable Alkaline Protease from Rhizopus oryzae

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Zareena Mushtaq

2015-04-01

Full Text Available In this study, extracellular alkaline protease was produced from Rhizopus oryzae in submerged fermentation using dairy waste (whey as a substrate. Fermentation kinetics was studied and various parameters were optimized. The strain produced maximum protease at initial medium pH of 6.0 medium depth of 26 mm, inoculum size of 2% at incubation temperature of 35ºC for 168 h of fermentation. Alkaline protease was purified to homogeneity by ammonium sulphate fractionation followed by sephadex G-100 chromatography. The molecular mass of alkaline protease was 69 kDa determined by 10% SDS-PAGE. The optimum pH and temperature of alkaline protease was 9.0 and 40ºC, respectively. Metal profile of the enzyme showed that the enzyme was non-metallic in nature. The Km , Kcat , Vmax and Kcat/Km values of purified protease were 7.0 mg/mL, 3.8 x102S-1, 54.30 µmol/min and 54.28 s-1mg -1.mL respectively, using casein as substrate. The purified alkaline protease had stability with commercial detergents.

Construction of the yeast whole-cell Rhizopus oryzae lipase biocatalyst with high activity.

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Chen, Mei-ling; Guo, Qin; Wang, Rui-zhi; Xu, Juan; Zhou, Chen-wei; Ruan, Hui; He, Guo-qing

2011-07-01

Surface display is effectively utilized to construct a whole-cell biocatalyst. Codon optimization has been proven to be effective in maximizing production of heterologous proteins in yeast. Here, the cDNA sequence of Rhizopus oryzae lipase (ROL) was optimized and synthesized according to the codon bias of Saccharomyces cerevisiae, and based on the Saccharomyces cerevisiae cell surface display system with α-agglutinin as an anchor, recombinant yeast displaying fully codon-optimized ROL with high activity was successfully constructed. Compared with the wild-type ROL-displaying yeast, the activity of the codon-optimized ROL yeast whole-cell biocatalyst (25 U/g dried cells) was 12.8-fold higher in a hydrolysis reaction using p-nitrophenyl palmitate (pNPP) as the substrate. To our knowledge, this was the first attempt to combine the techniques of yeast surface display and codon optimization for whole-cell biocatalyst construction. Consequently, the yeast whole-cell ROL biocatalyst was constructed with high activity. The optimum pH and temperature for the yeast whole-cell ROL biocatalyst were pH 7.0 and 40 °C. Furthermore, this whole-cell biocatalyst was applied to the hydrolysis of tributyrin and the resulted conversion of butyric acid reached 96.91% after 144 h.

Study of energetic-particle-irradiation induced biological effect on Rhizopus oryzae through synchrotron-FTIR micro-spectroscopy

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Liu, Jinghua; Qi, Zeming; Huang, Qing; Wei, Xiaoli; Ke, Zhigang; Fang, Yusheng; Tian, Yangchao; Yu, Zengliang

2013-01-01

Energetic particles exist ubiquitously and cause varied biological effects such as DNA strand breaks, lipid peroxidation, protein modification, cell apoptosis or death. An emerging biotechnology based on ion-beam technique has been developed to serve as an effective tool for mutation breeding of crops and microbes. In order to improve the effectiveness of ion-beam biotechnology for mutation breeding, it is indispensible to gain a better understanding of the mechanism of the interactions between the energetic ions and biological systems which is still elusive. A new trend is to conduct more comprehensive research which is based on micro-scaled observation of the changes of the cellular structures and compositions under the interactions. For this purpose, advanced synchrotron FTIR (s-FTIR) microscopy was employed to monitor the cellular changes of single fungal hyphae under irradiation of α-particles from 241Am. Intracellular contents of ROS, MDA, GSSG/GSH and activities of CAT and SOD were measured via biochemical assay. Ion-irradiation on Rhizopus oryzae causes localized vacuolation, autolysis of cell wall and membrane, lipid peroxidation, DNA damage and conformational changes of proteins, which have been clearly revealed by the s-FTIR microspectroscopy. The different changes of cell viability, SOD and CAT activities can be explained by the ROS-involved chemical reactions. Evidently, the elevated level of ROS in hyphal cells upon irradiation plays the key role in the caused biological effect. This study demonstrates that s-FTIR microspectroscopy is an effective tool to study the damage of fungal hyphae caused by ionizing radiation and it facilitates the exploit of the mechanism for the interactions between the energetic ions and biological systems.

Biodiesel production from Nannochloropsis gaditana lipids through transesterification catalyzed by Rhizopus oryzae lipase.

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Navarro López, Elvira; Robles Medina, Alfonso; González Moreno, Pedro Antonio; Esteban Cerdán, Luis; Martín Valverde, Lorena; Molina Grima, Emilio

2016-03-01

Biodiesel (fatty acid methyl esters, FAMEs) was produced from saponifiable lipids (SLs) extracted from wet Nannochloropsis gaditana biomass using methanolysis catalyzed by Rhizopus oryzae intracellular lipase. SLs were firstly extracted with ethanol to obtain 31 wt% pure SLs. But this low SL purity also gave a low biodiesel conversion (58%). This conversion increased up to 80% using SLs purified by crystallization in acetone (95 wt% purity). Polar lipids play an important role in decreasing the reaction velocity - using SLs extracted with hexane, which have lower polar lipid content (37.4% versus 49.0% using ethanol), we obtained higher reaction velocities and less FAME conversion decrease when the same lipase batch was reused. 83% of SLs were transformed to biodiesel using a 70 wt% lipase/SL ratio, 11:1 methanol/SL molar ratio, 10 mL t-butanol/g SLs after 72 h. The FAME conversion decreased to 71% after catalyzing three reactions with the same lipase batch. Copyright © 2015 Elsevier Ltd. All rights reserved.

Isolation and optimization of pectinase enzyme production one of useful industrial enzyme in Aspergillus niger, Rhizopus oryzae, Penicilium chrysogenum

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akram songol

2016-06-01

Full Text Available Introduction: Pectinase enzyme is one of the most important industrial enzymes which isolated from a wide variety of microorganisms such as bacteria and filamentous fungi. This enzyme has been usually used in the fruit and textile industry. In this study, the isolation and optimization of pectinase-producing fungi on decaying rotten fruits were studied. Materials and methods: Isolation and screening of pectinase producing fungi performed through plate culture on pectin medium and staining with Lugol's iodine solution. The best strains were identified by ITS1, 4 sequencing as Aspergillus fumigatus, Rhizopus oryzae, Penicilium chrysogenum. The enzyme production was optimized by application of the five factorial design, each at three levels. These factors are carbon sources (whey, glucose and stevia, ammonium sulfate, manganese sulfate, temperature, and pH. Pectinase concentration was measured by the Miller method. Results: The results indicate that optimum condition for enzyme production for three fungi strains was obtained at 32 °C, pH = 6, 3g / L manganese sulfate, 2.75g / L of ammonium sulfate and 10g / L of each carbon source. The best experiment in obtaining the optimum enzyme contained 1.328 mg / ml of glucose for Aspergillus niger 1.284 and 1.039 mg / ml of whey for Rhizopus oryzae and Penicilium chrysogenum. Molecular weight of enzyme was about 40 and 37 kDa which was obtained by SDS- PAGE. Discussion and conclusion: The results indicate that three strains could grow in a wide range of carbon source, pH and temperature, which could be a good candidate for industrial application.

The influence of inoculum composition on selected bioactive and nutritional parameters of grass pea tempeh obtained by mixed-culture fermentation with Rhizopus oligosporus and Aspergillus oryzae strains.

Science.gov (United States)

Starzynska-Janiszewska, A; Stodolak, B; Dulinski, R; Mickowska, B

2012-04-01

Tempeh is a popular Indonesian product obtained from legume seeds by solid-state fermentation with Rhizopus sp. The aim of this research was to study the effect of simultaneous mixed-culture fermentation of grass pea seeds on selected parameters of products as compared to traditional tempeh. The inoculum contained different ratios of Rhizopus oligosporus and Aspergillus oryzae spores. The simultaneous fermentation of grass pea seeds with inoculum consisting of 1.2 × 10(6) R. oligosporus and 0.6 × 10(6) A. oryzae spores (per 60 g of seeds) resulted in a product of improved quality, as compared with traditionally made tempeh (obtained after inoculation with 1.2 × 10(6) R. oligosporus spores), at the same fermentation time. This product had radical scavenging ability 70% higher than the one obtained with pure R. oligosporus culture and contained 2.23 g/kg dm of soluble phenols. The thiamin and riboflavin levels were above three (340 µg/g dm) and two (50.50 µg/g dm) folds higher than in traditionally made tempeh, respectively. The product had 65% in vitro bioavailability of proteins and 33% in vitro bioavailability of sugars. It also contained 40% less 3-N-oxalyl-L-2, 3-diaminopropionic acid (0.074 g/kg dm), as compared to traditional tempeh.

Extra-cellular isoamylase production by Rhizopus oryzae in solid-state fermentation of agro wastes

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Barnita Ghosh

2011-10-01

Full Text Available Extra-cellular isoamylase was produced by Rhizopus oryzae PR7 in solid-state fermentations of various agro wastes, among which millet, oat, tapioca, and arum (Colocasia esculenta showed promising results. The highest amount of enzyme production was obtained after 72 h of growth at 28°C. The optimum pH for enzyme production was - 8.0. Among the various additives tested, enzyme production increased with ions such as Ca2+, Mg2+ and also with cysteine, GSH, and DTT. The enzyme synthesis was reduced in the presence of thiol inhibitors like Cu2+ and pCMB. The surfactants like Tween-40, Tween-80 and Triton X-100 helped in enhancing the enzyme activity. The production could be further increased by using the combinations of substrates. The ability to produce high amount of isoamylase within a relatively very short period and the capability of degrading wastes could make the strain suitable for commercial production of the enzyme.

The impact of Rhizopus oryzae cultivation on rice bran: Gamma-oryzanol recovery and its antioxidant properties.

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Massarolo, Kelly Cristina; Denardi de Souza, Taiana; Collazzo, Carolina Carvalho; Badiale Furlong, Eliana; Souza Soares, Leonor Almeida de

2017-08-01

This study evaluated the effect of the solid state cultivation (SSC) time of rice bran by Rhizopus oryzae on γ-oryzanol recovery and its antioxidant properties. Gamma-oryzanol was extracted with organic solvents and its extracts were characterized by GC-FID and HPLC-UV. The antioxidant capacity was assessed by DPPH and ABTS + assays, β-carotene/linoleic acid system, and reduction of oxidation in lipid system. The biomass showed the γ-oryzanol recovery increased by 51.5% (20.52mg/g), and 5.7% in polyunsaturated fatty acids. The γ-oryzanol major components changing in their profile. The γ-oryzanol extract from biomass (72h) showed the greatest DPPH inhibition (59.0%), while 90.5% inhibition of oxidation of β-carotene/linoleic acid system, and 30% reduction of the indicators of oxidation in olive oil was observed in the one cultivated at 96h, these behaviors were confirmed by PCA analyses. SSC provides an increase in the γ-oryzanol recovery followed by improving of the functional properties of rice bran. Copyright © 2017 Elsevier Ltd. All rights reserved.

One-pot conjugated linoleic acid production from castor oil by Rhizopus oryzae lipase and resting cells of Lactobacillus plantarum.

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Khaskheli, Abid Ali; Talpur, Farah Naz; Cebeci Aydin, Aysun; Jawaid, Sana; Surhio, Muhammad Ali; Afridi, Hassan Imran

2017-10-01

Conjugated linoleic acid (CLA) has attracted as novel type of fatty acids having unusual health-promoting properties such as anticarcinogenic and antiobesitic effects. The present work employed castor oil as substrate for one-pot production of CLA using washed cells of Lactobacillus plantarum (L. plantarum) and lipases as catalysts. Among the screened lipases, the lipase Rhizopus oryzae (ROL) greatly assisted resting cells to produce CLA. Mass spectral analysis of the product showed that two major isomers of CLA were produced in the reaction mixture i.e. cis-9, trans-11 56.55% and trans-10, cis-12 43.45%. Optimum factors for CLA synthesis were found as substrate concentration (8 mg/mL), pH (6.5), washed cell concentration (12% w/v), and incubation time of 20 h. Hence, the combination of ROL with L. plantarum offers one pot production of CLA selectively using castor oil as a cost-effective substrate.

The production of corn kernel miso based on rice-koji fermented by Aspergillus oryzae and Rhizopus oligosporus

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Diah Ratnaningrum

2018-04-01

Full Text Available The suitability of corn kernel as raw material to produce miso fermented by rice-koji containing Aspergillus oryzae and Rhizopus oligosporus has been investigated. The optimization was conducted on two important factors in miso production namely mold composition in rice-koji and salt concentration. The mold composition was prepared by inoculating the spores of 2% A. oryzae, 2% R. oligosporus, and 2% the mixture of both in a ratio of 1:1, 2:1, and 1:2 (v/v into different rice media. The mold composition was optimized to produce rice-koji with high α-amylase and protease activity. Different NaCl concentrations of 10%, 15%, and 20% were subjected to optimization process and added to each mixture after five days of fermentation. The salt concentration was also optimized to produce corn kernel miso with high glucose and high dissolved protein concentration. The result showed that rice-koji containing A. oryzae and R. oligosporus in the ratio of 1:1 had the highest α-amylase and protease activity of 0.42 U/mL and 0.45 U/mL respectively. In addition, the presence of 10% NaCl in corn kernel miso fermented by A. oryzae and R. oligosporus in the ratio of 1:1 exhibited the highest glucose and dissolved protein concentration of 0.64 mg/mL and 8.80 mg/mL respectively. The optimized corn kernel miso by A. oryzae and R. oligosporus in the ratio of 1:1 with 10% NaCl was subjected to nutrient content analysis and compared to the result before the corn kernel was fermented. The nutrient content analysis showed nutrient enhancement after corn kernel was fermented and transformed into a miso. Glucose, dissolved protein, and fat content increased 6.74, 1.34, 7.63 times respectively. This study concludes corn kernel could be utilized to produce a novel corn kernel miso for dietary diversification and for improving nutritional and health status.

Production of α-amylase by solid state fermentation by Rhizopus ...

African Journals Online (AJOL)

2015-02-18

Feb 18, 2015 ... However, only a few strains of fungi and bacteria meet the criteria for production of ... amylase production, but solid-state fermentation (SSF) is emerging as a ..... synthesis of lactic acid in R. oryzae and Rhizopus arrhizus using ...

Purification and characterization of an extracellular lipase from a thermophilic Rhizopus oryzae strain isolated from palm fruit.

Science.gov (United States)

Hiol; Jonzo; Rugani; Druet; Sarda; Comeau

2000-03-01

We have isolated a lipolytic strain from palm fruit that was identified as a Rhizopus oryzae. Culture conditions were optimized and highest lipase production amounting to 120 U/ml was achieved after 4 days of cultivation. The extracellular lipase was purified 1200-fold by ammonium sulfate precipitation, sulphopropyl-Sepharose chromatography, Sephadex G 75 gel filtration and a second sulphopropyl-Sepharose chromatography. The specific activity of the purified enzyme was 8800 U/mg. The lipolytic enzyme has a molecular mass of 32 kDa by SDS-polyacrylamide gel electrophoresis and gel filtration. The enzyme exhibited a single band in active polyacrylamide gel electrophoresis and its isoelectric point was 7.6. Analysis of Rhizopus oryzae lipase by RP-HPLC confirmed the homogeneity of the enzyme preparation. Determination of the N-terminal sequence over 19 amino acid residues showed a high homology with lipases of the same genus. The optimum pH for enzyme activity was 7.5. Lipase was stable in the pH range from 4.5 to 7.5. The optimum temperature for lipase activity was 35 degrees C and about 65% of its activity was retained after incubation at 45 degrees C for 30 min. The lipolytic enzyme was inhibited by Triton X100, SDS, and metal ions such as Fe(3+), Cu(2+), Hg(2+) and Fe(2+). Lipase activity against triolein was enhanced by sodium cholate or taurocholate. The purified lipase had a preference for the hydrolysis of saturated fatty acid chains (C(8)-C(18)) and a 1, 3-position specificity. It showed a good stability in organic solvents and especially in long chain-fatty alcohol. The enzyme poorly hydrolyzed triacylglycerols containing n-3 polyunsaturated fatty acids, and appeared as a suitable biocatalyst for selective esterification of sardine free fatty acids with hexanol as substrate. About 76% of sardine free fatty acids were esterified after 30 h reaction whereas 90% of docosahexaenoic acid (DHA) was recovered in the unesterified fatty acids.

Variation in accumulation of isoflavonoids in Phaseoleae seedlings elicited by Rhizopus

NARCIS (Netherlands)

Aisyah, Siti; Gruppen, Harry; Andini, Silvia; Bettonvil, Monique; Severing, Eduard; Vincken, Jean Paul

2016-01-01

Seeds from seven species of tribe Phaseoleae, i.e. Phaseolus, Vigna, Lablab and Psophocarpus, were investigated for inducibility of isoflavonoids by germination with or without subsequent elicitation with Rhizopus oryzae. Germination alone poorly induced isoflavonoid production (in the range of

Estudio de la producción heteróloga de una lipasa del hongo Rhizopus oryzae en la levadura metilotrófica Pichia pastoris

OpenAIRE

Serrano González, Alicia

2004-01-01

Consultable des del TDX Títol obtingut de la portada digitalitzada El trabajo recoge los principales aspectos de la producción recombinante de una lipasa del hongo Rhizopus oryzae (ROL) expresada en la levadura Pichia pastoris. El sistema de expresión empleado está basado en la utilización del promotor de la alcohol oxidasa (PAOX1) de la levadura y, por lo tanto es dependiente de la utilización de metanol como substrato inductor. El trabajo recoge una serie de estudios de producción en ...

Proteolysis in tempeh-type products obtained with Rhizopus and Aspergillus strains from grass pea (Lathyrus sativus) seeds.

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Starzyńska-Janiszewska, Anna; Stodolak, Bożena; Wikiera, Agnieszka

2015-01-01

Tempeh is a food product obtained from legumes by means of solid-state fermentation with Rhizopus sp. Our previous research proved that mixed-culture inoculum may also be successfully applied. The objective of present research was to study the proteolytic activity of R. microsporus var. chinensis and A. oryzae during tempeh-type fermentation of grass pea seeds, and the effect of inoculum composition on the protein level and in vitro protein bioavailability in products. Fermentation substrate were soaked and cooked grass pea seeds. Material was mixed with single- or mixed-culture inoculum, and incubated in perforated plastic bags at 30°C for 32 hrs. In the products, the proteolytic activity (pH 3, 5 and 7), glucosamine, total protein and free amino acids levels, as well as protein in vitro bioavailability and degree of protein hydrolysis were obtained. The significant correlation was found between glucosamine content and proteolytic activity in grass pea seeds fermented with Rhizopus or Aspergillus. The activities of Rhizopus proteases were higher than Aspergillus ones, which corresponded with the degree of seed protein hydrolysis. Both strains showed the highest activity of protease at pH 3. Tempeh made with pure culture of Rhizopus had 37% protein of 69% in-vitro bioavailability. Mixed-culture fermentation improved nutritional parameters of products only when the dose of Aspergillus spores in the inoculum was equal and lower than that of Rhizopus. This process resulted in higher in-vitro bioavailability of protein, slightly more efficient protein hydrolysis and higher level of free amino acids, as compared to standard tempeh. The activity of A. oryzae in tempeh-type fermentation is beneficial as long as it does not dominate the activity and/or growth of Rhizopus strain.

Chitosan production by psychrotolerant Rhizopus oryzae in non-sterile open fermentation conditions.

Science.gov (United States)

Tasar, Ozden Canli; Erdal, Serkan; Taskin, Mesut

2016-08-01

A new chitosan producing fungus was locally isolated from soil samples collected around Erzurum, Turkey and identified as Rhizopus oryzae PAS 17 (GenBank accession number KU318422.1). Cultivation in low cost non-sterile conditions was achieved by exploiting its ability to grow at low temperature and pH, thus, undesired microbial contamination could be eliminated when appropriate culture conditions (incubation temperature as 15°C and initial pH of the medium as 4.5) were selected. Medium composition and culture conditions were optimized using Taguchi orthogonal array (OA) design of experiment (DOE). An OA layout of L16 (4(5)) was constructed with five most influensive factors at four levels on chitosan production like, carbon source (molasses), metal ion (Mg(2+)), inoculum amount, agitation speed and incubation time. The optimal combinations of factors (molasses, 70ml/l; MgSO4·7H2O, 0.5g/l; inoculum, 6.7×10(6) spores/disc; agitation speed, 150rpm and incubation time, 8days) obtained from the proposed DOE methodology was further validated by analysis of variance (ANOVA) test and the results revealed the increment of chitosan and biomass yields of 14.45 and 8.58 folds from its unoptimized condition, respectively. Copyright © 2016 Elsevier B.V. All rights reserved.

Optimization of L(+)-Lactic Acid Fermentation Without Neutralisation of Rhizopus Oryzae Mutant RK02 by Low-Energy Ion Implantation

International Nuclear Information System (INIS)

Li Wen; Wang Tao; Yang Yingge; Liu Dan; Fan Yonghong; Wang Dongmei; Yang Qian; Yao Jianming; Zheng Zhiming; Yu Zengliang

2008-01-01

In order to get an industrial strain which can yield a high concentration of lactic acid for ISPR (in situ product removal), the original strain Rhizopus oryzae RE3303 was mutated by low-energy ion beam implantation. A mutant RK02 was screened, and the factors such as the substrate concentration, nitrogen source concentration, inoculum size, seed age, aeration and temperature that affect the production of lactic acid were studied in detail. Under optimal conditions, the maximum concentration of L(+)-lactic acid reached 34.85 g/L after 30 h shake-flask cultivation without adding any neutralisation (5% Glucose added), which was a 146% increase in lactic acid production after ion implantation compared with the original strain. It was also shown that RK02 can be used in ISPR to reduce the number of times of separation.

Species boundaries and nomenclature of Rhizopus arrhizus (syn. R. oryzae)

NARCIS (Netherlands)

Dolatabadi, Somayeh; de Hoog, G Sybren; Meis, Jacques F; Walther, Grit

2014-01-01

Rhizopus arrhizus (Mucorales, Mucoromycotina) is the prevalent opportunist worldwide among the mucoralean species causing human infections. On the other hand the species has been used since ancient times to ferment African and Asian traditional foods and condiments based on ground soybeans. As

Response surface optimization for the transesterification of karanja oil using immobilized whole cells of Rhizopus oryzae in n-hexane system

Energy Technology Data Exchange (ETDEWEB)

Ganesan, Devanesan; Rajendran, Aravindan; Thangavelu, Viruthagiri [Annamalai University, Department of Chemical Engineering, Faculty of Engineering and Technology, Biochemical Engineering Laboratory, Annamalai Nagar, Tamil Nadu (India)

2012-03-15

Non-edible oils represent one of the most viable alternative feed stocks for the production of large volumes of biodiesel at cheaper cost in tropical countries. The objective of the present study is to investigate the ability of the immobilized whole cells of Rhizopus oryzae MTCC 262 to catalyze the biodiesel production from karanja oil in n-hexane system. Response surface methodology was employed to evaluate the effects of synthesis parameters, such as molar ratio of oil to alcohol, reaction temperature and reaction time on percentage biodiesel (methyl esters) yield. Transesterification was performed in shake flasks containing immobilized cells in the reaction mixture with 10% oil weight of n-hexane. The quadratic effects of molar ratio of oil to alcohol and reaction time proved to be the significant at 1% and 5% levels, respectively. The optimum synthesis conditions were found to be: molar ratio of oil to alcohol 1:2.73, reaction temperature 41.39 C and reaction time 73.97 h. Biodiesel yield (methyl ester) was 75.98 (wt.%) under the optimal conditions and the subsequent verification experiments with biodiesel yield of 78.0 (wt.%) confirmed the validity of the proposed model. (orig.)

Ethanol production by Mucor indicus and Rhizopus oryzae from rice straw by separate hydrolysis and fermentation

Energy Technology Data Exchange (ETDEWEB)

Abedinifar, Sorahi [Department of Chemical Engineering, Isfahan University of Technology, Isfahan 84156-83111 (Iran); Karimi, Keikhosro [Department of Chemical Engineering, Isfahan University of Technology, Isfahan 84156-83111 (Iran); School of Engineering, University of Boraas, SE-501 90 Boraas (Sweden); Khanahmadi, Morteza [Isfahan Agriculture and Natural Resources Research Centre, Isfahan (Iran); Taherzadeh, Mohammad J. [School of Engineering, University of Boraas, SE-501 90 Boraas (Sweden)

2009-05-15

Rice straw was successfully converted to ethanol by separate enzymatic hydrolysis and fermentation by Mucor indicus, Rhizopus oryzae, and Saccharomyces cerevisiae. The hydrolysis temperature and pH of commercial cellulase and {beta}-glucosidase enzymes were first investigated and their best performance obtained at 45 C and pH 5.0. The pretreatment of the straw with dilute-acid hydrolysis resulted in 0.72 g g{sup -1} sugar yield during 48 h enzymatic hydrolysis, which was higher than steam-pretreated (0.60 g g{sup -1}) and untreated straw (0.46 g g{sup -1}). Furthermore, increasing the concentration of the dilute-acid pretreated straw from 20 to 50 and 100 g L{sup -1} resulted in 13% and 16% lower sugar yield, respectively. Anaerobic cultivation of the hydrolyzates with M. indicus resulted in 0.36-0.43 g g{sup -1} ethanol, 0.11-0.17 g g{sup -1} biomass, and 0.04-0.06 g g{sup -1} glycerol, which is comparable with the corresponding yields by S. cerevisiae (0.37-0.45 g g{sup -1} ethanol, 0.04-0.10 g g{sup -1} biomass and 0.05-0.07 glycerol). These two fungi produced no other major metabolite from the straw and completed the cultivation in less than 25 h. However, R. oryzae produced lactic acid as the major by-product with yield of 0.05-0.09 g g{sup -1}. This fungus had ethanol, biomass and glycerol yields of 0.33-0.41, 0.06-0.12, and 0.03-0.04 g g{sup -1}, respectively. (author)

High-Level Expression of Pro-Form Lipase from Rhizopus oryzae in Pichia pastoris and Its Purification and Characterization

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Jian-Rong Wang

2013-12-01

Full Text Available A gene encoding Rhizopus oryzae lipase containing prosequence (ProROL was cloned into the pPICZαA and electrotransformed into the Pichia pastoris X-33 strain. The lipase was functionally expressed and secreted in Pichia pastoris with a molecular weight of 35 kDa. The maximum lipase activity of recombinant lipase (rProROL was 21,000 U/mL, which was obtained in a fed-batch cultivation after 168 h induction with methanol in a 50-L bioreactor. After fermentation, the supernatant was concentrated by ultrafiltration with a 10 kDa cut off membrane and purified with ion exchange chromatography using SP Sepharose Fast Flow chromatography. The optimum pH and temperature of the rProROL were pH 9.0 and 40 °C, respectively. The lipase was stable from pH 4.0 to 9.0 and from 25 to 55 °C. The enzyme activity was enhanced by Ca2+ and inhibited by Hg2+ and Ag+. The lipase showed high activity toward triglyceride-Tripalmitin (C16:0 and triglyceride-Trilaurin (C12:0.

Optimization of lactic acid production by pellet-form Rhizopus oryzae in 3-L airlift bioreactor using response surface methodology.

Science.gov (United States)

Maneeboon, Thanapoom; Vanichsriratana, Wirat; Pomchaitaward, Chaiyaporn; Kitpreechavanich, Vichien

2010-05-01

The influence of two key environmental factors, pH and oxygen transfer coefficient (k(L)a), was evaluated on the lactic acid production as the main answer and, on the size of cell pellets of the fungal strain Rhizopus oryzae KPS106, as second dependant answer by response surface methodology using a central composite design. The results of the analysis of variance and modeling demonstrated that pH and k(L)a had a significant effect on lactic acid production by this strain. However, no interaction was observed between these two experimental factors. pH and k(L)a had no significant influence on the pellet size. Optimal pH and k(L)a of the fermentation medium for lactic acid production from response surface analysis was 5.85 and of 3.6 h(-1), respectively. The predicted and experimental lactic acid maximal values were 75.4 and 72.0 g/l, respectively, with pellets of an average of 2.54 +/- 0.41 mm. Five repeated batches in series were conducted with a mean lactic acid production of 77.54 g/l. The productivity was increased from 0.75 in the first batch to 0.99 g/l h in the last fifth batch.

L-lactic acid production by Aspergillus brasiliensis overexpressing the heterologous ldha gene from Rhizopus oryzae.

Science.gov (United States)

Liaud, Nadège; Rosso, Marie-Noëlle; Fabre, Nicolas; Crapart, Sylvaine; Herpoël-Gimbert, Isabelle; Sigoillot, Jean-Claude; Raouche, Sana; Levasseur, Anthony

2015-05-03

Lactic acid is the building block of poly-lactic acid (PLA), a biopolymer that could be set to replace petroleum-based plastics. To make lactic acid production cost-effective, the production process should be carried out at low pH, in low-nutrient media, and with a low-cost carbon source. Yeasts have been engineered to produce high levels of lactic acid at low pH from glucose but not from carbohydrate polymers (e.g. cellulose, hemicellulose, starch). Aspergilli are versatile microbial cell factories able to naturally produce large amounts of organic acids at low pH and to metabolize cheap abundant carbon sources such as plant biomass. However, they have never been used for lactic acid production. To investigate the feasibility of lactic acid production with Aspergillus, the NAD-dependent lactate dehydrogenase (LDH) responsible for lactic acid production by Rhizopus oryzae was produced in Aspergillus brasiliensis BRFM103. Among transformants, the best lactic acid producer, A. brasiliensis BRFM1877, integrated 6 ldhA gene copies, and intracellular LDH activity was 9.2 × 10(-2) U/mg. At a final pH of 1.6, lactic acid titer reached 13.1 g/L (conversion yield: 26%, w/w) at 138 h in glucose-ammonium medium. This extreme pH drop was subsequently prevented by switching nitrogen source from ammonium sulfate to Na-nitrate, leading to a final pH of 3 and a lactic acid titer of 17.7 g/L (conversion yield: 47%, w/w) at 90 h of culture. Final titer was further improved to 32.2 g/L of lactic acid (conversion yield: 44%, w/w) by adding 20 g/L glucose to the culture medium at 96 h. This strain was ultimately able to produce lactic acid from xylose, arabinose, starch and xylan. We obtained the first Aspergillus strains able to produce large amounts of lactic acid by inserting recombinant ldhA genes from R. oryzae into a wild-type A. brasiliensis strain. pH regulation failed to significantly increase lactic acid production, but switching nitrogen source and changing culture feed

Kinetic properties of two Rhizopus exo-polygalacturonase enzymes hydrolyzing galacturonic acid oligomers using isothermal titration calorimetry

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The kinetic characteristics of two Rhizopus oryzae exo-polygalacturonases acting on galacturonic acid oligomers (GalpA) were determined using isothermal titration calorimetry (ITC). RPG15 hydrolyzing (GalpA)2 demonstrated a Km of 55 uM and kcat of 10.3 s^-1^ while RPG16 was shown to have greater af...

Identification of prenylated pterocarpans and other isoflavonoids in Rhizopus spp. elicited soya bean seedlings by electrospray ionisation mass spectrometry

NARCIS (Netherlands)

Simons, R.; Vincken, J.P.; Bohin, M.C.; Kuijpers, T.F.M.; Verbruggen, M.A.; Gruppen, H.

2011-01-01

Phytoalexins from soya are mainly characterised as prenylated pterocarpans, the glyceollins. Extracts of non-soaked and soaked soya beans, as well as that of soya seedlings, grown in the presence of Rhizopus microsporus var. oryzae, were screened for the presence of prenylated flavonoids with a

Immobilized Rhizopus oryzae lipase catalyzed synthesis of palm stearin and cetyl alcohol wax esters: Optimization by Response Surface Methodology

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Gargouri Youssef

2011-06-01

Full Text Available Abstract Background Waxes are esters of long-chain fatty acids and long-chain alcohols. Their principal natural sources are animals (sperm whale oil and vegetables (jojoba which are expensive and not easily available. Wax esters synthesized by enzymatic transesterification, using palm stearin as raw material, can be considered as an alternative to natural ones. Results Palm stearin is a solid fraction obtained by fractionation of palm oil. Palm stearin was esterified with cetyl alcohol to produce a mixture of wax esters. A non-commercial immobilized lipase from Rhizopus oryzae was used as biocatalyst. Response surface methodology was employed to determine the effects of the temperature (30-50°C, the enzyme concentration (33.34-300 IU/mL, the alcohol/palm stearin molar ratio (3-7 mol/mol and the substrate concentration (0.06-0.34 g/mL on the conversion yield of palm stearin. Under optimal conditions (temperature, 30°C; enzyme concentration, 300 IU/mL; molar ratio 3 and substrate concentration 0.21 g/mL a high conversion yield of 98.52% was reached within a reaction time of 2 h. Conclusions Response surface methodology was successfully applied to determine the optimum operational conditions for synthesis of palm stearin based wax esters. This study may provide useful tools to develop economical and efficient processes for the synthesis of wax esters.

The effects of irradiated fungi to produce glucose from sago starch

International Nuclear Information System (INIS)

Andini, L.S.; Sjarief, Sri Hariani; Sumartono, Agustin; Sudradjat, Dadang; Anastasia, S.D.

1994-01-01

The effects of irradiated fungi to produce glucose from sago starch had been studied. Fungi used in this experiment were Aspergillus niger K-23, Rhizopus oryzae K-22, and Rhizopus oligosporus K-18. The experiment was carried out to find the optimum doses of irradiation for glucose production of sago starch. Irradiation dose at 0; 0.25; 0.5; and 0.75 kGy respectively from cobalt-60 gamma-cell 220 at a dose rate of 0.42 kGy/h. Concentration of sago starch is 6%. Total glucose and pH were measured on the 3rd and 6th day after incubation at 30 Celcius degrees in shaker incubator. The optimum irradiation doses obtained were 0.25 kGy for Rhizopus oryzae K-22, and 0.75 kGy for Aspergillus niger K-23, and Rhizopus oligosporus K-18. Higher dose of irradiation decreased the pH value of sago substrat, for Aspergillus niger K-23 and Rhizopus oligosporus K-18, but the opposite result was obtained for Rhizopus oryzae K-22. The glucose content at 3 days incubation was higher than at 6 days. (author). 13 refs, 5 tabs, 3 figs

Low Lactose Milk Production of Soybean by Fermentation Technique Using Rhizopus oligosporus

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Farid Salahudin

2013-06-01

Full Text Available Milk is an important food for baby that contains lactose. Normally, a baby could produce lactase enzyme that digest lactose, but in the diarrhea case lactose could not be digested. So, Low Lactose Milk is needed. Low Lactose Milk usually produced from rice or almonds that have low protein. Soybean (Glycine max is the commodity with rich of protein and also contains raffinose and stachyose, which can lead flatulence. Raffinose and stachyose could be reduced by Rhizopus oryzae at tempe process from lamtoro beans.  So the aim in this research is to know the optimum time of soybean fermentation with R. oryzae to reduce stachyiose  and raffinose. The research was done with innoculation of R. oryzae isolate in the soybeans fermentation for 72 hours. N index, raffinose and stachyose level was tested. The result shows that optimum fermentation time is 48 hour and using 5% skim milk as filler.

Antifungal effect of phenolic extract of fermented rice bran with Rhizopus oryzae and its potential use in loaf bread shelf life extension.

Science.gov (United States)

Denardi-Souza, Taiana; Luz, Carlos; Mañes, Jordi; Badiale-Furlong, Eliana; Meca, Giuseppe

2018-03-30

In this study the antifungal potential of a phenolic extract obtained from rice bran fermented with Rhizopus oryzae CECT 7560 and its application in the elaboration of bread was assessed. Eighteen compounds with antifungal potential were identified by LC-ESI-qTOF-MS in the extract: organic acids, gallates and gallotannins, flavonoids, ellagic acid and benzophenone derivatives. The extract was active against strains of Fusarium, Aspergillus and Penicillium, with minimum inhibitory concentration ranging from 390 to 3100 µg mL -1 and minimum fungicidal concentration variable from 780 to 6300 µg mL -1 . The strains that were most sensitive to the phenolic extract were F. graminearum, F. culmorum, F. poae, P. roqueforti, P. expansum and A. niger. The phenolic extract added at 5 and 1 g kg -1 concentrations in the preparation of bread loaves contaminated with P. expansum produced a reduction of 0.6 and 0.7 log CFU g -1 . The bread loaves treated with calcium propionate and 10 g kg -1 of the phenolic extract evidenced an improvement in their shelf lives of 3 days. The phenolic extract assessed in this study could be considered as an alternative for inhibiting toxigenic fungi and as a substitute for synthetic compounds in food preservation. © 2018 Society of Chemical Industry. © 2018 Society of Chemical Industry.

Eco-friendly methodology for efficient synthesis and scale-up of 2-ethylhexyl-p-methoxycinnamate using Rhizopus oryzae lipase and its biological evaluation.

Science.gov (United States)

Kumar, Vinod; Jahan, Firdaus; Kameswaran, Karthikeya; Mahajan, Richi V; Saxena, Rajendra Kumar

2014-06-01

Lipase-mediated synthesis of phenolic acid esters is a green and economical alternative to current chemical methods. Octyl methoxycinnamate, an important UVB-absorbing compound, was synthesized by the esterification of p-methoxycinnamic acid with 2-ethyl hexanol using Rhizopus oryzae lipase. A molar ratio of 1:2 of p-methoxycinnamic acid and 2-ethyl hexanol was found to give an optimum yield using cyclo-octane (50 ml) as reaction solvent, at a temperature of 45 °C, and 750 U of lipase, resulting in a yield of 91.3 % in 96 h. This reaction was successfully scaled up to 400-ml reaction size where 88.6 %bioconversion was achieved. The synthesized compound was found to have superior antioxidant activity as compared to ascorbic acid. The synthesized compound also exhibited good antimicrobial activity against Escherichia coli, Klebsiella pneumonia, Salmonella typhi, Staphylococcus aures, Candida albicans (yeast), Aspergillus niger, Alternaria solani, and Fussarium oxysporum by well diffusion method in terms of zone of inhibitions (in mm).

Hydrolysis of Soybean Protein by Aspergillus Sojae, a. Oryzae, and Rhizpus Oligosporus

OpenAIRE

Rahayu, Endang Sutriswati

1991-01-01

Three species of molds, i.e. Aspergillus sojae. A. oryzae and Rhizopus oligosporus were used to hydrolyze soybean proteins. Whole soybeans were soaked overnight and cooked in boiling water for an hour, drained, sterilized at 121°C for 15 minutes, then cooled and inoculated with A. oryzae. A. sojae, and R. oligosporus. As a control treatment another batch of soybeans was prepared for spontaneous fermentation. Fermentation tasted for five days. Based on the colony forming units (CFU), A. sojae,...

Two unique ligand-binding clamps of Rhizopus oryzae starch binding domain for helical structure disruption of amylose.

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Ting-Ying Jiang

Full Text Available The N-terminal starch binding domain of Rhizopus oryzae glucoamylase (RoSBD has a high binding affinity for raw starch. RoSBD has two ligand-binding sites, each containing a ligand-binding clamp: a polyN clamp residing near binding site I is unique in that it is expressed in only three members of carbohydrate binding module family 21 (CBM21 members, and a Y32/F58 clamp located at binding site II is conserved in several CBMs. Here we characterized different roles of these sites in the binding of insoluble and soluble starches using an amylose-iodine complex assay, atomic force microscopy, isothermal titration calorimetry, site-directed mutagenesis, and structural bioinformatics. RoSBD induced the release of iodine from the amylose helical cavity and disrupted the helical structure of amylose type III, thereby significantly diminishing the thickness and length of the amylose type III fibrils. A point mutation in the critical ligand-binding residues of sites I and II, however, reduced both the binding affinity and amylose helix disruption. This is the first molecular model for structure disruption of the amylose helix by a non-hydrolytic CBM21 member. RoSBD apparently twists the helical amylose strands apart to expose more ligand surface for further SBD binding. Repeating the process triggers the relaxation and unwinding of amylose helices to generate thinner and shorter amylose fibrils, which are more susceptible to hydrolysis by glucoamylase. This model aids in understanding the natural roles of CBMs in protein-glycan interactions and contributes to potential molecular engineering of CBMs.

Effect of dietary supplementation with Rhizopus oryzae or Chrysonilia crassa on growth performance, blood profile, intestinal microbial population, and carcass traits in broilers exposed to heat stress

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S. Sugiharto

2017-09-01

Full Text Available Dietary supplementation of additives has recently been part of strategies to deal with the detrimental effects of heat stress (HS on the performance and carcass traits in broiler chicks. This study aimed to investigate the effect of dietary supplementation with the fungi Rhizopus oryzae or Chrysonilia crassa on growth, blood profile, intestinal microbial population and carcass traits in broiler chicks subjected to HS. R. oryzae and C. crassa are filamentous fungi isolated from the ileum of indigenous Indonesian chickens which exhibited probiotic and antioxidant properties. Two hundred and forty 21-day-old male broiler chicks were randomly allotted into six groups, including birds reared under normal temperature (28 ± 2 °C (CONT, birds reared under HS conditions (35 ± 2 °C (HS-CONT, birds reared under HS and provided with commercial anti-stress formula (HS-VIT, birds reared under HS and provided with R. oryzae (HS-RO, birds reared under HS and provided with C. crassa (HS-CC and birds reared under HS and provided with rice bran (HS-RB. Body weight gain was highest (P < 0. 01 and lowest (P < 0. 01 in CONT and HS-CONT birds, respectively. The heart was heavier (P < 0. 05 in CONT than in HS-CONT and HS-VIT birds. CONT birds had heavier duodenum (P < 0. 05 and jejunum (P < 0. 01 than other birds. Eosinophils was higher (P < 0. 05 in HS-CC than in other birds. Low-density lipoprotein (LDL was higher (P < 0. 05 in HS-CONT than in CONT, HS-VIT and HS-CC birds. Total triglyceride was highest (P < 0. 05 and lowest (P < 0. 05 in HS-RB and HS-RO birds, respectively. Alanine aminotransferase (ALT was higher (P < 0. 05 in HS-CONT than in other HS birds. Total protein was lowest and highest (P < 0. 05 in CONT and HS-CONT birds, respectively. Albumin was higher (P < 0. 05 in HS-CONT and HS-VIT than in HS-RO birds. Globulin was lower (P < 0. 05 in CONT than in HS

Transcriptional response of P. pastoris in fed-batch cultivations to Rhizopus oryzae lipase production reveals UPR induction

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Valero Francisco

2007-07-01

Full Text Available Abstract Background The analysis of transcriptional levels of the genes involved in protein synthesis and secretion is a key factor to understand the host organism's responses to recombinant protein production, as well as their interaction with the cultivation conditions. Novel techniques such as the sandwich hybridization allow monitoring quantitatively the dynamic changes of specific RNAs. In this study, the transcriptional levels of some genes related to the unfolded protein response (UPR and central metabolism of Pichia pastoris were analysed during batch and fed-batch cultivations using an X-33-derived strain expressing a Rhizopus oryzae lipase under control of the formaldehyde dehydrogenase promoter (FLD1, namely the alcohol oxidase gene AOX1, the formaldehyde dehydrogenase FLD1, the protein disulfide isomerase PDI, the KAR2 gene coding for the BiP chaperone, the 26S rRNA and the R. oryzae lipase gene ROL. Results The transcriptional levels of the selected set of genes were first analysed in P. pastoris cells growing in shake flask cultures containing different carbon and nitrogen sources combinations, glycerol + ammonium, methanol + methylamine and sorbitol + methylamine. The transcriptional levels of the AOX1 and FLD1 genes were coherent with the known regulatory mechanism of C1 substrates in P. pastoris, whereas ROL induction lead to the up-regulation of KAR2 and PDI transcriptional levels, thus suggesting that ROL overexpression triggers the UPR. This was further confirmed in fed-batch cultivations performed at different growth rates. Transcriptional levels of the analysed set of genes were generally higher at higher growth rates. Nevertheless, when ROL was overexpressed in a strain having the UPR constitutively activated, significantly lower relative induction levels of these marker genes were detected. Conclusion The bead-based sandwich hybridization assay has shown its potential as a reliable instrument for quantification of

Possible pulmonary Rhizopus oryzae infection in a previously healthy child after a near-drowning incident.

Science.gov (United States)

Gerlach, Magdalena M; Lippmann, Norman; Kobelt, Louise; Petzold-Quinque, Stefanie; Ritter, Lutz; Kiess, Wieland; Siekmeyer, Manuela

2016-06-01

This article reports on a previously healthy 17-month-old boy who developed pulmonary mucormycosis after a near-drowning incident in a goose pond. The patient survived without neurological sequelae and recovered, under treatment with amphotericin B, from the rare and often invasive fungal infection with Rhizopus spp., usually occurring in immunodeficient patients.

Biocatalytic Behaviour of Immobilized Rhizopus oryzae Lipase in the 1,3-Selective Ethanolysis of Sunflower Oil to Obtain a Biofuel Similar to Biodiesel

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Carlos Luna

2014-08-01

Full Text Available A new biofuel similar to biodiesel was obtained in the 1,3-selective transesterification reaction of sunflower oil with ethanol using as biocatalyst a Rhizopus oryzae lipase (ROL immobilized on Sepiolite, an inorganic support. The studied lipase was a low cost powdered enzyme preparation, Biolipase-R, from Biocon-Spain, a multipurpose additive used in food industry. In this respect, it is developed a study to optimize the immobilization procedure of these lipases on Sepiolite. Covalent immobilization was achieved by the development of an inorganic-organic hybrid linker formed by a functionalized hydrocarbon chain with a pendant benzaldehyde, bonded to the AlPO4 support surface. Thus, the covalent immobilization of lipases on amorphous AlPO4/sepiolite (20/80 wt % support was evaluated by using two different linkers (p-hydroxybenzaldehyde and benzylamine-terephthalic aldehyde, respectively. Besides, the catalytic behavior of lipases after physical adsorption on the demineralized sepiolite  was also evaluated. Obtained results indicated that covalent immobilization with the p-hydroxybenzaldehyde linker gave the best biocatalytic behavior. Thus, this covalently immobilized lipase showed a remarkable stability as well as an excellent capacity of reutilization (more than five successive reuses without a significant loss of its initial catalytic activity. This could allow a more efficient fabrication of biodiesel minimizing the glycerol waste production.

Fob1 and Fob2 proteins are virulence determinants of Rhizopus oryzae via facilitating iron uptake from ferrioxamine

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Dialysis patients with chronic renal failure receiving deferoxamine for treating iron overload are uniquely predisposed for mucormycosis. Although not secreted by Mucorales, previous studies established that Rhizopus species utilize iron from ferrioxamine (iron-rich form of deferoxamine). Here we de...

Efficient heterologous expression and secretion in Aspergillus oryzae of a llama variable heavy-chain antibody fragment V(HH) against EGFR.

Science.gov (United States)

Okazaki, Fumiyoshi; Aoki, Jun-ichi; Tabuchi, Soichiro; Tanaka, Tsutomu; Ogino, Chiaki; Kondo, Akihiko

2012-10-01

We have constructed a filamentous fungus Aspergillus oryzae that secretes a llama variable heavy-chain antibody fragment (V(HH)) that binds specifically to epidermal growth factor receptor (EGFR) in a culture medium. A major improvement in yield was achieved by fusing the V(HH) with a Taka-amylase A signal sequence (sTAA) and a segment of 28 amino acids from the N-terminal region of Rhizopus oryzae lipase (N28). The yields of secreted, immunologically active anti-EGFR V(HH) reached 73.8 mg/1 in a Sakaguchi flask. The V(HH) fragments were released from the sTAA or N28 proteins by an indigenous A. oryzae protease during cultivation. The purified recombinant V(HH) fragment was specifically recognized and could bind to the EGFR with a high affinity.

Phylogenetic and Phylogenomic Definition of Rhizopus Species

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Andrii P. Gryganskyi

2018-06-01

Full Text Available Phylogenomic approaches have the potential to improve confidence about the inter-relationships of species in the order Mucorales within the fungal tree of life. Rhizopus species are especially important as plant and animal pathogens and bioindustrial fermenters for food and metabolite production. A dataset of 192 orthologous genes was used to construct a phylogenetic tree of 21 Rhizopus strains, classified into four species isolated from habitats of industrial, medical and environmental importance. The phylogeny indicates that the genus Rhizopus consists of three major clades, with R. microsporus as the basal species and the sister lineage to R. stolonifer and two closely related species R. arrhizus and R. delemar. A comparative analysis of the mating type locus across Rhizopus reveals that its structure is flexible even between different species in the same genus, but shows similarities between Rhizopus and other mucoralean fungi. The topology of single-gene phylogenies built for two genes involved in mating is similar to the phylogenomic tree. Comparison of the total length of the genome assemblies showed that genome size varies by as much as threefold within a species and is driven by changes in transposable element copy numbers and genome duplications.

Degradação de deoxinivalenol (DON) e a atividade da enzima peroxidase durante fermentação submersa

OpenAIRE

Garda-Buffon, Jaqueline; Kupski, Larine; Badiale-Furlong, Eliana

2011-01-01

This work aims to evaluate deoxynivalenol degradation by Aspergillus oryzae and Rhizopus oryzae in a submerged fermentation system and to correlate it to the activity of oxydo-reductase enzymes. The submerged medium consisted of sterile distilled water contaminated with 50 μg of DON and 4 × 10(6) spore.mL-1 inoculum of Aspergillus oryzae and Rhizopus oryzae species, respectively in each experiment. Sampling was performed every 24 hours for monitoring the peroxidase specific activity, and ever...

SOLID-STATE FERMENTATIVE PRODUCTION AND BIOACTIVITY OF FUNGAL CHITOSAN

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Barry Aigbodion Omogbai

2013-10-01

Full Text Available Chitosan production was investigated using a laboratory-scale solid substrate fermentation (SSF technique with four species of fungi: Penicillium expansum, Aspergillus niger, Rhizopus oryzae and Fusarium moniliforme.The peak growth for the organisms was after 16 days. Aspergillus niger had the highest growth with a maximal dry cell biomass of 15.8g/kg after 16 days cultivation on corn straw under solid substrate fermentation. This was closely followed by Rhizopus oryzae (14.6g/kg, Penicillium expansum (13.8g/kg and Fusarium moniliforme (10.6g/kg respectively. The fungus Rhizopus oryzae had the highest chitosan production with a maximum of 8.57g/kg in 16 days under solid substrate fermentation (SSF with a medium containing corn straw. Aspergillus niger showed a modest chitosan yield of 6.8g/kg. Penicillium expansum and Fusarium moniliforme had low chitosan yields of 4.31g/kg and 3.1g/kg respectively. The degree of deacetylation of fungal chitosans ranged between 75.3-91.5% with a viscosity of 3.6-7.2 centipoises (Cp.Chitosan extracted from Rhizopus oryzae was found to have antibacterial activity on some bacterial isolates. At a concentration of 50mg/L, Rhizopus oryzae chitosan paralleled crab chitosan in susceptibility testing against some food-borne bacterial pathogens. Escherichia coli, Salmonella typhi, Pseudomonas aeruginosa and Bacillus subtilis showed inhibition rates of 83.2%, 67.9%, 63.8% and 62.4% respectively in response to 50mg/l Rhizopus oryzae chitosan in 24 h. The rate of inhibition (% increased with increase in chitosan concentration.

Isolation of Mucorales from processed maize (Zea mays L. and screening for protease activity Isolamento de Mucorales de milho processado (Zea mays L. e seleção quanto à atividade proteásica

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André Luiz Cabral Monteiro de Azevedo Santiago

2008-12-01

Full Text Available Mucoraleswere isolated from maize flour, corn meal and cooked cornflakes using surface and depth plate methods. Rhizopus oryzae, Circinella muscae, Mucor subtilissimus,Mucor hiemalis f. hiemalis, Syncephalastrum racemosum, Rhizopus microsporus var. chinensis and Absidia cylindrospora showed protease activity.Mucorales foram isolados da farinha de milho, fubá e flocos de milho pré-cozidos pelos métodos de plaqueamento em superfície e em profundidade. Rhizopus oryzae, Circinella muscae, Mucor subtilissimus,Mucor hiemalis f. hiemalis, Syncephalastrum racemosum, Rhizopus microsporus var. chinensis e Absidia cylindrospora exibiram atividade proteásica.

Sperathe effects of solid-state fermentation in the functional properties of defatted rice bran and wheat bran

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Cristina Moreira da Silveira

2009-12-01

Full Text Available Functional properties of fermented bran produced by Aspergillus oryzae and Rhizopus sp. in a solid-state fermentation system were determined, with an aim to evaluate their application in food formulation. The defatted rice bran and wheat bran were inoculated with the spores of the cultures and incubated at 30º C for 72 h. Samples were withdrawn at 0, 24, 48 and 72 h. Protein content, protein solubility, in-vitro digestibility, gelation and water holding capacity were determined in bran with or without fermentation. Rhizopus sp. increased significantly the protein content (69.0 and 56.0%, respectively, for defatted rice bran and wheat bran; protein solubility (28.5 and 36.2 and water holding capacity (11.4% for wheat bran. When A. oryzae was used all these properties were modified significantly after fermentation.O objetivo do presente trabalho foi investigar as propriedades funcionais de farelo de arroz desengordurado e farelo de trigo, submetidos à fermentação em estado sólido pelos fungos Aspergillus oryzae e Rhizopus sp., para avaliar seu potencial de aplicação em formulações alimentícias. O farelo de arroz desengordurado e o farelo de trigo foram inoculados com esporos de Rhizopus sp. e Aspergillus oryzae (4x10(6 esporos/grama de meio e incubados durante 72 horas a 30º C. Amostras foram coletadas em 0, 24, 48 e 72 horas de fermentação. Foram determinados o conteúdo protéico, a solubilidade protéica, a digestibilidade in-vitro, a capacidade de formação de gel e a capacidade de retenção de água nos farelos fermentados. Quando Rhizopus sp. foi utilizado, as seguintes propriedades aumentaram significativamente: o conteúdo protéico 69,0 e 56,0%, respectivamente, para farelo de arroz e farelo de trigo, a solubilidade protéica (28,5 e 36,2% e a capacidade de retenção de água (11,4% para farelo de trigo. Quando Aspergillus oryzae foi empregado, as propriedades não foram modificadas de maneira significativa pelo processo

Antibiotic effective against Saccharomyces produced by Aspergillus oryzae

Energy Technology Data Exchange (ETDEWEB)

Nakata, H.; Sakai, T.; Takeda, M.; Tsukahara, T.

1980-01-01

Production of an antibiotic effective against Saccharomyces cerevisiae was investigated in 85 strains of Aspergillus oryzae, isolated from commercial koji molds. The antibiotic was produced by 50 strains. A. oryzae was cultivated at 30 degrees for 15-20 days in koji extract. The crude preparation was obtained by precipitation from the culture filtrate with EtOH, MeOH, or Me/sub 2/CO.

Effect of cadmium salts on some growth and metabolic activities of selected fungi from the rhizosphere of Glycine max and Zea mays

Energy Technology Data Exchange (ETDEWEB)

Naguib, M.I.; Ali, M.I.; Haikal, N.Z.; Sharaf, E.F. (Cairo Univ. (Egypt))

1986-12-01

Six fungi, isolated from the rhizosphere of Glycine max and Zea Mays, were selected for their variable resistance to soil application of cadmium nitrate and cadmium sulfate, at the range of 100-1000 ppm concentration. These were (a) Aspergillus terreus and Rhizopus oryzae; (b) Paecilomyces divaricata, Curvularia tetramera; (c) Fusarium oxysporum and (d) Cladosporium herbarum. Fifty ppm cadmium salts exerted no effect on spore germination of R. oryzae, F. oxysporum or C. tetramera, stimulated A. terreus and P. divaricata and suppressed C. herbarum. 100 ppm dose exerted variable effects dependent on the organism and/or accompanying anion. 1000 ppm cadmium arrested spore germination of all the tested fungi except F. oxysporum where marked suppression were observed. Cadmium arrested growth of C. herbarum and P. divaricata at 50 ppm, initiated that of A. terreus, C. tetramera and F. oxysporum but seemed without effect on R. oryzae. Larger doses seemed to be inhibitory to all organisms, R. oryzae being least affected. Results show that nitrogen secretion was not a function of tolerance of the fungi to cadmium applied to the soil but was largely controlled by the effect of cadmium on the permeability of the cells of each individual fungus. Cadmium seemed to stimulate nitrogen metabolism of Aspergillus and Fusarium, had little or no effect on that of Rhizopus but suppressed that of Curvularia. Most accumulated nitrogen was in the form of amino and other soluble nitrogen. Cadmium inhibited the incorporation of absorbed phosphorus into energy-rich compounds. This was concomitant with the failure of amino acid incorporation into proteins.

Rhizopus in Kalanchoë

NARCIS (Netherlands)

Paternotte, S.J.

2001-01-01

In 2001 is door PPO-glastuinbouw onderzoek gedaan aan Rhizopus stolonifer in Kalanchoë ( verslag PPO 533). Naar aanleiding van dit onderzoek en na gesprekken met de begeleidingscommissie en telers van Kalanchoë moerplanten is de conclusie getrokken dat vooral grote schommelingen in watergehalte van

IN VITRO BIOACTIVITY OF CREOSOTE BUSH EXTRACTS (LARREA TRIDENTATA ON THE INHIBITION OF POSTHARVEST FUNGI: PENICILLIUM POLONICUM, ASPERGILLUS NIGER, RHIZOPUS ORYZAE Y ALTERNARIA TENUISSIMA

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O. Peñuelas-Rubio

2015-11-01

Full Text Available En el presente estudio se evaluó la eficiencia de extractos vegetales deLarrea tridentataobtenidos con diclorometano, etanol, metanol y agua, sobre el crecimiento radial in vitro de cuatro hongos fitopatógenos, los cuales primeramente fueron identificados en género y especie empleando claves taxonómicas y técnicas moleculares. Para los bioensayosin vitrose aplicaron diseños completamente al azar con cuatro tratamientos y tres repeticiones en cada hongo, utilizando las concentraciones: 0, 250, 500 y 750 ppm paraAlternaria sp.; 0, 2000, 2500 y 3000 paraAspergillus sp.; 0, 1500, 1750 y 2000 paraPenicillium sp. y 0, 150, 200 y 250 ppm paraRhizopus sp. Cada tratamiento tuvo tres repeticiones. El análisis molecular determinó la especie tenuissima paraAlternaria,nigerparaApergillus,polonicumparaPenicilliumyoryzaeparaRhizopus. En cuanto a las pruebasin vitro, se determinaron inhibiciones del 100% para tres de los hongos en estudio:Alternaria tenuissimacon extracto EtOH a 750 ppm;Aspergillus nigercon extracto DCM a 3000 ppm yRhizopus oryzaea partir de 150 ppm y 250 ppm de los extractos DCM y EtOH respectivamente. Se presentó una inhibición del 82% a 2000 ppm paraPenicillium polonicum. Se concluye que a pesar de las diferencias en susceptibilidad entre las especies fúngicas, los extractos deLarrea tridentataobtenidos con etanol y dicloromentano son efectivos para el control de los hongos fitopatógenos bajo estudioin vitro.

POTENSI DARI KAPANG Aspergilus niger, Rhizophus oryzae DAN Neurospora sitophila SEBAGAI PENGHASIL EZIM FITASE DAN AMILASE PADA SUBSTRATE AMPAS TAHU

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Atit - Kanti

2017-02-01

Full Text Available Penambahan enzim hidrolisis untuk pakan ternak dapat meningkatkan nilai nutrisi pakan. Penelitian bertujuan untuk mendapatkan kondisi optimal untuk produksi enzim amilase dan fitase pada media ampas tahu menggunakan Aspergilus niger, Rhizophus oryzae dan Neurospora sitophila. Uji kemampuan produksi enzim fitase dan amilase oleh Aspergilus niger, Rhizophus oryzae dan Neurospora sitophila dilakukan menggunakan media ampas tahu yang disterilisasi. Pemilihan ketiga isolat ini diawali dengan uji produksi enzim amilase pada kultur cair yang mengandung 2 % pati, dan uji fitase dilakukan pada media yang mengandung 0.5 % sodium fitat. Hasil uji pada medium cair selanjutnya digunakan untuk uji produksi enzim fitase dan fitase pada sistem fermentasi padat (SSF menggunakan ampas tahu sebagai media fermentasi. Untuk mendapatkan produksi enzim yang tinggi dilakukan melalui optimasi waktu inkubasi, suhu inkubasi dan pH media. Fitase dan amilase dapat diproduksi dengan media ampas tahu oleh R. oryzae, A. niger dan N. sitophila. Kondisi optimum untuk produksi fitase, yaitu waktu inkubasi pada hari keempat untuk ketiga kapang, suhu 25 °C untuk R. oryzae dan A. niger, suhu 30°C untuk N. sitophila, pH 8 untuk R. oryzae, pH 6 untuk Aspergillus niger dan N. Sitophila. Neurospora sitophila menghasilkan amilase optimum pada suhu 35°C, sedangkan Aspergillus niger dan Rhizopus oryzae optimum pada suhu 30°C. Penurunan aktivitas produksi amilase menurun oleh R. oryzae pada suhu 40°C. Amilase diproduksi optimal pada pH 6-7. Pakan ternak yang mengandung asam fitat mampu dihidrolisis oleh fitase pada kondisi optimum. Ketiga kapang juga menghasilkan enzim amilase pada media ampas tahu mengindikasikan bahwa ampas tahu merupakan susbtrat yang baik untuk produksi enzim hidrolisis yang berguna untuk meningkatkan nilai nutrisi pakan ternak. (Kata kunci: Amilase, Aspergilus niger, Neurospora sitophila, phytase, Rhizophus oryzae

Identification of prenylated pterocarpans and other isoflavonoids in Rhizopus spp. elicited soya bean seedlings by electrospray ionisation mass spectrometry.

Science.gov (United States)

Simons, Rudy; Vincken, Jean-Paul; Bohin, Maxime C; Kuijpers, Tomas F M; Verbruggen, Marian A; Gruppen, Harry

2011-01-15

Phytoalexins from soya are mainly characterised as prenylated pterocarpans, the glyceollins. Extracts of non-soaked and soaked soya beans, as well as that of soya seedlings, grown in the presence of Rhizopus microsporus var. oryzae, were screened for the presence of prenylated flavonoids with a liquid chromatography/mass spectrometry (LC/MS)-based screening method. The glyceollins I-III and glyceollidins I-II, belonging to the isoflavonoid subclass of the pterocarpans, were tentatively assigned. The formation of these prenylated pterocarpans was accompanied by that of other prenylated isoflavonoids of the subclasses of the isoflavones and the coumestans. It was estimated that approx. 40% of the total isoflavonoid content in Rhizopus-challenged soya bean seedlings were prenylated pterocarpans, whereas 7% comprised prenylated isoflavones and prenylated coumestans. The site of prenylation (A-ring or B-ring) of the prenylated isoflavones was tentatively annotated using positive-ion mode MS by comparing the (1,3) A(+) retro-Diels-Alder (RDA) fragments of prenylated and non-prenylated isoflavones. Furthermore, the fragmentation pathways of the five pterocarpans in negative-ion (NI) mode were proposed, which involved the cleavage of the C-ring and/or D-ring. The absence of the ring-closed prenyl (pyran or furan) gave exclusively -H(2) O(x,y) RDA fragments, whereas its presence gave predominantly the common RDA fragments. Copyright © 2010 John Wiley & Sons, Ltd.

CatB is Critical for Total Catalase Activity and Reduces Bactericidal Effects of Phenazine-1-Carboxylic Acid on Xanthomonas oryzae pv. oryzae and X. oryzae pv. oryzicola.

Science.gov (United States)

Pan, Xiayan; Wu, Jian; Xu, Shu; Duan, Yabing; Zhou, Mingguo

2017-02-01

Rice bacterial leaf blight, caused by Xanthomonas oryzae pv. oryzae, and rice bacterial leaf streak, caused by X. oryzae pv. oryzicola, are major diseases of rice. Phenazine-1-carboxylic acid (PCA) is a natural product that is isolated from Pseudomonas spp. and is used to control many important rice diseases in China. We previously reported that PCA disturbs the redox balance, which results in the accumulation of reactive oxygen species in X. oryzae pv. oryzae. In this study, we found that PCA significantly upregulated the transcript levels of catB and katE, which encode catalases, and that PCA sensitivity was reduced when X. oryzae pvs. oryzae and oryzicola were cultured with exogenous catalase. Furthermore, catB deletion mutants of X. oryzae pvs. oryzae and oryzicola showed dramatically decreased total catalase activity, increased sensitivity to PCA, and reduced virulence in rice. In contrast, deletion mutants of srpA and katG, which also encode catalases, exhibited little change in PCA sensitivity. The results indicate that catB in both X. oryzae pvs. oryzae and oryzicola encodes a catalase that helps protect the bacteria against PCA-induced stress.

Comparative analysis for the production of fatty acid alkyl esterase using whole cell biocatalyst and purified enzyme from Rhizopus oryzae on waste cooking oil (sunflower oil).

Science.gov (United States)

Balasubramaniam, Bharathiraja; Sudalaiyadum Perumal, Ayyappasamy; Jayaraman, Jayamuthunagai; Mani, Jayakumar; Ramanujam, Praveenkumar

2012-08-01

The petroleum fuel is nearing the line of extinction. Recent research and technology have provided promising outcomes to rely on biodiesel as the alternative and conventional source of fuel. The use of renewable source - vegetable oil constitutes the main stream of research. In this preliminary study, Waste Cooking Oil (WCO) was used as the substrate for biodiesel production. Lipase enzyme producing fungi Rhizopus oryzae 262 and commercially available pure lipase enzyme were used for comparative study in the production of Fatty Acid Alkyl Esters (FAAE). The whole cell (RO 262) and pure lipase enzyme (PE) were immobilized using calcium alginate beads. Calcium alginate was prepared by optimizing with different molar ratios of calcium chloride and different per cent sodium alginate. Entrapment immobilization was done for whole cell biocatalyst (WCB). PE was also immobilized by entrapment for the transesterification reaction. Seven different solvents - methanol, ethanol, n-propanol, n-butanol, iso-propanol, iso-butanol and iso-amyl alcohol were used as the acyl acceptors. The reaction parameters like temperature (30°C), molar ratio (1:3 - oil:solvent), reaction time (24 h), and amount of enzyme (10% mass ratio to oil) were also optimized for methanol alone. The same parameters were adopted for the other acyl acceptors too. Among the different acyl acceptors - methanol, whose reaction parameters were optimized showed maximum conversion of triglycerides to FAAE-94% with PE and 84% with WCB. On the whole, PE showed better catalytic converting ability with all the acyl acceptor compared to WCB. Gas chromatography analysis (GC) was done to determine the fatty acid composition of WCO (sunflower oil) and FAAE production with different acyl acceptors. Copyright © 2012 Elsevier Ltd. All rights reserved.

Effect of solid-state fermentation with Rhizopus oligosporus on bioactive compounds and antioxidant capacity of raw and roasted buckwheat groats

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Wronkowska Małgorzata

2015-12-01

Full Text Available The effect of solid-state fermentation with Rhizopus oligosporus on the changes in the total phenolic compounds, rutin, vitamin B and C, tocopherol, phytic acid and antioxidant capacity of raw and roasted buckwheat groats was studied. The roasted groats contained reduced level of studied bioactive compounds as compared to raw groats. In this study was evidenced that the solidstate fermentation with Rhizopus oligosporus enhanced water soluble vitamins (thiamine, pyridoxine and L-ascorbic acid as well as tocopherols contents. In contrast the decrease of the inositol hexaphosphate, phenolic compounds, the rutin content and antioxidant capacity determined by ACL and ABTS methods was noticed.

Biosorptive removal of Hg(II) ions by Rhizopus oligosporus ...

African Journals Online (AJOL)

In this study, corn processing wastewater was used as a new low-cost substrate to produce Rhizopus oligosporus. Dried biomass of R. oligosporus was evaluated as a biosorbent for treatment of synthetically contaminated waters with Hg(II) ions. The biosorption process was carried out in a batch process and the effects of ...

21 CFR 184.1420 - Lipase enzyme preparation derived from Rhizopus niveus.

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Lipase enzyme preparation derived from Rhizopus... GENERALLY RECOGNIZED AS SAFE Listing of Specific Substances Affirmed as GRAS § 184.1420 Lipase enzyme preparation derived from Rhizopus niveus. (a) Lipase enzyme preparation contains lipase enzyme (CAS Reg. No...

Effect of Seed Quality and Combination Fungicide-Trichoderma spp. Seed Treatments on Pre- and Postemergence Damping-Off in Cotton.

Science.gov (United States)

Howell, Charles R

2007-01-01

ABSTRACT Good quality seeds of cotton cultivars often escaped pre-emergence damping-off incited by Pythium spp. and Rhizopus oryzae, and they were resistant to postemergence damping-off incited by Rhizoctonia solani. Poor quality seeds, however, were highly susceptible to both phases of seedling disease and required seed treatment in order to survive. Pre-emergence damping-off incited by Pythium spp. and Rhizopus oryzae could be controlled by seed treatment with biocontrol preparations of a number of Trichoderma spp., but these treatments were much less effective in controlling postemergence disease incited by Rhizoctonia solani. Postemergence seedling disease can be controlled by fungicides, but they were much less effective in controlling the pre-emergence phase of the disease. Combination seed treatments of poor quality cotton seeds with fungicides and Trichoderma spp. preparations, followed by planting in pathogen-infested soil, indicated that this technique will control both phases of seedling disease. Seed treatment with either the fungicides or the biocontrol agents alone did not achieve this goal. The optimum combination treatment for disease control was that of chloroneb plus Trichoderma spp., followed by chloroneb plus metalaxyl (Deltacoat AD) plus T. virens strain G-6.

Effect of culture medium on polymer production and temperature on recovery of polymer produced from newly identified Rhyzopus oryzae ST29

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Tipparat Hongpattarakere

2008-04-01

Full Text Available Thermotolerant fungal isolate ST29 was identified by observing on cell morphology and molecular technique based on internal transcribed spacer (ITS gene to be Rhizopus oryzae. Among four culture media tested, the strain exhibited the highest growth in yeast malt extract (YM medium (4.87 g/l, followed by Sabouraud dextrose broth (SDB (4.25 g/l, potato dextrose broth (PDB (4.10 g/l and palm oil mill effluent (POME (3.29 g/l, respectively, after 4 days cultivation at 45oC. However, the strain was found to produce polymer only in POME medium at 45oC, but not in the three synthetic media tested. Effect of temperature on separation of the biopolymer produced by this fungal strain was studied by incubating the culture broth in water bath with temperatures in the range of room temperature to 70oC. The biopolymer was recovered by filtration, centrifugation, and precipitation by adding 4 volumes of 95% ethanol, then freeze-drying. These temperatures therefore had no influence on the biopolymer yields (5.58-5.78 g/l or on biomass yields (2.90-3.29 g/l.

Effects of aqueous extract of Cinnamomum verum on growth of bread spoilage fungi

OpenAIRE

Monir Doudi; Mahbubeh Setorki; Zahra Rezayatmand

2016-01-01

Food waste has been identified as a considerable problem and bread is the most wasted food. This study aimed to evaluate In-vitro anti-fungal activity of cinnamon extract on bread spoilage fungi and to determine its anti-fungal effect in the bread slices. At first, the MIC and MFC values of the extract were determined against Aspergillus niger, Aspergillus flavus, Aspergillus fumigatus, Penicillium chrysogenum, Penicillium notatum and Rhizopus oryzae. Then, Aspergillus sp was sele...

Isolation of Mucorales from processed maize (Zea mays L.) and screening for protease activity

OpenAIRE

de Azevedo Santiago, Andr? Luiz Cabral Monteiro; de Souza Motta, Cristina Maria

2008-01-01

Mucorales were isolated from maize flour, corn meal and cooked cornflakes using surface and depth plate methods. Rhizopus oryzae, Circinella muscae, Mucor subtilissimus, Mucor hiemalis f. hiemalis, Syncephalastrum racemosum, Rhizopus microsporus var. chinensis and Absidia cylindrospora showed protease activity.

Isolation of Mucorales from processed maize (Zea mays L.) and screening for protease activity

Science.gov (United States)

de Azevedo Santiago, André Luiz Cabral Monteiro; de Souza Motta, Cristina Maria

2008-01-01

Mucorales were isolated from maize flour, corn meal and cooked cornflakes using surface and depth plate methods. Rhizopus oryzae, Circinella muscae, Mucor subtilissimus, Mucor hiemalis f. hiemalis, Syncephalastrum racemosum, Rhizopus microsporus var. chinensis and Absidia cylindrospora showed protease activity. PMID:24031292

Cyanide removal by combined adsorption and biodegradation process

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R. Roshan Dash, Ch. Balomajumder, A. Kumar

2006-04-01

Full Text Available Investigation of the effectiveness of simultaneous adsorption and biodegradation (SAB process over individual processes by using microbes Rhizopus oryzae and Stemphylium loti with granular activated carbon (GAC as adsorbent was carried out. The maximum removal efficiency of cyanide had been achieved by biodegradation alone was 83% by R. oryzae, while it was 90% by S. loti at initial pH of 5.6 and 7.2 respectively and at initial CN- concentration of 150 mg/L. In the combined process efficiency of R. oryzae closer to S. loti (95.3% and 98.6% respectively

Rheology, microstructure and baking characteristics of frozen dough containing Rhizopus chinensis lipase and transglutaminase

Science.gov (United States)

The beneficial effects of a new recombinant lipase (Rhizopus chinensis lipase, RCL) and transglutaminase (TG) were investigated on frozen dough systems and their breadmaking quality. Rheological properties and microstructure of doughs were measured using a dynamic rheometer, rheofermentometer F3, an...

Autolysis of Aspergillus oryzae Mycelium and Effect on Volatile Flavor Compounds of Soy Sauce.

Science.gov (United States)

Xu, Ning; Liu, Yaqi; Hu, Yong; Zhou, Mengzhou; Wang, Chao; Li, Dongsheng

2016-08-01

The autolyzed mycelia of Aspergillus oryzae are rich in proteins, nucleic acids, sugar, and other biomacromolecules, and are one of the main contributors to the flavor profile of commercially important fermented goods, including soy sauce and miso. We induced autolysis of the mycelia of A. oryzae over 1 to 10 d, and found that the maximum dissolved amounts of total protein and nucleic acid ratio accounted for 28.63% and 88.93%, respectively. The organic acid content, such as citric acid, tartaric acid, succinic acid, lactic acid, and acetic acid, initially increased and then decreased as autolysis progressed, corresponding to changes in pH levels. The main characteristic flavor compounds in soy sauce, namely, ethanol, 2-phenylethanol, and 2-methoxy-4-vinylphenol, were all detected in the autolysate. Subsequently, we tested the effect of adding mycelia of A. oryzae during the fermentation process of soy sauce for 60 d, and found that addition of 1.2‰ A. oryzae mycelia provided the richest flavor. Overall, our findings suggest that compounds found in the autolysate of A. oryzae may promote the flavor compounds of soy sauce, such as alcohols, aldehydes, phenols, and esters. © 2016 Institute of Food Technologists®

Antifungal properties of Musa paradisiaca (Plantain) peel and stalk ...

African Journals Online (AJOL)

Effect of plantain (Musa paradisiaca (L) AAB genomic group) peel and stalk extracts were investigated using percentage inhibition test. Complete inhibition of growth (100%) was observed for Aspergillusniger, Aspergillus oryzae and Rhizopus stolonifer at 1.0 mg/ml concentration of stalk extract. Peel extract inhibited A.

Lethal and mutagenic effects of ion beams and γ-rays in Aspergillus oryzae

International Nuclear Information System (INIS)

Toyoshima, Yoshiyuki; Takahashi, Akemi; Tanaka, Hisaki; Watanabe, Jun; Mogi, Yoshinobu; Yamazaki, Tatsuo; Hamada, Ryoko; Iwashita, Kazuhiro; Satoh, Katsuya; Narumi, Issay

2012-01-01

Highlights: ► We investigated the effects of different LET radiation in A. oryzae. ► Both γ-rays and ion beams induced base substitutions, frameshifts, deletions. ► Both γ-rays and ion beams induced genome-wide large-scale mutations in A. oryzae. ► Some differences in the types and frequencies of mutations were found. ► Our results provide new basic insights into the mutation breeding of A. oryzae. - Abstract: Aspergillus oryzae is a fungus that is used widely in traditional Japanese fermentation industries. In this study, the lethal and mutagenic effects of different linear energy transfer (LET) radiation in freeze-dried conidia of A. oryzae were investigated. The lethal effect, which was evaluated by a 90% lethal dose, was dependent on the LET value of the ionizing radiation. The most lethal ionizing radiation among that tested was 12 C 5+ ion beams with an LET of 121 keV/μm. The 12 C 5+ ion beams had a 3.6-times higher lethal effect than low-LET (0.2 keV/μm) γ-rays. The mutagenic effect was evaluated by the frequency of selenate resistant mutants. 12 C 6+ ion beams with an LET of 86 keV/μm were the most effective in inducing selenate resistance. The mutant frequency following exposure to 12 C 6+ ion beams increased with an increase in dose and reached 3.47 × 10 −3 at 700 Gy. In the dose range from 0 to 700 Gy, 12 C 5+ ion beams were the second most effective in inducing selenate resistance, the mutant frequency of which reached a maximum peak (1.67 × 10 −3 ) at 400 Gy. To elucidate the characteristics of mutation induced by ionizing radiation, mutations in the sulphate permease gene (sB) and ATP sulfurylase gene (sC) loci, the loss of function of which results in a selenate resistant phenotype, were compared between 12 C 5+ ion beams and γ-rays. We detected all types of transversions and transitions. For frameshifts, the frequency of a +1 frameshift was the highest in all cases. Although the incidence of deletions >2 bp was generally low

Bio-synthesis and hydrolysis of ethyl phenylacetate and ethyl 2-phenylpropionate in organic solvent by lyophilized mycelia Biossíntese e hidrólise de fenilacetato de etila e 2-fenilpropionato de etila em solvente orgânico por meio de micélios liofilizados

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Paolo Torre

2007-06-01

Full Text Available To select the best biocatalysts for ethanol acylations with phenylacetic and 2-phenylpropionic acids, lyophilized mycelia of Aspergillus oryzae CBS 10207, A. oryzae MIM, Rhizopus oryzae CBS 11207, R. oryzae CBS 39134, R. oryzae CBS 26028 and R. oryzae CBS 32847 were tested in this study. The carboxylesterase activities of A. oryzae MIM and R. oryzae 11207, which revealed to be the best biocatalysts, were investigated either in 0.1 M phosphate buffer or in n-heptane to catalyze the hydrolysis or the synthesis of ethyl esters of these acids, respectively. A. oryzae proved more effective than R. oryzae, probably due to more favorable microenvironment conditions and thermodynamic scenario. The results in terms of product formation and substrate consumption versus time were used to estimate the maximum conversion yields, the equilibrium constants and the times needed to reach half maximum conversion, thus providing sufficient information about these equilibria.Micélios liofilizados de Aspergillus oryzae CBS 10207, A. oryzae MIM, Rhizopus oryzae CBS 11207, R. oryzae CBS 39134, R. oryzae CBS 26028 e R. oryzae CBS 32847 foram testados neste estudo com vista à seleção do melhor biocatalisador para efetuar a acilação de etanol com ácidos fenilacético e 2-fenilpropiônico. As atividades carboxilesterásicas de A. oryzae MIM e R. oryzae 11207, que resultaram ser os melhores biocatalisadores, foram investigadas tanto em tampão fosfato 0,1 M como em n-heptano para catalisar a hidrólise ou a síntese dos ésteres etílicos destes ácidos. A. oryzae pareceu ser mais eficaz que R. oryzae, provavelmente devido a condições micro-ambientais e a um cenário termodinâmico mais favoráveis. Os resultados obtidos em termos de formação do produto e consumo dos substratos em função do tempo foram usados para a estimativa dos rendimentos de conversão máximos, as constantes de equilíbrio e os tempos necessários para alcançar metade da conversão m

Molecular detection of Xanthomonas oryzae pv. oryzae, Xanthomonas oryzae pv. oryzicola, and Burkholderia glumae in infected rice seeds and leaves

Science.gov (United States)

Polymerase chain reaction (PCR) is particularly useful for plant pathogen detection. In the present study, multiplex PCR and SYBR green real-time PCR were developed to facilitate simultaneous detection of three important rice pathogens, Xanthomonas oryzae pv. oryzae, X. oryzae pv. oryzicola, and Bur...

Improving the Yield of Glucoamylase and α-amylase in Solid-state Co-culture

OpenAIRE

Takefuji Haruka; Ninomiya Junko; Morita Hiroshi

2016-01-01

Steamed rice inoculated with Aspergillus oryzae, called Koji, is an essential ingredient for making amazake or brewing sake. However, A. oryzae usually offers low enzyme yield, especially in case of glucoamylase. Co-culture of Aspergillus and Rhizopus strains in Koji increased glucoamylase and α-amylase activities. The ratio of initial spore counts of A. oryzae and R. oryzae influenced the activity of amylolytic enzymes. When this ratio was 1:1, α-amylase showed maximum activity (573 U/g-subs...

27 CFR 24.247 - Materials authorized for the treatment of distilling material.

Science.gov (United States)

2010-04-01

... fermentable carbohydrates The amylase enzyme activity shall be derived from Aspergillus niger, Aspergillus... starches to fermentable carbohydrates The amylase enzyme actvity shall be derived from Aspergillus niger or Aspergillus oryzae per FDA advisory opinion dated 8/18/83 or from Rhizopus oryzae per 21 CFR 173.130 or from...

Lethal and mutagenic effects of ion beams and γ-rays in Aspergillus oryzae

Energy Technology Data Exchange (ETDEWEB)

Toyoshima, Yoshiyuki, E-mail: toyoshima@yamasa.com [Soy Sauce Laboratory, Yamasa Corporation, 2-10-1 Araoicho, Choshi, Chiba 288-0056 (Japan); Takahashi, Akemi; Tanaka, Hisaki; Watanabe, Jun; Mogi, Yoshinobu; Yamazaki, Tatsuo [Soy Sauce Laboratory, Yamasa Corporation, 2-10-1 Araoicho, Choshi, Chiba 288-0056 (Japan); Hamada, Ryoko; Iwashita, Kazuhiro [Fundamental Research Division, National Research Institute of Brewing, 3-7-1 Kagamiyama, Higashihiroshima, Hiroshima 739-0046 (Japan); Satoh, Katsuya; Narumi, Issay [Ion Beam Mutagenesis Research Group, Quantum Beam Science Directorate, Japan Atomic Energy Agency, 1233 Watanuki, Takasaki, Gunma 370-1292 (Japan)

2012-12-15

Highlights: ► We investigated the effects of different LET radiation in A. oryzae. ► Both γ-rays and ion beams induced base substitutions, frameshifts, deletions. ► Both γ-rays and ion beams induced genome-wide large-scale mutations in A. oryzae. ► Some differences in the types and frequencies of mutations were found. ► Our results provide new basic insights into the mutation breeding of A. oryzae. - Abstract: Aspergillus oryzae is a fungus that is used widely in traditional Japanese fermentation industries. In this study, the lethal and mutagenic effects of different linear energy transfer (LET) radiation in freeze-dried conidia of A. oryzae were investigated. The lethal effect, which was evaluated by a 90% lethal dose, was dependent on the LET value of the ionizing radiation. The most lethal ionizing radiation among that tested was {sup 12}C{sup 5+} ion beams with an LET of 121 keV/μm. The {sup 12}C{sup 5+} ion beams had a 3.6-times higher lethal effect than low-LET (0.2 keV/μm) γ-rays. The mutagenic effect was evaluated by the frequency of selenate resistant mutants. {sup 12}C{sup 6+} ion beams with an LET of 86 keV/μm were the most effective in inducing selenate resistance. The mutant frequency following exposure to {sup 12}C{sup 6+} ion beams increased with an increase in dose and reached 3.47 × 10{sup −3} at 700 Gy. In the dose range from 0 to 700 Gy, {sup 12}C{sup 5+} ion beams were the second most effective in inducing selenate resistance, the mutant frequency of which reached a maximum peak (1.67 × 10{sup −3}) at 400 Gy. To elucidate the characteristics of mutation induced by ionizing radiation, mutations in the sulphate permease gene (sB) and ATP sulfurylase gene (sC) loci, the loss of function of which results in a selenate resistant phenotype, were compared between {sup 12}C{sup 5+} ion beams and γ-rays. We detected all types of transversions and transitions. For frameshifts, the frequency of a +1 frameshift was the highest in all

OryzaGenome: Genome Diversity Database of Wild Oryza Species

KAUST Repository

Ohyanagi, Hajime

2015-11-18

The species in the genus Oryza, encompassing nine genome types and 23 species, are a rich genetic resource and may have applications in deeper genomic analyses aiming to understand the evolution of plant genomes. With the advancement of next-generation sequencing (NGS) technology, a flood of Oryza species reference genomes and genomic variation information has become available in recent years. This genomic information, combined with the comprehensive phenotypic information that we are accumulating in our Oryzabase, can serve as an excellent genotype-phenotype association resource for analyzing rice functional and structural evolution, and the associated diversity of the Oryza genus. Here we integrate our previous and future phenotypic/habitat information and newly determined genotype information into a united repository, named OryzaGenome, providing the variant information with hyperlinks to Oryzabase. The current version of OryzaGenome includes genotype information of 446 O. rufipogon accessions derived by imputation and of 17 accessions derived by imputation-free deep sequencing. Two variant viewers are implemented: SNP Viewer as a conventional genome browser interface and Variant Table as a textbased browser for precise inspection of each variant one by one. Portable VCF (variant call format) file or tabdelimited file download is also available. Following these SNP (single nucleotide polymorphism) data, reference pseudomolecules/ scaffolds/contigs and genome-wide variation information for almost all of the closely and distantly related wild Oryza species from the NIG Wild Rice Collection will be available in future releases. All of the resources can be accessed through http://viewer.shigen.info/oryzagenome/.

Fungi and some mycotoxins contaminating rice ( Oryza Sativa ) in ...

African Journals Online (AJOL)

The major fungal genera contaminating rice were Aspergillus, Penicillium, Fusarium, Alternaria, Mucor, Rhizopus, Trichoderma, Curvularia, elminthosporium and Cladosporium. The most prevalent fungal species on rice were .Penicillium spp., A. flavus, A. parasiticus, A. niger, Mucor spp., Rhizopus spp. and Alternaria spp.

Upgrading the antioxidant potential of cereals by their fungal fermentation under solid-state cultivation conditions.

Science.gov (United States)

Bhanja Dey, T; Kuhad, R C

2014-11-01

Solid-state fermentation (SSF) at 30°C for 72 h with four generally recognized as safe (GRAS) filamentous fungi (Aspergillus oryzae NCIM 1212, Aspergillus awamori MTCC No. 548, Rhizopus oligosporus NCIM 1215 and Rhizopus oryzae RCK2012) showed high efficiency for the improvement of water-soluble total phenolic content (TPC) and antioxidant properties including ABTS(●+) [2,2'-azinobis (3-ethylbenzothiazoline-6-sulphonic acid)] and DPPH(●) (2,2'-diphenyl-1-picrylhydrazyl) scavenging capacities of four whole grain cereals, namely wheat, brown rice, maize and oat. A maximum 14-fold improvement in TPC (11·61 mg gallic acid equivalent g(-1) grain) was observed in A. oryzae fermented wheat, while extract of R. oryzae fermented wheat (ROFW) showed maximum of 6·6-fold and fivefold enhancement of DPPH(●) scavenging property (8·54 μmol Trolox equivalent g(-1) grain) and ABTS(●+) scavenging activity (19·5 μmol Trolox equivalent g(-1) grain), respectively. The study demonstrates that SSF is an efficient method for the improvement of antioxidant potentials of cereals and R. oryzae RCK2012 fermented wheat can be a powerful source of natural antioxidants. Antioxidant-rich food products are getting popularity day by day. In this study, potential of solid-state fermentation (SSF) has been studied for the improvement of antioxidant potential of different cereals by GRAS micro-organisms. The comparative evaluation of the antioxidant potential of various fungal fermented products derived from whole grain cereals, such as wheat, brown rice, oat and maize, has been carried out. Among these, Rhizopus oryzae RCK2012-fermented wheat was observed as a potent source of natural antioxidants. A diet containing fermented cereals would be useful for the prevention of free radical-mediated diseases. © 2014 The Society for Applied Microbiology.

Radiation resistance of Rhizopus stolonifer

International Nuclear Information System (INIS)

Robbertse, P.J.; Du Toit, T.L.; Van der Merwe, L.J.; Koekemoer, M.L.; Eilers, I.M.I.

1983-01-01

A problem encountered with the irradiation of food is that certain micro-organisms are highly resistant to gamma rays. This includes the fungus, Rhizopus stolonifer, associated with most fruits. The Nuclear Development Corporation of South Africa (NUCOR) has found that a combination of radiation and mild heat treatment reduces the radiation dose necessary to kill 90% of R. stolonifer by approximately half. Treatment at 50 degrees Celsius for 10 minutes or at 55 degrees Celsius for five minutes is sufficient. The article discusses the mechanism of radiation resistance in R. stolonifer and the way in which heating affects this resistance

Differentiation of clinically relevant Mucorales Rhizopus microsporus and R. arrhizus by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS).

Science.gov (United States)

Dolatabadi, Somayeh; Kolecka, Anna; Versteeg, Matthijs; de Hoog, Sybren G; Boekhout, Teun

2015-07-01

This study addresses the usefulness of matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) MS for reliable identification of the two most frequently occurring clinical species of Rhizopus, namely Rhizopus arrhizus with its two varieties, arrhizus and delemar, and Rhizopus microsporus. The test-set comprised 38 isolates of clinical and environmental origin previously identified by internal transcribed spacer (ITS) sequencing of rDNA. Multi-locus sequence data targeting three gene markers (ITS, ACT, TEF ) showed two monophylic clades for Rhizopus arrhizus and Rhizopus microsporus (bootstrap values of 99 %). Cluster analysis confirmed the presence of two distinct clades within Rhizopus arrhizus representing its varieties arrhizus and delemar. The MALDI Biotyper 3.0 Microflex LT platform (Bruker Daltonics) was used to confirm the distinction between Rhizopus arrhizus and Rhizopus microsporus and the presence of two varieties within the species Rhizopus arrhizus. An in-house database of 30 reference main spectra (MSPs) was initially tested for correctness using commercially available databases of Bruker Daltonics. By challenging the database with the same strains of which an in-house database was created, automatic identification runs confirmed that MALDI-TOF MS is able to recognize the strains at the variety level. Based on principal component analysis, two MSP dendrograms were created and showed concordance with the multi-locus tree; thus, MALDI-TOF MS is a useful tool for diagnostics of mucoralean species.

The effect of different phosphate ion concentrations and ph of the phosphate buffer on lipase bioproduction by rhizopus oligosporus

International Nuclear Information System (INIS)

Haq, I.; Ali, S.; Awan, U.F.; Javed, W.; Mirza, S.

2005-01-01

In the present investigation, we report the effect of phosphate ion concentration and different ph of the phosphate buffer (as diluent) on lipase bioproduction by Rhizopus oligosporus. For this purpose, solid state fermentation was employed. Different agricultural by-products such as wheat bran, rice husk, almond meal, soybean meal and sunflower meal were used as substrate. The maximum lipase activity (72.60 U/g) was observed with the almond meal. Addition of phosphate ions (K/sub 2/HPO/sub 4/) influenced the lipase production. The ph of the phosphate buffer (7.0) was found to be effective for higher yield of lipase. (author)

Gamma radiation effects on rice, substrate for Sitophilus oryzae (L.) rearing

International Nuclear Information System (INIS)

Wiendl, F.M.; Arthur, V.; Walder, J.M.M.; Domarco, R.E.

1987-01-01

The gamma radiation effects (800 Krad, 60 Co) on rice are studied. Degraded substances by the radiation and how this degradation affects the biology of Sitophilus oryzae (L.) are searched. The vitamins, proteins and stard of the rice are affected by the radiation. (M.A.C.) [pt

Isolation and identification of fungi responsible for leaf spots disease ...

African Journals Online (AJOL)

The diseased plant leaves were taken to the laboratory for culture, isolation, and ... These included; Alternaria longipes, Aspergillus fumigatus, Aspergillus niger, ... Phomopsis mangiferae, Pseudofusicoccum spp. and Rhizopus oryzae.

Assessment of genetic diversity of Xanthomonas oryzae pv. oryzae

Institute of Scientific and Technical Information of China (English)

无

2000-01-01

@@Bacterial blight of rice, caused by Xanthomonas oryzae pv. Oryzae(Xoo. ), is one of the major rice diseases in China. Making clear the shift of genetic diversity of the pathogen will provide important information for rice breeding. Strains collected from 11 provinces located in Southern region of the Changjiang River in China were assessed by using inoculation method and IS-PCR(Insertion Sequence-Based Polymerase Chain Reaction) analysis.

FERMENTASI BUNGKIL KOPRA DENGAN Rhizopus sp. DAN PEMANFAATANNYA DALAM PAKAN PEMBESARAN IKAN BANDENG DI TAMBAK

Directory of Open Access Journals (Sweden)

Usman Usman

2014-12-01

Full Text Available Bungkil kopra (BK hasil fermentasi dengan Rhizopus sp. memiliki kandungan protein yang lebih tinggi dan lemak yang lebih rendah dibandingkan yang tidak difermentasi, sehingga memiliki potensi dan perlu dimanfaatkan sebagai sumber protein dalam pakan ikan-ikan herbivora-omnivora seperti ikan bandeng. Penelitian ini bertujuan mengamati pemanfaatan bungkil kopra hasil fermentasi dengan Rhizopus sp. sebagai sumber protein dalam pakan untuk pembesaran ikan bandeng di tambak. Penelitian dilakukan dengan menggunakan empat petak tambak masing-masing berukuran 2.500 m2/petak. Ikan uji yang digunakan adalah yuwana ikan bandeng berukuran awal rata-rata 10 g/ekor yang ditebar dengan kepadatan 1.500 ekor/petak, dan diaplikasikan pakan uji pada saat ikan berukuran rata-rata 65 g. Pakan uji yang digunakan adalah pakan buatan yang berbahan baku utama (A tepung bungkil kopra tanpa fermentasi, dan (B tepung bungkil kopra hasil fermentasi dengan Rhizopus sp. Hasil penelitian menunjukkan bahwa laju pertumbuhan spesifik ikan, rasio konversi pakan, dan sintasan ikan tidak berbeda nyata (P>0,05 di antara kedua perlakuan. Namun ikan yang diberi pakan uji B memiliki produksi yang lebih tinggi (P<0,05 daripada yang diberi pakan uji A. Bungkil kopra yang difermentasi dengan Rhizopus sp. dapat dimanfaatkan sebagai sumber protein dalam pakan ikan bandeng.

Optimization of marine waste based-growth media for microbial lipase production using mixture design methodology.

Science.gov (United States)

Sellami, Mohamed; Kedachi, Samiha; Frikha, Fakher; Miled, Nabil; Ben Rebah, Faouzi

2013-01-01

Lipase production by Staphylococcus xylosus and Rhizopus oryzae was investigated using a culture medium based on a mixture of synthetic medium and supernatants generated from tuna by-products and Ulva rigida biomass. The proportion of the three medium components was optimized using the simplex-centroid mixture design method (SCMD). Results indicated that the experimental data were in good agreement with predicted values, indicating that SCMD was a reliable method for determining the optimum mixture proportion of the growth medium. Maximal lipase activities of 12.5 and 23.5 IU/mL were obtained with a 50:50 (v:v) mixture of synthetic medium and tuna by-product supernatant for Staphylococcus xylosus and Rhizopus oryzae, respectively. The predicted responses from these mixture proportions were also validated experimentally.

Toxic effects of essential plant oils in adult Sitophilus oryzae (Linnaeus (Coleoptera, Curculionidae

Directory of Open Access Journals (Sweden)

Andréa Roveré Franz

2011-03-01

Full Text Available Toxic effects of essential plant oils in adult Sitophilus oryzae (Linnaeus (Coleoptera, Curculionidae. Stored grains are subject to losses in quality nutritional value and in sanitation from the time they are stored to the time they are consumed. Botanical insecticides may offer an alternative solution for pest control. The objective was to test the insecticidal properties of the essential oils of Cymbopogon citratus (leaf, Zingiber officinale (root and Mentha sp. (leaf. The efficacy of these oils was tested to control the rice weevil, S. oryzae, using hydrodistillation. Chemical analysis of the essential oils was carried out by gas chromatography. Major components of C. citratus were geranial (48% and neral (31%, of Z. officinale were α-zingibereno (13%, geranial (16%, neral (10% and α-farneseno (5% and of Mentha sp. was menthol (92%. Bioassays were carried out by fumigation and topical application. In topical application assays, the essential oil of C. citratus had greater toxicity (LC50 0.027 µL mL-1 and shorter exposure time than the oils of the other two plants. After 24 h and 48 h, 70% and 100% mortality of S. oryzae occurred, respectively. In fumigation assays, essential oil of Z. officinale had a lower LC50 (1.18 µL cm-2 and 70% mortality after 24 h exposure. Therefore, we recommend the use of essential oils of C. citratus and Z. officinale to control the rice weevil S. oryzae.

Utilization of biological control agents for the management of postharvest pathogens of tomato

International Nuclear Information System (INIS)

Zafar, M.U.; Ansari, S.U.

2016-01-01

Twenty five isolates of Trichoderma, Bacillus and Pseudomonas spp. were obtained from rhizosphere of tomato growing fields using soil dilution technique on potato dextrose agar (PDA) and nutrient agar (NA) medium. Screening of these isolates were done against Geotrichum candidum, Trichothecium roseum and Rhizopus oryzae, causal agents of sour rot, pink mold rot and Rhizopus soft rot of tomato under the laboratory conditions. One promising isolate of each Trichoderma harzianum, Bacillus spp. and Pseudomonas fluorescens from the twenty five isolates were chosen and further evaluated as potential biological control agents (BCAs) against three important postharvest pathogens of tomato. Dual culture and spore concentration assay revealed that all three isolates inhibited radial growth of G. candidum, T. roseum and R. oryzae. Tomato fruits were inoculated with 25 micro L suspension of 10/sup 8/ cfu mL-1 for T. harzianum and 10/sup 8/cfu mL-1for each Bacillus sp. and P. fluorescens. Twenty four hours later the treated fruits were inoculated with 25 micro L of 105 conidia/mL of each of three postharvest pathogens. The results showed that P. fluorescens provided good control (78.1%) of G. candidum and (82.2%) R. oryzae, while, T. harzianum proved less effective to control all three pathogens. Bacillus spp. was only effective (88.4%) against T. roseum. Hence, our results depicted that Bacillus spp. and P. fluorescens proved to be a potential antagonist of T. roseum and R. oryzae however, all the tested BCAs were not consistent in their action against three postharvest pathogens of tomato. (author)

Improving the Yield of Glucoamylase and α-amylase in Solid-state Co-culture

Directory of Open Access Journals (Sweden)

Takefuji Haruka

2016-01-01

Full Text Available Steamed rice inoculated with Aspergillus oryzae, called Koji, is an essential ingredient for making amazake or brewing sake. However, A. oryzae usually offers low enzyme yield, especially in case of glucoamylase. Co-culture of Aspergillus and Rhizopus strains in Koji increased glucoamylase and α-amylase activities. The ratio of initial spore counts of A. oryzae and R. oryzae influenced the activity of amylolytic enzymes. When this ratio was 1:1, α-amylase showed maximum activity (573 U/g-substrate, 95 h, and when this ratio was 200:1, glucoamylase showed maximum activity (180 U/g-substrate, 95 h.

PRODUCTION AND CHARACTERIZATION OF CELLULOLYTIC ENZYMES BY ASPERGILLUS NIGER AND RHIZOPUS SP . BY SOLID STATE FERMENTATION OF PRICKLY PEAR

Directory of Open Access Journals (Sweden)

TAMIRES CARVALHO DOS SANTOS

2016-01-01

Full Text Available Prickly palm cactus husk was used as a solid - state fermentation support substrate for the production of cellulolytic enzymes using Aspergillus niger and Rhizopus sp. A Box - Behnken design was used to evaluate the effects of water activity, fermentation time and temperature on endoglucanase and total cellulase production. Response Surface Methodology showed that optimum conditions for endoglucanase production were achieved at after 70.35 h of fermentation at 29.56°C and a water activity of 0.875 for Aspergillus niger and after 68.12 h at 30.41°C for Rhizopus sp. Optimum conditions for total cellulase production were achieved after 74.27 h of fermentation at 31.22°C for Aspergillus niger and after 72.48 h and 27.86°C for Rhizopus sp . Water activity had a significant effect on Aspergillus niger endoglucanase production only. In industrial applications, enzymatic characterization is important for optimizing variables such as temperature and pH. In this study we showed that endoglucanase and total cellulase had a high level of thermostability and pH stability in all the enzymatic extracts. Enzymatic deactivation kinetic experiments indicated that the enzymes remained active after the freezing of the crude extract. Based on the results, bioconversion of cactus is an excellent alternative for the production of thermostable enzymes.

PENGUJIAN BEBERAPA JENIS INSEKTISIDA NABATI TERHADAP KUTU BERAS (Sitophilus oryzae L

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Muhammad Isnaini

2015-08-01

Full Text Available The purpose of this study first to determine whether or not capable of  natural insecticides to kill Sitophilus oryzae L and to determine effective of  natural insecticides to kill Sitophilus oryzae L. This type of research is an experimental research with quantitative descriptive and The data collection techniques used is the observation and documentation. The data analyzed with ANOVA analysis techniques followed by BJND test. Based on the comparison of the calculated F value contained in the ANOVA analysis is greater than the F table , either at the significant level of 5 % and the 1% significance level (79.57 > 2.67 / 4:43. Thus stated that H1 accepted and rejected H0. The results showed that the first, the natural insecticide able to kill Sitophilus oryzae L and mortality rate highest in Cymbopogon citratus treatment that is equal to 13.2 with the total percentage of mortality of 66%, both effective plant-based insecticides to kill Sitophilus oryzae L that leaves just Cymbopogon citratus (66% and Morinda citrifolia L (60%.The conclusions obtained from this study: First, some kind of  natural insecticide able to kill Sitophilus oryzae L,  second based on the results of the ANOVA analysis of all types of insecticides to kill Sitophilus oryzae L effectively, but if effectiveness was seen by the number of mortality up to 50 % or more, then just Cymbopogon citratus and Morinda citrifolia L.

Interaction between Pyricularia oryzae, four Helminthosporium species and Curvularia lunata in rice leaves

Directory of Open Access Journals (Sweden)

M. Bahous

2003-08-01

Full Text Available The interaction between six fungal parasites of rice: Pyricularia oryzae, Helminthosporium oryzae, H. sativum, H. spiciferum, H. australiensis and Curvularia lunata was studied quantitatively by a modified plant ecology technique known as the de Wit replacement series. Each fungus was inoculated alone or in combination with one of the other five fungi in various proportions into rice plants under experimental conditions. Leaves developing lesions were harvested and incubated in a moist chamber. The yield of each fungus was its conidial production on the rice leaves. The artificial inoculations indicated that interactions between the pathogens in the mixture could be beneficial, antagonistic, or null. Interspecific interaction (i.e. antagonism occurred in the majority of paired combinations (H. oryzae + P. oryzae; H. sativum + H. spiciferum, H. australiensis, C. lunata or P. oryzae; H. australiensis + H. spiciferum, C. lunata or P. oryzae; and P. oryzae + C. lunata. The relative yield total (RYT lines were significantly lower than the expected value, which is 1. The RYT lines were concave upward, revealing a beneficial effect of one or both pathogens on the other, when H. oryzae was in mixture with H. sativum or H. spiciferum. A null effect between fungi occurred in four combinations (H. oryzae + H. australiensis or C. lunata; H. spiciferum + C. lunata; and P. oryzae + H. spiciferum showing that with these combinations inter- and intraspecific competitions were equal in intensity. Thus, the de Wit replacement series technique indicated that it was possible to quantify the interaction between all the pathogenic fungi tested.

Production of Cellulases by Rhizopus stolonifer from Glucose-Containing Media Based on the Regulation of Transcriptional Regulator CRE.

Science.gov (United States)

Zhang, Yingyiing; Tang, Bin; Du, Guocheng

2017-03-28

Carbon catabolite repression is a crucial regulation mechanism in microorganisms, but its characteristic in Rhizopus is still unclear. We extracted a carbon regulation gene, cre , that encoded a carbon catabolite repressor protein (CRE) from Rhizopus stolonifer TP-02, and studied the regulation of CRE by real-time qPCR. CRE responded to glucose in a certain range, where it could significantly regulate part of the cellulase genes ( eg, bg, and cbh2 ) without cbh1 . In the comparison of the response of cre and four cellulase genes to carboxymethylcellulose sodium and a simple carbon source (lactose), the effect of CRE was only related to the concentration of reducing sugars. By regulating the reducing sugars to range from 0.4% to 0.6%, a glucose-containing medium with lactose as the inducer could effectively induce cellulases without the repression of CRE. This regulation method could potentially reduce the cost of enzymes produced in industries and provide a possible solution to achieve the large-scale synthesis of cellulases.

Mucorales (Zygomycotina da Mata Atlântica da Reserva Biológica do Alto da Serra de Paranapiacaba, Santo André, SP Mucorales (Zygomycotina of the Atlantic Rainforest in the "Reserva Biológica do Alto da Serra de Paranapiacaba", Santo André, SP, Brazil

Directory of Open Access Journals (Sweden)

Iracema Helena Schoenlein-Crusius

1997-07-01

Full Text Available Treze táxons de Mucorales (Zygomycotina, distribuídos num total de 266 registros, foram isolados de folhas de Alchornea triplinervia (Spreng. Müll. Arg. colocadas em ambientes terrestre e aquático, e de amostras de solo e de água de riacho, coletadas mensalmente, de julho de 1988 a maio de 1990 na Reserva Biológica do Alto da Serra de Paranapiacaba, no município de Santo André, SP. As espécies que apresentaram os maiores números de registro de ocorrência foram Mucor hiemalis Wehmer (78 registros, Mucor circinelloides van Tieghem f. janssenii (Ledner Schipper (42 registros e Rhizopus arrhizus Fischer (30 registros. Cinqüenta por cento dos Mucorales encontrados na região são constituídos por espécies que estão sendo citadas pela primeira vez para a Mata Atlântica: Mucor hiemalis Wehmer f. silvaticus (Hagem Schipper, Rhizopus arrhizus Fischer, Rhizopus oligosporus Fischer, Rhizopus oryzae (Went. & Prinsen Geerlings, Zygorhynchus japonicus Vuill. e Zygorhynchns macrocarpus Ling-Young.Thirteen taxa of Mucorales (Zygomycotina, distributed in 266 occurrences were isolated from leaves of Alchornea triplinervia (Spreng. Müll. Arg. disposed in the terrestrial and aquatic environment, and of soil and stream water samples, monthly taken from July of 1988 to May of 1990, in the "Reserva Biológica do Alto da Serra de Paranapiacaba", in the municipality of Santo André, São Paulo State, Brazil. Mucor hiemalis Wehmer (78 records, Mucor circinelloides van Tieghem f. janssenii (Ledner Schipper (42 records and Rhizopus arrhizus Fischer (30 records were the most frequent species. Fifty percent of the Mucorales found are formed by species that are mentioned for the first time in the Atlantic Rainforest: Mucor hiemalis Wehmer f. silvaticus (Hagem Schipper, Rhizopus arrhizus Fischer, Rhizopus oligosporus Fischer, Rhizopus oryzae (Went. & Prinsen Geerlings, Zygorhynchus japonicus Vuill. and Zygorhynchus macrocarpus Ling-Young.

Bioconversion of D-psicose to D-tagatose and D-talitol by Mucoraceae fungi.

Science.gov (United States)

Yoshihara, Kazutoshi; Shinohara, Yoshihiro; Hirotsu, Takahiro; Izumori, Ken

2006-03-01

Rhizopus oryzae MYA-2483, which cannot utilize D-psicose as a sole source of carbon, converted D-psicose to two other compounds. These compounds were identified by NMR and IR as D-tagatose and D-talitol. In this study, we describe for the first time the bioconversion of D-psicose to D-tagatose. Various strains of Mucoraceae fungi, to which R. oryzae MYA-2483 belongs, exhibited conversion activity similar to that of R. oryzae MYA-2483. There is the possibility that a considerable number of fungi belonging to Mucoraceae possess such D-psicose conversion activity.

Genome Sequences of Oryza Species

KAUST Repository

Kumagai, Masahiko

2018-02-14

This chapter summarizes recent data obtained from genome sequencing, annotation projects, and studies on the genome diversity of Oryza sativa and related Oryza species. O. sativa, commonly known as Asian rice, is the first monocot species whose complete genome sequence was deciphered based on physical mapping by an international collaborative effort. This genome, along with its accurate and comprehensive annotation, has become an indispensable foundation for crop genomics and breeding. With the development of innovative sequencing technologies, genomic studies of O. sativa have dramatically increased; in particular, a large number of cultivars and wild accessions have been sequenced and compared with the reference rice genome. Since de novo genome sequencing has become cost-effective, the genome of African cultivated rice, O. glaberrima, has also been determined. Comparative genomic studies have highlighted the independent domestication processes of different rice species, but it also turned out that Asian and African rice share a common gene set that has experienced similar artificial selection. An international project aimed at constructing reference genomes and examining the genome diversity of wild Oryza species is currently underway, and the genomes of some species are publicly available. This project provides a platform for investigations such as the evolution, development, polyploidization, and improvement of crops. Studies on the genomic diversity of Oryza species, including wild species, should provide new insights to solve the problem of growing food demands in the face of rapid climatic changes.

Genome Sequences of Oryza Species

KAUST Repository

Kumagai, Masahiko; Tanaka, Tsuyoshi; Ohyanagi, Hajime; Hsing, Yue-Ie C.; Itoh, Takeshi

2018-01-01

This chapter summarizes recent data obtained from genome sequencing, annotation projects, and studies on the genome diversity of Oryza sativa and related Oryza species. O. sativa, commonly known as Asian rice, is the first monocot species whose complete genome sequence was deciphered based on physical mapping by an international collaborative effort. This genome, along with its accurate and comprehensive annotation, has become an indispensable foundation for crop genomics and breeding. With the development of innovative sequencing technologies, genomic studies of O. sativa have dramatically increased; in particular, a large number of cultivars and wild accessions have been sequenced and compared with the reference rice genome. Since de novo genome sequencing has become cost-effective, the genome of African cultivated rice, O. glaberrima, has also been determined. Comparative genomic studies have highlighted the independent domestication processes of different rice species, but it also turned out that Asian and African rice share a common gene set that has experienced similar artificial selection. An international project aimed at constructing reference genomes and examining the genome diversity of wild Oryza species is currently underway, and the genomes of some species are publicly available. This project provides a platform for investigations such as the evolution, development, polyploidization, and improvement of crops. Studies on the genomic diversity of Oryza species, including wild species, should provide new insights to solve the problem of growing food demands in the face of rapid climatic changes.

Screening of systemic fungicides and biochemicals against seed ...

African Journals Online (AJOL)

Jane

2011-07-18

Jul 18, 2011 ... Aspergillus flavus showed highest percentage that is, 27.3% followed by Rhizopus stolonifer 17.98%,. Alternaria .... characteristics were cultured on PDA medium. ..... Fungi of Rice (Oryzae sativa L) Variety Faro 15 In Vitro.

Identification and characterization of milk-clotting proteases ...

African Journals Online (AJOL)

SAM

2014-03-12

Mar 12, 2014 ... Two strains of fungi were isolated and identified as Aspergillus tamarii and Penicillium pinophilum ... for milk clotting enzymes by fermentation at acid pH on culture medium containing whey ..... Rhizopus oryzae NBRC 4749.

Inkorporasi Kromium pada Khamir dan Kapang dengan Substrat Dasar Singkong yang Diberi Kromium Anorganik

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W. D. Astuti

2006-08-01

Full Text Available Organic-chromium (Cr has higher availability for animals than inorganic-Cr sources. One of organic-Cr sources known is fungi contained high Cr. This organic-Cr was incorporated to fungi protein. Information about the best fungi or microorganisms used in organic-Cr production is still limited. The objective of this experiment was to study organic-Cr production using different kinds of fungi as Cr carrier. Organic-Cr production was conducted in a 4x3 factorial completely randomized experimental design with 3 replications. Four fungi used as carriers in organic-Cr production as the first factor were Saccharomyces cerevisiae, Aspergillus oryzae, Rhizopus oryzae and “ragi tape”. The second factor was levels of Cr addition (500, 1000, 1500 mg/kg. Addition of 1000 mg Cr/kg substrate gave the highest Cr incorporation to fungi protein (484 mg/kg and Cr incorporation efficiency (21.1%. The best fungi used as carrier in organic-Cr production was Rhizopus oryzae which gave the highest Cr incorporation (488 mg/kg and incorporation efficiency (24.7%.

Mucorales isolados do solo de mineração de cobre e produção de amilase e inulinase Mucorales isolated from copper-mining soils and production of amylase and inulinase

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André Luiz Cabral Monteiro Azevedo Santiago

2006-09-01

different species of Mucorales were obtained by the serial-dilution and soil-plate methods. Absidia blakesleeana Lendner, A. cylindrospora Hagem, A. hialospora (Saito Lendn., Cunninghamella elegans Lendner, Rhizopus microsporus V. Thieghen, R. oryzae Went. & Prinsen Geerl. and Syncephalastrum racemosum (Cohn. Schroet. were isolated in the dry season. Absidia blakesleeana and A. hialospora were isolated only in the wet season. Rhizopus oryzae had the greatest number of isolates (30.43% followed by A. blakesleeana (26.09% and C. elegans (21.74%. The genus Absidia had the greatest number of species followed by Rhizopus. The number of isolates obtained from each sample site was significantly different, proving the negative effects of environmental impact on Mucorales abundance in the affected soils. All species were capable of degrading starch. However, C. elegans was the only species that also degraded inulin. Isolates of the same species from different areas showed the same behavior as regards capacity to degrade starch and inulin.

Biosorption of Acid Blue 290 (AB 290) and Acid Blue 324 (AB 324) dyes on Spirogyra rhizopus

International Nuclear Information System (INIS)

Ozer, Ayla; Akkaya, Goenuel; Turabik, Meral

2006-01-01

In this study, the biosorption of Acid Blue 290 and Acid Blue 324 on Spirogyra rhizopus, a green algae growing on fresh water, was studied with respect to initial pH, temperature, initial dye concentration and biosorbent concentration. The optimum initial pH and temperature values for AB 290 and AB 324 biosorption were found to be 2.0, 30 deg. C and 3.0, 25 deg. C, respectively. It was observed that the adsorbed AB 290 and AB 324 amounts increased with increasing the initial dye concentration up to 1500 and 750 mg/L, respectively. The Langmuir, Freundlich, Redlich-Peterson and Koble-Corrigan isotherm models were applied to the experimental equilibrium data and the isotherm constants were determined by using Polymath 4.1 software. The monolayer coverage capacities of S. rhizopus for AB 290 and AB 324 dyes were found as 1356.6 mg/g and 367.0 mg/g, respectively. The intraparticle diffusion model and the pseudo-second order kinetic model were applied to the experimental data in order to describe the removal mechanism of these acidic dyes by S. rhizopus. The pseudo-second order kinetic model described very well the biosorption kinetics of AB 290 and AB 324 dyes. Thermodynamic studies showed that the biosorption of AB 290 and AB 324 on S. rhizopus was exothermic in nature

Effect of gamma irradiation on the long headed flour beetle latheticus oryzae

Energy Technology Data Exchange (ETDEWEB)

Ahmed, Z A

1986-12-31

The application of gamma radiation for controlling insects infesting grain and grain products appears promising and has some advantages over conventional insect control methods. The results arrived at from these studies could be summarized in the following points: A- Effect of gamma irradiation on the different developmental stages of L. oryzae.B-Effect of gamma irradiation on mating competitiveness.C- The effect of gamma radiation on restoration of sperm viability. 16 tabs., 10 figs., 92 refs.

Biosynthesis and Characterization of Zearalenone-14-Sulfate, Zearalenone-14-Glucoside and Zearalenone-16-Glucoside Using Common Fungal Strains

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Antje Borzekowski

2018-03-01

Full Text Available Zearalenone (ZEN and its phase II sulfate and glucoside metabolites have been detected in food and feed commodities. After consumption, the conjugates can be hydrolyzed by the human intestinal microbiota leading to liberation of ZEN that implies an underestimation of the true ZEN exposure. To include ZEN conjugates in routine analysis, reliable standards are needed, which are currently not available. Thus, the aim of the present study was to develop a facilitated biosynthesis of ZEN-14-sulfate, ZEN-14-glucoside and ZEN-16-glucoside. A metabolite screening was conducted by adding ZEN to liquid fungi cultures of known ZEN conjugating Aspergillus and Rhizopus strains. Cultivation conditions and ZEN incubation time were varied. All media samples were analyzed for metabolite formation by HPLC-MS/MS. In addition, a consecutive biosynthesis was developed by using Fusarium graminearum for ZEN biosynthesis with subsequent conjugation of the toxin by utilizing Aspergillus and Rhizopus species. ZEN-14-sulfate (yield: 49% is exclusively formed by Aspergillus oryzae. ZEN-14-glucoside (yield: 67% and ZEN-16-glucoside (yield: 39% are formed by Rhizopus oryzae and Rhizopus oligosporus, respectively. Purities of ≥73% ZEN-14-sulfate, ≥82% ZEN-14-glucoside and ≥50% ZEN-16-glucoside were obtained by 1H-NMR. In total, under optimized cultivation conditions, fungi can be easily utilized for a targeted and regioselective synthesis of ZEN conjugates.

Genetic variation and population structure in Oryza ...

Indian Academy of Sciences (India)

Oryza malampuzhaensis Krish. et Chand. ( 2 n = 4 x = 48 ; Poaceae, Oryza) is endemic to Western Ghats, South India, and shows a highly localized distribution over a small geographical area in this region. This is the most poorly understood taxon in genus Oryza and is often misidentified as O. officinalis owing to their close ...

Effect of yeast culture and Aspergillus oryzae fermentation extract on ruminal characteristics and nutrient digestibility.

Science.gov (United States)

Wiedmeier, R D; Arambel, M J; Walters, J L

1987-10-01

Four nonpregnant and nonlactating Holstein cows fitted with ruminal fistulas were assigned to each of four diets in a 4 X 4 Latin square design. Dietary treatments were 1) basal diet containing 50% concentrate; 2) basal diet plus 90 g/d yeast culture; 3) basal diet plus 2.63 g/d Aspergillus oryzae fermentation extract; 4) basal diet plus 90 g/d of A. oryzae fermentation extract and yeast culture. Cows were fed diets at a rate of 86 g DM/kg BW.75 for 14 d adaptation followed by an 8-d collection period. Digestibility of dry matter was increased by A. oryzae and A. oryzae and yeast culture combination treatments. Digestibility of CP was increased regardless of fungal culture addition. Hemicellulose digestibility, percent ruminal cellulolytic organisms, and acetate to propionate ratio were increased by the addition of fungal supplements.

Aspergillus oryzae-based cell factory for direct kojic acid production from cellulose.

Science.gov (United States)

Yamada, Ryosuke; Yoshie, Toshihide; Wakai, Satoshi; Asai-Nakashima, Nanami; Okazaki, Fumiyoshi; Ogino, Chiaki; Hisada, Hiromoto; Tsutsumi, Hiroko; Hata, Yoji; Kondo, Akihiko

2014-05-18

Kojic acid (5-Hydroxy-2-(hydroxymethyl)-4-pyrone) is one of the major secondary metabolites in Aspergillus oryzae. It is widely used in food, pharmaceuticals, and cosmetics. The production cost, however, is too high for its use in many applications. Thus, an efficient and cost-effective kojic acid production process would be valuable. However, little is known about the complete set of genes for kojic acid production. Currently, kojic acid is produced from glucose. The efficient production of kojic acid using cellulose as an inexpensive substrate would help establish cost-effective kojic acid production. A kojic acid transcription factor gene over-expressing the A. oryzae strain was constructed. Three genes related to kojic acid production in this strain were transcribed in higher amounts than those found in the wild-type strain. This strain produced 26.4 g/L kojic acid from 80 g/L glucose. Furthermore, this strain was transformed with plasmid harboring 3 cellulase genes. The resultant A. oryzae strain successfully produced 0.18 g/L of kojic acid in 6 days of fermentation from the phosphoric acid swollen cellulose. Kojic acid was produced directly from cellulose material using genetically engineered A. oryzae. Because A. oryzae has efficient protein secretion ability and secondary metabolite productivity, an A. oryzae-based cell factory could be a platform for the production of various kinds of bio-based chemicals.

Antagonism of rice phylloplane fungi against Cercospora oryzae

Science.gov (United States)

Mardani, A.; Hadiwiyono

2018-03-01

Narrow brown leaf spot (NBLS) caused by Cercospora oryzae Miyake is one of the important obstacle in rice cultivation that can decrease the productivity up to 40%. It has been known well that some phylloplane fungi are antagonistic to some leaf diseases. Phylloplane fungi of rice however haven’t been studied much and poorly understood as biological control agent of rice pathogen such C. oryzae. The research aimed to study the antagonism of some phylloplane fungi of rice against C. oryzae. At least 14 isolates of phylloplane fungi were collected which consisted of six pathogenic and eight nonpathogenic variants. All of nonpathogenic isolates were antagonistic against C. oryzae both in vitro and only one isolate could not inhibit the infection of the pathogen in vivo. Some isolates were identified as Aspergillus, Mucor, Penicillium, Fusarium, and Trichoderma. The isolate of Mucor and Fusarium could inhibit the highest growth of pathogen on potato dextrose medium that were at 36.0% and 35.5% respectively. Whereas on artificial inoculation on rice, some isolates such Penicillium and Fusarium could inhibit most effectively and were significantly different to Mencozeb application with dosage 5g L-1.

Inhibitory effect of gamma radiation and Nigella sativa seeds oil on growth, spore germination and toxin production of fungi

International Nuclear Information System (INIS)

El-Bazza, Z.E.; Hala, A.F.; El-Fouly, M.E.Z.; El-Tablawy, S.Y.M.

2001-01-01

Twenty samples of Nigella sativa seeds (Black cumin) were purchased from different localities in Egypt. The mold viable count ranged from 1.7x10 1 to 9.8x10 3 c.f.u. Sixty six molds were isolated belonging to six genera Aspergillus, Penicillium, Rhizopus, Mucor, Alternaria and Fusarium. Exposure of seeds samples to different radiation doses showed that a dose level of 6.0 kGy could be considered as a sufficient dose for decontamination of the tested samples. Seven radioresistant isolates were identified as Rhizopus oryzae, Rhizopus stolonifer, Penicillium chrysogenum and Penicillium corylophillum. All the herb samples were found to be free from aflatoxins B 1 , B 2 , G 1 , G 2 and ochratoxin A. One mold isolate was identified as Aspergillus flavus could produce aflatoxin B 1 and G 1 . None of the isolated radioresistant strains could produce mycotoxins. The water activities of seeds were slightly decreased by the storage time and the seeds needed to be stored at relative humidity not more than 85%. The addition of extract volatile and fixed oil from tested seeds to the medium stimulated the growth of isolated Aspergillus sp. (author)

Inhibitory effect of gamma radiation and Nigella sativa seeds oil on growth, spore germination and toxin production of fungi

Science.gov (United States)

Zeinab, E. M. EL-Bazza; Hala, A. Farrag; Mohie, E. D. Z. EL-Fouly; Seham, Y. M. EL-Tablawy

2001-02-01

Twenty samples of Nigella sativa seeds (Black cumin) were purchased from different localities in Egypt. The mold viable count ranged from 1.7×10 1 to 9.8×10 3 c.f.u. Sixty six molds were isolated belonging to six genera Aspergillus, Penicillium, Rhizopus, Mucor, Alternaria and Fusarium. Exposure of seeds samples to different radiation doses showed that a dose level of 6.0 kGy could be considered as a sufficient dose for decontamination of the tested samples. Seven radioresistant isolates were identified as Rhizopus oryzae, Rhizopus stolonifer, Penicillium chrysogenum and Penicillium corylophillum. All the herb samples were found to be free from aflatoxins B 1, B 2, G 1, G 2 and ochratoxin A. One mold isolate was identified as Aspergillus flavus could produce aflatoxin B 1 and G 1. None of the isolated radioresistant strains could produce mycotoxins. The water activities of seeds were slightly decreased by the storage time and the seeds needed to be stored at relative humidity not more than 85%. The addition of extract volatile and fixed oil from tested seeds to the medium stimulated the growth of isolated Aspergillus sp.

Crystallization and preliminary crystallographic studies of LipA, a secretory lipase/esterase from Xanthomonas oryzae pv. oryzae

Energy Technology Data Exchange (ETDEWEB)

Aparna, Gudlur; Chatterjee, Avradip; Jha, Gopaljee; Sonti, Ramesh V.; Sankaranarayanan, Rajan, E-mail: sankar@ccmb.res.in [Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad 500 007 (India)

2007-08-01

The crystallization and preliminary crystallographic studies of LipA, a lipase/esterase secreted by X. oryzae pv. oryzae during its infection of rice plants, are reported. Xanthomonas oryzae pv. oryzae is the causal agent of bacterial leaf blight, a serious disease of rice. Several enzymes that are secreted through the type II secretion system of this bacterium play an important role in the plant–microbe interaction, being important for virulence and also being able to induce potent host defence responses. One of these enzymes is a secretory lipase/esterase, LipA, which shows a very weak homology to other bacterial lipases and gives a positive tributyrin plate assay. In this study, LipA was purified from the culture supernatant of an overexpressing clone of X. oryzae pv. oryzae and two types of crystals belonging to space group C2 but with two different unit-cell parameters were obtained using the hanging-drop vapour-diffusion method. Type I crystals diffract to a maximum resolution of 1.89 Å and have unit-cell parameters a = 93.1, b = 62.3, c = 66.1 Å, β = 90.8°. Type II crystals have unit-cell parameters a = 103.6, b = 54.6, c = 66.3 Å, β = 92.6° and diffract to 1.86 Å. Solvent-content analysis shows one monomer in the asymmetric unit in both the crystal forms.

Rhinocerebral zygomycosis in a diabetic patient

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Mirella Alves da Cunha

2011-04-01

Full Text Available Rhinocerebral zygomycosis is the most frequent form of fungal infection caused by members of the Zygomycetes class. A fatal case of rhinocerebral zygomycosis caused by Rhizopus (oryzae arrhizus with histopathological and mycological diagnosis is reported in a diabetic patient.

Analysis of microbial diversity in Shenqu with different fermentation times by PCR-DGGE

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Tengfei Liu

Full Text Available Abstract Shenqu is a fermented product that is widely used in traditional Chinese medicine (TCM to treat indigestion; however, the microbial strains in the fermentation process are still unknown. The aim of this study was to investigate microbial diversity in Shenqu using different fermentation time periods. DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis profiles indicated that a strain of Pediococcus acidilactici (band 9 is the predominant bacteria during fermentation and that the predominant fungi were uncultured Rhizopus, Aspergillus oryzae, and Rhizopus oryzae. In addition, pathogenic bacteria, such as Enterobacter cloacae, Klebsiella oxytoca, Erwinia billingiae, and Pantoea vagan were detected in Shenqu. DGGE analysis showed that bacterial and fungal diversity declined over the course of fermentation. This determination of the predominant bacterial and fungal strains responsible for fermentation may contribute to further Shenqu research, such as optimization of the fermentation process.

Elusive Origins of the Extra Genes in Aspergillus oryzae

Science.gov (United States)

Khaldi, Nora; Wolfe, Kenneth H.

2008-01-01

The genome sequence of Aspergillus oryzae revealed unexpectedly that this species has approximately 20% more genes than its congeneric species A. nidulans and A. fumigatus. Where did these extra genes come from? Here, we evaluate several possible causes of the elevated gene number. Many gene families are expanded in A. oryzae relative to A. nidulans and A. fumigatus, but we find no evidence of ancient whole-genome duplication or other segmental duplications, either in A. oryzae or in the common ancestor of the genus Aspergillus. We show that the presence of divergent pairs of paralogs is a feature peculiar to A. oryzae and is not shared with A. nidulans or A. fumigatus. In phylogenetic trees that include paralog pairs from A. oryzae, we frequently find that one of the genes in a pair from A. oryzae has the expected orthologous relationship with A. nidulans, A. fumigatus and other species in the subphylum Eurotiomycetes, whereas the other A. oryzae gene falls outside this clade but still within the Ascomycota. We identified 456 such gene pairs in A. oryzae. Further phylogenetic analysis did not however indicate a single consistent evolutionary origin for the divergent members of these pairs. Approximately one-third of them showed phylogenies that are suggestive of horizontal gene transfer (HGT) from Sordariomycete species, and these genes are closer together in the A. oryzae genome than expected by chance, but no unique Sordariomycete donor species was identifiable. The postulated HGTs from Sordariomycetes still leave the majority of extra A. oryzae genes unaccounted for. One possible explanation for our observations is that A. oryzae might have been the recipient of many separate HGT events from diverse donors. PMID:18725939

Elusive origins of the extra genes in Aspergillus oryzae.

Directory of Open Access Journals (Sweden)

Nora Khaldi

Full Text Available The genome sequence of Aspergillus oryzae revealed unexpectedly that this species has approximately 20% more genes than its congeneric species A. nidulans and A. fumigatus. Where did these extra genes come from? Here, we evaluate several possible causes of the elevated gene number. Many gene families are expanded in A. oryzae relative to A. nidulans and A. fumigatus, but we find no evidence of ancient whole-genome duplication or other segmental duplications, either in A. oryzae or in the common ancestor of the genus Aspergillus. We show that the presence of divergent pairs of paralogs is a feature peculiar to A. oryzae and is not shared with A. nidulans or A. fumigatus. In phylogenetic trees that include paralog pairs from A. oryzae, we frequently find that one of the genes in a pair from A. oryzae has the expected orthologous relationship with A. nidulans, A. fumigatus and other species in the subphylum Eurotiomycetes, whereas the other A. oryzae gene falls outside this clade but still within the Ascomycota. We identified 456 such gene pairs in A. oryzae. Further phylogenetic analysis did not however indicate a single consistent evolutionary origin for the divergent members of these pairs. Approximately one-third of them showed phylogenies that are suggestive of horizontal gene transfer (HGT from Sordariomycete species, and these genes are closer together in the A. oryzae genome than expected by chance, but no unique Sordariomycete donor species was identifiable. The postulated HGTs from Sordariomycetes still leave the majority of extra A. oryzae genes unaccounted for. One possible explanation for our observations is that A. oryzae might have been the recipient of many separate HGT events from diverse donors.

Suppressive effects of mycoviral proteins encoded by Magnaporthe oryzae chrysovirus 1 strain A on conidial germination of the rice blast fungus.

Science.gov (United States)

Urayama, Syun-Ichi; Kimura, Yuri; Katoh, Yu; Ohta, Tomoko; Onozuka, Nobuya; Fukuhara, Toshiyuki; Arie, Tsutomu; Teraoka, Tohru; Komatsu, Ken; Moriyama, Hiromitsu

2016-09-02

Magnaporthe oryzae chrysovirus 1 strain A (MoCV1-A) is the causal agent of growth repression and attenuated virulence (hypovirulence) of the rice blast fungus, Magnaporthe oryzae. We previously revealed that heterologous expression of the MoCV1-A ORF4 protein resulted in cytological damage to the yeasts Saccharomyces cerevisiae and Cryptococcus neoformans. Since the ORF4 protein is one of the components of viral particles, we evaluated the inhibitory effects of the purified virus particle against the conidial germination of M. oryzae, and confirmed its suppressive effects. Recombinant MoCV1-A ORF4 protein produced in Pichia pastoris was also effective for suppression of conidial germination of M. oryzae. MoCV1-A ORF4 protein sequence showed significant similarity to 6 related mycoviral proteins; Botrysphaeria dothidea chrysovirus 1, two Fusarium graminearum viruses, Fusarium oxysporum f. sp. dianthi mycovirus 1, Penicillium janczewski chrysovirus and Agaricus bisporus virus 1 in the Chrysoviridae family. Multiple alignments of the ORF4-related protein sequences showed that their central regions (210-591 aa in MoCV1-A ORF4) are relatively conserved. Indeed, yeast transformants expressing the conserved central region of MoCV1-A ORF4 protein (325-575 aa) showed similar impaired growth phenotypes as those observed in yeasts expressing the full-length MoCV1-A ORF4 protein. These data suggest that the mycovirus itself and its encoded viral protein can be useful as anti-fungal proteins to control rice blast disease caused by M. oryzae and other pathogenic fungi. Copyright © 2016 Elsevier B.V. All rights reserved.

Improvement of heterologous protein production in Aspergillus oryzae by RNA interference with alpha-amylase genes.

Science.gov (United States)

Nemoto, Takashi; Maruyama, Jun-ichi; Kitamoto, Katsuhiko

2009-11-01

Aspergillus oryzae RIB40 has three alpha-amylase genes (amyA, amyB, and amyC), and secretes alpha-amylase abundantly. However, large amounts of endogenous secretory proteins such as alpha-amylase can compete with heterologous protein in the secretory pathway and decrease its production yields. In this study, we examined the effects of suppression of alpha-amylase on heterologous protein production in A. oryzae, using the bovine chymosin (CHY) as a reporter heterologous protein. The three alpha-amylase genes in A. oryzae have nearly identical DNA sequences from those promoters to the coding regions. Hence we performed silencing of alpha-amylase genes by RNA interference (RNAi) in the A. oryzae CHY producing strain. The silenced strains exhibited a reduction in alpha-amylase activity and an increase in CHY production in the culture medium. This result suggests that suppression of alpha-amylase is effective in heterologous protein production in A. oryzae.

Lethal and mutagenic effects of ion beams and γ-rays in Aspergillus oryzae.

Science.gov (United States)

Toyoshima, Yoshiyuki; Takahashi, Akemi; Tanaka, Hisaki; Watanabe, Jun; Mogi, Yoshinobu; Yamazaki, Tatsuo; Hamada, Ryoko; Iwashita, Kazuhiro; Satoh, Katsuya; Narumi, Issay

2012-12-01

Aspergillus oryzae is a fungus that is used widely in traditional Japanese fermentation industries. In this study, the lethal and mutagenic effects of different linear energy transfer (LET) radiation in freeze-dried conidia of A. oryzae were investigated. The lethal effect, which was evaluated by a 90% lethal dose, was dependent on the LET value of the ionizing radiation. The most lethal ionizing radiation among that tested was (12)C(5+) ion beams with an LET of 121keV/μm. The (12)C(5+) ion beams had a 3.6-times higher lethal effect than low-LET (0.2keV/μm) γ-rays. The mutagenic effect was evaluated by the frequency of selenate resistant mutants. (12)C(6+) ion beams with an LET of 86keV/μm were the most effective in inducing selenate resistance. The mutant frequency following exposure to (12)C(6+) ion beams increased with an increase in dose and reached 3.47×10(-3) at 700Gy. In the dose range from 0 to 700Gy, (12)C(5+) ion beams were the second most effective in inducing selenate resistance, the mutant frequency of which reached a maximum peak (1.67×10(-3)) at 400Gy. To elucidate the characteristics of mutation induced by ionizing radiation, mutations in the sulphate permease gene (sB) and ATP sulfurylase gene (sC) loci, the loss of function of which results in a selenate resistant phenotype, were compared between (12)C(5+) ion beams and γ-rays. We detected all types of transversions and transitions. For frameshifts, the frequency of a +1 frameshift was the highest in all cases. Although the incidence of deletions >2bp was generally low, deletions >20bp were characteristic for (12)C(5+) ion beams. γ-rays had a tendency to generate mutants carrying a multitude of mutations in the same locus. Both forms of radiation also induced genome-wide large-scale mutations including chromosome rearrangements and large deletions. These results provide new basic insights into the mutation breeding of A. oryzae using ionizing radiation. Crown Copyright © 2012. Published

Xanthomonas oryzae pv oryzae the Causal Agent of Bacterial Leaf Blight of rice: Isolation, Characterization, and Study of Transposon Mutagenesis

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Abdjad Asih Nawangsih

2011-04-01

Full Text Available Xanthomonas oryzae pv oryzae the Causal Agent of Bacterial Leaf Blight of rice: Isolation, Characterization, and Study of Transposon Mutagenesis. X. oryzae pv. oryzae (Xoo causes bacterial leaf blight (BLB of rice (Oryza sativa L., a major disease that constrains production of the staple crop in many countries of the world. Identification of X. oryzae pv. oryzae (Xoo was conducted based on the disease symptoms, pathogenicity, morphological, physiological, and genetic characteristics of bacterial cultures isolated from the infected plants. Fifty bacterial isolates predicted as Xoo have been successfully isolated. They are aerobic, rod shaped, and Gram negative bacteria. The isolates were evaluated for their hypersensitivity in tobacco and pathogenicity in rice plant. Fifty isolates induced hypersensitive reaction in tobacco and showed pathogenicity symptom in rice in different length. Based on physiological test, hypersensitivity and pathogenicity reactions, three bacterial isolates strongly predicted as Xoo, i.e. STG21, STG42, and STG46, were non indole formation, non pigment fluorescent, hydrolyzed casein, catalase activity positive, but negative oxidase. Partial sequencing of 16S rRNA genes of STG21 and STG42 showed 80% and 82% homology with X. oryzae, respectively, while STG46 showed 84% homology with X. campestris. Mini-Tn5 transposon mutagenesis of STG21 generated one of the mutants (M5 lossed it’s ability to induce hypersensitive reaction in tobacco plant and deficient in pathogenicity on rice. The lesion length of rice leaf caused by the mutant M5 decreased up to 80%.

Subsite binding energies of an exo-polygalacturonase using isothermal titration calorimetry

Science.gov (United States)

Thermodynamic parameters for binding of a series of galacturonic acid oligomers to an exo-polygalacturonase, RPG16 from Rhizopus oryzae, were determined by isothermal titration calorimetry. Binding of oligomers varying in chain length from two to five galacturonic acid residues is an exothermic proc...

Control of Blumeria graminis f.sp. hordei by treatment with mycelial extracts from cultured fungi

DEFF Research Database (Denmark)

Haugaard, H.; Jørgensen, H.J.L.; Lyngkjær, M.F.

2001-01-01

, Trichoderma harzianum, Pythium ultimum and Rhizopus stolonifer). Three mycelial extracts from the taxonomically different fungi B. oryzae, A ultimum and R. stolonifer were selected for detailed study. In general the number of colonies formed was reduced by 70-98% compared with controls. Furthermore, the few...

Gamma radiation effects in Sitophilus oryzae (Linne, 1763) (Coleoptera, Curculionidae)

International Nuclear Information System (INIS)

Pacheco, J.M.

1973-01-01

Lethal effects were studied for the egg, larval and pupal stages as well as sterilization of adults of Sitophilus oryzae (L.) by the application of gamma radiation. Irradiation of pupae with an estimated dosis of 7887 ± 158 rad inhibits adult emergency. Hypotheses have been suggested that the apparent density and (or) nutrient value of food substrates decisively influence the sterilizing dosis for this insect. Thus, in rice and maize which are apparently denser and (or) of higher nutrient value than maccaroni, insects are sterilized by a dosis of 7 krad, while in maccaroni the sterilizing dosis decreases to 5 krad. (author)

A novel non-thermostable deuterolysin from Aspergillus oryzae.

Science.gov (United States)

Maeda, Hiroshi; Katase, Toru; Sakai, Daisuke; Takeuchi, Michio; Kusumoto, Ken-Ichi; Amano, Hitoshi; Ishida, Hiroki; Abe, Keietsu; Yamagata, Youhei

2016-09-01

Three putative deuterolysin (EC 3.4.24.29) genes (deuA, deuB, and deuC) were found in the Aspergillus oryzae genome database ( http://www.bio.nite.go.jp/dogan/project/view/AO ). One of these genes, deuA, was corresponding to NpII gene, previously reported. DeuA and DeuB were overexpressed by recombinant A. oryzae and were purified. The degradation profiles against protein substrates of both enzymes were similar, but DeuB showed wider substrate specificity against peptidyl MCA-substrates compared with DeuA. Enzymatic profiles of DeuB except for thermostability also resembled those of DeuA. DeuB was inactivated by heat treatment above 80° C, different from thermostable DeuA. Transcription analysis in wild type A. oryzae showed only deuB was expressed in liquid culture, and the addition of the proteinous substrate upregulated the transcription. Furthermore, the NaNO3 addition seems to eliminate the effect of proteinous substrate for the transcription of deuB.

Improved annotation through genome-scale metabolic modeling of Aspergillus oryzae

DEFF Research Database (Denmark)

Vongsangnak, Wanwipa; Olsen, Peter; Hansen, Kim

2008-01-01

Background: Since ancient times the filamentous fungus Aspergillus oryzae has been used in the fermentation industry for the production of fermented sauces and the production of industrial enzymes. Recently, the genome sequence of A. oryzae with 12,074 annotated genes was released but the number...... to a genome scale metabolic model of A. oryzae. Results: Our assembled EST sequences we identified 1,046 newly predicted genes in the A. oryzae genome. Furthermore, it was possible to assign putative protein functions to 398 of the newly predicted genes. Noteworthy, our annotation strategy resulted...... model was validated and shown to correctly describe the phenotypic behavior of A. oryzae grown on different carbon sources. Conclusion: A much enhanced annotation of the A. oryzae genome was performed and a genomescale metabolic model of A. oryzae was reconstructed. The model accurately predicted...

Evaluation of Rhizopus oligosporus in nursery pig diets on growth performance and nutrient digestibility

Science.gov (United States)

The thin stillage leftover from ethanol production contains biodegradable organic compounds and sufficient micronutrients that are ideal for fungal cultivation of Rhizopus oligosporus (RO). This fungus removes about 60% of the organic material, including the suspended solids and even more of some sp...

The lipolytic activity of Rhizopus microsporus var rhizopodiformis isolat UICC No. 6

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Doni Prayudi

2002-12-01

Full Text Available A study was carried out to examine the effect of peptone concentration (2,5% or 5%, inoculum concentration (0,1%, 0,5% or 1% and incubation period (0 – 96 hours, interval 12 hours on the lipolytic activity of Rhizopus microsporus var rhizopodiformis isolat UICC No. 6. Fermentation was done using the basal medium from Samad and the lipolytic activity on olive oil substrate was measured employing titration method with 0,05M NaOH. Lipolytic activity is expressed as unit/ml and one unit is defined as 1 Ωmol fatty acid liberated per minute. Results show that optimum lipolytic activity was obtained from 1 % inoculum, 5% peptone after 72 hours incubation period.

Characterization of Microbial Communities in Chinese Rice Wine Collected at Yichang City and Suzhou City in China.

Science.gov (United States)

Lü, Yucai; Gong, Yanli; Li, Yajie; Pan, Zejiang; Yao, Yi; Li, Ning; Guo, Jinling; Gong, Dachun; Tian, Yihong; Peng, Caiyun

2017-08-28

Two typical microbial communities from Chinese rice wine fermentation collected in Yichang city and Suzhou city in China were investigated. Both communities could ferment glutinous rice to rice wine in 2 days. The sugar and ethanol contents were 198.67 and 14.47 mg/g, respectively, for rice wine from Yichang city, and 292.50 and 12.31 mg/g, respectively, for rice wine from Suzhou city. Acetic acid and lactic acid were the most abundant organic acids. Abundant fungi and bacteria were detected in both communities by high-throughput sequencing. Saccharomycopsis fibuligera and Rhizopus oryzae were the dominant fungi in rice wine from Suzhou city, compared with R. oryzae , Wickerhamomyces anomalus, Saccharomyces cerevisiae, Mucor indicus , and Rhizopus microsporus in rice wine from Yichang city. Bacterial diversity was greater than fungal diversity in both communities. Citrobacter was the most abundant genus. Furthermore, Exiguobacterium, Aeromonas, Acinetobacter, Pseudomonas, Enterobacter, Bacillus , and Lactococcus were highly abundant in both communities.

Characterization of two thermostable inulinases from Rhizopus oligosporus NRRL 2710

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Saleh A. Mohamed

2015-06-01

Full Text Available Two inulinases (Inu2 and Inu3 were purified from Rhizopus oligosporus NRRL 2710 by chromatography on DEAE-Sepharose and Sephacryl S-200 columns. The molecular weight of Inu2 and Inu3 were determined to be 76 and 30 kDa, respectively. Inu2 and Inu3 had the same pH optimum at 5.0, temperature optimum at 50 and 60 °C, and thermal stability up to 60 and 70 °C for 1 h, respectively. Inu2 and Inu3 had low km values (0.93 and 0.70 mM, respectively indicating the high affinity toward inulin. Mg2+, Ca2+, Zn2+ and EDTA did not significantly influence the enzyme activity. Ni2+, Cu2+, Fe2+ and Co2+ showed a partial inhibitory effect, and Hg2+ had a strong inhibitory effect. p-Chloromercuribenzoate had a partial inhibitory effect on Inu2. From these findings, R. oligosporus inulinases can be beneficial enzymes for industrial enzymatic production of high fructose syrup.

Silver nanoparticle production by Rhizopus stolonifer and its antibacterial activity against extended spectrum β-lactamase producing (ESBL) strains of Enterobacteriaceae

International Nuclear Information System (INIS)

Banu, Afreen; Rathod, Vandana; Ranganath, E.

2011-01-01

Highlights: → Silver nanoparticle production by using Rhizopus stolonifer. → Antibacterial activity of silver nanoparticles against extended spectrum β-lactamase producing (ESBL) strains of Enterobacteriaceae. → Synergistic effect of antibiotics with silver nanoparticles towards ESBL-strains. → Characterization of silver nanoparticles made by UV-vis spectra, scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier transformed infrared (FTIR) spectroscopy, atomic force microscopy (AFM). -- Abstract: This report focuses on the synthesis of silver nanoparticles using the fungus, Rhizopus stolonifer and its antimicrobial activity. Research in nanotechnology highlights the possibility of green chemistry pathways to produce technologically important nanomaterials. Characterization of newly synthesized silver nanoparticles was made by UV-visible absorption spectroscopy, scanning electron microscope (SEM), transmission electron microscope (TEM), Fourier transform infrared (FTIR) spectroscopy and atomic force microscope (AFM). TEM micrograph revealed the formation of spherical nanoparticles with size ranging between 3 and 20 nm. The biosynthesized silver nanoparticles (AgNPs) showed excellent antibacterial activity against ESBL-strains which includes E. coli, Proteus. sp. and Klebsiella sp.

Silver nanoparticle production by Rhizopus stolonifer and its antibacterial activity against extended spectrum {beta}-lactamase producing (ESBL) strains of Enterobacteriaceae

Energy Technology Data Exchange (ETDEWEB)

Banu, Afreen [Department of Microbiology, Gulbarga University, Gulbarga 585106, Karnataka (India); Rathod, Vandana, E-mail: drvandanarathod@rediffmail.com [Department of Microbiology, Gulbarga University, Gulbarga 585106, Karnataka (India); Ranganath, E. [Department of Microbiology, Gulbarga University, Gulbarga 585106, Karnataka (India)

2011-09-15

Highlights: {yields} Silver nanoparticle production by using Rhizopus stolonifer. {yields} Antibacterial activity of silver nanoparticles against extended spectrum {beta}-lactamase producing (ESBL) strains of Enterobacteriaceae. {yields} Synergistic effect of antibiotics with silver nanoparticles towards ESBL-strains. {yields} Characterization of silver nanoparticles made by UV-vis spectra, scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier transformed infrared (FTIR) spectroscopy, atomic force microscopy (AFM). -- Abstract: This report focuses on the synthesis of silver nanoparticles using the fungus, Rhizopus stolonifer and its antimicrobial activity. Research in nanotechnology highlights the possibility of green chemistry pathways to produce technologically important nanomaterials. Characterization of newly synthesized silver nanoparticles was made by UV-visible absorption spectroscopy, scanning electron microscope (SEM), transmission electron microscope (TEM), Fourier transform infrared (FTIR) spectroscopy and atomic force microscope (AFM). TEM micrograph revealed the formation of spherical nanoparticles with size ranging between 3 and 20 nm. The biosynthesized silver nanoparticles (AgNPs) showed excellent antibacterial activity against ESBL-strains which includes E. coli, Proteus. sp. and Klebsiella sp.

Inhibitory effect of gamma radiation and Nigella sativa seeds oil on growth, spore germination and toxin production of fungi

Energy Technology Data Exchange (ETDEWEB)

El-Bazza, Z.E.; Hala, A.F. E-mail: hfarragmassoud@hotmail.com; El-Fouly, M.E.Z.; El-Tablawy, S.Y.M

2001-02-01

Twenty samples of Nigella sativa seeds (Black cumin) were purchased from different localities in Egypt. The mold viable count ranged from 1.7x10{sup 1} to 9.8x10{sup 3} c.f.u. Sixty six molds were isolated belonging to six genera Aspergillus, Penicillium, Rhizopus, Mucor, Alternaria and Fusarium. Exposure of seeds samples to different radiation doses showed that a dose level of 6.0 kGy could be considered as a sufficient dose for decontamination of the tested samples. Seven radioresistant isolates were identified as Rhizopus oryzae, Rhizopus stolonifer, Penicillium chrysogenum and Penicillium corylophillum. All the herb samples were found to be free from aflatoxins B{sub 1}, B{sub 2}, G{sub 1}, G{sub 2} and ochratoxin A. One mold isolate was identified as Aspergillus flavus could produce aflatoxin B{sub 1} and G{sub 1}. None of the isolated radioresistant strains could produce mycotoxins. The water activities of seeds were slightly decreased by the storage time and the seeds needed to be stored at relative humidity not more than 85%. The addition of extract volatile and fixed oil from tested seeds to the medium stimulated the growth of isolated Aspergillus sp. (author)

Effect of Meadowsweet Flower Extract-Pullulan Coatings on Rhizopus Rot Development and Postharvest Quality of Cold-Stored Red Peppers

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Alicja Synowiec

2014-08-01

Full Text Available The study involved an examination of the antifungal activity on red peppers of pullulan coating (P and pullulan coating containing either water-ethanol (P + eEMF or ethanol extract of meadowsweet flowers (P + eEMF. Pullulan was obtained from a culture of Aureobasidium pullulans B-1 mutant. Both non-inoculated peppers and those artificially inoculated with Rhizopus arrhizus were coated and incubated at 24 °C for 5 days. The intensity of the decay caused by Rhizopus arrhizus in the peppers with P and P + eEMF coatings was nearly 3-fold lower, and in the case of P + weEMF 5-fold lower, than that observed in the control peppers. Additionally, the P + weEMF coating decreased, almost two-fold the severity of pepper decay compared to other samples. The influence of coating of pepper postharvest quality was examined after 30 days of storage at 6 °C and 70%–75% RH. All coatings formed a thin and well-attached additional layer of an intensified gloss. During storage, color, total soluble solid content and weight loss of coated peppers were subject to lower changes in comparison with uncoated ones. The results indicate the possibility of the application of pullulan coatings containing MFEs as an alternative to the chemical fungicides used to combat pepper postharvest diseases.

Cell biology of the Koji mold Aspergillus oryzae.

Science.gov (United States)

Kitamoto, Katsuhiko

2015-01-01

Koji mold, Aspergillus oryzae, has been used for the production of sake, miso, and soy sauce for more than one thousand years in Japan. Due to the importance, A. oryzae has been designated as the national micro-organism of Japan (Koku-kin). A. oryzae has been intensively studied in the past century, with most investigations focusing on breeding techniques and developing methods for Koji making for sake brewing. However, the understanding of fundamental biology of A. oryzae remains relatively limited compared with the yeast Saccharomyces cerevisiae. Therefore, we have focused on studying the cell biology including live cell imaging of organelles, protein vesicular trafficking, autophagy, and Woronin body functions using the available genomic information. In this review, I describe essential findings of cell biology of A. oryzae obtained in our study for a quarter of century. Understanding of the basic biology will be critical for not its biotechnological application, but also for an understanding of the fundamental biology of other filamentous fungi.

Gamma radiation effects in Sitophilus oryzae (Linne, 1763) (Coleoptera, Curculionidae); Efeitos da radiacao gama em Sitophilus oryzae (Linne, 1763) (Coleoptera, Curculionidae)

Energy Technology Data Exchange (ETDEWEB)

Pacheco, J M

1974-12-31

Lethal effects were studied for the egg, larval and pupal stages as well as sterilization of adults of Sitophilus oryzae (L.) by the application of gamma radiation. Irradiation of pupae with an estimated dosis of 7887 {+-} 158 rad inhibits adult emergency. Hypotheses have been suggested that the apparent density and (or) nutrient value of food substrates decisively influence the sterilizing dosis for this insect. Thus, in rice and maize which are apparently denser and (or) of higher nutrient value than maccaroni, insects are sterilized by a dosis of 7 krad, while in maccaroni the sterilizing dosis decreases to 5 krad. (author).

Screening of gamma radiation-induced pathogen resistance rice lines against Xanthomonas oryzae pv. oryzae

Energy Technology Data Exchange (ETDEWEB)

Lim, Chan Ju; Lee, Ha Yeon; Kim, Woong Bom; Ahmad, Raza; Moon, Jae Sun; Kwon, Suk Yoon [Korea Research Institute of Beoscience and Biotechnology, Daejeon (Korea, Republic of); Kim, Dong Sub [Korea Atomic Energy Research Institute, Jeongeup (Korea, Republic of)

2010-09-15

Bacterial blight is one of the most serious diseases of rice (Oryza sativa L.), and it has been known that Xanthomonas oryzae pv. oryzae (Xoo) causes this disease symptom. To develop resistance rice cultivars against Xoo, 3,000 lines of M{sub 3}, which were irradiated with gamma ray, were tested by 'scissor-dip method' primarily, and 191 putative resistant lines were selected. In M{sub 4} generation, these lines were screened again with various ways such as measuring of symptom of bacterial blight in leaf, number of tiller, fresh weight, and phenotypic segregation ratio in next generation. Finally, six resistance lines were selected. RT-PCR analysis revealed that these lines displayed high level of R-genes such as Xa21, Pi36, and Pi-ta. These results indicate that mutations by gamma ray cause disruptions of regulatory signal transduction systems of these R-genes. Furthermore, these selected mutants could be useful for the development of rice cultivar resistant to Xoo.

Efficient formation of heterokaryotic sclerotia in the filamentous fungus Aspergillus oryzae.

Science.gov (United States)

Wada, Ryuta; Jin, Feng Jie; Koyama, Yasuji; Maruyama, Jun-ichi; Kitamoto, Katsuhiko

2014-01-01

Heterokaryon formation by hyphal fusion occurs during a sexual/parasexual cycle in filamentous fungi, and therefore, it is biotechnologically important for crossbreeding. In the industrial filamentous fungus Aspergillus oryzae, a parasexual cycle has been reported, and it was recently suggested that sexual reproduction should be possible. However, as A. oryzae enters into hyphal fusion with a much lower frequency than Neurospora crassa, the process of heterokaryon formation has not been extensively characterized in A. oryzae. Here, we developed a detection system for heterokaryon formation by expressing red or green fluorescent proteins in nuclei and conferring uridine/uracil or adenine auxotrophy to MAT1-1 and MAT1-2 strains of A. oryzae. The heterokaryon formation of A. oryzae was investigated in paired culture using the genetically modified strains. No sclerotial formation was observed in the hyphal contact regions of the two strains with the same auxotrophy, whereas numerous sclerotia were formed between the strains with different auxotrophies. In most of the formed sclerotia, the uridine/uracil and adenine auxotrophies were complemented, and both red and green fluorescence were detected, indicating that heterokaryotic fusants were formed by hyphal fusion before or during sclerotial formation. Moreover, overexpressing the sclR gene, which encodes a transcription factor promoting sclerotial formation, increased the number of heterokaryotic sclerotia formed between the two auxotrophic strains. Notably, these effects in sclerotial formation of heterokaryotic fusants were observed independently of the mating type pairing combinations. Taken together, these findings demonstrated that paring of different auxotrophs and sclR overexpression promote the formation of heterokaryotic sclerotia in A. oryzae.

KAJIAN PENGARUH KONSENTRASI Rhizopus sp. SEBAGAI AGEN PENGIKIS PROTEIN TERHADAP MUTU KULIT IKAN GURAMI TERSAMAK

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Ratri Nur Hayati

2013-06-01

Full Text Available The study aimed to assess the effect of protease activity of Rhizopus sp. as a bating agent on the tanned carp leather quality, and their use for commercial leather products. The study was designed with a completely randomized design with four treatment, namely: the concentration of protease from Rhizopus sp. 0.5% (a1, 1.0% (a2, 1.5% (a3 and palkobat 1.0% (a4/control with 3 replications. Parameters were observed that is (1 protease activity of Rhizopus sp and palcobat; (2 Thickness (mm, tensile strength (N/cm2, tear strength (N/cm, elongation (%, enervation (mm, shrinkage temperature (ºC, fat/oil content (% of tanned leather; and (3 level of consumer acceptance (% and added valuead of comercial leather product. The data of tanned leather quality is then compared with SNI 06-4586-1998 concenring freshwater snake skin leather and chrome Burk’s Bay (1996 on the raw material quality leather products. The test results of protease activity of Rhizopus sp. = 7.97 mg/50mg/ jam and palkobat = 13.62 mg/50 mg/hour. The results of the analysis of tanned leather carp qulity sample from each treatment and comparison with the SNI as follows: (1 Thickness: 0.41 (a1, 0.38 (a2, 0.43 (a3, 0.41mm (a4 and ≥ 0.22 (SNI 06-4586-1998. Tensile strength: 1682.67; 1818.17; 1195.57; 1670.55 and 1,000N/cm2 (SNI. Tear strength: 314.87; 310.95; 332.06; 462.22, and 150.00 N/cm (SNI. Elongation: 93.33; 62.67; 56.67; 97.33, and <30.00% (SNI. Enervation: 1.87; 2.20; 1.63, 2.49, and ≥ 2.0 mm (Burk’s Bay, 1996. Shrinkage temperature: 74.00; 95.33; 96.33; 97.33, and ≥ 70.33ºC (SNI. The fat/oil content: 7.84; 8.17; 5.25; 8.49, and 2.00 to 6.00% (SNI. All treatments were tested has met the SNI quality, except elongation and fat/oil content. The level of consumer acceptance of the Oval HP Cover that is (1 aspects of the display (76.67% like and 23.33% dislike, (2 the use of convenience (66.67% like and 33.33% dislike , and (3 price (63.33% accepted and 23.67 reject. The

Some factors affecting reproduction in the Rice weevil Sitophilus Oryzae (L.)

International Nuclear Information System (INIS)

Hasaballa, Z.A.; Abdelkawy, F.K.

1992-01-01

Laboratory investigations on the effects or radiation, type of food and population density on the reproductive potential of the rice weevil. Sitophilus Oryzae were conducted at 30 degree C. and 75% R.H. The results indicate that the survival number of adult weevils infesting wheat grains increased markedly after 45 and 90 days of infestation. The survival and reproductive potential of the rice weevils declined significantly after exposure to gamma rays. This decline was more pronounced after 90 days and appeared to be markedly dose dependent. It was noticed that the rate of reproduction of the rice weevils was greatly influenced by insect crowding, science adults of S. Oryzae reared under crowded conditions failed to increase in numbers for 3 months as was expected. The reproduction of S. Oryzae was affected by the type of food. Wheat grains were more suitable than rice and maize grains as the average survival numbers of S. Oryzae reared on wheat grains were more after 45 and 90 days than those reared on rice and maize grains.3 tab

PENGGUNAAN PROTEASE ASPERGILLUS sp. DAN RHIZOPUS sp. DENGAN KONSENTRASI YANG BERBEDA DALAM TAHAPAN UNHAIRING TERHADAP KUALITAS FISIK DAN LIMBAH CAIR PADA PENYAMAKAN KULIT DOMBA

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Yunus Syafie

2013-10-01

Full Text Available Penelitian ini bertujuan untuk mengetahui aktivitas proteolitik yang dihasilkan jamur Aspergillus sp. dan Rhizopus sp. dalam tahapan unhairing (buang rambut pada proses penyamakan kulit domba serta pengaruh penggunaan dengan konsentrasi berbeda, terhadap kuat tarik, kemuluran, suhu kerut, dan kualitas limbah (pH, BOD, dan COD. Materi yang digunakan yaitu 15 lembar kulit domba awetan garam dibagi 2 bagian sepanjang garis lurus punggung sehingga diperoleh 30 lembar kulit, kulit dibagi secara acak menjadi 10 kelompok. Perlakuan terdiri dari dua belas kombinasi yaitu protease dari Aspergillus sp., Rhizopus sp. serta gabungan antara Aspergillus sp. dan Rhizopus sp. dengan konsentrasi protease 2% (P1, 2,5% (P2, 3% (P3, dan sebagai kontrol P0. Proses unhairing secara konvensional menggunakan bahan kimia Na2S (3% dan kapur Ca(OH2 6% dengan 3 ulangan. Sampel air limbah setelah proses unhairing diambil dan dibawa ke laboratorium untuk uji kualitas. Kulit diproses lebih lanjut menjadi kulit samak glazed. Data yang diperoleh dianalisis menggunakan Rancangan Acak Lengkap pola faktorial 3 x 4, apabila berbeda nyata dilakukan uji banding dengan uji Duncan’s new Multiple Range Test (DMRT. Hasil uji aktivitas proteolitik paling tinggi adalah gabungan antara protease dari Aspergillus sp. dan Rhizopus sp. yaitu sebesar 1.079,17 μM/ml/menit, sedangkan protease Aspergillus sp. dan Rhizopus sp. masing-masing memiliki aktivitas proteolitik sebesar 542,96 μM/ml/menit dan 392,89 μM/ml/menit. Hasil penelitian menunjukkan bahwa penggunaan protease dengan konsentrasi yang berbeda dapat memberikan efek yang positif terhadap kualitas fisik dan limbah cair proses unhairing kulit domba. Konsentrasi protease 2,5% dan 3% dapat meningkatkan nilai kuat tarik dan suhu kerut kulit domba serta menghasilkan kulit yang bersih tanpa ada rambut yang menempel dan struktur serabut kolagen terbuka. Perlakuan protease sangat potensial karena dapat menekan angka BOD dan COD limbah

Pathway and rate-limiting step of glyphosate degradation by Aspergillus oryzae A-F02.

Science.gov (United States)

Fu, Gui-Ming; Chen, Yan; Li, Ru-Yi; Yuan, Xiao-Qiang; Liu, Cheng-Mei; Li, Bin; Wan, Yin

2017-09-14

Aspergillus oryzae A-F02, a glyphosate-degrading fungus, was isolated from an aeration tank in a pesticide factory. The pathway and rate-limiting step of glyphosate (GP) degradation were investigated through metabolite analysis. GP, aminomethylphosphonic acid (AMPA), and methylamine were detected in the fermentation liquid of A. oryzae A-F02, whereas sarcosine and glycine were not. The pathway of GP degradation in A. oryzae A-F02 was revealed: GP was first degraded into AMPA, which was then degraded into methylamine. Finally, methylamine was further degraded into other products. Investigating the effects of the exogenous addition of substrates and metabolites showed that the degradation of GP to AMPA is the rate-limiting step of GP degradation by A. oryzae A-F02. In addition, the accumulation of AMPA and methylamine did not cause feedback inhibition in GP degradation. Results showed that degrading GP to AMPA was a crucial step in the degradation of GP, which determines the degradation rate of GP by A. oryzae A-F02.

Aflatoxin B1 Detoxification by Aspergillus oryzae from Meju, a Traditional Korean Fermented Soybean Starter.

Science.gov (United States)

Lee, Kyu Ri; Yang, Sun Min; Cho, Sung Min; Kim, Myunghee; Hong, Sung-Yong; Chung, Soo Hyun

2016-11-04

Aflatoxins are classified as Group 1 (carcinogenic to humans) by the International Agency for Research on Cancer (IARC). In this study, a total of 134 fungal strains were isolated from 65 meju samples, and two fungal isolates were selected as potential aflatoxin B₁ (AFB₁)-biodetoxification fungi. These fungi were identified as Aspergillus oryzae MAO103 and A. oryzae MAO104 by sequencing the beta-tubulin gene. The two A. oryzae strains were able to degrade more than 90% of AFB1 (initial concentration: 40 µg/L) in a culture broth in 14 days. The mutagenic effects of AFB₁ treated with A. oryzae MAO103 and MAO104 significantly decreased to 5.7% and 6.4%, respectively, in the frame-shift mutation of Ames tests using Salmonella typhimurium TA 98. The base-substituting mutagenicity of AFB₁ was also decreased by the two fungi. Moreover, AFB1 production by A. flavus was significantly decreased by the two A. oryzae strains on soybean-based agar plates. Our data suggest that the two AFB1-detoxification A. oryzae strains have potential application to control AFB₁ in foods and feeds.

Evaluation of Oryza sativa x O. glaberrima derived progenies for ...

African Journals Online (AJOL)

USER

2010-06-28

Jun 28, 2010 ... The genus Oryza has two cultivated species, Asian rice (Oryza sativa L.) and African rice (Oryza glaberrima Steud.) and 22 wild species. O. glaberrima is low yielding but has useful genes for resistance to biotic and abiotic stresses. Introgression lines derived from backcrossing of O. sativa x O. glaberrima,.

Fermentative intensity of L-lactic acid production using self ...

African Journals Online (AJOL)

Tuoyo Aghomotsegin

2016-05-25

May 25, 2016 ... Full Length Research Paper. Fermentative ... This study investigated the medium compositions for .... shaker for 12 or 24 h (12 h for stirred tank fermentor, 24 h for flask) at 32°C, with ..... was about 5% of relative error when compared with .... Rhizopus Oryzae in 3-L airlift bioreactor using response surface.

Dicty_cDB: Contig-U15589-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available DV288387 ) NAAHZ89TF Aedes aegypti - Fat Bodies Normalized (... 111 7e-31 3 ( DW611127 ) CLJ261-B09.y1d-s S...251 Rhizopus oryzae Company Rhizopus oryz... 84 2e-29 3 ( DV279302 ) NAAF564TF Aedes aegypti - Fat Bodies No...2 ( DV281619 ) NAAGT04TF Aedes aegypti - Fat Bodies Normalized (... 74 2e-27 4 ( ...404141 ) 293792124 Nasonia vitripennis Female Pupae Nasoni... 105 3e-27 2 ( DV288878 ) NAAHH95TF Aedes aegypti - Fat Bodie...-25 7 ( DV279303 ) NAAF564TR Aedes aegypti - Fat Bodies Normalized (... 119 5e-25 2 ( GE353884 ) 289177336 N

Dicty_cDB: Contig-U15493-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available rongyloides ratti L2 SL1 TOPO v2 Str... 100 3e-33 2 ( FE846057 ) CAFI528.fwd CAFI Pichia stipitis aerobic dextrose..... 62 2e-32 6 ( FE846193 ) CAFI603.fwd CAFI Pichia stipitis aerobic dextrose... 62 2e-32 6 ( GE907061 ) 9545...erobic dextros... 62 3e-32 6 ( FE846897 ) CAFI976.rev CAFI Pichia stipitis aerobic dextrose... 62 3e-32 6 ( ...Pichia stipitis aerobic dextrose... 62 4e-33 7 ( FG074107 ) UI-FF-IF0-abh-b-20-0-...Pichia stipitis aerobic dextrose... 62 3e-32 6 ( EE006326 ) ROE00003876 Rhizopus oryzae Company Rhizopus ory

Genetic analysis of conidiation regulatory pathways in koji-mold Aspergillus oryzae.

Science.gov (United States)

Ogawa, Masahiro; Tokuoka, Masafumi; Jin, Feng Jie; Takahashi, Tadashi; Koyama, Yasuji

2010-01-01

Conidia of koji-mold Aspergillus oryzae are often used as starters in the fermented food industry. However, little is known about conidiation regulation in A. oryzae. To improve the productivity of conidia in A. oryzae, it is necessary to understand conidiation regulation in the strain. Therefore, we analyzed the conidiation regulatory system in A. oryzae using 10 kinds of conidiation regulatory gene disruptants. The phenotypes of AorfluG, AorflbA, AorflbB, AorflbC, AorflbD, AorflbE, AorbrlA, AorabaA, AorwetA, and AorfadA mutants are almost identical to those of the corresponding mutants in Aspergillus nidulans. The results indicated that the functions of conidiation regulatory genes are almost conserved between A. oryzae and A. nidulans. However, the severely reduced conidiation phenotype of the AorfluG disruptant in A. oryzae differs from the phenotype of the corresponding mutant in Aspergillus fumigatus in air-exposed culture conditions. These results suggest that A. oryzae, A. nidulans, and A. fumigatus have a G-protein signaling pathway and brlA orthologs in common, and only A. fumigatus has particular brlA activation pathways that are independent of the fluG ortholog. Furthermore, the analyses of AorflbA disruptant and AorfadA dominant-active mutants implicated that AorFadA-mediated G-protein signaling suppresses vegetative growth of A. oryzae.

Deletion of creB in Aspergillus oryzae increases secreted hydrolytic enzyme activity.

Science.gov (United States)

Hunter, A J; Morris, T A; Jin, B; Saint, C P; Kelly, J M

2013-09-01

Aspergillus oryzae has been used in the food and beverage industry for centuries, and industrial strains have been produced by multiple rounds of selection. Targeted gene deletion technology is particularly useful for strain improvement in such strains, particularly when they do not have a well-characterized meiotic cycle. Phenotypes of an Aspergillus nidulans strain null for the CreB deubiquitinating enzyme include effects on growth and repression, including increased activity levels of various enzymes. We show that Aspergillus oryzae contains a functional homologue of the CreB deubiquitinating enzyme and that a null strain shows increased activity levels of industrially important secreted enzymes, including cellulases, xylanases, amylases, and proteases, as well as alleviated inhibition of spore germination on glucose medium. Reverse transcription-quantitative PCR (RT-qPCR) analysis showed that the increased levels of enzyme activity in both Aspergillus nidulans and Aspergillus oryzae are mirrored at the transcript level, indicating transcriptional regulation. We report that Aspergillus oryzae DAR3699, originally isolated from soy fermentation, has a similar phenotype to that of a creB deletion mutant of the RIB40 strain, and it contains a mutation in the creB gene. Collectively, the results for Aspergillus oryzae, Aspergillus nidulans, Trichoderma reesei, and Penicillium decumbens show that deletion of creB may be broadly useful in diverse fungi for increasing production of a variety of enzymes.

Production of extracellular lipases by Rhizopus oligosporus in a stirred fermentor

OpenAIRE

Iftikhar, Tehreema; Niaz, Mubashir; Zia, Muhammad Anjum; Haq, Ikram ul

2010-01-01

The present investigation deals with the kinetics of submerged extracellular lipases fermentation by both wild and mutant strains of Rhizopus oligosporus var. microsporus in a laboratory scale stirred fermentor. Other parameters studied were inoculum size, pH, agitation and rate of aeration. It was found that the growth and lipases production was increased gradually and reached its maximum 9.07± 0.42ª U mL-1 (W) and 42.49 ± 3.91ª U mL-1 (M) after 30h of fermentation for both wild and mutant s...

Bioconversion of Capsaicin by Aspergillus oryzae.

Science.gov (United States)

Lee, Minji; Cho, Jeong-Yong; Lee, Yu Geon; Lee, Hyoung Jae; Lim, Seong-Il; Park, So-Lim; Moon, Jae-Hak

2015-07-08

This study identified metabolites of capsaicin bioconverted by Aspergillus oryzae, which is generally used for mass production of gochujang prepared by fermenting red pepper powder in Korea. A. oryzae was incubated with capsaicin in potato dextrose broth. Capsaicin decreased depending on the incubation period, but new metabolites increased. Five capsaicin metabolites purified from the ethyl acetate fraction of the capsaicin culture were identified as N-vanillylcarbamoylbutyric acid, N-vanillyl-9-hydroxy-8-methyloctanamide, ω-hydroxycapsaicin, 8-methyl-N-vanillylcarbamoyl-6(E)-octenoic acid, and 2-methyl-N-vanillylcarbamoyl-6(Z)-octenoic acid by nuclear magnetic resonance (NMR) and mass spectrometry (MS). The capsaicin metabolites in gochujang were confirmed and quantitated by selective multiple reaction monitoring detection after liquid chromatography electrospray ionization MS using the isolated compounds as external standards. On the basis of the structures of the capsaicin metabolites, it is proposed that capsaicin metabolites were converted by A. oryzae by ω-hydroxylation, alcohol oxidation, hydrogenation, isomerization, and α- and/or β-oxidation.

Comparative proteomics of Rhizopus delemar ATCC 20344 unravels the role of amino acid catabolism in fumarate accumulation

NARCIS (Netherlands)

Odoni, Dorett I.; Tamayo-Ramos, Juan A.; Sloothaak, Jasper; Heck, van Ruben; Martins dos Santos, Vitor A.P.; Graaff, de Leo H.; Suarez-Diez, Maria; Schaap, Peter J.

2017-01-01

The filamentous fungus Rhizopus delemar naturally accumulates relatively high amounts of fumarate. Although the culture conditions that increase fumarate yields are well established, the network underlying the accumulation of fumarate is not yet fully understood. We set out to increase the

Morphological and molecular characterization of fungal pathogen, Magnaphorthe oryzae

International Nuclear Information System (INIS)

Hasan, Nor’Aishah; Rafii, Mohd Y.; Rahim, Harun A.; Ali, Nusaibah Syd; Mazlan, Norida; Abdullah, Shamsiah

2016-01-01

Rice is arguably the most crucial food crops supplying quarter of calories intake. Fungal pathogen, Magnaphorthe oryzae promotes blast disease unconditionally to gramineous host including rice species. This disease spurred an outbreaks and constant threat to cereal production. Global rice yield declining almost 10-30% including Malaysia. As Magnaphorthe oryzae and its host is model in disease plant study, the rice blast pathosystem has been the subject of intense interest to overcome the importance of the disease to world agriculture. Therefore, in this study, our prime objective was to isolate samples of Magnaphorthe oryzae from diseased leaf obtained from MARDI Seberang Perai, Penang, Malaysia. Molecular identification was performed by sequences analysis from internal transcribed spacer (ITS) region of nuclear ribosomal RNA genes. Phylogenetic affiliation of the isolated samples were analyzed by comparing the ITS sequences with those deposited in the GenBank database. The sequence of the isolate demonstrated at least 99% nucleotide identity with the corresponding sequence in GenBank for Magnaphorthe oryzae. Morphological observed under microscope demonstrated that the structure of conidia followed similar characteristic as M. oryzae. Finding in this study provide useful information for breeding programs, epidemiology studies and improved disease management

Morphological and molecular characterization of fungal pathogen, Magnaphorthe oryzae

Science.gov (United States)

Hasan, Nor'Aishah; Rafii, Mohd Y.; Rahim, Harun A.; Ali, Nusaibah Syd; Mazlan, Norida; Abdullah, Shamsiah

2016-02-01

Rice is arguably the most crucial food crops supplying quarter of calories intake. Fungal pathogen, Magnaphorthe oryzae promotes blast disease unconditionally to gramineous host including rice species. This disease spurred an outbreaks and constant threat to cereal production. Global rice yield declining almost 10-30% including Malaysia. As Magnaphorthe oryzae and its host is model in disease plant study, the rice blast pathosystem has been the subject of intense interest to overcome the importance of the disease to world agriculture. Therefore, in this study, our prime objective was to isolate samples of Magnaphorthe oryzae from diseased leaf obtained from MARDI Seberang Perai, Penang, Malaysia. Molecular identification was performed by sequences analysis from internal transcribed spacer (ITS) region of nuclear ribosomal RNA genes. Phylogenetic affiliation of the isolated samples were analyzed by comparing the ITS sequences with those deposited in the GenBank database. The sequence of the isolate demonstrated at least 99% nucleotide identity with the corresponding sequence in GenBank for Magnaphorthe oryzae. Morphological observed under microscope demonstrated that the structure of conidia followed similar characteristic as M. oryzae. Finding in this study provide useful information for breeding programs, epidemiology studies and improved disease management.

Transposable element distribution, abundance and role in genome size variation in the genus Oryza.

Science.gov (United States)

Zuccolo, Andrea; Sebastian, Aswathy; Talag, Jayson; Yu, Yeisoo; Kim, HyeRan; Collura, Kristi; Kudrna, Dave; Wing, Rod A

2007-08-29

The genus Oryza is composed of 10 distinct genome types, 6 diploid and 4 polyploid, and includes the world's most important food crop - rice (Oryza sativa [AA]). Genome size variation in the Oryza is more than 3-fold and ranges from 357 Mbp in Oryza glaberrima [AA] to 1283 Mbp in the polyploid Oryza ridleyi [HHJJ]. Because repetitive elements are known to play a significant role in genome size variation, we constructed random sheared small insert genomic libraries from 12 representative Oryza species and conducted a comprehensive study of the repetitive element composition, distribution and phylogeny in this genus. Particular attention was paid to the role played by the most important classes of transposable elements (Long Terminal Repeats Retrotransposons, Long interspersed Nuclear Elements, helitrons, DNA transposable elements) in shaping these genomes and in their contributing to genome size variation. We identified the elements primarily responsible for the most strikingly genome size variation in Oryza. We demonstrated how Long Terminal Repeat retrotransposons belonging to the same families have proliferated to very different extents in various species. We also showed that the pool of Long Terminal Repeat Retrotransposons is substantially conserved and ubiquitous throughout the Oryza and so its origin is ancient and its existence predates the speciation events that originated the genus. Finally we described the peculiar behavior of repeats in the species Oryza coarctata [HHKK] whose placement in the Oryza genus is controversial. Long Terminal Repeat retrotransposons are the major component of the Oryza genomes analyzed and, along with polyploidization, are the most important contributors to the genome size variation across the Oryza genus. Two families of Ty3-gypsy elements (RIRE2 and Atlantys) account for a significant portion of the genome size variations present in the Oryza genus.

A novel blast resistance gene, Pi54rh cloned from wild species of rice, Oryza rhizomatis confers broad spectrum resistance to Magnaporthe oryzae.

Science.gov (United States)

Das, Alok; Soubam, D; Singh, P K; Thakur, S; Singh, N K; Sharma, T R

2012-06-01

The dominant rice blast resistance gene, Pi54 confers resistance to Magnaporthe oryzae in different parts of India. In our effort to identify more effective forms of this gene, we isolated an orthologue of Pi54 named as Pi54rh from the blast-resistant wild species of rice, Oryza rhizomatis, using allele mining approach and validated by complementation. The Pi54rh belongs to CC-NBS-LRR family of disease resistance genes with a unique Zinc finger (C(3)H type) domain. The 1,447 bp Pi54rh transcript comprises of 101 bp 5'-UTR, 1,083 bp coding region and 263 bp 3'-UTR, driven by pathogen inducible promoter. We showed the extracellular localization of Pi54rh protein and the presence of glycosylation, myristoylation and phosphorylation sites which implicates its role in signal transduction process. This is in contrast to other blast resistance genes that are predicted to be intracellular NBS-LRR-type resistance proteins. The Pi54rh was found to express constitutively at basal level in the leaves, but upregulates 3.8-fold at 96 h post-inoculation with the pathogen. Functional validation of cloned Pi54rh gene using complementation test showed high degree of resistance to seven isolates of M. oryzae collected from different geographical locations of India. In this study, for the first time, we demonstrated that a rice blast resistance gene Pi54rh cloned from wild species of rice provides broad spectrum resistance to M. oryzae hence can be used in rice improvement breeding programme.

Genomics of Aspergillus oryzae: Learning from the History of Koji Mold and Exploration of Its Future

Science.gov (United States)

Machida, Masayuki; Yamada, Osamu; Gomi, Katsuya

2008-01-01

At a time when the notion of microorganisms did not exist, our ancestors empirically established methods for the production of various fermentation foods: miso (bean curd seasoning) and shoyu (soy sauce), both of which have been widely used and are essential for Japanese cooking, and sake, a magical alcoholic drink consumed at a variety of ritual occasions, are typical examples. A filamentous fungus, Aspergillus oryzae, is the key organism in the production of all these traditional foods, and its solid-state cultivation (SSC) has been confirmed to be the secret for the high productivity of secretory hydrolases vital for the fermentation process. Indeed, our genome comparison and transcriptome analysis uncovered mechanisms for effective degradation of raw materials in SSC: the extracellular hydrolase genes that have been found only in the A. oryzae genome but not in A. fumigatus are highly induced during SSC but not in liquid cultivation. Also, the temperature reduction process empirically adopted in the traditional soy-sauce fermentation processes has been found to be important to keep strong expression of the A. oryzae-specific extracellular hydrolases. One of the prominent potentials of A. oryzae is that it has been successfully applied to effective degradation of biodegradable plastic. Both cutinase, responsible for the degradation of plastic, and hydrophobin, which recruits cutinase on the hydrophobic surface to enhance degradation, have been discovered in A. oryzae. Genomic analysis in concert with traditional knowledge and technology will continue to be powerful tools in the future exploration of A. oryzae. PMID:18820080

Efeito da irradiação gama nas características físico-químicas e sensoriais do arroz (Oryza sativa L. e no desenvolvimento de Sitophilus oryzae L. Effect of gamma irradiation on physico-chemical and sensorial characteristics of rice (Oryza sativa L. and on the development of Sitophilus oryzae L.

Directory of Open Access Journals (Sweden)

Cíntia Fernanda Pedroso Zanão

2009-03-01

Full Text Available A pesquisa teve como objetivo verificar a viabilidade da radiação gama como método de conservação do arroz polido (Oryza sativa L.. As amostras foram irradiadas com doses 0,5; 1,0; 3,0; e 5,0 kGy. Foram realizadas análises da quebra do grão no beneficiamento, da longevidade e reprodução do Sitophilus oryzae L., a composição centesimal, o conteúdo de amilose aparente e propriedades de pasta dos amidos, e análises para cor (instrumental e análise sensorial do arroz cru e cozido. Foi utilizado teste de Tukey (p The objective of this research was to verify the viability of the gamma radiation as polished rice (Oryza sativa L. conservation method. The samples were irradiated with doses of 0.5; 1.0; 3.0; and 5.0 kGy. Analysis of the grain breakage during the enriching process, longevity and reproduction of the Sitophilus oryzae L., centesimal composition, apparent amylose content, starch paste properties, color (instrumental, and the sensorial evaluation of raw and cooked rice were performed. It was verified that the irradiation did not change the percentage of grain breakage during the enrichment process, and it caused a negative effect on the development of insects. The irradiation did not change significantly the centesimal composition and the apparent amylase content. The Tukey test (p < 0.05 was conducted to verify the differences between the treatments. Gamma irradiation affected the pasting properties of the rice flour. Pasting parameters as temperature, peak, final viscosity, and setback values showed decreasing values with irradiation doses. Differences were detected in the sensorial aspect among the samples, and the sample irradiated with the dose of 1.0 kGy presented greater averages. Regarding the instrumental color parameter, it was observed the difference in the values b* indicating that the rice changed the white color for yellowish with the increase in the irradiation dose. The irradiation dose of 1.0 kGy proved the best

Secretome of Aspergillus oryzae in Shaoxing rice wine koji.

Science.gov (United States)

Zhang, Bo; Guan, Zheng-Bing; Cao, Yu; Xie, Guang-Fa; Lu, Jian

2012-04-16

Shaoxing rice wine is the most famous and representative Chinese rice wine. Aspergillus oryzae SU16 is used in the manufacture of koji, the Shaoxing rice wine starter culture. In the current study, a comprehensive analysis of the secretome profile of A. oryzae SU16 in Shaoxing rice wine koji was performed for the first time. The proteomic analysis for the identification of the secretory proteins was done using two-dimensional electrophoresis combined with matrix-assisted laser desorption/ionization-tandem time of flight mass spectrometry based on the annotated A. oryzae genome sequence. A total of 41 unique proteins were identified from the secretome. These proteins included 17 extracellular proteins following the classical secretory pathway, and 10 extracellular proteins putatively secreted by the non-classical secretory pathway. The present secretome profile greatly differed from previous reports on A. oryzae growing in other solid-state nutrient sources. Several new secretory or putative secretory proteins were also found. These proteomic data will significantly aid the advancement of research on the secretome of A. oryzae, especially in solid-state cultures, and in elucidating the production process mechanism of Shaoxing rice wine koji. The findings may promote the technological development and innovation of the Shaoxing rice wine industry. Copyright © 2012 Elsevier B.V. All rights reserved.

Antimicrobial effects of Turkish propolis, pollen, and laurel on spoilage and pathogenic food-related microorganisms.

Science.gov (United States)

Erkmen, Osman; Ozcan, Mehmet Musa

2008-09-01

The antimicrobial activities of propolis extract, pollen extract, and essential oil of laurel (Laurus nobilis L.) at concentrations from 0.02% to 2.5% (vol/vol) were investigated on bacteria (Bacillus cereus, Bacillus subtilis, Escherichia coli, Salmonella typhimurium, Staphylococcus aureus, Yersinia enterocolitica, Enterococcus faecalis, and Listeria monocytogenes), yeasts (Saccharomyces cerevisiae and Candida rugosa), and molds (Aspergillus niger and Rhizopus oryzae). Pollen has no antimicrobial effects on the bacteria and fungi tested in the concentrations used. Propolis showed a bactericidal effect at 0.02% on B. cereus and B. subtilis, at 1.0% on S. aureus and E. faecalis, and at 0.2% on L. monocytogenes. The minimum inhibitory concentration of propolis for fungi was 2.5%. Propolis and laurel were ineffective against E. coli and S. typhimurium at the concentrations tested. The results showed that the antimicrobial activity were concentration dependent. Propolis and essential oil of laurel may be used as biopreservative agents in food processing and preservation.

Self-induction system for cellulase production by cellobiose produced from glucose in Rhizopus stolonifer

OpenAIRE

Zhang, Yingying; Tang, Bin; Du, Guocheng

2017-01-01

Cellulolytic fungi have evolved a sophisticated genetic regulatory network of cellulase synthesis to adapt to the natural environment. Even in the absence of lignocellulose, it still secretes low levels of ?constitutive? cellulase for standby application. However, the mechanisms of this constitutive expression remain incompletely understood. Here we identified a cellobiose synthetase (CBS) from Rhizopus stolonifer, which has the capacity to catalyse the synthesis of cellobiose from uridine di...

Enhanced Production of Bovine Chymosin by Autophagy Deficiency in the Filamentous Fungus Aspergillus oryzae

Science.gov (United States)

Maruyama, Jun-ichi; Kitamoto, Katsuhiko

2013-01-01

Aspergillus oryzae has been utilized as a host for heterologous protein production because of its high protein secretory capacity and food-safety properties. However, A. oryzae often produces lower-than-expected yields of target heterologous proteins due to various underlying mechanisms, including degradation processes such as autophagy, which may be a significant bottleneck for protein production. In the present study, we examined the production of heterologous protein in several autophagy (Aoatg) gene disruptants of A. oryzae. We transformed A. oryzae gene disruptants of Aoatg1, Aoatg13, Aoatg4, Aoatg8, or Aoatg15, with a bovine chymosin (CHY) expression construct and found that the production levels of CHY increased up to three fold compared to the control strain. Notably, however, conidia formation by the Aoatg gene disruptants was significantly reduced. As large amounts of conidia are necessary for inoculating large-scale cultures, we also constructed Aoatg gene-conditional expression strains in which the promoter region of the Aoatg gene was replaced with the thiamine-controllable thiA promoter. Conidiation by the resultant transformants was clearly enhanced in the absence of thiamine, while autophagy remained repressed in the presence of thiamine. Moreover, these transformants displayed increased CHY productivity, which was comparable to that of the Aoatg gene disruptants. Consequently, we succeeded in the construction of A. oryzae strains capable of producing high levels of CHY due to defects in autophagy. Our finding suggests that the conditional regulation of autophagy is an effective method for increasing heterologous protein production in A. oryzae. PMID:23658635

Characterization and selection of Bacillus thuringiensis isolates effective against Sitophilus oryzae Caracterização e seleção de isolados de Bacillus thuringiensis efetivos contra Sitophilus oryzae

Directory of Open Access Journals (Sweden)

Najara da Silva

2010-08-01

Full Text Available The entomopathogenic bacterium Bacillus thuringiensis is a control agent with toxic and environmental characteristics that allows the control of pest insects according to the Integrate Pest Management (IPM precepts. In order to find new strains, potentially toxic to Sitophilus oryzae L. 1763 (Coleoptera: Curculinidae, 1.073 strains of B. thuringiensis from parts of Brazil were used. Genetic material was extracted with InstaGene Matrix kit, used for the amplification of sequences in Polymerase chain reaction (PCR, and viewed in 1.5% agarose gel. The gene cry35Ba class was represented by 60 B. thuringiensis isolates (5.6%, which were then subjected to bioassays with S. oryzae larvae. Among the isolates studied, four caused more than 50% mortality in pathogenicity tests, and the isolates 544 and 622 were the most virulent, as determined by CL50 estimates. The four toxic isolates had spherical, bi-pyramidal and cuboid crystals, and a 44-kDa protein was found in sodium dodecyl sulphate - polyacrylamide gel electrophoresis (SDS-PAGE, which coded for the product of cry35Ba genes. These data demonstrate the potential of B. thuringiensis for the management of S. oryzae larvae.A bactéria entomopatogênica Bacillus thuringiensis (Bt é um agente de controle com características tóxicas e ambientais que permitem o controle de insetos-praga de acordo com as premissas do Manejo integrado de pragas (MIP. Com o objetivo de buscar novas linhagens potencialmente tóxicas para Sitophilus oryzae L. 1763 (Coleoptera: Curculinidae, caracterizaram-se molecularmente 1,073 isolados de B. thuringiensis de regiões do Brasil. O material genético foi extraído através do kit InstaGene Matrix, utilizado para a amplificação das seqüências através da técnica de Polymerase chain reaction PCR, sendo os resultados visualizados em gel de agarose 1,5%. A classe do gene cry35Ba foi representada por 60 isolados (5,6% de Bt, os quais foram submetidos a bioensaio com larvas

Agmatine Production by Aspergillus oryzae is Elevated by Low pH During Solid-State Cultivation.

Science.gov (United States)

Akasaka, Naoki; Kato, Saori; Kato, Saya; Hidese, Ryota; Wagu, Yutaka; Sakoda, Hisao; Fujiwara, Shinsuke

2018-05-25

Sake (rice wine) produced by multiple parallel fermentation (MPF) involving Aspergillus oryzae (strain RW) and Saccharomyces cerevisiae under solid-state cultivation conditions contained 3.5 mM agmatine, while that produced from enzymatically saccharified rice syrup by S. cerevisiae contained oryzae under solid-state cultivation (3.1 mM) but not under submerged cultivation, demonstrating that A. oryzae in solid-state culture produces agmatine. The effect of cultivation conditions on agmatine production was examined. Agmatine production was boosted at 30°C and reached the highest level (6.3 mM) at pH 5.3. The addition of l-lactic, succinic, and citric acids reduced the initial culture pH to 3.0, 3.5, and 3.2, respectively, resulting in further increase in agmatine accumulation (8.2, 8.7, and 8.3 mM, respectively). Homogenate from a solid-state culture exhibited a maximum l-arginine decarboxylase (ADC) activity (74 pmol min -1 μg -1 ) at pH 3.0 at 30°C; that from a submerged culture exhibited an extremely low activity (oryzae , even though A. oryzae lacks ADC orthologs and, instead, possesses four ornithine decarboxylases (ODC1-4). Recombinant ODC1 and ODC2 exhibited no ADC activity at acidic pH (pH 4.0>), suggesting that other decarboxylases or an unidentified ADC is involved in agmatine production. IMPORTANCE It has been speculated that, in general, fungi do not synthesize agmatine from l-arginine because they do not possess genes encoding for arginine decarboxylase. Numerous preclinical studies have shown that agmatine exerts pleiotropic effects on various molecular targets, leading to an improved quality of life. In the present study, we first demonstrated that l-arginine was a feasible substrate for agmatine production by the fungus Aspergillus oryzae RW. We observed that the productivity of agmatine by A. oryzae RW was elevated at low pH only during solid-state cultivation. A. oryzae is utilized in the production of various oriental fermented foods. The

In-vitro evaluation of fungicides, plant extracts and bio-control agents against rice blast pathogen magnaporthe oryzae couch

International Nuclear Information System (INIS)

Hajano, J.U.D.; Lodhi, M.; Pathan, M.A.; Khanzada, A.; Shah, G.S.

2012-01-01

Among 5 fungicides viz., Thiophanate-methyl, Carbendazim, Fosetyl-aluminium, Mancozeb and Copper oxychloride, used against the Magnaporthe oryzae, only Mancozeb appeared as the highly effective fungicide that completely inhibited the mycelial growth of the fungus. All other fungicides showed little effect at higher concentrations. The extracts of garlic (Allium sativum L.), neem (Azadirachta indica L.) and calatropis (Calotropis procera L.) when used against M. oryzae by food poisoning method, only higher dose of garlic completely inhibited the mycelial growth of the test fungus. Six bio-control agents viz., Trichoderma harzianum, Trichoderma polysporum, Trichoderma pseudokoningii, Gliocladium virens, Paecilomyces variotii and Paecilomyces lilacinus were used. Maximum mycelial inhibition of M. oryzae was provided by P. lilacinus followed by Trichoderma spp. (author)

In planta gene expression analysis of Xanthomonas oryzae pathovar oryzae, African strain MAI1

Directory of Open Access Journals (Sweden)

Verdier Valérie

2010-06-01

Full Text Available Abstract Background Bacterial leaf blight causes significant yield losses in rice crops throughout Asia and Africa. Although both the Asian and African strains of the pathogen, Xanthomonas oryzae pv. oryzae (Xoo, induce similar symptoms, they are nevertheless genetically different, with the African strains being more closely related to the Asian X. oryzae pv. oryzicola (Xoc. Results Changes in gene expression of the African Xoo strain MAI1 in the susceptible rice cultivar Nipponbare were profiled, using an SSH Xoo DNA microarray. Microarray hybridization was performed comparing bacteria recovered from plant tissues at 1, 3, and 6 days after inoculation (dai with bacteria grown in vitro. A total of 710 bacterial genes were found to be differentially expressed, with 407 up-regulated and 303 down-regulated. Expression profiling indicated that less than 20% of the 710 bacterial transcripts were induced in the first 24 h after inoculation, whereas 63% were differentially expressed at 6 dai. The 710 differentially expressed genes were one-end sequenced. 535 sequences were obtained from which 147 non-redundant sequences were identified. Differentially expressed genes were related to metabolism, secretion and transport, pathogen adherence to plant tissues, plant cell-wall degradation, IS elements, and virulence. In addition, various other genes encoding proteins with unknown function or showing no similarity to other proteins were also induced. The Xoo MAI1 non-redundant set of sequences was compared against several X. oryzae genomes, revealing a specific group of genes that was present only in MAI1. Numerous IS elements were also found to be differentially expressed. Quantitative real-time PCR confirmed 86% of the identified profile on a set of 14 genes selected according to the microarray analysis. Conclusions This is the first report to compare the expression of Xoo genes in planta across different time points during infection. This work shows that

First report of wheat blast caused by magnaporthe oryzae pathotype triticum in Bangladesh

Science.gov (United States)

Wheat blast or ‘brusone’, caused by the ascomycetous fungus Magnaporthe oryzae B.C. Couch (synonym Pyricularia oryzae Cavara), was first identified in 1985 in Brazil. M. oryzae is composed of a range of morphologically identical but genetically different host-specific pathotypes that are specialized...

Rice (Oryza) hemoglobins

Science.gov (United States)

Hemoglobins (Hbs) corresponding to non-symbiotic (nsHb) and truncated (tHb) Hbs have been identified in rice (Oryza). This review discusses the major findings from the current studies on rice Hbs. At the molecular level, a family of the nshb genes, consisting of hb1, hb2, hb3, hb4 and hb5, and a sin...

Growth Simulation and Discrimination of Botrytis cinerea, Rhizopus stolonifer and Colletotrichum acutatum Using Hyperspectral Reflectance Imaging.

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Ye Sun

Full Text Available This research aimed to develop a rapid and nondestructive method to model the growth and discrimination of spoilage fungi, like Botrytis cinerea, Rhizopus stolonifer and Colletotrichum acutatum, based on hyperspectral imaging system (HIS. A hyperspectral imaging system was used to measure the spectral response of fungi inoculated on potato dextrose agar plates and stored at 28°C and 85% RH. The fungi were analyzed every 12 h over two days during growth, and optimal simulation models were built based on HIS parameters. The results showed that the coefficients of determination (R2 of simulation models for testing datasets were 0.7223 to 0.9914, and the sum square error (SSE and root mean square error (RMSE were in a range of 2.03-53.40×10(-4 and 0.011-0.756, respectively. The correlation coefficients between the HIS parameters and colony forming units of fungi were high from 0.887 to 0.957. In addition, fungi species was discriminated by partial least squares discrimination analysis (PLSDA, with the classification accuracy of 97.5% for the test dataset at 36 h. The application of this method in real food has been addressed through the analysis of Botrytis cinerea, Rhizopus stolonifer and Colletotrichum acutatum inoculated in peaches, demonstrating that the HIS technique was effective for simulation of fungal infection in real food. This paper supplied a new technique and useful information for further study into modeling the growth of fungi and detecting fruit spoilage caused by fungi based on HIS.

Influence of the Oil Pequi in the Growth Mycelial and Sporulation of Colletotrichum Musae and Growth Micelial Of Rhizopus sp. in vitro.

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B. L. Mendes

2013-07-01

Full Text Available Abstract: The anthracnose caused by Colletotrichum musae and soft rot caused by Rhizopus sp are post-harvest diseases that influenced quality of exported fruits. To control these diseases fungicides are used, but in light of these waste products in the fruit and the environment, alternative control methods such as using biofungicides have been studied. The objective of this study was to evaluate the effect of pequi oil on mycelia growth and sporulation of Colletotrichum musae and mycelia growth of Rhizopus sp. in vitro. The oil was added in aliquots of 5, 10, 15, 20, 25, and 50L in the center of Petri dishes containing approximately 15mL of Potato-Dextrose-ar (PDA and distributed over the surface of the medium with Drigalskys handle. After 24 hours, a disc of 8mm diameter from mycelium of fungi was peaked to the center of the plates. As a control, were used Petri dishes containing only PDA. Five replicates were used for each aliquot. After analysis, it was concluded that the pequi oil is ineffective for controlling these pathogens because induced mycelium growth and sporulation.Key Words: Phytopathogens, alternative control, anthracnose, soft rot

Comparative Genome Analysis Between Aspergillus oryzae Strains Reveals Close Relationship Between Sites of Mutation Localization and Regions of Highly Divergent Genes among Aspergillus Species

OpenAIRE

Umemura, Myco; Koike, Hideaki; Yamane, Noriko; Koyama, Yoshinori; Satou, Yuki; Kikuzato, Ikuya; Teruya, Morimi; Tsukahara, Masatoshi; Imada, Yumi; Wachi, Youji; Miwa, Yukino; Yano, Shuichi; Tamano, Koichi; Kawarabayasi, Yutaka; Fujimori, Kazuhiro E.

2012-01-01

Aspergillus oryzae has been utilized for over 1000 years in Japan for the production of various traditional foods, and a large number of A. oryzae strains have been isolated and/or selected for the effective fermentation of food ingredients. Characteristics of genetic alterations among the strains used are of particular interest in studies of A. oryzae. Here, we have sequenced the whole genome of an industrial fungal isolate, A. oryzae RIB326, by using a next-generation sequencing system and ...

Identification of Pectin Degrading Enzymes Secreted by Xanthomonas oryzae pv. oryzae and Determination of Their Role in Virulence on Rice

OpenAIRE

Tayi, Lavanya; Maku, Roshan V.; Patel, Hitendra Kumar; Sonti, Ramesh V.

2016-01-01

Xanthomonas oryzae pv.oryzae (Xoo) causes the serious bacterial blight disease of rice. Xoo secretes a repertoire of plant cell wall degrading enzymes (CWDEs) like cellulases, xylanases, esterases etc., which act on various components of the rice cell wall. The major cellulases and xylanases secreted by Xoo have been identified and their role in virulence has been determined. In this study, we have identified some of the pectin degrading enzymes of Xoo and assessed their role in virulence. Bi...

Growth and α-amylase production by strains of Lactobacillus ...

African Journals Online (AJOL)

A cassava starch medium was used to analyse the dynamics of batch growth and α-amylase production of strains of Lactobacillus plantarum and Rhizopus oryzae isolated from cassava dried chips. The strains displayed a growth of 0.5h-1 and 0.55 h-1, a biomass yield on cassava starch of 0.49g/g and 0.5g/g, a maximum ...

Could abiotic stress tolerance in wild relatives of rice be used to improve Oryza sativa?

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Atwell, Brian J; Wang, Han; Scafaro, Andrew P

2014-02-01

Oryza sativa and Oryza glaberrima have been selected to acquire and partition resources efficiently as part of the process of domestication. However, genetic diversity in cultivated rice is limited compared to wild Oryza species, in spite of 120,000 genotypes being held in gene banks. By contrast, there is untapped diversity in the more than 20 wild species of Oryza, some having been collected from just a few coastal locations (e.g. Oryza schlechteri), while others are widely distributed (e.g. Oryza nivara and Oryza rufipogon). The extent of DNA sequence diversity and phenotypic variation is still being established in wild Oryza, with genetic barriers suggesting a vast range of morphologies and function even within species, such as has been demonstrated for Oryza meridionalis. With increasing climate variability and attempts to make more marginal land arable, abiotic and biotic stresses will be managed over the coming decades by tapping into the genetic diversity of wild relatives of O. sativa. To help create a more targeted approach to sourcing wild rice germplasm for abiotic stress tolerance, we have created a climate distribution map by plotting the natural occurrence of all Oryza species against corresponding temperature and moisture data. We then discuss interspecific variation in phenotype and its significance for rice, followed by a discussion of ways to integrate germplasm from wild relatives into domesticated rice. Crown Copyright © 2013. Published by Elsevier Ireland Ltd. All rights reserved.

Survey of the transcriptome of Aspergillus oryzae via massively parallel mRNA sequencing.

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Wang, Bin; Guo, Guangwu; Wang, Chao; Lin, Ying; Wang, Xiaoning; Zhao, Mouming; Guo, Yong; He, Minghui; Zhang, Yong; Pan, Li

2010-08-01

Aspergillus oryzae, an important filamentous fungus used in food fermentation and the enzyme industry, has been shown through genome sequencing and various other tools to have prominent features in its genomic composition. However, the functional complexity of the A. oryzae transcriptome has not yet been fully elucidated. Here, we applied direct high-throughput paired-end RNA-sequencing (RNA-Seq) to the transcriptome of A. oryzae under four different culture conditions. With the high resolution and sensitivity afforded by RNA-Seq, we were able to identify a substantial number of novel transcripts, new exons, untranslated regions, alternative upstream initiation codons and upstream open reading frames, which provide remarkable insight into the A. oryzae transcriptome. We were also able to assess the alternative mRNA isoforms in A. oryzae and found a large number of genes undergoing alternative splicing. Many genes and pathways that might be involved in higher levels of protein production in solid-state culture than in liquid culture were identified by comparing gene expression levels between different cultures. Our analysis indicated that the transcriptome of A. oryzae is much more complex than previously anticipated, and these results may provide a blueprint for further study of the A. oryzae transcriptome.

Enhanced accumulation of U(VI) by Aspergillus oryzae mutant generated by dielectric barrier discharge air plasma

International Nuclear Information System (INIS)

Wencheng Song; North China Electric Power University, Beijing; Xiangxue Wang; Soochow University, Suzhou; Wen Tao; Hongqing Wang; Tasawar Hayat; Quaid-I-Azam University, Islamabad; Xiangke Wang; Soochow University, Suzhou; King Abdulaziz University, Jeddah

2016-01-01

Aspergillus oryzae was isolated from radionuclides' contaminated soils, and dielectric barrier discharge plasma was used to mutate A. oryzae to improve bioremediation capability of U(VI) pollution. The maximum accumulation capacities of U(VI) on mutated A.oryzae was 627.4 mg/g at T = 298 K and pH = 5.5, which was approximately twice than that of raw A.oryzae. XPS analysis indicated that U(VI) accumulation on mutated A. oryzae was largely attributable to nitrogen- and oxygen-containing functional groups on fungal mycelia. The mutated A. oryzae can be harnessed as bioremediation agents for radionuclides pollution. (author)

Enhanced production of fructosyltransferase in Aspergillus oryzae by genome shuffling.

Science.gov (United States)

Wang, Shenghai; Duan, Mengjie; Liu, Yalan; Fan, Sen; Lin, Xiaoshan; Zhang, Yi

2017-03-01

To breed Aspergillus oryzae strains with high fructosyltransferase (FTase) activity using intraspecific protoplast fusion via genome-shuffling. A candidate library was developed using UV/LiCl of the conidia of A. oryzae SBB201. By screening for enzyme activity and cell biomass, two mutants (UV-11 and UV-76) were chosen for protoplast fusion and subsequent genome shuffling. After three rounds of genome recombination, a fusion mutant RIII-7 was obtained. Its FTase activity was 180 U g -1 , approximately double that of the original strain, and RIII-7 was genetically stable. In fermentation culture, FTase activity of the genome-shuffled strain reached a maximum of 353 U g -1 using substrate-feeding method, and this value was approximately 3.4-times higher than that of the original strain A. oryzae SBB201. Intraspecific protoplast fusion of A. oryzae significantly enhanced FTase activity and generated a potentially useful strain for industrial production.

Dicty_cDB: Contig-U05205-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available ormalization and subtr... 64 4e-09 2 ( FK758356 ) av02131e10r1.1 Symbiotic sea anemone (Anemonia vi... 38 7e...-09 4 ( EE000082 ) ROE00011310 Rhizopus oryzae Company Rhizopus oryz... 54 8e-09 2 ( FK729955 ) av01023i17r1.1 Symbiotic...ncorhynchus mykiss clone omyk-evo-505-080 Charge... 56 1e-08 3 ( FK744942 ) av02063f01r1.1 Symbiotic sea ane...mone (Anemonia vi... 38 1e-08 4 ( FK726627 ) av01045a04r1.1 Symbiotic sea anemone...rus cDNA clone IM... 40 2e-05 3 ( FK744316 ) av01008j21r1.1 Symbiotic sea anemone

LABORATORY TESTS ON INSECTICIDAL EFFECTIVENESS OF DISODIUM OCTABORATE TETRAHYDRATE, DIATOMACEOUS EARTH AND AMORPHOUS SILICA GEL AGAINST Sitophilus oryzae (L. AND THEIR EFFECT ON WHEAT BULK DENSITY

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Zlatko Korunić

2017-01-01

Full Text Available We examined insecticide effectiveness of three different inert dusts: disodium octaborate tetrahydrate (DOT and diatomaceous earth (DE Celatom® Mn 51 applied as powder and suspension, and silica gel Sipernat® 50 S applied as powder against Sitophilus oryzae (L. and their reduction of grain bulk density. Sipernat® 50 S was the most effective dust with a very fast initial effectiveness. DE and DOT generated similar effectiveness against S. oryzae. DOT generated low initial effectiveness but after prolonged exposure time of 8 and especially after 21 days, the mortality was very high (100%, similar to the effectiveness of DE. The similar order of dusts was obtained in the reduction of wheat bulk density. Applied at dose of 200 and 500 ppm, the lowest bulk density difference in regard to bulk density of untreated wheat had DOT (-1.1 and -1.9 kg hl-1, respectively, followed by Celatom® Mn 51 (-3.5 and -4.3 kg hl-1, respectively and Sipernat® 50 S (-5.2 and -5.5 kg hl-1, respectively. Due to the effect on wheat bulk density DOT belongs to the group with the least negative effect on bulk density and therefore, it is a promising dust to control stored grain insect pests.

Random Mutagenesis of the Aspergillus oryzae Genome Results in Fungal Antibacterial Activity

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Cory A. Leonard

2013-01-01

Full Text Available Multidrug-resistant bacteria cause severe infections in hospitals and communities. Development of new drugs to combat resistant microorganisms is needed. Natural products of microbial origin are the source of most currently available antibiotics. We hypothesized that random mutagenesis of Aspergillus oryzae would result in secretion of antibacterial compounds. To address this hypothesis, we developed a screen to identify individual A. oryzae mutants that inhibit the growth of Methicillin-resistant Staphylococcus aureus (MRSA in vitro. To randomly generate A. oryzae mutant strains, spores were treated with ethyl methanesulfonate (EMS. Over 3000 EMS-treated A. oryzae cultures were tested in the screen, and one isolate, CAL220, exhibited altered morphology and antibacterial activity. Culture supernatant from this isolate showed antibacterial activity against Methicillin-sensitive Staphylococcus aureus, MRSA, and Pseudomonas aeruginosa, but not Klebsiella pneumonia or Proteus vulgaris. The results of this study support our hypothesis and suggest that the screen used is sufficient and appropriate to detect secreted antibacterial fungal compounds resulting from mutagenesis of A. oryzae. Because the genome of A. oryzae has been sequenced and systems are available for genetic transformation of this organism, targeted as well as random mutations may be introduced to facilitate the discovery of novel antibacterial compounds using this system.

Random Mutagenesis of the Aspergillus oryzae Genome Results in Fungal Antibacterial Activity

Science.gov (United States)

Leonard, Cory A.; Brown, Stacy D.; Hayman, J. Russell

2013-01-01

Multidrug-resistant bacteria cause severe infections in hospitals and communities. Development of new drugs to combat resistant microorganisms is needed. Natural products of microbial origin are the source of most currently available antibiotics. We hypothesized that random mutagenesis of Aspergillus oryzae would result in secretion of antibacterial compounds. To address this hypothesis, we developed a screen to identify individual A. oryzae mutants that inhibit the growth of Methicillin-resistant Staphylococcus aureus (MRSA) in vitro. To randomly generate A. oryzae mutant strains, spores were treated with ethyl methanesulfonate (EMS). Over 3000 EMS-treated A. oryzae cultures were tested in the screen, and one isolate, CAL220, exhibited altered morphology and antibacterial activity. Culture supernatant from this isolate showed antibacterial activity against Methicillin-sensitive Staphylococcus aureus, MRSA, and Pseudomonas aeruginosa, but not Klebsiella pneumonia or Proteus vulgaris. The results of this study support our hypothesis and suggest that the screen used is sufficient and appropriate to detect secreted antibacterial fungal compounds resulting from mutagenesis of A. oryzae. Because the genome of A. oryzae has been sequenced and systems are available for genetic transformation of this organism, targeted as well as random mutations may be introduced to facilitate the discovery of novel antibacterial compounds using this system. PMID:23983696

Random Mutagenesis of the Aspergillus oryzae Genome Results in Fungal Antibacterial Activity.

Science.gov (United States)

Leonard, Cory A; Brown, Stacy D; Hayman, J Russell

2013-01-01

Multidrug-resistant bacteria cause severe infections in hospitals and communities. Development of new drugs to combat resistant microorganisms is needed. Natural products of microbial origin are the source of most currently available antibiotics. We hypothesized that random mutagenesis of Aspergillus oryzae would result in secretion of antibacterial compounds. To address this hypothesis, we developed a screen to identify individual A. oryzae mutants that inhibit the growth of Methicillin-resistant Staphylococcus aureus (MRSA) in vitro. To randomly generate A. oryzae mutant strains, spores were treated with ethyl methanesulfonate (EMS). Over 3000 EMS-treated A. oryzae cultures were tested in the screen, and one isolate, CAL220, exhibited altered morphology and antibacterial activity. Culture supernatant from this isolate showed antibacterial activity against Methicillin-sensitive Staphylococcus aureus, MRSA, and Pseudomonas aeruginosa, but not Klebsiella pneumonia or Proteus vulgaris. The results of this study support our hypothesis and suggest that the screen used is sufficient and appropriate to detect secreted antibacterial fungal compounds resulting from mutagenesis of A. oryzae. Because the genome of A. oryzae has been sequenced and systems are available for genetic transformation of this organism, targeted as well as random mutations may be introduced to facilitate the discovery of novel antibacterial compounds using this system.

The biosynthesis, structure and gelatinization properties of starches from wild and cultivated African rice species (Oryza barthii and Oryza glaberrima).

Science.gov (United States)

Wang, Kai; Wambugu, Peterson W; Zhang, Bin; Wu, Alex Chi; Henry, Robert J; Gilbert, Robert G

2015-09-20

The molecular structure and gelatinization properties of starches from domesticated African rice (Oryza glaberrima) and its wild progenitor (Oryza barthii) are determined and comparison made with Asian domesticated rice (Oryza sativa), the commonest commercial rice. This suggests possible enzymatic processes contributing to the unique traits of the African varieties. These have similar starch structures, including smaller amylose molecules, but larger amounts of amylose chains across the whole amylose chain-length distribution, and higher amylose contents, than O. sativa. They also show a higher proportion of two- and three-lamellae spanning amylopectin branch chains (degree of polymerization 34-100) than O. sativa, which contributes to their higher gelatinization temperatures. Fitting amylopectin chain-length distribution with a biosynthesis-based mathematical model suggests that the reason for this difference might be because O. glaberrima and O. barthii have more active SSIIIa and/or less active SBEIIb enzymes. Copyright © 2015 Elsevier Ltd. All rights reserved.

Highly efficient gene targeting in Aspergillus oryzae industrial strains under ligD mutation introduced by genome editing: Strain-specific differences in the effects of deleting EcdR, the negative regulator of sclerotia formation.

Science.gov (United States)

Nakamura, Hidetoshi; Katayama, Takuya; Okabe, Tomoya; Iwashita, Kazuhiro; Fujii, Wataru; Kitamoto, Katsuhiko; Maruyama, Jun-Ichi

2017-07-11

Numerous strains of Aspergillus oryzae are industrially used for Japanese traditional fermentation and for the production of enzymes and heterologous proteins. In A. oryzae, deletion of the ku70 or ligD genes involved in non-homologous end joining (NHEJ) has allowed high gene targeting efficiency. However, this strategy has been mainly applied under the genetic background of the A. oryzae wild strain RIB40, and it would be laborious to delete the NHEJ genes in many A. oryzae industrial strains, probably due to their low gene targeting efficiency. In the present study, we generated ligD mutants from the A. oryzae industrial strains by employing the CRISPR/Cas9 system, which we previously developed as a genome editing method. Uridine/uracil auxotrophic strains were generated by deletion of the pyrG gene, which was subsequently used as a selective marker. We examined the gene targeting efficiency with the ecdR gene, of which deletion was reported to induce sclerotia formation under the genetic background of the strain RIB40. As expected, the deletion efficiencies were high, around 60~80%, in the ligD mutants of industrial strains. Intriguingly, the effects of the ecdR deletion on sclerotia formation varied depending on the strains, and we found sclerotia-like structures under the background of the industrial strains, which have never been reported to form sclerotia. The present study demonstrates that introducing ligD mutation by genome editing is an effective method allowing high gene targeting efficiency in A. oryzae industrial strains.

Evidence for biotrophic lifestyle and biocontrol potential of dark septate endophyte Harpophora oryzae to rice blast disease.

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Zhen-Zhu Su

Full Text Available The mutualism pattern of the dark septate endophyte (DSE Harpophora oryzae in rice roots and its biocontrol potential in rice blast disease caused by Magnaporthe oryzae were investigated. Fluorescent protein-expressing H. oryzae was used to monitor the colonization pattern. Hyphae invaded from the epidermis to the inner cortex, but not into the root stele. Fungal colonization increased with root tissue maturation, showing no colonization in the meristematic zone, slight colonization in the elongation zone, and heavy colonization in the differentiation zone. H. oryzae adopted a biotrophic lifestyle in roots accompanied by programmed cell death. Real-time PCR facilitated the accurate quantification of fungal growth and the respective plant response. The biocontrol potential of H. oryzae was visualized by inoculation with eGFP-tagged M. oryzae in rice. H. oryzae protected rice from M. oryzae root invasion by the accumulation of H2O2 and elevated antioxidative capacity. H. oryzae also induced systemic resistance against rice blast. This systemic resistance was mediated by the OsWRKY45-dependent salicylic acid (SA signaling pathway, as indicated by the strongly upregulated expression of OsWRKY45. The colonization pattern of H. oryzae was consistent with the typical characteristics of DSEs. H. oryzae enhanced local resistance by reactive oxygen species (ROS and high antioxidative level and induced OsWRKY45-dependent SA-mediated systemic resistance against rice blast.

Expression, crystallization and preliminary X-ray crystallographic analysis of Xoo0352, d-alanine-d-alanine ligase A, from Xanthomonas oryzae pv. oryzae

International Nuclear Information System (INIS)

Doan, Thanh Thi Ngoc; Kim, Jin-Kwang; Kim, Hyesoon; Ahn, Yeh-Jin; Kim, Jeong-Gu; Lee, Byoung-Moo; Kang, Lin-Woo

2008-01-01

Xoo0352, which encodes d-alanine-d-alanine ligase A (DdlA), from X. oryzae pv. oryzae was cloned, purified and crystallized. Preliminary X-ray crystallographic analysis of DdlA crystals was performed. Xanthomonas oryzae pv. oryzae (Xoo) causes bacterial blight (BB), which is one of the most devastating diseases of rice in most rice-growing countries. d-Alanine-d-alanine ligase A (DdlA), coded by the Xoo0352 gene, was expressed, purified and crystallized. DdlA is an enzyme that is involved in d-alanine metabolism and the biosynthesis of an essential bacterial peptidoglycan precursor, in which it catalyzes the formation of d-alanyl-d-alanine from two d-alanines, and is thus an attractive antibacterial drug target against Xoo. The DdlA crystals diffracted to 2.3 Å resolution and belonged to the primitive tetragonal space group P4 3 2 1 2, with unit-cell parameters a = b = 83.0, c = 97.6 Å. There is one molecule in the asymmetric unit, with a corresponding V M of 1.88 Å 3 Da −1 and a solvent content of 34.6%. The initial structure was determined by molecular replacement using d-alanine-d-alanine ligase from Staphylococcus aureus as a template model

A walk on the wild side: Oryza species as source for rice abiotic stress tolerance.

Science.gov (United States)

Menguer, Paloma Koprovski; Sperotto, Raul Antonio; Ricachenevsky, Felipe Klein

2017-01-01

Oryza sativa, the common cultivated rice, is one of the most important crops for human consumption, but production is increasingly threatened by abiotic stresses. Although many efforts have resulted in breeding rice cultivars that are relatively tolerant to their local environments, climate changes and population increase are expected to soon call for new, fast generation of stress tolerant rice germplasm, and current within-species rice diversity might not be enough to overcome such needs. The Oryza genus contains other 23 wild species, with only Oryza glaberrima being also domesticated. Rice domestication was performed with a narrow genetic diversity, and the other Oryza species are a virtually untapped genetic resource for rice stress tolerance improvement. Here we review the origin of domesticated Oryza sativa from wild progenitors, the ecological and genomic diversity of the Oryza genus, and the stress tolerance variation observed for wild Oryza species, including the genetic basis underlying the tolerance mechanisms found. The summary provided here is important to indicate how we should move forward to unlock the full potential of these germplasms for rice improvement.

Comparative Chemistry of Aspergillus oryzae (RIB40) and A. flavus (NRRL 3357)

Science.gov (United States)

Rank, Christian; Klejnstrup, Marie Louise; Petersen, Lene Maj; Kildgaard, Sara; Frisvad, Jens Christian; Gotfredsen, Charlotte Held; Larsen, Thomas Ostenfeld

2012-01-01

Aspergillus oryzae and A. flavus are important species in industrial biotechnology and food safety and have been some of the first aspergilli to be fully genome sequenced. Bioinformatic analysis has revealed 99.5% gene homology between the two species pointing towards a large coherence in the secondary metabolite production. In this study we report on the first comparison of secondary metabolite production between the full genome sequenced strains of A. oryzae (RIB40) and A. flavus (NRRL 3357). Surprisingly, the overall chemical profiles of the two strains were mostly very different across 15 growth conditions. Contrary to previous studies we found the aflatrem precursor 13-desoxypaxilline to be a major metabolite from A. oryzae under certain growth conditions. For the first time, we additionally report A. oryzae to produce parasiticolide A and two new analogues hereof, along with four new alkaloids related to the A. flavus metabolites ditryptophenalines and miyakamides. Generally the secondary metabolite capability of A. oryzae presents several novel end products likely to result from the domestication process from A. flavus. PMID:24957367

A Model of the Effect of the Microbial Biomass on the Isotherm of the Fermenting Solids in Solid-State Fermentation

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Barbara Celuppi Marques

2006-01-01

Full Text Available We compare isotherms for soybeans and soybeans fermented with Rhizopus oryzae, showing that in solid-state fermentation the biomass affects the isotherm of the fermenting solids. Equations are developed to calculate, for a given overall water content of the fermenting solids, the water contents of the biomass and residual substrate, as well as the water activity. A case study, undertaken using a mathematical model of a well-mixed bioreactor, shows that if water additions are made on the basis of the assumption that fermenting solids have the same isotherm as the substrate itself, poor growth can result since the added water does not maintain the water activity at levels favorable for growth. We conclude that the effect of the microbial biomass on the isotherm of the fermenting solids must be taken into account in mathematical models of solid-state fermentation bioreactors.

An evaluation of aflatoxin and cyclopiazonic acid production in Aspergillus oryzae.

Science.gov (United States)

Kim, Nam Yeun; Lee, Jin Hee; Lee, Inhyung; Ji, Geun Eog

2014-06-01

To date, edible fungi such as Aspergillus flavus var. oryzae (A. oryzae) has been considered as safe. However, some strains can produce mycotoxins. Thus, the biosynthetic ability to produce mycotoxins should be reevaluated to determine the safety of edible fungi. We analyzed the production of aflatoxins and cyclopiazonic acid (CPA) from edible fungi such as A. oryzae isolated from various Korean foods using multiplex PCR, enzyme-linked immunosorbent assay, and high-performance liquid chromatography (HPLC). In the multiplex PCR analysis of aflatoxin biosynthetic genes omtB, aflR, ver-1, and omtA, 5 of 19 Aspergillus strains produced all PCR products. Among them, aflatoxin B1 and aflatoxin B2 were detected from only A. flavus KACC 41403 by HPLC. Aflatoxins were not detected from the other four strains that produced all positive PCR bands. Aflatoxin also was not detected from 12 strains that had PCR patterns without aflR or ver-1 and from 2 strains that did not produce any of the expected PCR products. Only the seven A. oryzae strains that produced all of the positive PCR bands including the CPA biosynthetic genes maoA, dmaT, and pks-nrps produced CPA. CPA and aflatoxin production must be evaluated before A. oryzae strains are used for the development of fermented foods.

Effects of gamma irradiation on long-storage seeds of Oryza sativa (cv. 2233) and their surface infecting fungal diversity

International Nuclear Information System (INIS)

Maity, J.P.; Kar, S.; Banerjee, S.; Chakraborty, A.; Santra, S.C.

2009-01-01

We have reported an effective 60 Co gamma irradiation method for the removal and long-time prevention of contaminating fungi of Oryza sativa cv-2233, without the losses of seed viabilities. The fungal growth and their population on gamma treated seeds was found to decrease significantly (p 50%) at 2 kGy, whereas total inhibition was noticed at 3 kGy after 1.5 month.

The normal mycoflora of commodities from Thailand. 1. Nuts and oilseeds.

Science.gov (United States)

Pitt, J I; Hocking, A D; Bhudhasamai, K; Miscamble, B F; Wheeler, K A; Tanboon-Ek, P

1993-12-01

A comprehensive study was carried out of the fungi occurring in commodities normally traded in Thailand. Samples of major commodities were obtained from farmers' stocks and middlemen in major producing areas throughout the country. Retail samples were obtained from outlets in and around Bankok. Samples were divided into two portions, one being examined in Bangkok, and the second in Sydney. After surface disinfection, fungi were enumerated by direct plating on dichloran rose bengal chloramphenicol agar, dichloran 18% glycerol agar, Aspergillus flavus and parasiticus agar and dichloran chloramphenicol peptone agar. Figures for percentage infection were calculated, and fungi were isolated and identified to species level. In all 602 samples were examined, and at North Ryde about 18,000 fungal isolates identified. Data obtained from 329 samples are reported here, comprising maize (154), peanuts (109), cashews (45) and copra (21). Major fungi in maize included Fusarium moniliforme (present in 97% of samples), Aspergillus flavus (85%), Penicillium citrinum (67%), Aspergillus niger (64%), Lasiodiplodia theobromae (58%) and Fusarium semitectum (45%). In peanuts, the major fungi were Aspergillus flavus (95% of samples), Aspergillus niger (86%), Rhizopus oryzae (60%), Eurotium rubrum (51%), Macromina phaseolina (49%), Penicillium citrinum (46%) and Eurotium chevalieri (46%). Invasion in cashews was lower, major fungi being Aspergillus flavus (60%), Nigrospora oryzae (58%), Aspergillus niger (53%), Chaetomium globosum (47%) and Eurotium chevalieri (40%). Aspergillus flavus (86% of samples) was again dominant in copra, with Rhizopus oryzae (52%), Aspergillus niger (43%), Eurotium chevalieri (43%) the only other species exceeding 40% infection. Aspergillus parasiticus was rarely seen, and Aspergillus nomius was reported from foods for the first time.

Physico-chemical and sensorial characteristics of rice (Oryza sativa L.) irradiated and the effect in Sitophilus oryzae L. development

International Nuclear Information System (INIS)

Zanao, Cintia Fernanda Pedroso

2007-01-01

Rice is exposed to injuries especially due to the attack of insects, which represent great qualitative and quantitative losses. The aim of the present research was to verify the viability of the gamma irradiation as rice (Oryza sativa L.) conservation method; its efficiency in the control of an important pest for stored grains Sitophilus oryzae L. and the effects that irradiation may present in irradiated grains in relation to the sensory aspect and starch alterations. Samples were composed of polished rice and the gamma irradiation dosages used were 0.5; 1.0; 3.0; and 5.0 kGy, and kept at room temperature. Analyses of the grain breakage percentages during processing and the longevity and reproduction of the rice weevil were performed. Analyses of the centesimal composition, phytic acid, apparent amylose content and starch paste properties were also conducted, as well as analyses of the rice color (instrumental) and acceptability (hedonic test), aimed at the determination of the raw and cooked rice sensory quality. It was verified that the gamma irradiation did not change the grain breakage percentage significantly and caused a negative effect on the insect development. Irradiation did not change the centesimal composition significantly. No significant values of phytic acid were found because during rice processing, the outer part (bran) containing 85-92% of total phytates was removed. Values from 17.33 to 18.44% for the apparent amylose content were found, and irradiation did not affect the rice starch amylose content significantly. The paste properties results were significantly changed, where reduction on the paste temperature, decrease on the time for the occurrence of the peak viscosity, reduction on the final viscosity values were observed. The retrogradation tendency became less intense in starches with increasing irradiation dosages. Statistical differences (p<0.05) were detected in the sensory aspect between irradiated and non-irradiated samples, and the

Fumaric acid production by Rhizopus oryzae and its facilitated ...

African Journals Online (AJOL)

Anupreet

2014-03-05

Mar 5, 2014 ... membrane' setup for a 'fumaric acid' source, with toluene as organic membrane and sodium hydroxide as strip phase. The liquid ... polymer films can be extended to include liquids. They are defined as Liquid .... Membrane solutions were prepared by dissolution of trioctylamine. (TOA) (Fluka A.G. ...

Fumaric acid production by Rhizopus oryzae and its facilitated ...

African Journals Online (AJOL)

Anupreet

2014-03-05

Mar 5, 2014 ... Currently, fumaric acid is produced from petroleum based derivative maleic ... best possible option that came up for the strip phase was an alkaline medium. .... Future directions of membrane gas separation technology. Indus.

Immobilization of Isolated Lipase From Moldy Copra (Aspergillus Oryzae)

OpenAIRE

Dali, Seniwati; Patong, A. B. D. Rauf; Jalaluddin, M. Noor; Pirman; Hamzah, Baharuddin

2011-01-01

Enzyme immobilization is a recovery technique that has been studied in several years, using support as a media to help enzyme dissolutions to the reaction substrate. Immobilization method used in this study was adsorption method, using specific lipase from Aspergillus oryzae. Lipase was partially purified from the culture supernatant of Aspergillus oryzae. Enzyme was immobilized by adsorbed on silica gel. Studies on free and immobilized lipase systems for determination of optimum pH, optimum ...

Effect of deletion of chitin synthase genes on mycelial morphology and culture viscosity in Aspergillus oryzae

DEFF Research Database (Denmark)

Müller, Christian; Hansen, K.; Szabo, Peter

2003-01-01

The objective of this study was to quantify the effect of disrupting two chitin synthases, chsB and csmA, on the morphology and rheology during batch cultivation of Aspergillus oryzae. The rheological properties were characterized in batch cultivations at different biomass concentrations (from 3...... broth was significantly affected by the biomass concentration, the morphology, and also by pH. The chsB disruption strain had lower consistency index K values for all biomass concentrations investigated, which is a desirable trait for industrial Aspergillus fermentations. (C) 2003 Wiley Periodicals, Inc....

Pathotype profile of Xanthomonas oryzae pv. oryzae isolates from North Sumatera

Science.gov (United States)

Noer, Z.; Hasanuddin; Lisnawita; Suryanto, D.

2018-02-01

The Bacterial blight disease caused by Xanthomonas oryzae pv. oryzae (Xoo) is one of the most important diseases and has caused crop failure in rice crops. This pathogen infects the leaves in all plant growth phases. The purpose of this study is to investigation 10 Xoo isolates pathotype obtained from North Sumatra based on their interactions with 10 near-isogenic rice lines (NIL) of IRRI. The results showed that there are 6 pathotypes of virulence in North Sumatra, they are; pathotype I with incompatible interaction to all Xa genes, pathotype II with compatible interaction to Xa1 and Xa3 genes, while it has incompatible interaction to other genes, pathotype III with compatible interaction to Xa1, Xa5, Xa7, Xa8, Xa10 and Xa11 genes, but it has incompatible interaction to other genes, pathotype IV with compatible interaction to all Xa genes, pathotype V with compatible interaction to Xa1 gene and incompatible interaction to other genes, and pathotype VI with compatible interaction to Xa3 gene and incompatible interaction to other genes. Based on the resistant genes in each individual Xa2, Xa4, and Xa21 genes are the combination of Xa genes which are most suitable for use in the development of rice cultivars in North Sumatra.

Utilization of Aspergillus oryzae to produce pectin lyase from various agro-industrial residues

OpenAIRE

Koser, Safia; Anwar, Zahid; Iqbal, Zafar; Anjum, Awais; Aqil, Tahir; Mehmood, Sajid; Irshad, Muhammad

2014-01-01

The present study was aimed to investigate the culture influence on pectin lyase production potential of fungal strain Aspergillus oryzae. The enzyme profile of A. oryzae showed highest activity of pectin lyase after 3rd day of incubation on lemon peel waste under solid state fermentation conditions. To induce the pectin lyase synthesis capability of A. oryzae at optimal level various culture variables including physical and nutritional parameters were optimized by adopting classical optimiza...

Survey of the transcriptome of Aspergillus oryzae via massively parallel mRNA sequencing

OpenAIRE

Wang, Bin; Guo, Guangwu; Wang, Chao; Lin, Ying; Wang, Xiaoning; Zhao, Mouming; Guo, Yong; He, Minghui; Zhang, Yong; Pan, Li

2010-01-01

Aspergillus oryzae, an important filamentous fungus used in food fermentation and the enzyme industry, has been shown through genome sequencing and various other tools to have prominent features in its genomic composition. However, the functional complexity of the A. oryzae transcriptome has not yet been fully elucidated. Here, we applied direct high-throughput paired-end RNA-sequencing (RNA-Seq) to the transcriptome of A. oryzae under four different culture conditions. With the high resoluti...

Impact of Engineered Nanoparticles on Virulence of Xanthomonas oryzae pv oryzae and on Rice Sensitivity at its Infection

Directory of Open Access Journals (Sweden)

Giuliano Degrassi

2014-12-01

Full Text Available The present work of nanocotoxicity wants to propose a new plant model starting from the rice plant. The model takes into consideration the impact of engineered nanoparticles (Ag, Co, Ni, CeO2, Fe3O4, TiO2 on rice plants that were weakened by infections of Xanthomonas oryzae pv oryzae bacteria. The results indicate that some NPs increase the rice sensitivity to the pathogen while others decrease the virulence of the pathogen towards rice. No-enrichment in component metal concentration is detected in above organs of rice, with exception of Ni-NPs treatment. An imbalance of major elements in infected rice crops treated with NPs was investigated.

Molecular diversity of lactic acid bacteria on ileum broiler chicken fed by bran and bran fermentation

Science.gov (United States)

Baniyah, Laelatul; Nur Jannah, Siti; Rukmi, Isworo; Sugiharto

2018-05-01

Lactic Acid Bacteria (LAB) is a digestive tract microflora that have a positive role in poultry health. The number and diversity of LAB in the digestive tract affected by several factors, among them was the kind of feed. The purpose of this research was to know diversity of Lactic Acid Bacteria (LAB) ileum broiler’s after feeding with prebiotic bran and Rhizopus oryzae fermented bran which was added to commercial feed. As much as 15 broilers were used to determine the diversity of LAB. All broilers were fed using commercial feed. The control used commercial feed no addition of bran or fermented bran, and commercial feed with fermented bran and nonfermented bran were as a treatment. To determine the diversity of LAB, T-RFLP method was applied. The Hae III and Msp I were used as restriction enzymes. The number of phylotype, relative abundance, Shannon diversity index (H '), evenness (E), and Dominance (D) were examined. The results indicated that the addition of prebiotic bran on commercial feed showed a higher diversity of lactic acid bacteria on broiler’s ileum, compared with control and addition of Rhizopus oryzae fermented bran. LAB group that dominates in the ileum is Lactobacillus sp. and L. delbruecii subs bulgaricus.

Simultaneous Saccharification and Fermentation of Lactic Acid from Empty Fruit Bunch at High Solids Loading

Directory of Open Access Journals (Sweden)

Nursia Hassan

2016-03-01

Full Text Available The production of value-added chemicals from the bioconversion of lignocellulose biomass has been considered a promising venture. In this study, microwave, alkali-pretreated empty fruit bunch (EFB was used as the substrate, utilizing pelletized filamentous Rhizopus oryzae NRRL 395 and cellulolytic enzymes for lactic acid production in a fed-batch simultaneous saccharification and fermentation (SSF process. Insoluble solids generally do not affect the SSF process until a certain concentration is exceeded. To achieve a high lactic acid concentration in the broth, a high solids loading was required to allow a higher rate of glucose conversion. However, the results revealed a decrease in the final lactic acid yield when running SSF at a massive insoluble solids level. High osmotic pressure in the medium led to poor cellular performance and caused the Rhizopus oryzae pellets to break down, affecting the lactic acid production. To improve the process performance, a fed-batch operation mode was used. The fed-batch operation was shown to facilitate higher lactic acid yield, compared with the SSF batch mode. Enzyme feeding, as well as substrate feeding, was also investigated as a means of enabling a higher dry matter content, with a high glucose conversion in SSF of cellulose-rich EFB.

Selection of Suitable Microorganism for Biocatalytic Oxidation Reaction of Racemic Propranolol

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Rahime SONGÜR

2017-12-01

Full Text Available Propranolol is one of the β-blockers which are pharmaceutically important, especially used for treatment of cardiovasculer disease. In this study, the production of enantiomerically pure propranolol was aimed via biocatalytic deracemization including tandem oxidation-reduction reactions of racemic propranolol. Within this content, firstly suitable microorganism for the oxidation of racemic propranolol was investigated. Alcohol dehydrogenase (ADH enzyme for oxidation of propranolol and NADH oxidase enzyme for cofactor regeneration were necessary for the oxidation reactions. For this reason, ADH and NADH oxidase enzymes activities of different microorganisms were measured to select the microorganism for using as enzyme source. These microorganisms are Lactobacillus kefir NRRL B-1839, Rhodotorula glutunis DSM 70398, Rhizopus oryzae CBS 111718, Rhizopus arhizus. The highest ADH and NADH oxidase activities were obtained for L. kefir.

Effect of atmospheric plasma treatment on seed germination of rice (Oryza sativa L.)

Science.gov (United States)

Penado, Keith Nealson M.; Mahinay, Christian Lorenz S.; Culaba, Ivan B.

2018-01-01

Multiple methods of improving plant development have been utilized over the past decades. Despite these improvements, there still exists a need for better planting methods due to the increasing population of a global community. Studies have reported that plasma treatment affects the growth and germination of a variety of plant species, including a multitude of grains which often takes the bulk in the diet of the average human being. This study explores the effect of atmospheric air plasma jet treatment on the seed germination of rice (Oryza sativa L.). The seeds were treated using an atmospheric air plasma jet for 1, 2, and 3 s. The effect of plasma exposure shows a reduction of trichomes on the surface of the seed. This caused a possible increase in wettability which significantly affected the seed germ length but did not affect the seed germination count after the germination period of 72 h.

Extracellular biosynthesis of silver nanoparticles using Rhizopus stolonifer

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Khalid AbdelRahim

2017-01-01

Full Text Available Synthesis of silver nanoparticles (AgNPs has become a necessary field of applied science. Biological method for synthesis of AgNPs by Rhizopus stolonifer aqueous mycelial extract was used. The AgNPs were identified by UV–visible spectrometry, X-ray diffraction (XRD, transmission electron microscopy (TEM and Fourier transform infrared spectrometry (FT-IR. The presence of surface plasmon band around 420 nm indicates AgNPs formation. The characteristic of the AgNPs within the face-centered cubic (fcc structure are indicated by the peaks of the X-ray diffraction (XRD pattern corresponding to (111, (200 and (220 planes. Spherical, mono-dispersed and stable AgNPs with diameter around 9.47 nm were prepared and affirmed by high-resolution transmission electron microscopy (HR-TEM. Fourier Transform Infrared (FTIR shows peaks at 1426 and 1684 cm−1 that affirm the presence of coat covering protein the AgNPs which is known as capping proteins. Parameter optimization showed the smallest size of AgNPs (2.86 ± 0.3 nm was obtained with 10−2 M AgNO3 at 40 °C. The present study provides the proof that the molecules within aqueous mycelial extract of R. stolonifer facilitate synthesis of AgNPs and highlight on value-added from R. stolonifer for cost effectiveness. Also, eco-friendly medical and nanotechnology-based industries could also be provided. Size of prepared AgNPs could be controlled by temperature and AgNO3 concentration. Further studies are required to study effect of more parameters on size and morphology of AgNPs as this will help in the control of large scale production of biogenic AgNPs.

Comparative Genome Analysis Between Aspergillus oryzae Strains Reveals Close Relationship Between Sites of Mutation Localization and Regions of Highly Divergent Genes among Aspergillus Species

Science.gov (United States)

Umemura, Myco; Koike, Hideaki; Yamane, Noriko; Koyama, Yoshinori; Satou, Yuki; Kikuzato, Ikuya; Teruya, Morimi; Tsukahara, Masatoshi; Imada, Yumi; Wachi, Youji; Miwa, Yukino; Yano, Shuichi; Tamano, Koichi; Kawarabayasi, Yutaka; Fujimori, Kazuhiro E.; Machida, Masayuki; Hirano, Takashi

2012-01-01

Aspergillus oryzae has been utilized for over 1000 years in Japan for the production of various traditional foods, and a large number of A. oryzae strains have been isolated and/or selected for the effective fermentation of food ingredients. Characteristics of genetic alterations among the strains used are of particular interest in studies of A. oryzae. Here, we have sequenced the whole genome of an industrial fungal isolate, A. oryzae RIB326, by using a next-generation sequencing system and compared the data with those of A. oryzae RIB40, a wild-type strain sequenced in 2005. The aim of this study was to evaluate the mutation pressure on the non-syntenic blocks (NSBs) of the genome, which were previously identified through comparative genomic analysis of A. oryzae, Aspergillus fumigatus, and Aspergillus nidulans. We found that genes within the NSBs of RIB326 accumulate mutations more frequently than those within the SBs, regardless of their distance from the telomeres or of their expression level. Our findings suggest that the high mutation frequency of NSBs might contribute to maintaining the diversity of the A. oryzae genome. PMID:22912434

Comparative genome analysis between Aspergillus oryzae strains reveals close relationship between sites of mutation localization and regions of highly divergent genes among Aspergillus species.

Science.gov (United States)

Umemura, Myco; Koike, Hideaki; Yamane, Noriko; Koyama, Yoshinori; Satou, Yuki; Kikuzato, Ikuya; Teruya, Morimi; Tsukahara, Masatoshi; Imada, Yumi; Wachi, Youji; Miwa, Yukino; Yano, Shuichi; Tamano, Koichi; Kawarabayasi, Yutaka; Fujimori, Kazuhiro E; Machida, Masayuki; Hirano, Takashi

2012-10-01

Aspergillus oryzae has been utilized for over 1000 years in Japan for the production of various traditional foods, and a large number of A. oryzae strains have been isolated and/or selected for the effective fermentation of food ingredients. Characteristics of genetic alterations among the strains used are of particular interest in studies of A. oryzae. Here, we have sequenced the whole genome of an industrial fungal isolate, A. oryzae RIB326, by using a next-generation sequencing system and compared the data with those of A. oryzae RIB40, a wild-type strain sequenced in 2005. The aim of this study was to evaluate the mutation pressure on the non-syntenic blocks (NSBs) of the genome, which were previously identified through comparative genomic analysis of A. oryzae, Aspergillus fumigatus, and Aspergillus nidulans. We found that genes within the NSBs of RIB326 accumulate mutations more frequently than those within the SBs, regardless of their distance from the telomeres or of their expression level. Our findings suggest that the high mutation frequency of NSBs might contribute to maintaining the diversity of the A. oryzae genome.

Transcriptome analysis of WRKY gene family in Oryza officinalis Wall ex Watt and WRKY genes involved in responses to Xanthomonas oryzae pv. oryzae stress.

Science.gov (United States)

Jiang, Chunmiao; Shen, Qingxi J; Wang, Bo; He, Bin; Xiao, Suqin; Chen, Ling; Yu, Tengqiong; Ke, Xue; Zhong, Qiaofang; Fu, Jian; Chen, Yue; Wang, Lingxian; Yin, Fuyou; Zhang, Dunyu; Ghidan, Walid; Huang, Xingqi; Cheng, Zaiquan

2017-01-01

Oryza officinalis Wall ex Watt, a very important and special wild rice species, shows abundant genetic diversity and disease resistance features, especially high resistance to bacterial blight. The molecular mechanisms of bacterial blight resistance in O. officinalis have not yet been elucidated. The WRKY transcription factor family is one of the largest gene families involved in plant growth, development and stress response. However, little is known about the numbers, structure, molecular phylogenetics, and expression of the WRKY genes under Xanthomonas oryzae pv. oryzae (Xoo) stress in O. officinalis due to lacking of O. officinalis genome. Therefore, based on the RNA-sequencing data of O. officinalis, we performed a comprehensive study of WRKY genes in O. officinalis and identified 89 OoWRKY genes. Then 89 OoWRKY genes were classified into three groups based on the WRKY domains and zinc finger motifs. Phylogenetic analysis strongly supported that the evolution of OoWRKY genes were consistent with previous studies of WRKYs, and subgroup IIc OoWRKY genes were the original ancestors of some group II and group III OoWRKYs. Among the 89 OoWRKY genes, eight OoWRKYs displayed significantly different expression (>2-fold, pWRKY family of transcription factors in O.officinalis. Insight was gained into the classification, evolution, and function of the OoWRKY genes, revealing the putative roles of eight significantly different expression OoWRKYs in Xoo strains PXO99 and C5 stress responses in O.officinalis. This study provided a better understanding of the evolution and functions of O. officinalis WRKY genes, and suggested that manipulating eight significantly different expression OoWRKYs would enhance resistance to bacterial blight.

Induction of salicylic acid-mediated defense response in perennial ryegrass against infection by Magnaporthe oryzae.

Science.gov (United States)

Rahman, Alamgir; Kuldau, Gretchen A; Uddin, Wakar

2014-06-01

Incorporation of plant defense activators is an innovative approach to development of an integrated strategy for the management of turfgrass diseases. The effects of salicylic acid (SA), benzothiadiazole (BTH, chemical analog of SA), jasmonic acid (JA), and ethephon (ET, an ethylene-releasing compound) on development of gray leaf spot in perennial ryegrass (Lolium perenne L.) caused by Magnaporthe oryzae were evaluated. Gray leaf spot disease incidence and severity were significantly decreased when plants were treated prior to inoculation with SA, BTH, and partially by ET but not by JA. Accumulation of endogenous SA and elevated expression of pathogenesis-related (PR)-1, PR-3.1, and PR-5 genes were associated with inoculation of plants by M. oryzae. Treatment of plants with SA enhanced expression levels of PR-3.1 and PR-5 but did not affect the PR-1 level, whereas BTH treatment enhanced relative expression levels of all three PR genes. Microscopic observations of leaves inoculated with M. oryzae revealed higher frequencies of callose deposition at the penetration sites in SA- and BTH-treated plants compared with the control plants (treated with water). These results suggest that early and higher induction of these genes by systemic resistance inducers may provide perennial ryegrass with a substantial advantage to defend against infection by M. oryzae.

Rhizopus-associated soft tissue infection in an immunocompetent air-conditioning technician after a road traffic accident: A case report and review of the literature

Directory of Open Access Journals (Sweden)

Nada B. Rabie

2012-02-01

Full Text Available Summary: Rhinocerebral or sinopulmonary mucromycosis is a well-recognized human fungal infection found among immunocompromised and diabetic patients. However, the infection is rare among immunocompetent hosts. We are reporting the case of an adult immunocompetent male patient working as an air-conditioning technician. The patient was a victim of a road traffic accident (RTA and sustained multiple fractures in the proximal part of the left tibia, distal femur, and scapula. Two weeks postoperatively, Rhizopus microspores were isolated from an infected traumatic wound over the distal femur. Surgical debridement was performed, and the patient was started on amphotericin B. Occupational exposure history and workplace environmental sanitation are crucial for the prevention of this potentially fatal yet preventable infection. Keywords: Rhizopus, Immunocompetent, Air conditioning

Imobilização de lipases em filme de caseinato de sódio/glicerol: aplicação na síntese de ésteres Lipase immobilization in sodium caseinate/glycerol film: application in ester synthesis

OpenAIRE

Damianni Sebrão; Vanessa Dutra Silva; Maria da Graça Nascimento; Marcelo Alves Moreira

2007-01-01

Lipases from different sources were immobilized in sodium caseinate/glycerol film and used in the esterification reactions of aliphatic acids with alcohols in the presence of organic solvents. Lipases from Pseudomonas sp and Rhizopus oryzae were selected and the influence of several parameters was analyzed, including: lipase loading, organic solvent polarity, reaction temperature, chain length of alcohol and acid and enzyme/support reuse. For comparison, free enzymes were used under similar e...

Devastating posttraumatic primary cutaneous mucormycosis in a diabetic patient

Directory of Open Access Journals (Sweden)

Poongodi Lakshmi Santhana Kumarasamy

2016-01-01

Full Text Available Mucorales are saprophytic fungi causing mucormycosis, which is a life threatening infection manifested as rhinocerebral, pulmonary, gastrointestinal, cutaneous, and disseminated forms. The cutaneous form is further divided into primary and secondary forms. The major risk factors include uncontrolled diabetes mellitus with or without ketoacidosis, other forms of metabolic acidosis, and trauma. We report here a case of primary cutaneous mucormycosis caused by Rhizopus oryzae, in a diabetic after a road traffic accident.

Korean Ginseng Berry Fermented by Mycotoxin Non-producing Aspergillus niger and Aspergillus oryzae: Ginsenoside Analyses and Anti-proliferative Activities.

Science.gov (United States)

Li, Zhipeng; Ahn, Hyung Jin; Kim, Nam Yeon; Lee, Yu Na; Ji, Geun Eog

2016-01-01

To transform ginsenosides, Korean ginseng berry (KGB) was fermented by mycotoxin non-producing Aspergillus niger and Aspergillus oryzae. Changes of ginsenoside profile and anti-proliferative activities were observed. Results showed that A. niger tended to efficiently transform protopanaxadiol (PPD) type ginsenosides such as Rb1, Rb2, Rd to compound K while A. oryzae tended to efficiently transform protopanaxatriol (PPT) type ginsenoside Re to Rh1 via Rg1. Butanol extracts of fermented KGB showed high cytotoxicity on human adenocarcinoma HT-29 cell line and hepatocellular carcinoma HepG2 cell line while that of unfermented KGB showed little. The minimum effective concentration of niger-fermented KGB was less than 2.5 µg/mL while that of oryzae-fermented KGB was about 5 µg/mL. As A. niger is more inclined to transform PPD type ginsenosides, niger-fermented KGB showed stronger anti-proliferative activity than oryzae-fermented KGB.

Comparative proteomics of Rhizopus delemar ATCC 20344 unravels the role of amino acid catabolism in fumarate accumulation

OpenAIRE

Odoni, Dorett I.; Tamayo-Ramos, Juan A.; Sloothaak, Jasper; van Heck, Ruben G.A.; Martins dos Santos, Vitor A.P.; de Graaff, Leo H.; Suarez-Diez, Maria; Schaap, Peter J.

2017-01-01

The filamentous fungus Rhizopus delemar naturally accumulates relatively high amounts of fumarate. Although the culture conditions that increase fumarate yields are well established, the network underlying the accumulation of fumarate is not yet fully understood. We set out to increase the knowledge about fumarate accumulation in R. delemar. To this end, we combined a transcriptomics and proteomics approach to identify key metabolic pathways involved in fumarate production in R. delemar, and ...

Elimination of Sitophilus oryzae (L.) by irradiation

Energy Technology Data Exchange (ETDEWEB)

Castro, Fernanda Peixoto [Exercito Brasileiro, Brasilia, DF (Brazil). Diretoria de Suprimento. Dept. Logistico]. E-mail: peixotocastro@dlog.eb.mil.br; Vital, Helio de Carvalho [Centro Tecnologico do Exercito (CTEx), Rio de Janeiro, RJ (brazil)]. E-mail: vital@ctex.eb.br

2005-07-01

Food treatment by exposure to ionizing radiation, known as food irradiation, presents several attractive features such as: leaving no residues, posing no threat to consumer health, usually causing no damage to sensory or nutritional properties and acting uniformly throughout the volume of the products. This work investigated the efficiency of irradiation for eliminating Sitophilus oryzae (L.), also known as 'the rice weevil', a small beetle frequently found in infested grains. A total of 444 individuals of Sitophilus oryzae (L.) found in corn meal and noodles supplies were irradiated with gamma ray doses of 0, 0.6, 0.9, 1.0, 1.2, 1.5 and 2.0 kGy and then visually monitored for 4 days in order to determine the number of insects still alive. The least-squares fitting method was used to determine the survival curves as functions of post-irradiation time and dose. The living fraction of the irradiated population was found to decrease exponentially with time. The results indicated that doses of 2.0, 1.5 and 0.6 kGy cause immediate death, instantaneous immobility and death of the species within one week, respectively. The findings suggest that disinfestation of Sitophilus oryzae (L.) by irradiation is an interesting option to the dangerous use of toxic chemicals. (author)

Elimination of Sitophilus oryzae (L.) by irradiation

International Nuclear Information System (INIS)

Castro, Fernanda Peixoto

2005-01-01

Food treatment by exposure to ionizing radiation, known as food irradiation, presents several attractive features such as: leaving no residues, posing no threat to consumer health, usually causing no damage to sensory or nutritional properties and acting uniformly throughout the volume of the products. This work investigated the efficiency of irradiation for eliminating Sitophilus oryzae (L.), also known as 'the rice weevil', a small beetle frequently found in infested grains. A total of 444 individuals of Sitophilus oryzae (L.) found in corn meal and noodles supplies were irradiated with gamma ray doses of 0, 0.6, 0.9, 1.0, 1.2, 1.5 and 2.0 kGy and then visually monitored for 4 days in order to determine the number of insects still alive. The least-squares fitting method was used to determine the survival curves as functions of post-irradiation time and dose. The living fraction of the irradiated population was found to decrease exponentially with time. The results indicated that doses of 2.0, 1.5 and 0.6 kGy cause immediate death, instantaneous immobility and death of the species within one week, respectively. The findings suggest that disinfestation of Sitophilus oryzae (L.) by irradiation is an interesting option to the dangerous use of toxic chemicals. (author)

Rhizomucor miehei triglyceride lipase is processed and secreted from transformed Aspergillus oryzae.

Science.gov (United States)

Huge-Jensen, B; Andreasen, F; Christensen, T; Christensen, M; Thim, L; Boel, E

1989-09-01

The cDNA encoding the precursor of the Rhizomucor miehei triglyceride lipase was inserted in an Aspergillus oryzae expression vector. In this vector the expression of the lipase cDNA is under control of the Aspergillus oryzae alpha-amylase gene promoter and the Aspergillus niger glucoamylase gene terminator. The recombinant plasmid was introduced into Aspergillus oryzae, and transformed colonies were selected and screened for lipase expression. Lipase-positive transformants were grown in a small fermentor, and recombinant triglyceride lipase was purified from the culture broth. The purified enzymatically active recombinant lipase (rRML) secreted from A. oryzae was shown to have the same characteristics with respect to mobility on reducing SDS-gels and amino acid composition as the native enzyme. N-terminal amino acid sequencing indicated that approximately 70% of the secreted rRML had the same N-terminal sequence as the native Rhizomucor miehei enzyme, whereas 30% of the secreted rRML was one amino acid residue shorter in the N-terminal. The recombinant lipase precursor, which has a 70 amino acid propeptide, is thus processed in and secreted from Aspergillus oryzae. We have hereby demonstrated the utility of this organism as a host for the production of recombinant triglyceride lipases.

Recombinant expression and purification of an Oxysterol Binding Protein from Aspergillus oryzae 3.042

Directory of Open Access Journals (Sweden)

Zhang Xian

2017-01-01

Full Text Available A full-length cDNA encoding a candidate Oxysterol-binding protein(OSBP from Aspergillus oryzae (AoOSBP was cloned and expressed in Escherichia coli as a maltose-binding protein (MBP fusion protein. The MBP-AoOSBP protein from the importantly industrial fungus A. oryzae was purified by amylose resin and chromatography column. SDS-PAGE showed that MBP-AoOSBP has an estimated molecular weight of 182 kDa. OSBP and its homologues (ORPs own the affinity for oxysterols, cholesterol and glycerophospholipids. According to the superiority of A. oryzae in the fermented foods and also in food-grade productions pharmaceutical enzyme manufacture, it is meaningful to identify the biochemical properties of OSBP in A. oryzae.

PATHOTYPE GROUPING OF Xanthomonas oryzae pv. oryzae ISOLATES FROM SOUTH SULAWESI AND SOUTHEAST SULAWESI

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Andi Khaeruni

2013-06-01

Full Text Available Bacterial leaf blight caused by Xanthomonas oryzae pv. oryzae (Xoo is an important rice disease, and has caused significant economic losses. This research aimed to determine the pathotype grouping and the distribution of Xoo isolates of South and Southeast Sulawesi. In order to obtain the information, 61 Xoo isolates of South Sulawesi and 29 isolates of Southeast Sulawesi were evaluated for their pathotype grouping against 5 diffential varieties. Research results showed that in South Sulawesi there were 2 pathotype groups, namely pathotype IV (32.79% and pathotype VIII(67.21%. Pathotype VIII was widely distributed over the Western and Central areas of South Sulawesi, whereas pathotype IV was widely distributed over the Southern area. In Southeast Sulawesi, it was found 5 pathotypes, namely pathotypes IV (27.58%, VI (10.34%, VIII (13.79%, IX (20.68%, and X (27.58%, with a limited and scattered distribution pattern on several areas. These results indicate that Xoo pathotype groups in South Sulawesi and Southeast Sulawesi are varied and tend to sift to more virulent pathotypes.

L-lactic acid production from starch by simultaneous saccharification and fermentation in a genetically engineered Aspergillus oryzae pure culture.

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Wakai, Satoshi; Yoshie, Toshihide; Asai-Nakashima, Nanami; Yamada, Ryosuke; Ogino, Chiaki; Tsutsumi, Hiroko; Hata, Yoji; Kondo, Akihiko

2014-12-01

Lactic acid is a commodity chemical that can be produced biologically. Lactic acid-producing Aspergillus oryzae strains were constructed by genetic engineering. The A. oryzae LDH strain with the bovine L-lactate dehydrogenase gene produced 38 g/L of lactate from 100g/L of glucose. Disruption of the wild-type lactate dehydrogenase gene in A. oryzae LDH improved lactate production. The resulting strain A. oryzae LDHΔ871 produced 49 g/L of lactate from 100g/L of glucose. Because A. oryzae strains innately secrete amylases, A. oryzae LDHΔ871 produced approximately 30 g/L of lactate from various starches, dextrin, or maltose (all at 100 g/L). To our knowledge, this is the first report describing the simultaneous saccharification and fermentation of lactate from starch using a pure culture of transgenic A. oryzae. Our results indicate that A. oryzae could be a promising host for the bioproduction of useful compounds such as lactic acid. Copyright © 2014 Elsevier Ltd. All rights reserved.

Identification and toxigenic potential of the industrially important fungi, Aspergillus oryzae and Aspergillus sojae.

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Jørgensen, Thomas R

2007-12-01

Mold strains belonging to the species Aspergillus oryzae and Aspergillus sojae are highly valued as koji molds in the traditional preparation of fermented foods, such as miso, sake, and shoyu, and as protein production hosts in modern industrial processes. A. oryzae and A. sojae are relatives of the wild molds Aspergillus flavus and Aspergillus parasiticus. All four species are classified to the A. flavus group. Strains of the A. flavus group are characterized by a high degree of morphological similarity. Koji mold species are generally perceived of as being nontoxigenic, whereas wild molds are associated with the carcinogenic aflatoxins. Thus, reliable identification of individual strains is very important for application purposes. This review considers the pheno- and genotypic markers used in the classification of A. flavus group strains and specifically in the identification of A. oryzae and A. sojae strains. Separation of A. oryzae and A. sojae from A. flavus and A. parasiticus, respectively, is inconsistent, and both morphologic and molecular evidence support conspecificity. The high degree of identity is reflected by the divergent identification of reference cultures maintained in culture collections. As close relatives of aflatoxin-producing wild molds, koji molds possess an aflatoxin gene homolog cluster. Some strains identified as A. oryzae and A. sojae have been implicated in aflatoxin production. Identification of a strain as A. oryzae or A. sojae is no guarantee of its inability to produce aflatoxins or other toxic metabolites. Toxigenic potential must be determined specifically for individual strains. The species taxa, A. oryzae and A. sojae, are currently conserved by societal issues.

Characteristics of immobilized aminoacylase from Aspergillus oryzae on macroporous copolymers.

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He, B L; Jiang, P; Qiu, Y B

1990-01-01

Aminoacylase from Aspergillus oryzae was adsorbed on functionallized macroporous copolymers where the enzyme showed excellent catalyzing activity and operation stability. Various factors which effect the activity of the immobilized aminoacylase such as temperature, pH and ionic strength were investigated. The continuous operation of the enzyme immobilized on macroporous copolymers was compared with that of the enzyme immobilized on DEAE-Sephadex.

Correlation between temperature and phenology prediction error in rice (Oryza sativa L.)

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Oort, van P.A.J.; Zhang, T.; Vries, de M.E.; Heinemann, A.B.; Meinke, H.B.

2011-01-01

For rice (Oryza sativa L.), simulation models like ORYZA2000 and CERES-Rice have been used to explore adaptation options to climate change and weather-related stresses (drought, heat). Output of these models is very sensitive to accurate modelling of crop development, i.e. phenology. What has to

Utilization of Aspergillus oryzae to produce pectin lyase from various agro-industrial residues

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Safia Koser

2014-07-01

Full Text Available The present study was aimed to investigate the culture influence on pectin lyase production potential of fungal strain Aspergillus oryzae. The enzyme profile of A. oryzae showed highest activity of pectin lyase after 3rd day of incubation on lemon peel waste under solid state fermentation conditions. To induce the pectin lyase synthesis capability of A. oryzae at optimal level various culture variables including physical and nutritional parameters were optimized by adopting classical optimization technique. Therefore, through fermentation process optimization the production of pectin lyase was substantially induced up to the level of 875 U/mL, when fermentation medium of lemon peel waste inoculated with 5 mL spore suspension of A. oryzae. The optimal fermentation conditions for maximum pectin lyase yield were as: optimum pH 5, 70% moisture level and incubated at 40 °C in addition with 1% sterile glucose solution as readily available carbon source and 0.2% yeast extract as an inexpensive nitrogen supplement (1%. The results obtained in current investigation so far demonstrated that culture conditions have great influence on the pectin lyase production potential of A. oryzae.

Avaliação de inseticidas no controle de Sitophilus oryzae (Linnaeus (Coleoptera, Curculionidae e Rhyzopertha dominica (Fabricius (Coleoptera, Bostrichidae em trigo armazenado Efficacy of insecticides to control Sitophilus oryzae (Linnaeus (Coleoptera, Curculionidae and Rhyzopertha dominica (Fabricius (Coleoptera, Bostrichidae in stored wheat

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Rui S. Furiatti

1999-03-01

Full Text Available The efficiency of insecticides against Sitophilus oryzae (Linnaeus, 1763 (Coleoptera, Curculionidae and Rhyzopertha dominica (Fabricius, 1792 (Coleoptera, Bostrichidae was determined exposing them to treated wheat in laboratory. Each plot, which was composed by 1.2 kg of wheat grain, was treated with 5 ml of the concentration/kg of wheat using a propelling air atomizer. Fifteen days after treatment 50 g samples of the grain were taken, from each plot, placed in plastic jars into which 10 adult insects of each species were released. This was repeated every 30 days until 165 days after treatment. The number of dead insects was counted 15 days after each infestation thus the evaluations were made at 30, 60, 90, 120, 150 and 180 days after the insecticide application. Esfenvalerate+fenitrothion+pyperonil butoxide (0,375+7,5+3,0 and 0,5+10,0+4,0 g a.i./t, fenitrothion+esfen-valerate (7,5+0,375 and 10,0+0,375 g a.i./t, fenitrothion+synergised deltamethrin (7,5+0,375 g a.i./t and fenitrothion (7,5 g a.i./t were effective against Sitophilus oryzae until 180 days after treatment. Esfenvalerate+fenitrothion+pyperonil butoxide (0,5+10,0+4,0 g a.i./t, fenitrothion+esfenvalerate (7,5+0,375 and 10,0+0,375 g a.i./t, fenitrothion+synergised deltamethrin (7,5+0,375 g a.i./t were effective against R. dominica until 180 days after treatment. Synergised deltamethrin (0,375 g a.i./t was effective against S. oryzae until 150 days and against R. dominica until 120 days after treatment. Esfenvalerate (0,5 g a.i./t was ineffective against S. oryzae and effective until 60 days after treatment against R. dominica. Fenitrothion (7,5 g a.i./t was ineffective against R. dominica. The use of mixtures of organophosphorus plus pyrethroids were more effective against S. oryzae and R. dominica when compared with these insecticides used alone.

Biosorption of americium-241 by immobilized Rhizopus arrihizus

International Nuclear Information System (INIS)

Liao Jiali; Yang Yuanyou; Luo Shunzhong; Liu Ning; Jin Jiannan; Zhang Taiming; Zhao Pengji

2004-01-01

Rhizopus arrihizus (R. arrihizus), a fungus, which in previous experiments had shown encouraging ability to remove 241 Am from solutions, was immobilized by calcium alginate and other reagents. The various factors affecting 241 Am biosorption by the immobilized R. arrihizus were investigated. The results showed that not only can immobilized R. arrihizus adsorb 241 Am as efficiently as free R. arrihizus, but that also can be used repeatedly or continuously. The biosorption equilibrium was achieved within 2 h, and more than 94% of 241 Am was removed from 241 Am solutions of 1.08 MBq/l by immobilized R. arrihizu in the pH range 1-7. Temperature did not affect the adsorption on immobilized R. arrihizus in the range 15-45 deg. C. After repeated adsorption for 8 times, the immobilized R. arrihizus still adsorbed more than 97% of 241 Am. At this time, the total adsorption of 241 Am was more than 88.6 KBq/g, and had not yet reached saturation. Ninety-five percent of the adsorbed 241 Am was desorbed by saturated EDTA solution and 98% by 2 mol/l HNO 3

Purification and characterization of a thermostable hypothetical xylanase from Aspergillus oryzae HML366.

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He, Haiyan; Qin, Yongling; Li, Nan; Chen, Guiguang; Liang, Zhiqun

2015-03-01

In the current study, fermentation broth of Aspergillus oryzae HML366 in sugar cane bagasse was subjected to ultrafiltration and ion exchange chromatography, and two xylanases, XynH1 and XynH2, were purified. Time-of-flight mass spectrometry coupled with SDS-PAGE analysis revealed that XynH1 is identical to the hypothetical A. oryzae RIB40 protein XP_001826985.1, with a molecular weight of 33.671 kDa. Likewise, XynH2 was identified as xylanase XynF1 with a molecular weight of 35.402 kDa. Sequence analysis indicated that XynH1 belongs to glycosyl hydrolases family 10. The specific activity of XynH1 was measured at 476.9 U/mg. Optimal xylanase activity was observed at pH 6.0, and enzyme remained active within pH 4.0-10.0 and at a temperature below 70 °C. Mg(2+), Mn(2+), Ca(2+), and K(+) enhanced the XynH1 xylanase activity to 146, 122, 114, and 108%, respectively. XynH1 hydrolyzed Birchwood xylan and Larchwood xylan effectively. The K m and V max of XynH1 values determined were 1.16 mM and 336 μmol/min/mg with Birchwood xylan as the substrate. A. oryzae HML366 xylanase XynH1 showed superior heat and pH tolerance, therefore may have significant applications in paper and biofuel industries. These studies constitute the first investigation of the xylanase activities of the hypothetical protein XP_001826985.1 form A. oryzae.

Aspergillus oryzae NRRL 35191 from coffee, a non-toxigenic endophyte with the ability to synthesize kojic acid

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Aspergillus oryzae was isolated as an endophyte from coffee leaves and found to produce kojic acid in culture. When inoculated in cacao seedlings (Theobroma cacao L.), A. oryzae grew endophytically and synthesize kojic acid in planta. Cacao seedlings inoculated with A. oryzae produced higher levels...

ISOLATION OF FUNGI FROM THE GASTROINTESTINAL TRACT OF INDIGENOUS CHICKEN

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T. Yudiarti

2014-10-01

Full Text Available Gastrointestinal tract of chicken is a place in which many kinds of fungi can be found. The aim ofthe research was to isolate fungi from the gastrointestinal tract of the indigenous chicken (AyamKampung. The chicken samples were four days, one week and two months old and were sampled fromchicken farm located in Yogyakarta. Potato dextrose agar (PDA medium was used to grow the fungi.Fifty pure isolates of fungi were found from three different ages, those were four days, one week andtwo months old chicken were 5, 10 and 35 isolates respectively. The largest number of isolate was foundin ileum, then followed by caecum, jejenum and duodenum. The fifty isolate of fungi belonged to sevenspecies, those were Aspergillus fumigatus, Aspergillus niger, Chrysonilia crassa, Mucor circinelloides,Mucor sp, Rhizopus oligosporus and Rhizopus oryzae.

ISOLATION OF FUNGI FROM THE GASTROINTESTINAL TRACT OF INDIGENOUS CHICKEN

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E. Kusdiyantini

2012-06-01

Full Text Available Gastrointestinal tract of chicken is a place in which many kinds of fungi can be found. The aim of the research was to isolate fungi from the gastrointestinal tract of the indigenous chicken (Ayam Kampung. The chicken samples were four days, one week and two months old and were sampled from chicken farm located in Yogyakarta. Potato dextrose agar (PDA medium was used to grow the fungi. Fifty pure isolates of fungi were found from three different ages, those were four days, one week and two months old chicken were 5, 10 and 35 isolates respectively. The largest number of isolate was found in ileum, then followed by caecum, jejenum and duodenum. The fifty isolate of fungi belonged to seven species, those were Aspergillus fumigatus, Aspergillus niger, Chrysonilia crassa, Mucor circinelloides, Mucor sp, Rhizopus oligosporus and Rhizopus oryzae.

Imobilização de lipases em filme de caseinato de sódio/glicerol: aplicação na síntese de ésteres Lipase immobilization in sodium caseinate/glycerol film: application in ester synthesis

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Damianni Sebrão

2007-10-01

Full Text Available Lipases from different sources were immobilized in sodium caseinate/glycerol film and used in the esterification reactions of aliphatic acids with alcohols in the presence of organic solvents. Lipases from Pseudomonas sp and Rhizopus oryzae were selected and the influence of several parameters was analyzed, including: lipase loading, organic solvent polarity, reaction temperature, chain length of alcohol and acid and enzyme/support reuse. For comparison, free enzymes were used under similar experimental conditions.

Control del desarrollo de hongos contaminantes de alimentos mediante iones dosificados por materiales microporosos.

OpenAIRE

Chiericatti, Carolina Andrea Antonia; Chiericatti, Carolina Andrea Antonia

2014-01-01

Se estudió el efecto inhibitorio de Ag y Cu incorporado en matrices microporosas sobre el crecimiento de diferentes hongos, que son problemáticos en las industrias alimenticias de nuestra zona. Se estudiaron las especies fúngicas: Rhizopus oryzae, Mucor circinelloides, Geotrichum candidum, Saccharomyces cerevisiae, Debaryomyces hansenii y Zygosaccharomyces rouxii. Los Hongos filamentosos fueron aislados de productos lácteos que provocaron alteración a baja tensión de oxígeno y las levaduras f...

Activity of posaconazole and other antifungal agents against Mucorales strains identified by sequencing of internal transcribed spacers.

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Alastruey-Izquierdo, Ana; Castelli, Maria Victoria; Cuesta, Isabel; Monzon, Araceli; Cuenca-Estrella, Manuel; Rodriguez-Tudela, Juan Luis

2009-04-01

The antifungal susceptibility profiles of 77 clinical strains of Mucorales species, identified by internal transcribed spacer sequencing, were analyzed. MICs obtained at 24 and 48 h were compared. Amphotericin B was the most active agent against all isolates, except for Cunninghamella and Apophysomyces isolates. Posaconazole also showed good activity for all species but Cunninghamella bertholletiae. Voriconazole had no activity against any of the fungi tested. Terbinafine showed good activity, except for Rhizopus oryzae, Mucor circinelloides, and Rhizomucor variabilis isolates.

Endophytic Streptomyces spp. as Biocontrol Agents of Rice Bacterial Leaf Blight Pathogen (Xanthomonas oryzae pv. oryzae

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RATIH DEWI HASTUTI

2012-12-01

Full Text Available Xanthomonas oryzae pv. oryzae (Xoo, a causal agent of bacterial leaf blight (BLB, is one of the most important pathogens of rice. The effectiveness of ten Streptomyces spp. isolates in suppressing Xoo disease was assessed in planta and in vitro. In planta experiments were carried out in a greenhouse and arranged in a randomized completely block design (RCBD with three replications. Twenty treatments were tested which included plants inoculated with both Streptomyces spp. and Xoo, and plants inoculated with only Streptomyces spp. Plants inoculated with Xoo and sprayed with a chemical bactericide, and plants inoculated with only Xoo served as positive controls, whereas plants not inoculated with either Streptomyces spp. or Xoo were used as negative controls. The results showed that the effect of endophytic Streptomyces spp. on BLB disease expressed as area under disease progress curve (AUDPC was not significantly different to that on control plants (P > 0.05. However, plants inoculated with endophytic Streptomyces spp. were significantly taller and produced higher tiller number than control plants (P < 0.05. Streptomyces spp. isolate AB131-1 gave the highest plant height. In vitro studies on biocontrol mechanisms of selected Streptomyces spp. isolates showed that isolate LBR02 gave the highest inhibition activity on Xoo growth, followed by AB131-1 and AB131-2. Two isolates (AB131-1 and LBR02 were able to produce chitinase, phosphatase, and siderophore which included biocontrol characteristics. Morphological and colonization studies under SEM and light microscopy confirmed that the three isolates were endophytic Streptomyces spp. from different species. These studies found that the paddy plant which was inoculated with endophytic Streptomyces spp. AB131-1 and infected by Xoo could increase the height of plant and number of tillers.

Evaluation of commercial soy sauce koji strains of Aspergillus oryzae for γ-aminobutyric acid (GABA) production.

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Ab Kadir, Safuan; Wan-Mohtar, Wan Abd Al Qadr Imad; Mohammad, Rosfarizan; Abdul Halim Lim, Sarina; Sabo Mohammed, Abdulkarim; Saari, Nazamid

2016-10-01

In this study, four selected commercial strains of Aspergillus oryzae were collected from soy sauce koji. These A. oryzae strains designated as NSK, NSZ, NSJ and NST shared similar morphological characteristics with the reference strain (A. oryzae FRR 1675) which confirmed them as A. oryzae species. They were further evaluated for their ability to produce γ-aminobutyric acid (GABA) by cultivating the spore suspension in a broth medium containing 0.4 % (w/v) of glutamic acid as a substrate for GABA production. The results showed that these strains were capable of producing GABA; however, the concentrations differed significantly (P sauce production.

An ecogeographic analysis of Oryza series Sativae in Asia and the Pacific

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Banaticla-Hilario, M.C.N.

2012-01-01

The non-cultivated speciesof the genus Oryza can provide a genetic arsenal of useful traits for improving the widely cultivated and consumed Asian rice (O. sativa). The diversity of these valuable plant resources must be well understood to ensure their effective in- and ex-situ

The rice XA21 ectodomain fused to the Arabidopsis EFR cytoplasmic domain confers resistance to Xanthomonas oryzae pv. oryzae.

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Thomas, Nicholas C; Oksenberg, Nir; Liu, Furong; Caddell, Daniel; Nalyvayko, Alina; Nguyen, Yen; Schwessinger, Benjamin; Ronald, Pamela C

2018-01-01

Rice ( Oryza sativa ) plants expressing the XA21 cell-surface receptor kinase are resistant to Xanthomonas oryzae pv. oryzae (Xoo) infection. We previously demonstrated that expressing a chimeric protein containing the ELONGATION FACTOR Tu RECEPTOR (EFR) ectodomain and the XA21 endodomain (EFR:XA21) in rice does not confer robust resistance to Xoo . To test if the XA21 ectodomain is required for Xoo resistance, we produced transgenic rice lines expressing a chimeric protein consisting of the XA21 ectodomain and EFR endodomain (XA21:EFR) and inoculated these lines with Xoo . We also tested if the XA21:EFR rice plants respond to a synthetic sulfated 21 amino acid derivative (RaxX21-sY) of the activator of XA21-mediated immunity, RaxX. We found that five independently transformed XA21:EFR rice lines displayed resistance to Xoo as measured by lesion length analysis, and showed that five lines share characteristic markers of the XA21 defense response (generation of reactive oxygen species and defense response gene expression) after treatment with RaxX21-sY. Our results indicate that expression of the XA21:EFR chimeric receptor in rice confers resistance to Xoo . These results suggest that the endodomain of the EFR and XA21 immune receptors are interchangeable and the XA21 ectodomain is the key determinant conferring robust resistance to Xoo .

Comparison of the genomes and transcriptomes associated with the different protease secretions of Aspergillus oryzae 100-8 and 3.042.

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Zhao, Guozhong; Yao, Yunping; Hou, Lihua; Wang, Chunling; Cao, Xiaohong

2014-10-01

Aspergillus oryzae is used to produce traditional fermented foods and beverages. A. oryzae 3.042 produces a neutral protease and an alkaline protease but rarely an acid protease, which is unfavourable to soy-sauce fermentation. A. oryzae 100-8 was obtained by N(+) ion implantation mutagenesis of A. oryzae 3.042, and the protease secretions of these two strains are different. Sequencing the genome of A. oryzae 100-8 and comparing it to the genomes of A. oryzae 100-8 and 3.042 revealed some differences, such as single nucleotide polymorphisms, nucleotide deletion or insertion. Some of these differences may reflect the ability of A. oryzae to secrete proteases. Transcriptional sequencing and analysis of the two strains during the same growth processes provided further insights into the genes and pathways involved in protease secretion.

High-yields heterologous production of the novel Aspergillus fumigatus elastase inhibitor AFUEI in Aspergillus oryzae.

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Yamashita, Nobuo; Komori, Yumiko; Okumura, Yoshiyuki; Uchiya, Kei-Ichi; Matsui, Takeshi; Nishimura, Akira; Ogawa, Kenji; Nikai, Toshiaki

2011-08-01

AFUEI, an elastase inhibitor produced by Aspergillus fumigatus strongly inhibits the elastolytic activity of A. fumigatus etc. To purify AFUEI, we constructed a strain that overproduces AFUEI by introducing the gene encoding AFUEI (Genbank accession no. AB546725) under control of the amyB promoter into the heterologous host Aspergillus oryzae. A. oryzae TF-4 displayed strong elastase inhibitory activity and produced considerably more AFUEI than that of A. fumigatus. Furthermore, AFUEI could be purified using culture broth and single ultrafiltration (UF) treatment, allowing for the effective production of AFUEI for use in clinical trials. Copyright © 2011 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Kinetics of improved 1,4-alpha-D-glucan glucohydrolase biosynthesis from a newly isolated Aspergillus oryzae IIB-6 and parameter significance analysis by 2-factorial design.

Science.gov (United States)

Fatima, Bilqees; Ali, Sikander

2012-01-01

Sixteen different mould cultures viz. Aspergillus, Alternaria, Arthroderma, Trichoderma, Fusarium, Penicillium, Rhizopus and Chochliobolus were isolated from the soil samples of Qatar by serial dilution method. The preliminary screening of isolates was done by selecting initial colonies showing relatively bigger zones of starch hydrolysis on nutrient agar plates. The isolates were then subjected to secondary screening by submerged fermentation (SmF). The 1,4-α-D-glucan glucohydrolase (GGH) activity ranged from 1.906-12.675 U/ml/min. The product yield was analysed in dependence of mycelial morphology, biomass level and protein content. The isolate Aspergillus oryzae llB-6 which gave maximum enzyme production was incubated in M3 medium containing 20 g/l starch, 10 g/l lactose, 8.5 g/l yeast extract, 6 g/l corn steep liquor (CSL), 1.2 g/l MgSO4.7H2O, 1.3 g/l NH4Cl, 0.6 g/l CaCl2.2H2O, pH 5 at 30±2°C and 200 rpm. On the basis of kinetic variables, notably Qp (0.058±0.01(a) U/g/h), Yp/s (0.308±0.03(ab) U/g) and qp (0.210±0.032(abc) U/g fungal biomass/h), A. oryzae IIB-6 was found to be a hyper producer of GGH (LSD 0.0345) compared to A. kawachii IIB-2. A noticeable enhancement in enzyme activity of over 30% was observed (13.917±1.01 U/ml/min) when the process parameters viz. cultural conditions (pH 5, incubation period 72 h) and nutritional requirements (6 g/l CSL, 9.5 g/l yeast extract, 10 g/l starch, 20 g/l lactose) were further optimized using a 2-factorial Plackett-Burman design. The model terms were found to be highly significant (HS, p≤0.05), indicating the potential utility of the culture (dof~3).

Volatiles Mediated Interactions Between Aspergillus oryzae Strains Modulate Morphological Transition and Exometabolomes.

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Singh, Digar; Lee, Choong H

2018-01-01

Notwithstanding its mitosporic nature, an improbable morpho-transformation state i. e., sclerotial development (SD), is vaguely known in Aspergillus oryzae . Nevertheless an intriguing phenomenon governing mold's development and stress response, the effects of exogenous factors engendering SD, especially the volatile organic compounds (VOCs) mediated interactions (VMI) pervasive in microbial niches have largely remained unexplored. Herein, we examined the effects of intra-species VMI on SD in A. oryzae RIB 40, followed by comprehensive analyses of associated growth rates, pH alterations, biochemical phenotypes, and exometabolomes. We cultivated A. oryzae RIB 40 (S1 VMI : KACC 44967) opposite a non-SD partner strain, A. oryzae (S2: KCCM 60345), conditioning VMI in a specially designed "twin plate assembly." Notably, SD in S1 VMI was delayed relative to its non-conditioned control (S1) cultivated without partner strain (S2) in twin plate. Selectively evaluating A. oryzae RIB 40 (S1 VMI vs. S1) for altered phenotypes concomitant to SD, we observed a marked disparity for corresponding growth rates (S1 VMI S1) 7days , and biochemical characteristics viz ., protease (S1 VMI > S1) 7days , amylase (S1 VMI > nS1) 3-7 days , and antioxidants (S1 VMI > S1) 7days levels. The partial least squares-discriminant analysis (PLS-DA) of gas chromatography-time of flight-mass spectrometry (GC-TOF-MS) datasets for primary metabolites exhibited a clustered pattern (PLS1, 22.04%; PLS2, 11.36%), with 7 days incubated S1 VMI extracts showed higher abundance of amino acids, sugars, and sugar alcohols with lower organic acids and fatty acids levels, relative to S1. Intriguingly, the higher amino acid and sugar alcohol levels were positively correlated with antioxidant activity, likely impeding SD in S1 VMI . Further, the PLS-DA (PLS1, 18.11%; PLS2, 15.02%) based on liquid chromatography-mass spectrometry (LC-MS) datasets exhibited a notable disparity for post-SD (9-11 days) sample extracts

Volatiles Mediated Interactions Between Aspergillus oryzae Strains Modulate Morphological Transition and Exometabolomes

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Digar Singh

2018-04-01

Full Text Available Notwithstanding its mitosporic nature, an improbable morpho-transformation state i. e., sclerotial development (SD, is vaguely known in Aspergillus oryzae. Nevertheless an intriguing phenomenon governing mold's development and stress response, the effects of exogenous factors engendering SD, especially the volatile organic compounds (VOCs mediated interactions (VMI pervasive in microbial niches have largely remained unexplored. Herein, we examined the effects of intra-species VMI on SD in A. oryzae RIB 40, followed by comprehensive analyses of associated growth rates, pH alterations, biochemical phenotypes, and exometabolomes. We cultivated A. oryzae RIB 40 (S1VMI: KACC 44967 opposite a non-SD partner strain, A. oryzae (S2: KCCM 60345, conditioning VMI in a specially designed “twin plate assembly.” Notably, SD in S1VMI was delayed relative to its non-conditioned control (S1 cultivated without partner strain (S2 in twin plate. Selectively evaluating A. oryzae RIB 40 (S1VMI vs. S1 for altered phenotypes concomitant to SD, we observed a marked disparity for corresponding growth rates (S1VMI < S17days, media pH (S1VMI > S17days, and biochemical characteristics viz., protease (S1VMI > S17days, amylase (S1VMI > nS13–7days, and antioxidants (S1VMI > S17days levels. The partial least squares—discriminant analysis (PLS-DA of gas chromatography—time of flight—mass spectrometry (GC-TOF-MS datasets for primary metabolites exhibited a clustered pattern (PLS1, 22.04%; PLS2, 11.36%, with 7 days incubated S1VMI extracts showed higher abundance of amino acids, sugars, and sugar alcohols with lower organic acids and fatty acids levels, relative to S1. Intriguingly, the higher amino acid and sugar alcohol levels were positively correlated with antioxidant activity, likely impeding SD in S1VMI. Further, the PLS-DA (PLS1, 18.11%; PLS2, 15.02% based on liquid chromatography—mass spectrometry (LC-MS datasets exhibited a notable disparity for post-SD (9�

Measurement and analysis on optical characteristics of Aspergillus oryzae spores in infrared band

Science.gov (United States)

Li, Le; Hu, Yihua; Gu, Youlin; Chen, Wei; Xu, Shilong; Zhao, Xinying

2015-10-01

Spore is an important part of bioaerosols. The optical characteristics of spore is a crucial parameter for study on bioaerosols. The reflection within the waveband of 2.5 to15μm were measured by squash method. Based on the measured data, Complex refractive index of Aspergillus oryzae spores within the waveband of 3 to 5μm and 8 to 14 μm were calculated by using Krames-Kronig (K-K) relationship. Then,the mass extinction coefficient of Aspergillus oryzae spores within the waveband of 3 to 5μm and 8 to 14μm were obtained by utilizing Mie scattering theory, and the results were analyzed and discussed. The average mass extinction coefficient of Aspergillus oryzae spores is 0.51 m2/g in the range of 3 to 5μm and 0.48m2/g in the range of 8 to 14μm. Compared with common inorganic compounds, Aspergillus oryzae spores possesses a good extinction performance in infrared band.

MIKROFLORA PADA TEMPOYAK The Microflora of Tempoyak

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Hasanuddin Hasanuddin

2012-05-01

Full Text Available Tempoyak is a traditional Bengkulu fermented food prepared from spontaneous fermentation of durian (Durio zibethi- nus by wild microorganisms. Tempoyak is found not only in Bengkulu but also as long as Sumatera island with differ- ent names, and also in Malaysia. Tempoyak is a Melayu term meaning fermented durian. The research was conducted to identify microflora in tempoyak, and to select the species of microorganisms which were able to ferment durian ef- fectivelly. The data in this study were laboratory analysis. Samples were collected weekly in a month from productions centre of tempoyak and traditional markets, and analyzed microbiologically to determine and identify the bacteria, yeasts and molds in tempoyak. There were four specieses of lactic acid bacteria found in fermented durian namely Pediococcus acidilactici, Lactobacillus plantarum Lactobacillus curvatus and Leuconostoc mesentroides. While two specieses which were not lactic acid bacteria namely Staphylococcus saprophyticus and Micrococcus varians. One species of yeast was Kluyveromyces marxianus. The molds were identified as Rhizopus oryzae, Monilia sitophila, Mucor roxii, Aspergillus repens and Penicillium sp. Among these specieses three specieses were produced lactic acid namelly Rhizopus oryzae, Monilia sitophila,and Mucor roxii. While Aspergillus ripens and Penicillium sp did not pro- duce lactic acid so these specieses might not function in fermentation process. ABSTRAK Tempoyak adalah makanan khas Bengkulu, yang dibuat dari fermentasi spontan daging buah durian (Durio zibet- hinus oleh mikroorganisme liar. Tempoyak juga bisa dijumpai di berbagai daerah Indonesia di sepanjang pulau Sumatera dengan nama yang berbeda, bahkan sampai ke Malaysia. Istilah tempoyak berasal dari terminolgi Melayu yang artinya adalah durian fermentasi. Penelitian ini bertujuan untuk mengidentifikasi mikroorganisme yang terdapat pada tempoyak. Data dari penelitian ini adalah data primer dari analisa

Insecticidal Effect of Fruit Extracts from Xylopia aethiopica and Dennettia tripetala (Annonaceae against Sitophilus oryzae (Coleoptera: Curculionidae Efecto Insecticida de Extractos de Fruta de Xylopia aethiopica y Dennettia tripetala (Annonaceae contra Sitophilus oryzae (Coleoptera: Curculionidae

Directory of Open Access Journals (Sweden)

Donald A . Ukeh

2012-06-01

Full Text Available The insecticidal and repellent activities of fruit extracts of Xylopia aethiopica (Dunal A. Rich. and Dennettia tripetala (Baker f. G.E. Schatz belonging to the family Annonaceae was studied against Sitophilus oryzae (L., an economic, primary post-harvest pest of rice, and other cereal products. Infested rice grains (100 g treated with 1, 2, 3, 4, and 5% (w/w powders of both plants were evaluated for toxicity against S. oryzae every 24 h for 3 d, and during Fi progeny emergence. The essential oils of both plants were also applied at 0.5, 1, 1.5 and 2 mg cm-2 filter paper in Petri dishes for toxicity bioassays at 24 h exposure. Repellence bioassay with 10 μL solution of essential oils on filter paper was performed in a Y-Tube airflow olfactometer. Results indicate that powders of both plants significantly (P La actividad insecticida y repelente de los extractos frutales de Xylopia aethiopica (Dunal A. Rich. y Dennettia tripetala (Baker f. G.E. Schatz pertenecientes a la familia Annonaceae fueron evaluados contra Sitophilus oryzae (L., plaga primaria de importancia económica en poscosecha de arroz y otros cereales. Granos de arroz (100 g infestados tratados con polvos de ambas plantas al 1, 2, 3, 4, y 5% (p/p fueron evaluados para la toxicidad contra S. oryzae cada 24 h por 3 d y durante la emergencia de la progenie F1. Los aceites esenciales de ambas plantas también fueron aplicados en papel filtro a 0,5; 1; 1,5 y 2 mg cm-2 en cajas de Petri para bioensayos de toxicidad con exposición de 24 h. Bioensayos de repelencia con 10 μL de solución de los aceites esenciales impregnados en papel filtro fueron realizados en un olfatómetro de flujo de aire Y-Tube. Los resultados indican que los polvos de ambas plantas causan una mortalidad significativa de estos insectos (P < 0,001 y una reducción en la emergencia de la progenie F1 con relación al control. Los aceites esenciales también mostraron un efecto adulticida significativo (P < 0,001 despu

Genetic analysis of yield and yield components in Oryza sativa x ...

African Journals Online (AJOL)

... inheritance of yield and yield components and to estimate the heritabilities of important quantitative traits in rice (Oryza sativa L.). Six generations viz., P1, P2, F1, F2, BCP1 and BCP2 of a cross between IET6279 and IR70445-146-3-3 were used for the study. Generation mean analysis suggested that additive effects had a ...

Rise of a Cereal Killer: The Biology of Magnaporthe oryzae Biotrophic Growth.

Science.gov (United States)

Fernandez, Jessie; Orth, Kim

2018-01-23

The rice blast fungus, Magnaporthe oryzae, causes one of the most destructive diseases of cultivated rice in the world. Infections caused by this recalcitrant pathogen lead to the annual destruction of approximately 10-30% of the rice harvested globally. The fungus undergoes extensive developmental changes to be able to break into plant cells, build elaborate infection structures, and proliferate inside host cells without causing visible disease symptoms. From a molecular standpoint, we are still in the infancy of understanding how M. oryzae manipulates the host during this complex multifaceted infection. Here, we describe recent advances in our understanding of the cell biology of M. oryzae biotrophic interaction and key molecular factors required for the disease establishment in rice cells. Copyright © 2017 Elsevier Ltd. All rights reserved.

Comparative chemistry of Aspergillus oryzae (RIB40) and A. flavus (NRRL 3357)

DEFF Research Database (Denmark)

Rank, Christian; Klejnstrup, Marie Louise; Petersen, Lene Maj

2012-01-01

Aspergillus oryzae and A. flavus are important species in industrial biotechnology and food safety and have been some of the first aspergilli to be fully genome sequenced. Bioinformatic analysis has revealed 99.5% gene homology between the two species pointing towards a large coherence in the sec...... alkaloids related to the A. flavus metabolites ditryptophenalines and miyakamides. Generally the secondary metabolite capability of A. oryzae presents several novel end products likely to result from the domestication process from A. flavus.......Aspergillus oryzae and A. flavus are important species in industrial biotechnology and food safety and have been some of the first aspergilli to be fully genome sequenced. Bioinformatic analysis has revealed 99.5% gene homology between the two species pointing towards a large coherence...

Glycogen Metabolic Genes Are Involved in Trehalose-6-Phosphate Synthase-Mediated Regulation of Pathogenicity by the Rice Blast Fungus Magnaporthe oryzae

Science.gov (United States)

Wilson, Richard A.; Wang, Zheng-Yi; Kershaw, Michael J.; Talbot, Nicholas J.

2013-01-01

The filamentous fungus Magnaporthe oryzae is the causal agent of rice blast disease. Here we show that glycogen metabolic genes play an important role in plant infection by M. oryzae. Targeted deletion of AGL1 and GPH1, which encode amyloglucosidase and glycogen phosphorylase, respectively, prevented mobilisation of glycogen stores during appressorium development and caused a significant reduction in the ability of M. oryzae to cause rice blast disease. By contrast, targeted mutation of GSN1, which encodes glycogen synthase, significantly reduced the synthesis of intracellular glycogen, but had no effect on fungal pathogenicity. We found that loss of AGL1 and GPH1 led to a reduction in expression of TPS1 and TPS3, which encode components of the trehalose-6-phosphate synthase complex, that acts as a genetic switch in M. oryzae. Tps1 responds to glucose-6-phosphate levels and the balance of NADP/NADPH to regulate virulence-associated gene expression, in association with Nmr transcriptional inhibitors. We show that deletion of the NMR3 transcriptional inhibitor gene partially restores virulence to a Δagl1Δgph1 mutant, suggesting that glycogen metabolic genes are necessary for operation of the NADPH-dependent genetic switch in M. oryzae. PMID:24098112

A mutation in the aroE gene affects pigment production, virulence, and chemotaxis in Xanthomonas oryzae pv. oryzae.

Science.gov (United States)

Kim, Hong-Il; Noh, Tae-Hwan; Lee, Chang-Soo; Park, Young-Jin

2015-01-01

Xanthomonas oryzae pv. oryzae (Xoo) causes bacterial blight (BB) in rice. To study its function, a random insertion mutation library of Xoo was constructed using the Tn5 transposon. A mutant strain with decreased virulence against the susceptible rice cultivar IR24 was isolated from the library (aroE mutant), which also had extremely low pigment production. Thermal asymmetric interlaced-polymerase chain reaction (TAIL-PCR) and sequence analysis of the mutant revealed that the transposon was inserted into the aroE gene (encoding shikimate dehydrogenase). To investigate gene expression changes in the pigment- and virulence-deficient mutant, DNA microarray analysis was performed, which showed downregulation of 20 genes involved in the chemotaxis of Xoo. Our findings reveal that mutation of the aroE gene affects virulence and pigment production, as well as expression of genes involved in Xoo chemotaxis. Copyright © 2014 Elsevier GmbH. All rights reserved.

Identification of secreted proteins of Aspergillus oryzae associated with growth on solid cereal substrates

NARCIS (Netherlands)

Biesebeke, R. te; Boussier, A.; Biezen, N. van; Hondel, C.A.M.J.J. van den; Punt, P.J.

2006-01-01

Filamentous growth of Aspergillus oryzae on solid cereal substrates involves secretion of substrate converting enzymes and a solid substrate specific polarised hyphal growth phenotype. To identify proteins produced under these specific conditions, the extracts of A. oryzae grown on wheat-based media

Increased metabolite production by deletion of an HDA1-type histone deacetylase in the phytopathogenic fungi, Magnaporthe oryzae (Pyricularia oryzae) and Fusarium asiaticum.

Science.gov (United States)

Maeda, K; Izawa, M; Nakajima, Y; Jin, Q; Hirose, T; Nakamura, T; Koshino, H; Kanamaru, K; Ohsato, S; Kamakura, T; Kobayashi, T; Yoshida, M; Kimura, M

2017-11-01

Histone deacetylases (HDACs) play an important role in the regulation of chromatin structure and gene expression. We found that dark pigmentation of Magnaporthe oryzae (anamorph Pyricularia oryzae) ΔMohda1, a mutant strain in which an orthologue of the yeast HDA1 was disrupted by double cross-over homologous recombination, was significantly stimulated in liquid culture. Analysis of metabolites in a ΔMohda1 mutant culture revealed that the accumulation of shunt products of the 1,8-dihydroxynaphthalene melanin and ergosterol pathways were significantly enhanced compared to the wild-type strain. Northern blot analysis of the ΔMohda1 mutant revealed transcriptional activation of three melanin genes that are dispersed throughout the genome of M. oryzae. The effect of deletion of the yeast HDA1 orthologue was also observed in Fusarium asiaticum from the Fusarium graminearum species complex; the HDF2 deletion mutant produced increased levels of nivalenol-type trichothecenes. These results suggest that histone modification via HDA1-type HDAC regulates the production of natural products in filamentous fungi. Natural products of fungi have significant impacts on human welfare, in both detrimental and beneficial ways. Although HDA1-type histone deacetylase is not essential for vegetative growth, deletion of the gene affects the expression of clustered secondary metabolite genes in some fungi. Here, we report that such phenomena are also observed in physically unlinked genes required for melanin biosynthesis in the rice blast fungus. In addition, production of Fusarium trichothecenes, previously reported to be unaffected by HDA1 deletion, was significantly upregulated in another Fusarium species. Thus, the HDA1-inactivation strategy may be regarded as a general approach for overproduction and/or discovery of fungal metabolites. © 2017 The Society for Applied Microbiology.

A new and efficient approach for construction of uridine/uracil auxotrophic mutants in the filamentous fungus Aspergillus oryzae using Agrobacterium tumefaciens-mediated transformation.

Science.gov (United States)

Nguyen, Khuyen Thi; Ho, Quynh Ngoc; Do, Loc Thi Binh Xuan; Mai, Linh Thi Dam; Pham, Duc-Ngoc; Tran, Huyen Thi Thanh; Le, Diep Hong; Nguyen, Huy Quang; Tran, Van-Tuan

2017-06-01

Aspergillus oryzae is a filamentous fungus widely used in food industry and as a microbial cell factory for recombinant protein production. Due to the inherent resistance of A. oryzae to common antifungal compounds, genetic transformation of this mold usually requires auxotrophic mutants. In this study, we show that Agrobacterium tumefaciens-mediated transformation (ATMT) method is very efficient for deletion of the pyrG gene in different Aspergillus oryzae wild-type strains to generate uridine/uracil auxotrophic mutants. Our data indicated that all the obtained uridine/uracil auxotrophic transformants, which are 5- fluoroorotic acid (5-FOA) resistant, exist as the pyrG deletion mutants. Using these auxotrophic mutants and the pyrG selectable marker for genetic transformation via A. tumefaciens, we could get about 1060 transformants per 10 6 fungal spores. In addition, these A. oryzae mutants were also used successfully for expression of the DsRed fluorescent reporter gene under control of the A. oryzae amyB promoter by the ATMT method, which resulted in obvious red transformants on agar plates. Our work provides a new and effective approach for constructing the uridine/uracil auxotrophic mutants in the importantly industrial fungus A. oryzae. This strategy appears to be applicable to other filamentous fungi to develop similar genetic transformation systems based on auxotrophic/nutritional markers for food-grade recombinant applications.

Kadar Karbohidrat, Lemak, dan Protein pada Kecap dari Tempe

OpenAIRE

ARTINI PANGASTUTI; TJAHJADI PURWOKO,; YONA SEPTIANI

2004-01-01

Soy sauce as one of the soy fermented product could be produced through fermentation by molds and then fermentation by bacteria and yeasts, in salt solution (moromi). The aims of this research were to evaluate and to compare the value of carbohydrate), lipid, and protein in soy sauce which were produced from tempe by mixing Rhizopus oligosporus and R. oryzae with and without moromi fermentation; to compare the taste, aroma and color of soy sauce have been produced and commercial so...

Activity of Posaconazole and Other Antifungal Agents against Mucorales Strains Identified by Sequencing of Internal Transcribed Spacers▿

Science.gov (United States)

Alastruey-Izquierdo, Ana; Castelli, Maria Victoria; Cuesta, Isabel; Monzon, Araceli; Cuenca-Estrella, Manuel; Rodriguez-Tudela, Juan Luis

2009-01-01

The antifungal susceptibility profiles of 77 clinical strains of Mucorales species, identified by internal transcribed spacer sequencing, were analyzed. MICs obtained at 24 and 48 h were compared. Amphotericin B was the most active agent against all isolates, except for Cunninghamella and Apophysomyces isolates. Posaconazole also showed good activity for all species but Cunninghamella bertholletiae. Voriconazole had no activity against any of the fungi tested. Terbinafine showed good activity, except for Rhizopus oryzae, Mucor circinelloides, and Rhizomucor variabilis isolates. PMID:19171801

Overcoming the species hybridization barrier by ploidy manipulation in the genus Oryza.

Science.gov (United States)

Tonosaki, Kaoru; Sekine, Daisuke; Ohnishi, Takayuki; Ono, Akemi; Furuumi, Hiroyasu; Kurata, Nori; Kinoshita, Tetsu

2018-02-01

In most eudicot and monocot species, interspecific and interploidy crosses generally display abnormalities in the endosperm that are the major cause of a post-zygotic hybridization barrier. In some eudicot species, however, this type of hybridization barrier can be overcome by the manipulation of ploidy levels of one parental species, suggesting that the molecular mechanisms underlying the species hybridization barrier can be circumvented by genome dosage. We previously demonstrated that endosperm barriers in interspecific and interploidy crosses in the genus Oryza involve overlapping but different mechanisms. This result contrasts with those in the genus Arabidopsis, which shows similar outcomes in both interploidy and interspecific crosses. Therefore, we postulated that an exploration of pathways for overcoming the species hybridization barrier in Oryza endosperm, by manipulating the ploidy levels in one parental species, might provide novel insights into molecular mechanisms. We showed that fertile hybrid seeds could be produced by an interspecific cross of female tetraploid Oryza sativa and male diploid Oryza longistaminata. Although the rate of nuclear divisions did not return to normal levels in the hybrid endosperm, the timing of cellularization, nucellus degeneration and the accumulation of storage products were close to normal levels. In addition, the expression patterns of the imprinted gene MADS87 and YUCCA11 were changed when the species barrier was overcome. These results suggest that the regulatory machinery for developmental transitions and imprinted gene expression are likely to play a central role in overcoming species hybridization barriers by genome dosage in the genus Oryza. © 2017 The Authors The Plant Journal © 2017 John Wiley & Sons Ltd.

Production of L-lactic acid from Cassava peel wastes using single and mixed cultures of Rhizopus oligosporus and Lactobacillus plantarum

Directory of Open Access Journals (Sweden)

Nwokoro Ogbonnaya

2014-01-01

Full Text Available Production of L-lactic acid using cultures of Rhizopus oligosporus and Lactobacillus plantarum was investigated. Cassava peels were hydrolyzed by boiling for 1 h in either NaOH or HCl solutions followed by neutralization to a pH of 6.2. Reducing sugar produced from the hydrolysates increased with increasing concentrations of alkali or acid. Samples hydrolyzed with HCl produced a maximum reducing sugar concentration of 402 mg/g substrate while alkali hydrolyzed samples produced a maximum reducing sugar concentration of 213 mg/g substrate. Hydrolysates were amended with 0.5% ammonium sulphate solution and inoculated with either single or mixed cultures of Rhizopus oligosporus and Lactobacillus plantarum and incubated for 48 h for lactic acid production. The best lactic acid production of 50.2 g/100g substrate was observed in a mixed culture fermentation of acid hydrolyzed peels. Mixed culture fermentation of alkali hydrolyzed peels produced a maximum lactic acid concentration of 36.4 g/100g substrate. Un hydrolyzed cassava peels inoculated with a mixed culture of the microorganisms produced only 4.6 g/100g substrate. This work reports an efficient use of cassava peels for bio-product formation through microbial fermentation.

Aspergillus oryzae nrtA affects kojic acid production.

Science.gov (United States)

Sano, Motoaki

2016-09-01

We analyzed the role of the nitrate transporter-encoding gene (nrtA) of Aspergillus oryzae by gene disruption. Southern hybridization analysis indicated that homologous recombination occurred at the resident nrtA locus. Real-time PCR showed that the nrtA gene was strongly inducible by NaNO3. The nrtA disruptant did not exhibit normal growth when nitrate was available as the sole nitrogen source. These results indicate that NrtA is essential for nitrate uptake in A. oryzae. Kojic acid (KA) production was inhibited by the addition of a small amount of sodium nitrate. The nrtA-disrupted strain was deficient in the uptake of nitrate. As a result, KA production in this strain was not considerably affected by the presence of nitrate.

Selective effects of two systemic fungicides on soil fungi.

Science.gov (United States)

Abdel-Fattah, H M; Abdel-Kader, M I; Hamida, S

1982-08-20

BAS 317 00F was not toxic to the total count of fungi after 2 days but was regularly significantly toxic at the three doses after 5, 20 and 40 days and toxic at the low and the high doses after 80 days. In the agar medium, it was toxic to the counts of total fungi. Aspergillus, A. terreus, Rhizopus oryzae and Mucor racemosus at the high dose. Only the mycelial growth of Trichoderma viride which was significantly inhibited by the three doses when this fungicide was added to the liquid medium. Polyram-Combi induced two effects on the total population of soil fungi. One inhibitory and this was demonstrated almost regularly after 2, 10 and 40 days and the other stimulatory after 80 days of treatment with the low and the high doses. In the agar medium, this fungicide was very toxic to total fungi and to almost all fungal genera and species at the three doses. Several fungi could survive the high dose. In liquid medium, the test fungi showed variable degree of sensitivity and the most sensitive was Gliocladium roseum which was completely eradicated by the three doses.

The chemical heritage of Aspergillus flavus in A. oryzae RIB 40

OpenAIRE

Rank, Christian; Klejnstrup, Marie Louise; Petersen, Lene Maj; Frisvad, Jens Christian; Larsen, Thomas Ostenfeld

2011-01-01

Aspergillus oryzae is a very important species in biotechnology and has been used for centuries in traditional Asian fermentation. The RIB40 strain is particularly interesting as it was one of the first genome sequenced Aspergilli together with A. flavus, a prominent food and feed contaminant capable of producing aflatoxin. These species can be perceived as ecotypes. We have analyzed A. oryzae RIB40 and found that the chemical potential could be enhanced significantly under certain conditions...

Heterologous Production of a Novel Cyclic Peptide Compound, KK-1, in Aspergillus oryzae

Directory of Open Access Journals (Sweden)

Akira Yoshimi

2018-04-01

Full Text Available A novel cyclic peptide compound, KK-1, was originally isolated from the plant-pathogenic fungus Curvularia clavata. It consists of 10 amino acid residues, including five N-methylated amino acid residues, and has potent antifungal activity. Recently, the genome-sequencing analysis of C. clavata was completed, and the biosynthetic genes involved in KK-1 production were predicted by using a novel gene cluster mining tool, MIDDAS-M. These genes form an approximately 75-kb cluster, which includes nine open reading frames, containing a non-ribosomal peptide synthetase (NRPS gene. To determine whether the predicted genes were responsible for the biosynthesis of KK-1, we performed heterologous production of KK-1 in Aspergillus oryzae by introduction of the cluster genes into the genome of A. oryzae. The NRPS gene was split in two fragments and then reconstructed in the A. oryzae genome, because the gene was quite large (approximately 40 kb. The remaining seven genes in the cluster, excluding the regulatory gene kkR, were simultaneously introduced into the strain of A. oryzae in which NRPS had already been incorporated. To evaluate the heterologous production of KK-1 in A. oryzae, gene expression was analyzed by RT-PCR and KK-1 productivity was quantified by HPLC. KK-1 was produced in variable quantities by a number of transformed strains, along with expression of the cluster genes. The amount of KK-1 produced by the strain with the greatest expression of all genes was lower than that produced by the original producer, C. clavata. Therefore, expression of the cluster genes is necessary and sufficient for the heterologous production of KK-1 in A. oryzae, although there may be unknown factors limiting productivity in this species.

Heterologous Production of a Novel Cyclic Peptide Compound, KK-1, in Aspergillus oryzae.

Science.gov (United States)

Yoshimi, Akira; Yamaguchi, Sigenari; Fujioka, Tomonori; Kawai, Kiyoshi; Gomi, Katsuya; Machida, Masayuki; Abe, Keietsu

2018-01-01

A novel cyclic peptide compound, KK-1, was originally isolated from the plant-pathogenic fungus Curvularia clavata . It consists of 10 amino acid residues, including five N -methylated amino acid residues, and has potent antifungal activity. Recently, the genome-sequencing analysis of C. clavata was completed, and the biosynthetic genes involved in KK-1 production were predicted by using a novel gene cluster mining tool, MIDDAS-M. These genes form an approximately 75-kb cluster, which includes nine open reading frames, containing a non-ribosomal peptide synthetase (NRPS) gene. To determine whether the predicted genes were responsible for the biosynthesis of KK-1, we performed heterologous production of KK-1 in Aspergillus oryzae by introduction of the cluster genes into the genome of A. oryzae . The NRPS gene was split in two fragments and then reconstructed in the A. oryzae genome, because the gene was quite large (approximately 40 kb). The remaining seven genes in the cluster, excluding the regulatory gene kkR , were simultaneously introduced into the strain of A. oryzae in which NRPS had already been incorporated. To evaluate the heterologous production of KK-1 in A. oryzae , gene expression was analyzed by RT-PCR and KK-1 productivity was quantified by HPLC. KK-1 was produced in variable quantities by a number of transformed strains, along with expression of the cluster genes. The amount of KK-1 produced by the strain with the greatest expression of all genes was lower than that produced by the original producer, C. clavata . Therefore, expression of the cluster genes is necessary and sufficient for the heterologous production of KK-1 in A. oryzae , although there may be unknown factors limiting productivity in this species.

Isolation of industrial strains of Aspergillus oryzae lacking ferrichrysin by disruption of the dffA gene.

Science.gov (United States)

Watanabe, Hisayuki; Hatakeyama, Makoto; Sakurai, Hiroshi; Uchimiya, Hirofumi; Sato, Toshitsugu

2008-11-01

Based on studies using laboratory strains, the efficiency of gene disruption in Aspergillus oryzae, commonly known as koji mold, is low; thus, gene disruption has rarely been applied to the breeding of koji mold. To evaluate the efficiency of gene disruption in industrial strains of A. oryzae, we produced ferrichrysin biosynthesis gene (dffA) disruptants using three different industrial strains as hosts. PCR analysis of 438 pyrithiamine-resistant transformants showed dffA gene disruption efficiency of 42.9%-64.1%, which is much higher than previously reported. Analysis of the physiological characteristics of the disruptants indicated that dffA gene disruption results in hypersensitivity to hydrogen peroxide. To investigate the industrial characteristics of dffA gene disruptants, two strains were used to make rice koji and their properties were compared to those of the host strains. No differences were found between the dffA gene disruptants and the host strains, except that the disruptants did not produce ferrichrysin. Thus, this gene disruption technique is much more effective than conventional mutagenesis for A. oryzae breeding.

Differentiation of Clinically Relevant mucorales Rhizopus microsporus and R. arrhizus by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS)

NARCIS (Netherlands)

Dolatabadi, S.; Kolecka, A.; Versteeg, Matthijs; de Hoog, Sybren G; Boekhout, Teun

This study addresses the usefulness of Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) for reliable identification of the two most frequently occuring clinical species of Rhizopus, namely R. arrhizus with its two varieties arrhizus and delemar and R.

Identification of a Basic Helix-Loop-Helix-Type Transcription Regulator Gene in Aspergillus oryzae by Systematically Deleting Large Chromosomal Segments▿ †

OpenAIRE

Jin, Feng Jie; Takahashi, Tadashi; Machida, Masayuki; Koyama, Yasuji

2009-01-01

We previously developed two methods (loop-out and replacement-type recombination) for generating large-scale chromosomal deletions that can be applied to more effective chromosomal engineering in Aspergillus oryzae. In this study, the replacement-type method is used to systematically delete large chromosomal DNA segments to identify essential and nonessential regions in chromosome 7 (2.93 Mb), which is the smallest A. oryzae chromosome and contains a large number of nonsyntenic blocks. We con...

FY 2000 report on the results of the regional consortium R and D project - Regional consortium energy field. Final year report. R and D on the bio-fuel production by high functional bio-reactor; 2000 nendo chiiki consortium kenkyu kaihatsu jigyo - chiiki consortium energy bun'ya. Kokino bio reactor ni yoru bio nenryo seisan ni kansuru kenkyu kaihatsu (saishu nendo) seika hokokusho

Energy Technology Data Exchange (ETDEWEB)

NONE

2001-03-01

A system was developed for producing automobile fuel from the recycled paper and waste cooking oil using high functional intelligent yeast. Element technology is the functional yeast creation technology and the online intelligent control technology of the process into which the fixed bio-reactor was inserted. Studies were made on the following: 1) creation of high activity lipase production/ethanol production yeasts; 2) bio-fuel production by intelligent bio-reactor; 3) process optimization control technology by fuzzy control; 4) stabilization of bio-fuel production yeast; 5) comprehensive investigational study. In FY 2000, the results were obtained as written below: development of the stable lipase coming from rhizopus japonicus, fixed bacterium using rhizopus oryzae fungus body which can be used more than ten times, direct ethanol fermentation from starch by developing the multi-copy glucoamylase manifestation yeast, operation of a 20L capacity bench plant, etc. (NEDO)

Identification of molecular markers linked to rice bacterial blight resistance genes from Oryza meyeriana

Directory of Open Access Journals (Sweden)

Jing WANG,Chen CHENG,Yanru ZHOU,Yong YANG,Qiong MEI,Junmin LI,Ye CHENG,Chengqi YAN,Jianping CHEN

2015-09-01

Full Text Available Y73 is a progeny of asymmetric somatic hybridization between Oryza sativa cv. Dalixiang and the wild rice species Oryza meyeriana. Inoculation with a range of strains of Xanthomonas oryzae pv. oryzae showed that Y73 had inherited a high level of resistance to rice bacterial blight (BB from its wild parent. An F2 population of 7125 individuals was constructed from the cross between Y73 and a BB-susceptible cultivar IR24. After testing 615 SSR and STS markers covering the 12 rice chromosomes, 186 markers were selected that showed polymorphism between Y73 and IR24. Molecular markers linked to the BB resistance genes in Y73 were scanned using the F2 population and the polymorphic markers. The SSR marker RM128 on chromosome 1, the STS marker R03D159 on chromosome 3 and the STS marker R05D104 on chromosome 5 were found to be linked to the rice BB resistance genes in Y73.

ALPHA-AMYLASE PRODUCTION FROM Aspergillus oryzae M BY SUBMERGED FERMENTATION

Directory of Open Access Journals (Sweden)

Suleimenova

2016-08-01

Full Text Available The main goal of present study was implementation of the Aspergillus oryzae M strain improved technology using earlier developed method of microorganism selection. 8 pure strains of Aspergillus fungi were screened for the production of extra cellular alpha-amylase using agar medium with starch as a substrate and incubated for 72h at 30 ºС. Zone of clearance was observed for screening of the amylolytic fungi (in mm. Aspergillus oryzae M has demonstrated the highest zone of clearance. Aspergillus oryzae M was cultivated for 42 days in submerged conditions of growth using new method of fungal cultivation. This method based on immobilizing enzymes producers on solid career in submerged conditions of growth gives the way to improve quality of filtrates, which remain clear, does not require additional filtering and easily separated from the mycelium. Moreover, it allows to prolong the process of fungal cultivation and to maintain high enzymatic activity for a long period of time. Presented method allowed increasing alpha-amylase production from 321 U/ml (before immobilization to 502 U/ml (after immobilization.

An overproduction of astellolides induced by genetic disruption of chromatin-remodeling factors in Aspergillus oryzae.

Science.gov (United States)

Shinohara, Yasutomo; Kawatani, Makoto; Futamura, Yushi; Osada, Hiroyuki; Koyama, Yasuji

2016-01-01

The filamentous fungus Aspergillus oryzae is an important industrial mold. Recent genomic analysis indicated that A. oryzae has a large number of biosynthetic genes for secondary metabolites (SMs), but many of the SMs they produce have not been identified. For better understanding of SMs production by A. oryzae, we screened a gene-disruption library of transcription factors including chromatin-remodeling factors and found two gene disruptions that show similarly altered SM production profiles. One is a homolog of Aspergillus nidulans cclA, a component of the histone 3 lysine 4 (H3K4) methyltransferase complex of proteins associated with Set1 complex, and the other, sppA, is an ortholog of Saccharomyces cerevisiae SPP1, another component of a complex of proteins associated with Set1 complex. The cclA and sppA disruptions in A. oryzae are deficient in trimethylation of H3K4. Furthermore, one of the SMs that increased in the cclA disruptant was identified as astellolide F (14-deacetyl astellolide B). These data indicate that both cclA and sppA affect production of SMs including astellolides by affecting the methylation status of H3K4 in A. oryzae.

Random Mutagenesis of the Aspergillus oryzae Genome Results in Fungal Antibacterial Activity

OpenAIRE

Leonard, Cory A.; Brown, Stacy D.; Hayman, J. Russell

2013-01-01

Multidrug-resistant bacteria cause severe infections in hospitals and communities. Development of new drugs to combat resistant microorganisms is needed. Natural products of microbial origin are the source of most currently available antibiotics. We hypothesized that random mutagenesis of Aspergillus oryzae would result in secretion of antibacterial compounds. To address this hypothesis, we developed a screen to identify individual A. oryzae mutants that inhibit the growth of Methicillin-res...

Genome-scale analysis of the high-efficient protein secretion system of Aspergillus oryzae.

Science.gov (United States)

Liu, Lifang; Feizi, Amir; Österlund, Tobias; Hjort, Carsten; Nielsen, Jens

2014-06-24

The koji mold, Aspergillus oryzae is widely used for the production of industrial enzymes due to its particularly high protein secretion capacity and ability to perform post-translational modifications. However, systemic analysis of its secretion system is lacking, generally due to the poorly annotated proteome. Here we defined a functional protein secretory component list of A. oryzae using a previously reported secretory model of S. cerevisiae as scaffold. Additional secretory components were obtained by blast search with the functional components reported in other closely related fungal species such as Aspergillus nidulans and Aspergillus niger. To evaluate the defined component list, we performed transcriptome analysis on three α-amylase over-producing strains with varying levels of secretion capacities. Specifically, secretory components involved in the ER-associated processes (including components involved in the regulation of transport between ER and Golgi) were significantly up-regulated, with many of them never been identified for A. oryzae before. Furthermore, we defined a complete list of the putative A. oryzae secretome and monitored how it was affected by overproducing amylase. In combination with the transcriptome data, the most complete secretory component list and the putative secretome, we improved the systemic understanding of the secretory machinery of A. oryzae in response to high levels of protein secretion. The roles of many newly predicted secretory components were experimentally validated and the enriched component list provides a better platform for driving more mechanistic studies of the protein secretory pathway in this industrially important fungus.

Sustainable carbon sources for microbial organic acid production with filamentous fungi.

Science.gov (United States)

Dörsam, Stefan; Fesseler, Jana; Gorte, Olga; Hahn, Thomas; Zibek, Susanne; Syldatk, Christoph; Ochsenreither, Katrin

2017-01-01

The organic acid producer Aspergillus oryzae and Rhizopus delemar are able to convert several alternative carbon sources to malic and fumaric acid. Thus, carbohydrate hydrolysates from lignocellulose separation are likely suitable as substrate for organic acid production with these fungi. Before lignocellulose hydrolysate fractions were tested as substrates, experiments with several mono- and disaccharides, possibly present in pretreated biomass, were conducted for their suitability for malic acid production with A. oryzae. This includes levoglucosan, glucose, galactose, mannose, arabinose, xylose, ribose, and cellobiose as well as cheap and easy available sugars, e.g., fructose and maltose. A. oryzae is able to convert every sugar investigated to malate, albeit with different yields. Based on the promising results from the pure sugar conversion experiments, fractions of the organosolv process from beechwood ( Fagus sylvatica ) and Miscanthus giganteus were further analyzed as carbon source for cultivation and fermentation with A. oryzae for malic acid and R. delemar for fumaric acid production. The highest malic acid concentration of 37.9 ± 2.6 g/L could be reached using beechwood cellulose fraction as carbon source in bioreactor fermentation with A. oryzae and 16.2 ± 0.2 g/L fumaric acid with R. delemar . We showed in this study that the range of convertible sugars for A. oryzae is even higher than known before. We approved the suitability of fiber/cellulose hydrolysate obtained from the organosolv process as carbon source for A. oryzae in shake flasks as well as in a small-scale bioreactor. The more challenging hemicellulose fraction of F. sylvatica was also positively evaluated for malic acid production with A. oryzae .

Genome information of Methylobacterium oryzae, a plant-probiotic methylotroph in the phyllosphere.

Science.gov (United States)

Kwak, Min-Jung; Jeong, Haeyoung; Madhaiyan, Munusamy; Lee, Yi; Sa, Tong-Min; Oh, Tae Kwang; Kim, Jihyun F

2014-01-01

Pink-pigmented facultative methylotrophs in the Rhizobiales are widespread in the environment, and many Methylobacterium species associated with plants produce plant growth-promoting substances. To gain insights into the life style at the phyllosphere and the genetic bases of plant growth promotion, we determined and analyzed the complete genome sequence of Methylobacterium oryzae CBMB20T, a strain isolated from rice stem. The genome consists of a 6.29-Mb chromosome and four plasmids, designated as pMOC1 to pMOC4. Among the 6,274 coding sequences in the chromosome, the bacterium has, besides most of the genes for the central metabolism, all of the essential genes for the assimilation and dissimilation of methanol that are either located in methylotrophy islands or dispersed. M. oryzae is equipped with several kinds of genes for adaptation to plant surfaces such as defense against UV radiation, oxidative stress, desiccation, or nutrient deficiency, as well as high proportion of genes related to motility and signaling. Moreover, it has an array of genes involved in metabolic pathways that may contribute to promotion of plant growth; they include auxin biosynthesis, cytokine biosynthesis, vitamin B12 biosynthesis, urea metabolism, biosorption of heavy metals or decrease of metal toxicity, pyrroloquinoline quinone biosynthesis, 1-aminocyclopropane-1-carboxylate deamination, phosphate solubilization, and thiosulfate oxidation. Through the genome analysis of M. oryzae, we provide information on the full gene complement of M. oryzae that resides in the aerial parts of plants and enhances plant growth. The plant-associated lifestyle of M. oryzae pertaining to methylotrophy and plant growth promotion, and its potential as a candidate for a bioinoculant targeted to the phyllosphere and focused on phytostimulation are illuminated.

Production and biochemical characterization of an alkaline protease from Aspergillus oryzae CH93.

Science.gov (United States)

Salihi, Ahsan; Asoodeh, Ahmad; Aliabadian, Mansour

2017-01-01

In this study, Aspergillus oryzae CH93 was isolated from soil sample and examined using molecular analysis. Following culture of A. oryzae CH93 under optimal enzyme production, a 47.5kDa extracellular protease was purified using ammonium sulfate precipitation and Q-Sepharose chromatography. The optimal pH 8 and temperature of 50°C obtained for the isolated protease. Sodium dodecyl sulfate (SDS), cetyltrimethyl ammonium bromide (CTAB), H 2 O 2 decreased activity, while Triton X-100 and phenylmethanesulfonyl fluoride (PMSF) had no inhibitory effect on the enzyme activity; meanwhile, 2-mercaptoethanol and ethylenediaminetetraacetic acid (EDTA) declined the protease activity. Isoamyl alcohol and acetone (30%) enhanced activity whereas 2-propanol, isopropanol and dimethyl sulfoxide (DMSO) (30%) reduced protease activity. The enzyme exhibited a half-life of 100min at its optimum temperature. Among five substrates of bovine serum albumin (BSA), N-acetyl-l-tyrosine ethyl ester monohydrate (ATEE), casein, azocasein and gelatin results showed that casein is the best substrate with V max of 0.1411±0.004μg/min and K m of 2.432±0.266μg/ml. In conclusion, the extracted protease from A. oryzae CH93 as a fungal source possessed biochemical features which could be useful in some application usages. Copyright © 2016 Elsevier B.V. All rights reserved.

Gene Ontology annotation of the rice blast fungus, Magnaporthe oryzae

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Deng Jixin

2009-02-01

Full Text Available Abstract Background Magnaporthe oryzae, the causal agent of blast disease of rice, is the most destructive disease of rice worldwide. The genome of this fungal pathogen has been sequenced and an automated annotation has recently been updated to Version 6 http://www.broad.mit.edu/annotation/genome/magnaporthe_grisea/MultiDownloads.html. However, a comprehensive manual curation remains to be performed. Gene Ontology (GO annotation is a valuable means of assigning functional information using standardized vocabulary. We report an overview of the GO annotation for Version 5 of M. oryzae genome assembly. Methods A similarity-based (i.e., computational GO annotation with manual review was conducted, which was then integrated with a literature-based GO annotation with computational assistance. For similarity-based GO annotation a stringent reciprocal best hits method was used to identify similarity between predicted proteins of M. oryzae and GO proteins from multiple organisms with published associations to GO terms. Significant alignment pairs were manually reviewed. Functional assignments were further cross-validated with manually reviewed data, conserved domains, or data determined by wet lab experiments. Additionally, biological appropriateness of the functional assignments was manually checked. Results In total, 6,286 proteins received GO term assignment via the homology-based annotation, including 2,870 hypothetical proteins. Literature-based experimental evidence, such as microarray, MPSS, T-DNA insertion mutation, or gene knockout mutation, resulted in 2,810 proteins being annotated with GO terms. Of these, 1,673 proteins were annotated with new terms developed for Plant-Associated Microbe Gene Ontology (PAMGO. In addition, 67 experiment-determined secreted proteins were annotated with PAMGO terms. Integration of the two data sets resulted in 7,412 proteins (57% being annotated with 1,957 distinct and specific GO terms. Unannotated proteins

Efectividad biológica delazoxystrobin para el control de pyricularia oryzae cav. y cercospora oryzae miyake. en arroz de temporal en Veracruz, México

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Enrique Becerra

2001-01-01

Full Text Available El estado de Veracruz, México, siembra anualmente 22000 ha de arroz de temporal con un rendimiento medio de 3,5 t/ha, debido principalmente a problemas de sequía lo cual favorece la presencia de hongos como Pyricularia oryzae Cav. (quema del arroz y Cercospora oryzae Miyake. (Mancha angosta. Con el objeto de conocer el comportamiento de nuevos fungicidas se evaluó la eficacia del Azoxystrobin en el control de estos hongos. El experimento se estableció en el municipio de Tres Valles, Veracruz, durante el temporal de 1999, con semilla de Milagro Filipino Depurado. El diseño estadístico utilizado fue bloques al azar con cuatro repeticiones. Los tratamientos evaluados fueron: Azoxystrobin a 0,2, 0,4 y 0,6 l/ha vs Tecto60 a 0,5 kg/ha y un testigo sin aplicación. Estos fueron aplicados al presentarse los primeros síntomas de las enfermedades. Se evaluó la incidencia, número de lesiones en 20 plantas, índice de intensidad, el rendimiento de grano y fitotoxicidad. Se encontró que el Azoxystrobin obtuvo mejores resultados que el Tecto 60 y éste a su vez que el testigo no tratado. El mejor control para P. oryzae y C. oryzae y el mayor rendimiento de grano (4432 kg/ha se logró cuando se aplicó Azoxystrobin en dosis de 0,6 l/ha, aunque con 0,2 y 0,4 l/ha de este fungicida tuvo un buen control de estas enfermedades. Ninguno de los fungicidas causó toxicidad al arroz

Enhanced production of protease by mutagenized strain of aspergillus oryzae in solid substrate fermentation of rice bran

International Nuclear Information System (INIS)

Yousif, M.; Irfan, M.; Baig, S.; Iqbal, A.

2010-01-01

Neutral protease activity of parent strain of Aspergellus oryzae was enhanced by UV and chemical mutagenization with ethyl methane sulphonate (EMS). After screening, a hyper producing strain was isolated and found effective for tile production of neutral protease as compared to the parent strain of Aspergellus oryzae. Solid substrate fermentation was carried out in 250ml conical flask with 45 % initial moisture contents at a temperature of 30 deg. C for 72 flours. Under the optimum conditions maximum yield of neutral protease obtained was 662.61+-0.36 U/gds, Almost all the organic nitrogen supplements favored the enzyme production while sucrose proved as a best carbon source. (author)

Self-affine fractal growth front of Aspergillus oryzae

Science.gov (United States)

Matsuura, Shu; Miyazima, Sasuke

1992-12-01

Aspergillus oryzae have been grown in various environmental conditions and analyzed from the viewpoint of self-affinity. The growth behavior can be described by the Eden model in favorable conditions, and by DLA in unfavorable conditions.

Karakterisasi ekstrak kasar lipase Rhizopus stolonifer UICC 137

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Sri Sumiarsih

2001-12-01

Full Text Available There is an increasing commercial interest in enzymatic production of biologically active component, because there are a number of well-known advantages compared to chemical synthesis. One of the most valuable synthetic features of enzyme is their ability to discriminate between enantiomers of racemic substrates. Lipase have become of great interest to the chemical industries wing their usefulness in both hydrolytic and synthesis reactions. The aim of this work was to study the production of lipase by Rhizopus stolonifer UICC 137, and determine the crude lipase preparation characteristics. The lipolytic activity was determined by titrimetric method toward oil-arabic gum emultion as a substrate. The strain produced lipase at appreciable lipolytic when cultivated for 72 hours in medium containing 3% glucose and 1% olive oil. Our data suggest that the strain produced lipase since the exponential phase of its growth. Lipase with optimum lipolytic activity was obtained at late stationary phase. The optimum condition for lipolytic activity measurement were pH of 7.5 and temperature 37oC, the crude enzyme had a specific activity 20.2 unit/ mg protein, the Vmax was 15.1 mol/ min and KM was 12.5 mg/ ml. The crude enzyme retained 79.9%, 68.0% and 52.6% of its lipolytic activity, when incubated for 90 minutes at temperature of 40, 50, and 60oC respectively.

Evaluation of the catalytic activity of lipases immobilized on chrysotile for esterification

Energy Technology Data Exchange (ETDEWEB)

Silva, Jane E.S.; Jesus, Paulo C. [Universidade Regional de Blumenau, SC (Brazil). Dept. de Quimica]. E-mail: pcj@furb.rct-sc.br

2003-06-01

In the present work, the ester synthesis in organic media catalyzed by lipases immobilized on chrysotile was studied. Lipases of different sources (Mucor javanicus, Pseudomonas cepacia, Rhizopus oryzae, Aspergillus niger and Candida rugosa) were immobilized on chrysotile, an inexpensive magnesium silicate, and used for esterification of hexanoic, octanoic and lauric acid with methanol, ethanol, 1-butanol and 1-octanol at 25 deg C in hexane as solvent. The best results were obtained with Mucor javanicus lipase and lauric acid giving yields of 62-97% of ester. (author)

Evaluation of the catalytic activity of lipases immobilized on chrysotile for esterification

International Nuclear Information System (INIS)

Silva, Jane E.S.; Jesus, Paulo C.

2003-01-01

In the present work, the ester synthesis in organic media catalyzed by lipases immobilized on chrysotile was studied. Lipases of different sources (Mucor javanicus, Pseudomonas cepacia, Rhizopus oryzae, Aspergillus niger and Candida rugosa) were immobilized on chrysotile, an inexpensive magnesium silicate, and used for esterification of hexanoic, octanoic and lauric acid with methanol, ethanol, 1-butanol and 1-octanol at 25 deg C in hexane as solvent. The best results were obtained with Mucor javanicus lipase and lauric acid giving yields of 62-97% of ester. (author)

Breeding L(+)-lactic acid high productive mutant from xylose by nitrogen ions

International Nuclear Information System (INIS)

Yang Yingge; Li Wen; Liu Dan; Fan Yonghong; Wang Dongmei; Zheng Zhiming; Yu Zengliang

2007-01-01

In order to obtain higher L(+)-lactic acid yield strain fermentating from xylose, the original strain Rhizopus oryzae RLC41-6 was mutated by 10keV N + ion implantation. A mutant strain RQ4012 was obtained. After 72h shake-flask cultivation, the concentration of L(+)-lactic acid reached 74.37g/L, and the productivity was 1.03g/(L.h). Its lactic acid yield was 160% higher than that of the original one, and the mutant strain has high genetic stability. (authors)

Moderate expression of SEC16 increases protein secretion by Saccharomyces cerevisiae

DEFF Research Database (Denmark)

Bao, Jichen; Huang, Mingtao; Petranovic, Dina

2017-01-01

in yeast, by moderately overexpressing SEC16, which is involved in protein translocation from the endoplasmic reticulum to the Golgi apparatus. The moderate overexpression of SEC16 increased α-amylase secretion by generating more endoplasmic reticulum exit sites. The production of reactive oxygen species...... were observed. Finally, the moderate overexpression of SEC16 was shown to improve the secretion of two other recombinant proteins, Trichoderma reesei endoglucanase I and Rhizopus oryzae glucan-1,4-α-glucosidase, indicating that this mechanism is of general relevance....

Peran Direct Fed Microbials (DFM Saccharomyces cerevisiae dan Aspergillus oryzae terhadap Produktivitas Ternak Ruminansia : Review

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H. Suryani

2015-02-01

Full Text Available Mikroorganisme yang biasa digunakan dalam pakan ternak ruminansia biasanya berupa probiotik. Probiotik memiliki makna yang bersepadanan dengan Direct Fed Microbials (DFM. Penambahan DFM jenis Saccharomyces cerevisiae dan Aspergillus oryzae pada pakan ternak ruminansia mampu memanipulasi rumen dengan meningkatkan populasi bakteri pemecah serat sehingga dapat meningkatkan kecernaan dan meningkatkan bobot badan. Mekanisme kerja S. cerevisiae dan A. oryzae yang masuk kedalam tubuh ternak dan mempengaruhi pencernaan atau penyerapan, ada yang sudah diketahui secara jelas tetapi ada juga yang masih berupa hipotesa. Pemanfaatan DFM jenis S. cerevisiae dan A. oryzae secara tunggal maupun kombinasi sebagian telah diamati dan memberikan respon positif.

Direct suppression of a rice bacterial blight (Xanthomonas oryzae pv. oryzae) by monoterpene (S)-limonene.

Science.gov (United States)

Lee, Gun Woong; Chung, Moon-Soo; Kang, Mihyung; Chung, Byung Yeoup; Lee, Sungbeom

2016-05-01

Rice bacterial blight, caused by Xanthomonas oryzae pv. oryzae (Xoo), is a severe disease of rice plants. Upon pathogen infection, rice biosynthesizes phytoalexins, including diterpenoids such as momilactones, phytocassanes, and oryzalexins. However, information on headspace volatiles in response to Xoo infection is limited. We have examined headspace volatile terpenes, induced by the infection of Xoo, and investigated their biological roles in the rice plant. Monoterpenes α-thujene, α-pinene, sabinene, myrcene, α-terpene, and (S)-limonene and sesquiterpenes cyclosativene, α-copaene, and β-elemene were detected from 1-week-old Xoo-infected rice seedlings, by solid-phase microextraction-gas chromatography-mass spectrometry. All monoterpenes were constitutively released from rice seedlings before Xoo infection. However, (S)-limonene emission was further elicited after exposure of the seedlings to Xoo in coincidence with upregulation of limonene synthase gene (OsTPS20) transcripts. Only the stereospecific (S)-limonene [and not (R)-limonene or other monoterpenes] severely inhibited Xoo growth, as confirmed by disc diffusion and liquid culture assays. Rice seedlings showed suppressed pathogenic symptoms suggestive of resistance to Xoo infection after foliar treatment with (S)-limonene. Collectively, our findings suggest that (S)-limonene is a volatile phytoanticipin, which plays a significant role in suppressing Xoo growth in rice seedlings.

Magnesium-induced alterations in the photosynthetic performance and resistance of rice plants infected with Bipolaris oryzae

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Wiler Ribas Moreira

2015-08-01

Full Text Available Brown spot (BS, caused by the fungus Bipolaris oryzae, is one of the most important diseases contracted by rice. We investigated the effect of magnesium (Mg on the development of BS, caused by Bipolaris oryzae, and the effects of disease development on the photosynthetic performance of rice (Oryza sativa L. plants (cv. Metica-1 grown in nutrient solutions containing 0.25 or 4.0 mM of Mg. Assessments of BS severity, leaf Mg and pigment concentrations (total chlorophylls and carotenoids, were carried out at 120 h after inoculation, in addition to gas exchange parameters,. Higher leaf concentration of Mg was observed in plants supplied with 4.0 mM Mg than in those supplied with 0.25 mM. The increase in leaf Mg was accompanied by a decrease in BS severity, higher concentration of total chlorophyll and better photosynthetic performance. Plants supplied with 4.0 mM Mg had higher average values for carbon assimilation, stomatal conductance and internal leaf CO2 concentration when compared with plants supplied with 0.25 mM Mg. Conversely, the concentration of carotenoids was lower in plants supplied with the higher Mg rate. These results suggest that Mg suppresses disease severity and preserves photosynthetic performance by allowing for better stomatal conductance and, consequently, greater availability of CO2 at the carboxylation sites.

The chemical heritage of Aspergillus flavus in A. oryzae RIB 40

DEFF Research Database (Denmark)

Rank, Christian; Klejnstrup, Marie Louise; Petersen, Lene Maj

Aspergillus oryzae is a very important species in biotechnology and has been used for centuries in traditional Asian fermentation. The RIB40 strain is particularly interesting as it was one of the first genome sequenced Aspergilli together with A. flavus, a prominent food and feed contaminant...... capable of producing aflatoxin. These species can be perceived as ecotypes. We have analyzed A. oryzae RIB40 and found that the chemical potential could be enhanced significantly under certain conditions. Delicate analysis of their metabolic profiles allow for chemical insight on the transcription level...

Structural analysis of cerebrosides from Aspergillus fungi: the existence of galactosylceramide in A. oryzae.

Science.gov (United States)

Tani, Yasushi; Amaishi, Yasunori; Funatsu, Tori; Ito, Masahiro; Itonori, Saki; Hata, Yoji; Ashida, Hisashi; Yamamoto, Kenji

2014-12-01

Glucosylceramide and galactosylceramide were detected in three Aspergillus species: Aspergillus oryzae, Aspergillus sojae and Aspergillus. awamori, using borate-coated TLC. The cerebrosides from A. oryzae were further purified by ion exchange and iatrobeads column chromatographies with or without borate, and determined the composition of sugar, fatty acid and sphingoid base by GC/MS, MALDI-TOF/MS and (1)H-NMR. We identified them as β-glucosylceramide and β-galactosylceramide. The ceramide moiety of both cerebrosides consisted mainly of 2-hydroxystearic acid and either 9-methyl-octadeca-4, 8-sphingadienine or octadeca-4, 8-sphingadienine. To our knowledge, this is the first study to provide evidence for the presence of β-galactosylceramide in A. oryzae.

Comparative Chemistry of Aspergillus oryzae (RIB40) and A. flavus (NRRL 3357)

OpenAIRE

Rank, Christian; Klejnstrup, Marie Louise; Petersen, Lene Maj; Kildgaard, Sara; Frisvad, Jens Christian; Gotfredsen, Charlotte Held; Larsen, Thomas Ostenfeld

2012-01-01

Aspergillus oryzae and A. flavus are important species in industrial biotechnology and food safety and have been some of the first aspergilli to be fully genome sequenced. Bioinformatic analysis has revealed 99.5% gene homology between the two species pointing towards a large coherence in the secondary metabolite production. In this study we report on the first comparison of secondary metabolite production between the full genome sequenced strains of A. oryzae (RIB40) and A. flavus (NRRL 3357...

Pemanfaatan Ekstrak Tanaman Ketan Hitam (Oryza Sativa Glutinosa) sebagai Indikator Asam Basa

OpenAIRE

Hasibuan, Muhammad Arief; Amran, Elva Yasmi; ', Susilawati

2016-01-01

Research was conducted with the aims to determine pH range, percentage acid base indicator titration error, and stability of Oryza sativa Glutinosa extract as acid base indicator. Oryza sativa Glutinosa grains respectively extracted with maseration using ethanol (HCl 1%). pH range determined by added the extract into buffer solutions pH 1-9. Percentage acid base indicator titration error determined by applied the extract in strong acid-strong base, weak acid-strong base, strong acid-weak base...

Genome information of Methylobacterium oryzae, a plant-probiotic methylotroph in the phyllosphere.

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Min-Jung Kwak

Full Text Available Pink-pigmented facultative methylotrophs in the Rhizobiales are widespread in the environment, and many Methylobacterium species associated with plants produce plant growth-promoting substances. To gain insights into the life style at the phyllosphere and the genetic bases of plant growth promotion, we determined and analyzed the complete genome sequence of Methylobacterium oryzae CBMB20T, a strain isolated from rice stem. The genome consists of a 6.29-Mb chromosome and four plasmids, designated as pMOC1 to pMOC4. Among the 6,274 coding sequences in the chromosome, the bacterium has, besides most of the genes for the central metabolism, all of the essential genes for the assimilation and dissimilation of methanol that are either located in methylotrophy islands or dispersed. M. oryzae is equipped with several kinds of genes for adaptation to plant surfaces such as defense against UV radiation, oxidative stress, desiccation, or nutrient deficiency, as well as high proportion of genes related to motility and signaling. Moreover, it has an array of genes involved in metabolic pathways that may contribute to promotion of plant growth; they include auxin biosynthesis, cytokine biosynthesis, vitamin B12 biosynthesis, urea metabolism, biosorption of heavy metals or decrease of metal toxicity, pyrroloquinoline quinone biosynthesis, 1-aminocyclopropane-1-carboxylate deamination, phosphate solubilization, and thiosulfate oxidation. Through the genome analysis of M. oryzae, we provide information on the full gene complement of M. oryzae that resides in the aerial parts of plants and enhances plant growth. The plant-associated lifestyle of M. oryzae pertaining to methylotrophy and plant growth promotion, and its potential as a candidate for a bioinoculant targeted to the phyllosphere and focused on phytostimulation are illuminated.

Construction of a Shuttle Vector for Heterologous Expression of a Novel Fungal α-Amylase Gene in Aspergillus oryzae.

Science.gov (United States)

Yin, Yanchen; Mao, Youzhi; Yin, Xiaolie; Gao, Bei; Wei, Dongzhi

2015-07-01

The filamentous fungus Aspergillus oryzae is a well-known expression host used to express homologous and heterologous proteins in a number of industrial applications. To facilitate higher yields of proteins of interest, we constructed the pAsOP vector to express heterologous proteins in A. oryzae. pAsOP carries a selectable marker, pyrG, derived from Aspergillus nidulans, and a strong promoter and a terminator of the amyB gene derived from A. oryzae. pAsOP transformed A. oryzae efficiently via the PEG-CaCl2-mediated transformation method. As proof of concept, green fluorescent protein (GFP) was successfully expressed in A. oryzae transformed by pAsOP-GFP. Additionally, we identified a novel fungal α-amylase (PcAmy) gene from Penicillium sp. and cloned the gene into the vector. After transformation by pAsOPPcAmy, the α-amylase PcAmy from Penicillium sp. was successfully expressed in a heterologous host system for the first time. The α-amylase activity in the A. oryzae transformant was increased by 62.3% compared with the untransformed A. oryzae control. The PcAmy protein produced in the system had an optimum pH of 5.0 and optimum temperature of 30°C. As a cold-adapted enzyme, PcAmy shows potential value in industrial applications because of its high catalytic activity at low temperature. Furthermore, the expression vector reported in this study provides promising utility for further scientific research and biotechnological applications.

The late endosomal HOPS complex anchors active G-protein signaling essential for pathogenesis in magnaporthe oryzae.

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Ravikrishna Ramanujam

Full Text Available In Magnaporthe oryzae, the causal ascomycete of the devastating rice blast disease, the conidial germ tube tip must sense and respond to a wide array of requisite cues from the host in order to switch from polarized to isotropic growth, ultimately forming the dome-shaped infection cell known as the appressorium. Although the role for G-protein mediated Cyclic AMP signaling in appressorium formation was first identified almost two decades ago, little is known about the spatio-temporal dynamics of the cascade and how the signal is transmitted through the intracellular network during cell growth and morphogenesis. In this study, we demonstrate that the late endosomal compartments, comprising of a PI3P-rich (Phosphatidylinositol 3-phosphate highly dynamic tubulo-vesicular network, scaffold active MagA/GαS, Rgs1 (a GAP for MagA, Adenylate cyclase and Pth11 (a non-canonical GPCR in the likely absence of AKAP-like anchors during early pathogenic development in M. oryzae. Loss of HOPS component Vps39 and consequently the late endosomal function caused a disruption of adenylate cyclase localization, cAMP signaling and appressorium formation. Remarkably, exogenous cAMP rescued the appressorium formation defects associated with VPS39 deletion in M. oryzae. We propose that sequestration of key G-protein signaling components on dynamic late endosomes and/or endolysosomes, provides an effective molecular means to compartmentalize and control the spatio-temporal activation and rapid downregulation (likely via vacuolar degradation of cAMP signaling amidst changing cellular geometry during pathogenic development in M. oryzae.

Genetics of resistance to stored grain weevil (Sitophilus oryzae L. in maize

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Rajkumar Zunjare

2015-12-01

Full Text Available Stored grain weevil (Sitophilus oryzae has emerged as important storage grain pest of maize, causing substantial economic losses. Owing to high costs and environmental hazards of pesticides, host plant resistance holds promise for effective control of weevils. In the present study, a set of experimental maize hybrids generated using line × tester mating design were evaluated against S. oryzae. Significant variation for grain weight loss (GWL (6.0–49.1%, number of insect progeny emerged (NIP (17.8–203.3, grain hardness (GH (263.1–495.4 N, and pericarp thickness (PT (60.3–161.0 μm was observed. Strong positive association was observed between GWL and NIP. GH and PT did not show any correlation with GWL and NIP. Additive and non-additive gene actions were important for both GWL and NIP. Promising inbreds and experimental crosses identified can be effectively utilized in the resistance breeding programme. In majority of promising crosses having desirable SCA effects, one of the parents had desirable GCA effects, indicating that selection of inbred parents based on per se performance for generating resistant crosses may be possible. The commercial hybrid checks were highly susceptible compared to experimental hybrids. The inbreds and experimental hybrids identified hold promise in developing weevil resistant maize cultivars offering sustainable solution to management of weevils in maize.

Patogenicidade de Helminthosporium oryzae a algumas espécies de gramíneas Pathogenicity of Helminthosporium oryzae against some grass species

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V.H. Artigiani Filho

1995-04-01

Full Text Available O fungo Helminthosporium oryzae é um patógeno do arroz. Devido a sua variabilidade patogênica, foi investigada a possibilidade deste fungo infectar outras gramíneas. Através de inoculação artificial, ficou demonstrada a capacidade deste patógeno provocar infecção em aveia, cana, centeio, sorgo, trigo, Brachiaria decumbens e Panicum maximum. Assim, estas espécies vegetais podem ser consideradas potenciais hospedeiros do fungo na natureza.Helminthosporium oryzae is a rice pathogen. Due to its variability in pathogenicity, the possibility of this fungus Infecting other grasses was investigated. The capacity of this pathogen was demonstrated to be able to infect oat, sugar-cane, rye, sorghum, wheat, Brachiaria decumbens and Panicum maximum through artificial inoculations. Therefore, those plant species can be considered potencial hosts for the fungus in nature.

Ecogeographic variation in the morphology of two Asian wild rice species Oryza nivara and O. ruftipogon.

NARCIS (Netherlands)

Banaticla, M.C.N.; Sosef, M.S.M.; McNally, K.L.; Sackville Hamilton, R.; Berg, van den R.G.

2013-01-01

To search for variation patterns and diagnostic features between Asian wild rice species, several numerical methods were applied to phenotypic data obtained from 116 accessions representing sympatric populations of Oryza nivara and Oryza rufipogon from tropical continental Asia and O. rufipogon

Comparative effect of mutation of Aspergillus oryzae by gamma or ultraviolet irradiation

International Nuclear Information System (INIS)

Ito, Hitoshi; Nessa, A.

1994-01-01

Mutation frequency of Aspergillus oryzae IAM2630 was studied compared with gamma and ultraviolet irradiation. In this study, mutation frequency of morphological changes on survived colonies was increased up to 50% by irradiation of gamma-rays at survival fraction of 10 -3 to 10 -4 on potato-dextrose agar. On the contrary, mutation frequency of ultraviolet was obtained less than 17% at survival fraction of 10 -3 . Mutants with improvement of three-to-five hold production of α-amylase were isolated by irradiation of gamma-rays at 1.2 kGy. However, we could not isolate any mutants of higher production of α-amylase by ultraviolet irradiation. (author)

Comparative effect of mutation of Aspergillus oryzae by gamma or ultraviolet irradiation

Energy Technology Data Exchange (ETDEWEB)

Ito, Hitoshi; Nessa, A. [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment

1994-08-01

Mutation frequency of Aspergillus oryzae IAM2630 was studied compared with gamma and ultraviolet irradiation. In this study, mutation frequency of morphological changes on survived colonies was increased up to 50% by irradiation of gamma-rays at survival fraction of 10{sup -3} to 10{sup -4} on potato-dextrose agar. On the contrary, mutation frequency of ultraviolet was obtained less than 17% at survival fraction of 10{sup -3}. Mutants with improvement of three-to-five hold production of {alpha}-amylase were isolated by irradiation of gamma-rays at 1.2 kGy. However, we could not isolate any mutants of higher production of {alpha}-amylase by ultraviolet irradiation. (author).

Analysis of extracellular proteins of Aspergillus oryzae grown on soy sauce koji.

Science.gov (United States)

Liang, Yanchang; Pan, Li; Lin, Ying

2009-01-01

Aspergillus oryzae AS 3.951 is widely used in Chinese soy sauce manufacture, but little is known about the profiles of the extracellular proteins from the culture of soybean koji. In this study, we carried out MALDI-TOF/TOF MS analysis of extracellular proteins during koji culture. Besides well-known proteins (TAA and Oryzin), a variety of aminopeptidase and proteases were identical at the proteome level. This suggests that A. oryzae AS 3.951 has a powerful capacity to digest soybean protein.

Early detection of airborne inoculum of Magnaporthe oryzae in turfgrass fields using a quantitative LAMP assay

Science.gov (United States)

Gray leaf spot (GLS) is a destructive disease of perennial ryegrass caused by a host specific pathotype of the ascomycete Magnaporthe oryzae. Early diagnosis is crucial for effective disease management and the implementation of Integrated Pest Management practices. However, a rapid diagnostic protoc...

Sequence- and Structure-Based Functional Annotation and Assessment of Metabolic Transporters in Aspergillus oryzae: A Representative Case Study

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Nachon Raethong

2016-01-01

Full Text Available Aspergillus oryzae is widely used for the industrial production of enzymes. In A. oryzae metabolism, transporters appear to play crucial roles in controlling the flux of molecules for energy generation, nutrients delivery, and waste elimination in the cell. While the A. oryzae genome sequence is available, transporter annotation remains limited and thus the connectivity of metabolic networks is incomplete. In this study, we developed a metabolic annotation strategy to understand the relationship between the sequence, structure, and function for annotation of A. oryzae metabolic transporters. Sequence-based analysis with manual curation showed that 58 genes of 12,096 total genes in the A. oryzae genome encoded metabolic transporters. Under consensus integrative databases, 55 unambiguous metabolic transporter genes were distributed into channels and pores (7 genes, electrochemical potential-driven transporters (33 genes, and primary active transporters (15 genes. To reveal the transporter functional role, a combination of homology modeling and molecular dynamics simulation was implemented to assess the relationship between sequence to structure and structure to function. As in the energy metabolism of A. oryzae, the H+-ATPase encoded by the AO090005000842 gene was selected as a representative case study of multilevel linkage annotation. Our developed strategy can be used for enhancing metabolic network reconstruction.

Sequence- and Structure-Based Functional Annotation and Assessment of Metabolic Transporters in Aspergillus oryzae: A Representative Case Study.

Science.gov (United States)

Raethong, Nachon; Wong-Ekkabut, Jirasak; Laoteng, Kobkul; Vongsangnak, Wanwipa

2016-01-01

Aspergillus oryzae is widely used for the industrial production of enzymes. In A. oryzae metabolism, transporters appear to play crucial roles in controlling the flux of molecules for energy generation, nutrients delivery, and waste elimination in the cell. While the A. oryzae genome sequence is available, transporter annotation remains limited and thus the connectivity of metabolic networks is incomplete. In this study, we developed a metabolic annotation strategy to understand the relationship between the sequence, structure, and function for annotation of A. oryzae metabolic transporters. Sequence-based analysis with manual curation showed that 58 genes of 12,096 total genes in the A. oryzae genome encoded metabolic transporters. Under consensus integrative databases, 55 unambiguous metabolic transporter genes were distributed into channels and pores (7 genes), electrochemical potential-driven transporters (33 genes), and primary active transporters (15 genes). To reveal the transporter functional role, a combination of homology modeling and molecular dynamics simulation was implemented to assess the relationship between sequence to structure and structure to function. As in the energy metabolism of A. oryzae, the H(+)-ATPase encoded by the AO090005000842 gene was selected as a representative case study of multilevel linkage annotation. Our developed strategy can be used for enhancing metabolic network reconstruction.

Active-site-directed inactivation of Aspergillus oryzae beta-galactosidase with beta-D-galactopyranosylmethyl-p-nitrophenyltriazene.

Science.gov (United States)

Mega, T; Nishijima, T; Ikenaka, T

1990-04-01

beta-D-Galactopyranosylmethyl-p-nitrophenyltriazene (beta-GalMNT), a specific inhibitor of beta-galactosidase, was isolated as crystals by HPLC and its chemical and physicochemical characteristics were examined. Aspergillus oryzae beta-galactosidase was inactivated by the compound. We studied the inhibition mechanism in detail. The inhibitor was hydrolyzed by the enzyme to p-nitroaniline and an active intermediate (beta-galactopyranosylmethyl carbonium or beta-galactopyranosylmethyldiazonium), which inactivated the enzyme. The efficiency of inactivation of the enzyme (the ratio of moles of inactivated enzyme to moles of beta-GalMNT hydrolyzed by the enzyme) was 3%; the efficiency of Escherichia coli beta-galactosidase was 49%. In spite of the low efficiency, the rate of inactivation of A. oryzae enzyme was not very different from that of the E. coli enzyme, because the former hydrolyzed beta-GalMNT faster than the latter did. A. oryzae beta-galactosidase was also inactivated by p-chlorophenyl, p-tolyl, and m-nitrophenyl derivatives of beta-galactopyranosylmethyltriazene. However, E. coli beta-galactosidase was not inactivated by these triazene derivatives. The results showed that the inactivation of A. oryzae and E. coli beta-galactosidases by beta-GalMNT was an enzyme-activated and active-site-directed irreversible inactivation. The possibility of inactivation by intermediates produced nonenzymatically was ruled out for E. coli, but not for the A. oryzae enzyme.

Characterization of field isolates of Magnaporthe oryzae with mating type, DNA fingerprinting, and pathogenicity assays

Science.gov (United States)

Due to the harmful nature of the rice blast fungus, Magnaporthe oryzae, it is beneficial to characterize field isolates to help aid in the deployment of resistance (R) genes in rice. In the present study, 190 field isolates of M. oryzae, collected from rice fields of Yunnan province in China, were a...

Development of a genome editing technique using the CRISPR/Cas9 system in the industrial filamentous fungus Aspergillus oryzae.

Science.gov (United States)

Katayama, Takuya; Tanaka, Yuki; Okabe, Tomoya; Nakamura, Hidetoshi; Fujii, Wataru; Kitamoto, Katsuhiko; Maruyama, Jun-Ichi

2016-04-01

To develop a genome editing method using the CRISPR/Cas9 system in Aspergillus oryzae, the industrial filamentous fungus used in Japanese traditional fermentation and for the production of enzymes and heterologous proteins. To develop the CRISPR/Cas9 system as a genome editing technique for A. oryzae, we constructed plasmids expressing the gene encoding Cas9 nuclease and single guide RNAs for the mutagenesis of target genes. We introduced these into an A. oryzae strain and obtained transformants containing mutations within each target gene that exhibited expected phenotypes. The mutational rates ranged from 10 to 20 %, and 1 bp deletions or insertions were the most commonly induced mutations. We developed a functional and versatile genome editing method using the CRISPR/Cas9 system in A. oryzae. This technique will contribute to the use of efficient targeted mutagenesis in many A. oryzae industrial strains.

Comparative proteomics of Rhizopus delemar ATCC 20344 unravels the role of amino acid catabolism in fumarate accumulation

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Dorett I. Odoni

2017-03-01

Full Text Available The filamentous fungus Rhizopus delemar naturally accumulates relatively high amounts of fumarate. Although the culture conditions that increase fumarate yields are well established, the network underlying the accumulation of fumarate is not yet fully understood. We set out to increase the knowledge about fumarate accumulation in R. delemar. To this end, we combined a transcriptomics and proteomics approach to identify key metabolic pathways involved in fumarate production in R. delemar, and propose that a substantial part of the fumarate accumulated in R. delemar during nitrogen starvation results from the urea cycle due to amino acid catabolism.

Kinetic studies of acid inactivation of alpha-amylase from Aspergillus oryzae

DEFF Research Database (Denmark)

Carlsen, Morten; Nielsen, Jens Bredal; Villadsen, John

1996-01-01

The stability of alpha-amylase from Aspergillus oryzae has been studied at different pH. The enzyme is extremely stable at neutral pH (pH 5-8), whereas outside this pH-range a substantial loss of activity is observed. The acid-inactivation of alpha-amylase from A. oryzae was monitored on...... regains part of its activity, and the reactivation process also follows first-order kinetics. The irreversible loss of activity is found not to result from a protease contamination of the protein samples. A proposed model, where irreversibly inactivated a-amylase is formed both directly from the active...

Identification of growth phenotype-related genes in Aspergillus oryzae by heterologous macroarray and suppression subtractive hybridization

NARCIS (Netherlands)

Biesebeke, R. te; Levin, A.; Sagt, C.; Bartels, J.; Goosen, T.; Ram, A.; Hondel, C. van den; Punt, P.

2005-01-01

Aspergillus oryzae requires polarized growth for colonization of solid substrates, and this growth phenotype differs from that seen in liquid medium. Various experimental approaches were used to identify genes that are differentially expressed when A. oryzae is grown on wheat kernels and in a

Convenient enzymatic resolution of (R,S)-2-methylbutyric acid catalyzed by immobilized lipases.

Science.gov (United States)

Mittersteiner, Mateus; Linshalm, Bruna Luiza; Vieira, Ana Paula Furlan; Brondani, Patrícia Bulegon; Scharf, Dilamara Riva; de Jesus, Paulo Cesar

2018-01-01

The application of several immobilized lipases has been explored in the enantioselective esterification of (R,S)-2-methylbutyric acid, an insect pheromone precursor. With the use of Candida antarctica B, using hexane as solvent, (R)-pentyl 2-methylbutyrate was prepared in 2 h with c 40%, ee p 90%, and E = 35, while Thermomyces lanuginosus leads to c 18%, ee p 91%, and E = 26. The (S)-enantiomer was obtained by the use of Candida rugosa or Rhizopus oryzae (2-h reaction, c 34% and 35%, ee p 75 and 49%, and E = 10 and 4, respectively). Under optimal conditions, the effect of the solvent, the molar ratio, and the nucleophile were evaluated. © 2017 Wiley Periodicals, Inc.

Evaluation of the catalytic activity of lipases immobilized on chrysotile for esterification

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Silva Jane E. S.

2003-01-01

Full Text Available In the present work, the ester synthesis in organic media catalyzed by lipases immobilized on chrysotile was studied. Lipases of different sources (Mucor javanicus, Pseudomonas cepacia, Rhizopus oryzae, Aspergillus niger and Candida rugosa were immobilized on chrysotile, an inexpensive magnesium silicate, and used for esterification of hexanoic, octanoic and lauric acid with methanol, ethanol, 1-butanol and 1-octanol at 25ºC in hexane as solvent. The best results were obtained with Mucor javanicus lipase and lauric acid giving yields of 62-97% of ester.

Robustness and strategies of adaptation among farmer varieties of African Rice (Oryza glaberrima) and Asian Rice (Oryza sativa) across West Africa.

Science.gov (United States)

Mokuwa, Alfred; Nuijten, Edwin; Okry, Florent; Teeken, Béla; Maat, Harro; Richards, Paul; Struik, Paul C

2013-01-01

This study offers evidence of the robustness of farmer rice varieties (Oryza glaberrima and O. sativa) in West Africa. Our experiments in five West African countries showed that farmer varieties were tolerant of sub-optimal conditions, but employed a range of strategies to cope with stress. Varieties belonging to the species Oryza glaberrima - solely the product of farmer agency - were the most successful in adapting to a range of adverse conditions. Some of the farmer selections from within the indica and japonica subspecies of O. sativa also performed well in a range of conditions, but other farmer selections from within these two subspecies were mainly limited to more specific niches. The results contradict the rather common belief that farmer varieties are only of local value. Farmer varieties should be considered by breeding programmes and used (alongside improved varieties) in dissemination projects for rural food security.

Robustness and Strategies of Adaptation among Farmer Varieties of African Rice (Oryza glaberrima) and Asian Rice (Oryza sativa) across West Africa

Science.gov (United States)

Maat, Harro; Richards, Paul; Struik, Paul C.

2013-01-01

This study offers evidence of the robustness of farmer rice varieties (Oryza glaberrima and O. sativa) in West Africa. Our experiments in five West African countries showed that farmer varieties were tolerant of sub-optimal conditions, but employed a range of strategies to cope with stress. Varieties belonging to the species Oryza glaberrima – solely the product of farmer agency – were the most successful in adapting to a range of adverse conditions. Some of the farmer selections from within the indica and japonica subspecies of O. sativa also performed well in a range of conditions, but other farmer selections from within these two subspecies were mainly limited to more specific niches. The results contradict the rather common belief that farmer varieties are only of local value. Farmer varieties should be considered by breeding programmes and used (alongside improved varieties) in dissemination projects for rural food security. PMID:23536754

Bipolaris oryzae, a novel fungal opportunist causing keratitis

NARCIS (Netherlands)

Al-Hatmi, Abdullah

2015-01-01

We report a case of mycotic keratitis caused by Bipolaris oryzae with predisposing trauma from a foreign body. The fungus was identified by sequencing the internal transcribed spacer (ITS) region, translation elongation factor 1α (TEF1) gene and partial glyceraldehyde-3-phosphate dehydrogenase

Host-Induced Gene Silencing of Rice Blast Fungus Magnaporthe oryzae Pathogenicity Genes Mediated by the Brome Mosaic Virus.

Science.gov (United States)

Zhu, Lin; Zhu, Jian; Liu, Zhixue; Wang, Zhengyi; Zhou, Cheng; Wang, Hong

2017-09-26

Magnaporthe oryzae is a devastating plant pathogen, which has a detrimental impact on rice production worldwide. Despite its agronomical importance, some newly-emerging pathotypes often overcome race-specific disease resistance rapidly. It is thus desirable to develop a novel strategy for the long-lasting resistance of rice plants to ever-changing fungal pathogens. Brome mosaic virus (BMV)-induced RNA interference (RNAi) has emerged as a useful tool to study host-resistance genes for rice blast protection. Planta-generated silencing of targeted genes inside biotrophic pathogens can be achieved by expression of M. oryzae -derived gene fragments in the BMV-mediated gene silencing system, a technique termed host-induced gene silencing (HIGS). In this study, the effectiveness of BMV-mediated HIGS in M. oryzae was examined by targeting three predicted pathogenicity genes, MoABC1, MoMAC1 and MoPMK1 . Systemic generation of fungal gene-specific small interfering RNA (siRNA) molecules induced by inoculation of BMV viral vectors inhibited disease development and reduced the transcription of targeted fungal genes after subsequent M. oryzae inoculation. Combined introduction of fungal gene sequences in sense and antisense orientation mediated by the BMV silencing vectors significantly enhanced the efficiency of this host-generated trans-specific RNAi, implying that these fungal genes played crucial roles in pathogenicity. Collectively, our results indicated that BMV-HIGS system was a great strategy for protecting host plants against the invasion of pathogenic fungi.

Intraspecies Volatile Interactions Affect Growth Rates and Exometabolomes in Aspergillus oryzae KCCM 60345.

Science.gov (United States)

Singh, Digar; Lee, Choong Hwan

2018-02-28

Volatile organic compounds (VOCs) are increasingly been recognized as the chemical mediators of mold interactions, shaping their community dynamics, growth, and metabolism. Herein, we selectively examined the time-correlated (0 D-11 D, where D = incubation days) effects of intraspecies VOC-mediated interactions (VMI) on Aspergillus oryzae KCCM 60345 (S1), following co-cultivation with partner strain A. oryzae KACC 44967 (S2), in a specially designed twin plate assembly. The comparative evaluation of S1 VMI (S1 subjected to VMI with S2) and its control (S1 Con ) showed a notable disparity in their radial growth (S1 VMI S1 Con ) at 3-5 D, amylase activity (S1 VMI S1 Con ) at 3 D. Furthermore, we observed a distinct clustering pattern for gas chromatography-time of flight-mass spectrometry datasets from 5 D extracts of S1 VMI and S1 Con in principle component analysis (PC1: 30.85%; PC2: 10.31%) and partial least squares discriminant analysis (PLS-DA) (PLS1: 30.77; PLS2: 10.15%). Overall, 43 significantly discriminant metabolites were determined for engendering the metabolic variance based on the PLS-DA model (VIP > 0.7, p S1 Con ) at 5 D, organic acids (S1 VMI > S1 Con ) at 5 D, and kojic acid (S1 VMI < S1 Con ) at 5-7 D were observed. Examining the headspace VOCs shared between S1 and S2 in the twin plate for 5 D incubated samples, we observed the relatively higher abundance of C-8 VOCs (1-octen-3-ol, (5Z)-octa-1,5-dien-3-ol, 3-octanone, 1-octen-3-ol acetate) having known semiochemical functions. The present study potentially illuminates the effects of VMI on commercially important A. oryzae's growth and biochemical phenotypes with subtle details of altered metabolomes.

Transcriptome Analysis of Early Responsive Genes in Rice during Magnaporthe oryzae Infection

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Yiming Wang

2014-12-01

Full Text Available Rice blast disease caused by Magnaporthe oryzae is one of the most serious diseases of cultivated rice (Oryza sativa L. in most rice-growing regions of the world. In order to investigate early response genes in rice, we utilized the transcriptome analysis approach using a 300 K tilling microarray to rice leaves infected with compatible and incompatible M. oryzae strains. Prior to the microarray experiment, total RNA was validated by measuring the differential expression of rice defense-related marker genes (chitinase 2, barwin, PBZ1, and PR-10 by RT-PCR, and phytoalexins (sakuranetin and momilactone A with HPLC. Microarray analysis revealed that 231 genes were up-regulated (>2 fold change, p < 0.05 in the incompatible interaction compared to the compatible one. Highly expressed genes were functionally characterized into metabolic processes and oxidation-reduction categories. The oxidative stress response was induced in both early and later infection stages. Biotic stress overview from MapMan analysis revealed that the phytohormone ethylene as well as signaling molecules jasmonic acid and salicylic acid is important for defense gene regulation. WRKY and Myb transcription factors were also involved in signal transduction processes. Additionally, receptor-like kinases were more likely associated with the defense response, and their expression patterns were validated by RT-PCR. Our results suggest that candidate genes, including receptor-like protein kinases, may play a key role in disease resistance against M. oryzae attack.

Production of 8-hydroxydaidzein from soybean extract by Aspergillus oryzae KACC 40247.

Science.gov (United States)

Seo, Min-Ho; Kim, Bi-Na; Kim, Kyoung-Rok; Lee, Ki Won; Lee, Choong-Hwan; Oh, Deok-Kun

2013-01-01

Aspergillus oryzae KACC 40247 was selected from among 60 fungal strains as an effective 7,8,4'-trihydroxyisoflavone (8-hydroxydaidzein)-producing fungus. The optimal culture conditions for production by this strain in a 7-L fermentor were found to be 30 °C, pH 6, and 300 rpm. Under these conditions, A. oryzae KACC 40247 produced 62 mg/L of 8-hydroxydaidzein from soybean extract in 30 h, with a productivity of 2.1 mg/L/h. These are the highest production and productivity for 8-hydroxydaidzein ever reported. To increase production, several concentrations of daidzin and of daidzein as precursor were added at several culture times. The optimal addition time and concentration for daidzin were 12 h and 1,248 mg/L, and those for daidzein were 12 h and 254 mg/L respectively. Maximum production and productivity for 8-hydroxydaidzein with the addition of daidzein were 95 mg/L and 3.2 mg/L/h respectively, and those with the addition of daidzin were 160 mg/L and 4.4 mg/L/h respectively.

Deciphering the signaling mechanisms of the plant cell wall degradation machinery in Aspergillus oryzae

DEFF Research Database (Denmark)

Udatha, D. B. R. K. Gupta; Topakas, Evangelos; Salazar, Margarita Pena

2015-01-01

is required to rationally engineer filamentous fungi for more efficient bioconversion of different types of biomass. Results: In this study we focused on ten chemically defined inducers to drive expression of cellulases, hemicellulases and accessory enzymes in the model filamentous fungus Aspergillus oryzae....... oryzae genome were only partially explained by the chemical similarity of the enzyme inducers. Genes encoding enzymes that have attracted considerable interest such as cellobiose dehydrogenases and copper-dependent polysaccharide mono-oxygenases presented a substrate-specific induction. Several homology...... in 2360 reactions in the genome scale metabolic network of A. oryzae, was performed through a two-step molecular docking against the binding pockets of the transcription factors AoXlnR and AoAmyR. A total of six metabolites viz., sulfite (H2SO3), sulfate (SLF), uroporphyrinogen III (UPGIII), ethanolamine...

Establishment of a new method to quantitatively evaluate hyphal fusion ability in Aspergillus oryzae.

Science.gov (United States)

Tsukasaki, Wakako; Maruyama, Jun-Ichi; Kitamoto, Katsuhiko

2014-01-01

Hyphal fusion is involved in the formation of an interconnected colony in filamentous fungi, and it is the first process in sexual/parasexual reproduction. However, it was difficult to evaluate hyphal fusion efficiency due to the low frequency in Aspergillus oryzae in spite of its industrial significance. Here, we established a method to quantitatively evaluate the hyphal fusion ability of A. oryzae with mixed culture of two different auxotrophic strains, where the ratio of heterokaryotic conidia growing without the auxotrophic requirements reflects the hyphal fusion efficiency. By employing this method, it was demonstrated that AoSO and AoFus3 are required for hyphal fusion, and that hyphal fusion efficiency of A. oryzae was increased by depleting nitrogen source, including large amounts of carbon source, and adjusting pH to 7.0.

EFFECT OF GAMMA IRRADIATION AND ENVIRONMENTAL FACTORS ON -AMYLASE PRODUCTION BY ASPERGILLUS NIGER AND ASPERGILLUS ORYZAE FROM SOME AGRICULTURAL WASTES

International Nuclear Information System (INIS)

MATTAR, Z.A.

2008-01-01

Amylases are one of the most important and oldest industrial enzymes. The optimization of production of α -amylase from Aspergillus niger and Aspergillus oryzae fungi, using different agro-wastes as sole carbon sources, was performed. The highest productivity of α -amylase by the two organisms was recorded at pH 6 and incubation temperature at 30 0C when the two organisms were grown on potato peels (PPs) and/or wheat straw (Ws) after days of cultivation. Pre-treated PPs and Ws with 20 kGy gave the best enzyme productivity by the two organisms compared with untreated ones. Also, exposing the inoculums of A. niger and A.oryzae to 0.5 and 0.75 kGy, respectively, led to enhancement of α-amylase to 48 and 46 μ/ml, respectively

Construction of brewing-wine Aspergillus oryzae pyrG- mutant by pyrG gene deletion and its application in homology transformation.

Science.gov (United States)

Du, Yu; Xie, Guizhen; Yang, Chunfa; Fang, Baishan; Chen, Hongwen

2014-06-01

pyrG(-) host cells are indispensable for pyrG(-) based transformation system. Isolations of pyrG(-) host cells by random mutations are limited by time-consuming, unclear genetic background and potential interferences of homogenous recombination. The purpose of this study was to construct brewing-wine Aspergillus oryzae pyrG(-) mutant by site-directed mutation of pyrG gene deletion which would be used as a host for further transformation. pMD-pyrGAB, a vector carrying pyrG deletion cassette, was used to construct pyrG(-) mutant of A. oryzae. Three stable pyrG deletion mutants of A. oryzae were isolated by resistant to 5-fluoroorotic acid and confirmed by polymerase chain reaction analysis, indicating that pyrG was completely excised. The ΔpyrG mutants were applied as pyrG(-) host cells to disrupt xdh gene encoding xylitol dehydrogenase, which involves in xylitol production of A. oryzae. The xdh disruption mutants were efficiently constructed by transforming a pMD-pyrG-xdh disruption plasmid carrying pyrG, and the produced xylitol concentration of the Δxdh mutant was three times as much as that of the ΔpyrG recipient. Site-directed pyrG gene deletion is thus an effective way for the isolation of pyrG(-) host cells, and the established host-vector system could be applied in further functional genomics analysis and molecular breeding of A. oryzae. © The Author 2014. Published by ABBS Editorial Office in association with Oxford University Press on behalf of the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences.

Transcriptome and Proteome Expression Analysis of the Metabolism of Amino Acids by the Fungus Aspergillus oryzae in Fermented Soy Sauce

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Guozhong Zhao

2015-01-01

Full Text Available Amino acids comprise the majority of the flavor compounds in soy sauce. A portion of these amino acids are formed from the biosynthesis and metabolism of the fungus Aspergillus oryzae; however, the metabolic pathways leading to the formation of these amino acids in A. oryzae remain largely unknown. We sequenced the transcriptomes of A. oryzae 100-8 and A. oryzae 3.042 under similar soy sauce fermentation conditions. 2D gel electrophoresis was also used to find some differences in protein expression. We found that many amino acid hydrolases (endopeptidases, aminopeptidases, and X-pro-dipeptidyl aminopeptidase were expressed at much higher levels (mostly greater than double in A. oryzae 100-8 than in A. oryzae 3.042. Our results indicated that glutamate dehydrogenase may activate the metabolism of amino acids. We also found that the expression levels of some genes changed simultaneously in the metabolic pathways of tyrosine and leucine and that these conserved genes may modulate the function of the metabolic pathway. Such variation in the metabolic pathways of amino acids is important as it can significantly alter the flavor of fermented soy sauce.

Transcriptome and Proteome Expression Analysis of the Metabolism of Amino Acids by the Fungus Aspergillus oryzae in Fermented Soy Sauce.

Science.gov (United States)

Zhao, Guozhong; Yao, Yunping; Wang, Chunling; Tian, Fengwei; Liu, Xiaoming; Hou, Lihua; Yang, Zhen; Zhao, Jianxin; Zhang, Hao; Cao, Xiaohong

2015-01-01

Amino acids comprise the majority of the flavor compounds in soy sauce. A portion of these amino acids are formed from the biosynthesis and metabolism of the fungus Aspergillus oryzae; however, the metabolic pathways leading to the formation of these amino acids in A. oryzae remain largely unknown. We sequenced the transcriptomes of A. oryzae 100-8 and A. oryzae 3.042 under similar soy sauce fermentation conditions. 2D gel electrophoresis was also used to find some differences in protein expression. We found that many amino acid hydrolases (endopeptidases, aminopeptidases, and X-pro-dipeptidyl aminopeptidase) were expressed at much higher levels (mostly greater than double) in A. oryzae 100-8 than in A. oryzae 3.042. Our results indicated that glutamate dehydrogenase may activate the metabolism of amino acids. We also found that the expression levels of some genes changed simultaneously in the metabolic pathways of tyrosine and leucine and that these conserved genes may modulate the function of the metabolic pathway. Such variation in the metabolic pathways of amino acids is important as it can significantly alter the flavor of fermented soy sauce.

Robustness and Strategies of Adaptation among Farmer Varieties of African Rice (Oryza glaberrima) and Asian Rice (Oryza sativa) across West Africa

NARCIS (Netherlands)

Mokuwa, A.; Nuijten, H.A.C.P.; Okry, F.; Teeken, B.W.E.; Maat, H.; Richards, P.; Struik, P.C.

2013-01-01

This study offers evidence of the robustness of farmer rice varieties (Oryza glaberrima and O. sativa) in West Africa. Our experiments in five West African countries showed that farmer varieties were tolerant of sub-optimal conditions, but employed a range of strategies to cope with stress.

On-farm management practices against rice root weevil (Echinocnemus oryzae Marshall)

OpenAIRE

Rakesh Pandey; Ajit Kumar Chaturvedi; Rudal Prasad Chaudhary; Rajendra Prasad

2017-01-01

Rice is the staple food of over half the world's population and occupies almost one-fifth of the global cropland under cereals. The rice root weevil, Echinocnemus oryzae Marshall, (Coleoptera: Curculionidae) has posed a problem in paddy cultivation areas in India. The damage by this root weevil results in a significant decrease in root and shoot biomass and ultimately the yield of rice plants. Studies were conducted to test the effective management practices of rice root weevil using a seedli...

Control of brown spot pathogen of rice (Bipolaris oryzae using some phenolic antioxidants Controle da macha-parda do arroz (Bipolaris oryzae pelo emprego de antioxidantes fenólicos

Directory of Open Access Journals (Sweden)

Y.M. Shabana

2008-09-01

Full Text Available Bipolaris oryzae is the causal agent of rice brown spot disease and is responsible for significant economic losses. In order to control this disease, three phenolic antioxidants were tested (salicylic acid, benzoic acid and hydroquinone. The antifungal activity of the tested substances were investigated against B. oryzae at different concentrations in vitro, as well as the efficacy of their exogenous application in controlling rice brown spot disease under field conditions. In vitro, benzoic acid or salicylic acid at 9 mM completely inhibited the growth of B. oryzae. Under field conditions, spraying of benzoic acid at 20 mM led to a significant reduction in disease severity (DS and disease incidence (DI on the plant leaves, in addition to a significant increase in the grain yield and its components. Some biochemical responses were also detected, where the application of the previous treatment led to a significant increase in the total photosynthetic pigments (chlorophyll a and b and carotenoids in rice leaves and in the total carbohydrate and protein contents of the yielded grains.Bipolaris oryzae é o agente causador da doença mancha-parda do arroz e é responsável por significativas perdas econômicas. Três antioxidantes fenólicos (ácido salicílico, ácido benzóico e hidroquinona foram avaliados para o controle dessa doença do arroz. A atividade antifúngica destes compostos foi avaliada in vitro contra B. oryzae em diferentes concentrações e a eficiência de sua aplicação exógena no controle da mancha-parda foi avaliada em condições de campo. Nos ensaios in vitro, os ácidos benzóico e salicílico a 9 mM inibiram completamente a multiplicação de B. oryzae. Em condições de campo, a aspersão de ácido benzóico a 20 mM causou uma redução significativa na gravidade e incidência da doença na folhas da planta, além de aumentar significativamente o rendimento dos grãos e seus componentes. Algumas respostas bioqu

Synthesis of oligosaccharides derived from lactulose (OsLu using soluble and immobilized Aspergillus oryzae b-galactosidase

Directory of Open Access Journals (Sweden)

ALEJANDRA eCARDELLE COBAS

2016-03-01

Full Text Available b-galactosidase from Aspergillus oryzae offers a high yield for the synthesis of oligosaccharides derived from lactulose (OsLu by transgalactosylation. Oligosaccharides with degree of polymerization (DP ≥ 3 have shown to possess higher in vitro bifidogenic effect than di- and tetrasaccharides. Thus, in this work, an optimization of reaction conditions affecting the specific selectivity of A. oryzae b-galactosidase for synthesis of OsLu has been carried out to enhance OsLu with DP ≥ 3 production. Assays with b-galactosidase immobilized onto a glutaraldehyde-agarose support were also carried out with the aim of making the process cost-effective and industrially viable. Optimal conditions with both soluble and immobilized enzyme for the synthesis of OsLu with DP ≥ 3 were 50 °C, pH 6.5, 450 g/L of lactulose and 8 U/mL of enzyme, reaching yields of ca. 50% (w/v of total OsLu and ca. 20% (w/v of OsLu-3, being 6′-galactosyl-lactulose the major one, after a short reaction time. Selective formation of disaccharides, however, was favored at 60 °C, pH 4.5, 450 g/L of lactulose and 8 U/mL of enzyme. Immobilization increased the enzymatic stability to temperature changes and allowed to reuse the enzyme. We can conclude that the use, under determined optimal conditions, of the A. oryzae b-galactosidase immobilized on a support of glutaraldehyde-agarose constitutes an efficient and cost-effective alternative to the use of soluble b-galactosidases for the synthesis of prebiotic OsLu mixtures.

Identification and toxigenic potential of the industrially important fungi, Aspergillus oryzae and Aspergillus sojae

DEFF Research Database (Denmark)

Jørgensen, Thomas R

2007-01-01

Mold strains belonging to the species Aspergillus oryzae and Aspergillus sojae are highly valued as koji molds in the traditional preparation of fermented foods, such as miso, sake, and shoyu, and as protein production hosts in modern industrial processes. A. oryzae and A. sojae are relatives...... of the wild molds Aspergillus flavus and Aspergillus parasiticus. All four species are classified to the A. flavus group. Strains of the A. flavus group are characterized by a high degree of morphological similarity. Koji mold species are generally perceived of as being nontoxigenic, whereas wild molds...... are associated with the carcinogenic aflatoxins. Thus, reliable identification of individual strains is very important for application purposes. This review considers the pheno- and genotypic markers used in the classification of A. flavus group strains and specifically in the identification of A. oryzae and A...

Improved heterologous protein production by a tripeptidyl peptidase gene (AosedD) disruptant of the filamentous fungus Aspergillus oryzae.

Science.gov (United States)

Zhu, Lin; Nemoto, Takeshi; Yoon, Jaewoo; Maruyama, Jun-ichi; Kitamoto, Katsuhiko

2012-01-01

Proteolytic degradation is one of the serious bottlenecks limiting the yields of heterologous protein production by Aspergillus oryzae. In this study, we selected a tripeptidyl peptidase gene AosedD (AO090166000084) as a candidate potentially degrading the heterologous protein, and performed localization analysis of the fusion protein AoSedD-EGFP in A. oryzae. As a result, the AoSedD-EGFP was observed in the septa and cell walls as well as in the culture medium, suggesting that AoSedD is a secretory enzyme. An AosedD disruptant was constructed to investigate an effect of AoSedD on the production level of heterologous proteins and protease activity. Both of the total protease and tripeptidyl peptidase activities in the culture medium of the AosedD disruptant were decreased as compared to those of the control strain. The maximum yields of recombinant bovine chymosin (CHY) and human lysozyme (HLY) produced by the AosedD disruptants showed approximately 2.9- and 1.7-fold increases, respectively, as compared to their control strains. These results suggest that AoSedD is one of the major proteases involved in the proteolytic degradation of recombinant proteins in A. oryzae.

Comparison of volatile and non-volatile metabolites in rice wine fermented by Koji inoculated with Saccharomycopsis fibuligera and Aspergillus oryzae.

Science.gov (United States)

Son, Eun Yeong; Lee, Sang Mi; Kim, Minjoo; Seo, Jeong-Ah; Kim, Young-Suk

2018-07-01

This study investigated volatile and nonvolatile metabolite profiles of makgeolli (a traditional rice wine in Korea) fermented by koji inoculated with Saccharomycopsis fibuligera and/or Aspergillus oryzae. The enzyme activities in koji were also examined to determine their effects on the formation of metabolites. The contents of all 18 amino acids detected were the highest in makgeolli fermented by S. fibuligera CN2601-09, and increased after combining with A. oryzae CN1102-08, unlike the contents of most fatty acids. On the other hand, major volatile metabolites were fusel alcohols, acetate esters, and ethyl esters. The contents of most fusel alcohols and acetate esters were the highest in makgeolli fermented by S. fibuligera CN2601-09, for which the protease activity was the highest, leading to the largest amounts of amino acods. The makgeolli samples fermented only by koji inoculated with S. fibuligera could be discriminated on PCA plots from the makgeolli samples fermented in combination with A. oryzae. In the case of nonvolatile metabolites, all amino acids and some metabolites such as xylose, 2-methylbenzoic acid, and oxalic acid contributed mainly to the characteristics of makgeolli fermented by koji inoculated with S. fibuligera and A. oryzae. These results showed that the formations of volatile and nonvolatile metabolites in makgeolli can be significantly affected by microbial strains with different enzyme activities in koji. To our knowledge, this study is the first report on the effects of S. fibuligera strains on the formation of volatile and non-volatile metabolites in rice wine, facilitating their use in brewing rice wine. Copyright © 2018. Published by Elsevier Ltd.

The proportion of non-aflatoxigenic strains of the Aspergillus flavus/oryzae complex from meju by analyses of the aflatoxin biosynthetic genes.

Science.gov (United States)

Hong, Seung-Beom; Lee, Mina; Kim, Dae-Ho; Chung, Soo-Hyun; Shin, Hyeon-Dong; Samson, Robert A

2013-12-01

Strains of the Aspergillus flavus/oryzae complex are frequently isolated from meju, a fermented soybean product, that is used as the starting material for ganjang (soy sauce) and doenjang (soybean paste) production. In this study, we examined the aflatoxin producing capacity of A. flavus/oryzae strains isolated from meju. 192 strains of A. flavus/oryzae were isolated from more than 100 meju samples collected from diverse regions of Korea from 2008 to 2011, and the norB-cypA, omtA, and aflR genes in the aflatoxin biosynthesis gene cluster were analyzed. We found that 178 strains (92.7%) belonged to non-aflatoxigenic group (Type I of norB-cypA, IB-L-B-, IC-AO, or IA-L-B- of omtA, and AO type of aflR), and 14 strains (7.3%) belonged to aflatoxin-producible group (Type II of norB-cypA, IC-L-B+/B- or IC-L-B+ of omtA, and AF type of aflR). Only 7 strains (3.6%) in the aflatoxin-producible group produced aflatoxins on Czapek yeast-extract medium. The aflatoxin-producing capability of A. flavus/oryzae strains from other sources in Korea were also investigated, and 92.9% (52/56) strains from air, 93.9% (31/33) strains from rice straw, 91.7% (11/12) strains from soybean, 81.3% (13/16) strains from corn, 82% (41/50) strains from peanut, and 73.2% (41/56) strains from arable soil were included in the non-aflatoxigenic group. The proportion of non-aflatoxigenicity of meju strains was similar to that of strains from soybean, air and rice straw, all of which have an effect on the fermentation of meju. The data suggest that meju does not have a preference for non-aflatoxigenic or aflatoxin-producible strains of A. flavus/oryzae from the environment of meju. The non-aflatoxigenic meju strains are proposed to be named A. oryzae, while the meju strains that can produce aflatoxins should be referred to A. flavus in this study.

Gamma ray, EMS and sodium azide induced effectiveness and efficiency of chlorophyll mutations in basmati rice (Oryza sativa L.)

International Nuclear Information System (INIS)

Singh, Sanjeev; Singh, J.; Singh, R.K.

2001-01-01

The effectiveness and efficiency of gamma ray, EMS, sodium azide alone or in combination in relation to chlorophyll mutations in two varieties of Basmati rice (Oryza sativa L.) in M 2 generation were studied. The chlorophyll mutations were induced by all the doses of mutagens alone or in combination relatively at a fair frequency in both the varieties in M 2 generation. In general, it was found that combination treatments of gamma rays and EMS were observed to be more efficient in Taraori Basmati, while EMS alone and combination treatment of gamma rays and EMS were more efficient in Pusa Basmati 1 on sterility and growth injury basis both. Sodium azide at 0.5 mM was found as the most effective dose in both Taraori Basmati and Pusa Basmati 1 cultivars. (author)

Analysis of an acyl-CoA binding protein in Aspergillus oryzae that undergoes unconventional secretion.

Science.gov (United States)

Kwon, Hee Su; Kawaguchi, Kouhei; Kikuma, Takashi; Takegawa, Kaoru; Kitamoto, Katsuhiko; Higuchi, Yujiro

2017-11-04

Acyl-CoA binding protein (ACBP) plays important roles in the metabolism of lipids in eukaryotic cells. In the industrially important filamentous fungus Aspergillus oryzae, although we have previously demonstrated that the A. oryzae ACBP (AoACBP) localizes to punctate structures and exhibits long-range motility, which is dependent on autophagy-related proteins, the physiological role of AoACBP remains elusive. Here, we describe identification and characterization of another ACBP from A. oryzae; we named this ACBP as AoAcb2 and accordingly renamed AoACBP as AoAcb1. The deduced amino acid sequence of AoAcb2 lacked a signal peptide. Phylogenetic analysis classified AoAcb2 into a clade that was same as the ACBP Acb1 of the model yeast Saccharomyces cerevisiae, but was different from that of AoAcb1. In contrast to punctate localization of AoAcb1, AoAcb2 was found to be dispersedly distributed in the cytoplasm, as was previously observed for the S. cerevisiae Acb1. Since we could not generate an Aoacb2 disruptant, we created an Aoacb2 conditional mutant that exhibited less growth under Aoacb2-repressed condition, suggesting that Aoacb2 is an essential gene for growth. Moreover, we observed that A. oryzae AoAcb2, but not A. oryzae AoAcb1, was secreted under carbon-starved condition, suggesting that AoAcb2 might be secreted via the unconventional protein secretion (UPS) pathway, just like S. cerevisiae Acb1. We also demonstrated that the unconventional secretion of AoAcb2 was dependent on the t-SNARE AoSso1, but was independent of the autophagy-related protein AoAtg1, suggesting that the unconventional secretion of AoAcb2, unlike that of S. cerevisiae Acb1, via the UPS pathway, is not regulated by the autophagy machinery. Thus, the filamentous fungus A. oryzae harbors two types of ACBPs, one of which appears to be essential for growth and undergoes unconventional secretion. Copyright © 2017 Elsevier Inc. All rights reserved.

The study of mildew proof effect of 60Co γ irradiation on leather shoes

International Nuclear Information System (INIS)

Wu Caixuan; Deng Zhiqun; Yu Qin; Zeng Daonan; Deng Guili

1992-01-01

Leather products occupy an important place in the national economy. In order to prevent from mildewing of leather products in storage, transportation and sale, the packed shoes were irradiated with 60 Co γ ray. Test materials are cattle, sheep and pig leather shoes. The results shows that: 1. There are 11 types of molds which make leather shoes mildewed: Oospora sp, Rhizopus oryzae Went and Grrelings, Aspergillus flavus Link, Aspergillus terreus thom, Monilia sp, Botrytis sp, Aspergillus niger V. Tiegh, Penicillum Lividum Westling, Penicillum Corylophilum Dieckx, Penicillum Cyaneum (B. and S.) Biourge and Penicillum Verrulosum Peeyrone. 2. The three types of shoes packed with polyethylene film can be prevented from mildewing after irradiation at level of 12 kGy. 3. The irradiation at 30 kGy does not change the physical properties of the shoes. 4. The irradiated shoes packed with polyethylene film can prevent mildewing, but unpacked ones would be mildewed again

Preparation of Biodiesel with Liquid Synergetic Lipases from Rapeseed Oil Deodorizer Distillate.

Science.gov (United States)

Zeng, Leping; He, Yaojia; Jiao, Liangcheng; Li, Kai; Yan, Yunjun

2017-11-01

To reduce industrial production cost, cheap and easily available rapeseed oil deodorizer distillates were used as feedstock to prepare biodiesel in this study. As a result, liquid forms of Candida rugosa lipase and Rhizopus oryzae lipase (ROL) were functioned as new and effective catalysts with biodiesel yield of 92.63% for 30 h and 94.36% for 9 h, respectively. Furthermore, the synergetic effect between the two lipases was employed to enhance biodiesel yield with a result of 98.16% in 6 h under optimized conditions via response surface methodology. The obtained conversion rate surpassed both yields of the individual two lipases and markedly shortened the reaction time. The resultant optimal conditions were ROL ratio 0.84, water content 46 wt% (w/w), reaction temperature 34 °C, and reaction time 6 h.

Immobilization of Isolated Lipase From Moldy Copra (Aspergillus Oryzae

Directory of Open Access Journals (Sweden)

Seniwati Dali

2011-01-01

Full Text Available Enzyme immobilization is a recovery technique that has been studied in several years, using support as a media to help enzyme dissolutions to the reaction substrate. Immobilization method used in this study was adsorption method, using specific lipase from Aspergillus oryzae. Lipase was partially purified from the culture supernatant of Aspergillus oryzae. Enzyme was immobilized by adsorbed on silica gel. Studies on free and immobilized lipase systems for determination of optimum pH, optimum temperature, thermal stability and reusability were carried out. The results showed that free lipase had optimum pH 8,2 and optimum temperature 35 °C while the immobilized lipase had optimum 8,2 and optimum temperature 45 °C. The thermal stability of the immobilized lipase, relative to that of the free lipase, was markedly increased. The immobilized lipase can be reused for at least six times.

Isolation of a novel promoter for efficient protein expression by Aspergillus oryzae in solid-state culture.

Science.gov (United States)

Bando, Hiroki; Hisada, Hiromoto; Ishida, Hiroki; Hata, Yoji; Katakura, Yoshio; Kondo, Akihiko

2011-11-01

A novel promoter from a hemolysin-like protein encoding the gene, hlyA, was characterized for protein overexpression in Aspergillus oryzae grown in solid-state culture. Using endo-1,4-β-glucanase from A. oryzae (CelA) as the reporter, promoter activity was found to be higher than that of the α-amylase (amyA) and manganese superoxide dismutase (sodM) genes not only in wheat bran solid-state culture but also in liquid culture. Expression of the A. oryzae endoglucanase CelB and two heterologous endoglucanases (TrEglI and TrEglIII from Trichoderma reesei) under the control of the hlyA promoter were also found to be stronger than under the control of the amyA promoter in A. oryzae grown in wheat bran solid-state culture, suggesting that the hlyA promoter may be useful for the overproduction of other proteins as well. In wheat bran solid-state culture, the productivity of the hlyA promoter in terms of protein produced was high when the cultivation temperature was 30°C or 37°C, when the water content was 0.6 or 0.8 ml/g wheat bran, and from 48 to 72 h after inoculation. Because A. oryzae sporulated actively under these conditions and because hemolysin has been reported to play a role in fungal fruiting body formation, high-level expression of hlyA may be related to sporulation.

Protein enrichment of brewery spent grain from Rhizopus oligosporus by solid-state fermentation.

Science.gov (United States)

Canedo, Marianny Silva; de Paula, Fernanda Gomes; da Silva, Flávio Alves; Vendruscolo, Francielo

2016-07-01

Brewery spent grain represents approximately 85 % of total by-products generated in a brewery. Consisting of carbohydrates, fiber, minerals and low amounts of protein, the use of brewery spent grain is limited to the feeding of ruminants; however, its potential use should be investigated. The reuse of this by-product using microorganisms by solid-state fermentation process as the case of protein enrichment by single-cell protein incorporation is an alternative to ensure sustainability and generate commercially interesting products. In this context, the aim of this study was to grow Rhizopus oligosporus in brewery spent grain under different initial moisture contents and nitrogen sources to increase the protein content of the fermented material. After 7 days of fermentation, increase of 2-4 times in the crude protein and soluble protein content was verified, respectively, compared to unfermented brewery spent grain. The kinetics of protein enrichment demonstrated the possibility of application of this technique, which can be a great alternative for use in diets for animals.

Production of extracellular lipases by Rhizopus oligosporus in a stirred fermentor

Directory of Open Access Journals (Sweden)

Tehreema Iftikhar

2010-12-01

Full Text Available The present investigation deals with the kinetics of submerged extracellular lipases fermentation by both wild and mutant strains of Rhizopus oligosporus var. microsporus in a laboratory scale stirred fermentor. Other parameters studied were inoculum size, pH, agitation and rate of aeration. It was found that the growth and lipases production was increased gradually and reached its maximum 9.07± 0.42ª U mL-1 (W and 42.49 ± 3.91ª U mL-1 (M after 30h of fermentation for both wild and mutant strain. There is overall increase of 109% (W and 124% (M in the production of extracellular lipases as compared to shake flask. Another significant finding of the present study is that the fermentation period is reduced to 30 h in case of wild and 23 h in case of mutant from 48 h in shake flask studies. The specific productivity of mutant strain (qp = 377.3 U/g cells/h was several folds higher than wild strain. The specific production rate and growth coefficient revealed the hyperproducibility of extracellular lipases using mutant IIB-63NTG-7.

Effect of heavy atoms on the thermal stability of α-amylase from Aspergillus oryzae.

Directory of Open Access Journals (Sweden)

Michihiro Sugahara

Full Text Available Currently, there are no versatile and established methods for improving stability of proteins. In an entirely different approach from conventional techniques such as mutagenesis, we attempted to enhance enzyme stability of α-amylase from Aspergillus oryzae using a heavy-atom derivatization technique. We evaluated changes in stability using differential scanning calorimetry (DSC. Candidate heavy atoms were identified using the Heavy-Atom Database System HATODAS, a Web-based tool designed to assist in heavy-atom derivatization of proteins for X-ray crystallography. The denaturation temperature of α-amylase derivatized with gadolinium (Gd or samarium (Sm ions increased by 6.2 or 5.7°C, respectively, compared to that of the native protein (60.6°C. The binding of six Gd ions was confirmed by X-ray crystallography of the enzyme at 1.5 Å resolution. DSC and dynamic light-scattering data revealed a correlation between stability and the aggregation state upon addition of Gd ions. These results show that HATODAS search is an effective tool for selecting heavy atoms for stabilization of this protein.

Production of oat-based synbiotic beverage by two-stage fermentation with Rhizopus oryzae and Lactobacillus acidophilus

NARCIS (Netherlands)

Gao, F.; Cai, S.; Nout, M.J.R.; Wang, Y.; Xia, Y.; Li, Y.; Ji, B.

2012-01-01

Many studies have reported that oats could effectively reduce the serum cholesterol levels in humans, and the ß-glucan in oat is believed to be responsible for this physiologic effect. Probiotics are live microorganisms that can confer a healthy benefit on the host when administered in adequate

Chemometric analysis of in-line multi-wavelength fluorescence measurements obtained during cultivations with a lipase producing Aspergillus oryzae strain

DEFF Research Database (Denmark)

Haack, Martin Brian; Eliasson Lantz, Anna; Mortensen, P.P.

2007-01-01

The filamentous fungus, Aspergillus oryzae, was cultivated in batch and fed-batch cultivations in order to investigate the use of multi-wavelength fluorescence for monitoring course of events during filamentous fungi cultivations. The A. oryzae strain applied expressed a fungal lipase from...... Thermomyces lanuginosus. Spectra of multi-wavelength fluorescence were collected every 5 min with the BioView system (DELTA, Denmark) and both explorative and predictive models, correlating the fluorescence data with cell mass and lipase activity, were built. During the cultivations, A. oryzae displayed...

Using the concept of pseudo amino acid composition to predict resistance gene against Xanthomonas oryzae pv. oryzae in rice: an approach from chaos games representation.

Science.gov (United States)

Jingbo, Xia; Silan, Zhang; Feng, Shi; Huijuan, Xiong; Xuehai, Hu; Xiaohui, Niu; Zhi, Li

2011-09-07

To evaluate the possibility of an unknown protein to be a resistant gene against Xanthomonas oryzae pv. oryzae, a different mode of pseudo amino acid composition (PseAAC) is proposed to formulate the protein samples by integrating the amino acid composition, as well as the Chaos games representation (CGR) method. Some numerical comparisons of triangle, quadrangle and 12-vertex polygon CGR are carried to evaluate the efficiency of using these fractal figures in classifiers. The numerical results show that among the three polygon methods, triangle method owns a good fractal visualization and performs the best in the classifier construction. By using triangle + 12-vertex polygon CGR as the mathematical feature, the classifier achieves 98.13% in Jackknife test and MCC achieves 0.8462. Copyright © 2011 Elsevier Ltd. All rights reserved.

Fungicide sensitivity in the wild rice pathogen Bipolaris oryzae

Science.gov (United States)

In recent years the occurrence of fungal brown spot, caused by Bipolaris oryzae has increased in cultivated wild rice (Zizania palustris) paddies in spite of the use of fungicides. To implement an efficient integrated disease management system, we are exploring whether field isolates have developed ...

High Levels of Phytophenolics and Antioxidant Activities in Oryza ...

African Journals Online (AJOL)

Purpose: To investigate phenolic levels, phytophenolic profiles and total antioxidant activities of Oryza sativa, unpolished Thai rice. Methods: Unpolished Thai rice strains of Leum Phua, Klam, Hawm Nil and Black Rose were measured for antioxidant activity using 1,1-diphenyl-2-picrylhydrazyl (DPPH), ...

Biotransformation of L-tyrosine to Dopamine by a Calcium Alginate Immobilized Mutant Strain of Aspergillus oryzae.

Science.gov (United States)

Ali, Sikander; Nawaz, Wajeeha

2016-08-01

The present research work is concerned with the biotransformation of L-tyrosine to dopamine (DA) by calcium alginate entrapped conidiospores of a mutant strain of Aspergillus oryzae. Different strains of A. oryzae were isolated from soil. Out of 13 isolated strains, isolate-2 (I-2) was found to be a better DA producer. The wild-type I-2 was chemically improved by treating it with different concentrations of ethyl methyl sulfonate (EMS). Among seven mutant variants, EMS-6 exhibiting maximal DA activity of 43 μg/ml was selected. The strain was further exposed with L-cysteine HCl to make it resistant against diversion and environmental stress. The conidiospores of selected mutant variant A. oryzae EMS-6 strain were entrapped in calcium alginate beads. Different parameters for immobilization were investigated. The activity was further improved from 44 to 62 μg/ml under optimized conditions (1.5 % sodium alginate, 2 ml inoculum, and 2 mm bead size). The best resistant mutant variable exhibited over threefold increase in DA activity (62 μg/ml) than did wild-type I-2 (21 μg/ml) in the reaction mixture. From the results presented in the study, it was observed that high titers of DA activity in vitro could effectively be achieved by the EMS-induced mutagenesis of filamentous fungus culture used.

Effect of coupling agent on durian skin fibre nanocomposite reinforced polypropylene

Science.gov (United States)

Siti Nur E'zzati, M. A.; Anuar, H.; Siti Munirah Salimah, A. R.

2018-01-01

This paper reports on the development of a composite-based natural fiber to reduce the reliance on petroleum-based product in order to amplify environmental awareness. The production of Durian Skin Nanofiber (DSNF) was conducted using biological fermentation method via rhizopus oryzae in order to obtain the nano dimension of the particle size. Polypropylene (PP) and DSNF were produced using Haake internal mixer via melt blending technique. The significant effect of maleic anhydride grafted polypropylene (MAPP) on the properties of PP/DSNF nanocomposite was investigated to study its mechanical properties which are tensile strength and thermal stability using thermogravimetric (TGA) and differential scanning analysis (DSC). The tensile property of PP nanocomposites increased from 33 MPa to 38 MPa with the presence of MAPP. The addition of MAPP also increased the thermal stability of PP/DSNF nanocomposite where the char residue increased by 52%. Besides that, the thermal degradation of PP/DSNF and PP/DSNF-MAPP were higher than PP where they exerted higher amount of weight loss at an elevated temperature. The percentage of crystallinity, %Xc, of PP nanocomposites improved with the addition of MAPP by 35% based on the differential scanning calorimetry (DSC) result. The SEM analysis showed that the PP/DSNF-MAPP exerts ductile fracture while PP/DSNF exerts brittle fracture.

Equol, a Clinically Important Metabolite, Inhibits the Development and Pathogenicity of Magnaporthe oryzae, the Causal Agent of Rice Blast Disease

Directory of Open Access Journals (Sweden)

Jiaoyu Wang

2017-10-01

Full Text Available Equol, a metabolite of soybean isoflavone daidzein, has been proven to have various bioactivities related to human health, but little is known on its antifungal activity to plant fungal pathogens. Magnaporthe oryzae is a phytopathogenic fungus that causes rice blast, a devastating disease on rice. Here, we demonstrated that equol influences the development and pathogenicity of M. oryzae. Equol showed a significant inhibition to the mycelial growth, conidial generation and germination, and appressorial formation of M. oryzae. As a result, equol greatly reduced the virulence of M. oryzae on rice and barley leaves. The antifungal activity of equol was also found in several other plant fungal pathogens. These findings expand our knowledge on the bioactivities of equol.

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for rapid identification of fungal rhinosinusitis pathogens.

Science.gov (United States)

Huang, Yanfei; Wang, Jinglin; Zhang, Mingxin; Zhu, Min; Wang, Mei; Sun, Yufeng; Gu, Haitong; Cao, Jingjing; Li, Xue; Zhang, Shaoya; Lu, Xinxin

2017-03-01

Filamentous fungi are among the most important pathogens, causing fungal rhinosinusitis (FRS). Current laboratory diagnosis of FRS pathogens mainly relies on phenotypic identification by culture and microscopic examination, which is time consuming and expertise dependent. Although matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS has been employed to identify various fungi, its efficacy in the identification of FRS fungi is less clear. A total of 153 FRS isolates obtained from patients were analysed at the Clinical Laboratory at the Beijing Tongren Hospital affiliated to the Capital Medical University, between January 2014 and December 2015. They were identified by traditional phenotypic methods and Bruker MALDI-TOF MS (Bruker, Biotyper version 3.1), respectively. Discrepancies between the two methods were further validated by sequencing. Among the 153 isolates, 151 had correct species identification using MALDI-TOF MS (Bruker, Biot 3.1, score ≥2.0 or 2.3). MALDI-TOF MS enabled identification of some very closely related species that were indistinguishable by conventional phenotypic methods, including 1/10 Aspergillus versicolor, 3/20 Aspergillus flavus, 2/30 Aspergillus fumigatus and 1/20 Aspergillus terreus, which were misidentified by conventional phenotypic methods as Aspergillus nidulans, Aspergillus oryzae, Aspergillus japonicus and Aspergillus nidulans, respectively. In addition, 2/2 Rhizopus oryzae and 1/1 Rhizopus stolonifer that were identified only to the genus level by the phenotypic method were correctly identified by MALDI-TOF MS. MALDI-TOF MS is a rapid and accurate technique, and could replace the conventional phenotypic method for routine identification of FRS fungi in clinical microbiology laboratories.

EVALUACIÓN DE LA CONCENTRACIÓN DE LOS ÁCIDOS ACÉTICO, BUTÍRICO Y PROPIÓNICO EN EL CO-CULTIVO: ASPERGILLUS ORYZAE-BUTYRIVIBRIO FIBRISOLVENS. EVALUATION OF THE CONCENTRATION OF THE ACETIC, BUTYRIC AND PROPIONIC ACIDS IN THE CO-CULTURE: ASPERGILLUS ORYZAE-BUTYRIVIBRIO FIBRISOLVENS

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C. LARA MANTILLA

2008-12-01

Full Text Available Se realizó un estudio en co-cultivo entre el hongo Aspergillus oryzae y la bacteria ruminal celulolítica Butyrivibrio fibrisolvens, cuyo objetivo fue determinar "in vitro" el efecto del hongo sobre la producción de los ácidos acético, propiónico y butírico por parte de la bacteria. El medio de cultivo se preparó utilizando líquido ruminal filtrado, centrifugado, autoclavado y diluído al 40% con agua, y 0,05 p/v de pastos Angleton (Dichamthium aristatum (Córdoba, Colombia. Las condiciones de cultivo fueron en anaerobiosis, y el tiempo de incubación de 24 horas. A partir del sobrenadante fueron determinadas las concentraciones de los ácidos grasos volátiles por cromatografía de gases. Se estudiaron dos relaciones bacteria-hongo: 1:1 y 1:3. Como resultado se observó un efecto negativo de Aspergillus oryzae sobre Butyrivibrio fibrisolvens, que se reflejó en la disminución en la producción de ácidos grasos volátiles.A study in co-culture between Aspergillus oryzae with the cellulolytic ruminal bacteria Butyrivibrio fibrisolvens was carried out aiming the "in vitro" determination of the effect of the fungi on the production of acetic, propionic and butyric acids by the bacteria. The culture medium was prepared using filtered, centrifuged, autoclaved and ruminal liquid diluted to 40% with water, and 0,05 % p/v of Angleton grass [;Dichamthium aristatum]; [;Córdoba, Colombia];. Culture was performed in anaerobic conditions for 24 hours. The concentrations of volatile fatty acids in the supernatant were determined by gas chromatography. Two bacteria-fungi relations were studied: 1:1 and 1:3. The results showed a negative effect of Aspergillus oryzae on Butyrivibrio fibrisolvens which was reflected in a decrease in the production of volatile fatty acids.

Molecular characterization of intergeneric hybrid between Aspergillus oryzae and Trichoderma harzianum by protoplast fusion.

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Patil, N S; Patil, S M; Govindwar, S P; Jadhav, J P

2015-02-01

Protoplast fusion between Aspergillus oryzae and Trichoderma harzianum and application of fusant in degradation of shellfish waste. The filamentous chitinolytic fungal strains A. oryzae NCIM 1272 and T. harzianum NCIM 1185 were selected as parents for protoplast fusion. Viable protoplasts were released from fungal mycelium using enzyme cocktail containing 5 mg ml(-1) lysing enzymes from T. harzianum, 0.06 mg ml(-1) β-glucuronidase from Helix pomatia and 1 mg ml(-1) purified Penicillium ochrochloron chitinase in 0.8 mol l(-1) sorbitol as an osmotic stabilizer. Intergeneric protoplast fusion was carried out using 60% polyethylene glycol as a fusogen. At optimum conditions, the regeneration frequency of the fused protoplasts on colloidal chitin medium and fusion frequency were calculated. Fusant showed higher rate of growth pattern, chitinase activity and protein content than parents. Fusant formation was confirmed by morphological markers, viz. colony morphology and spore size and denaturation gradient gel electrophoresis (DGGE). This study revealed protoplast fusion between A. oryzae and T. harzianum significantly enhanced chitinase activity which ultimately provides potential strain for degradation of shellfish waste. Consistency in the molecular characterization results using DGGE is the major outcome of this study which can be emerged as a fundamental step in fusant identification. Now it is need to provide attention over effective chitin degradation to manage shrimp processing issues. In this aspect, ability of fusant to degrade shellfish waste efficiently in short incubation time revealed discovery of potential strain in the reclamation of seafood processing crustacean bio-waste. © 2014 The Society for Applied Microbiology.

Recombinant expression, purification, and characterization of an acyl-CoA binding protein from Aspergillus oryzae.

Science.gov (United States)

Hao, Qing; Liu, Xiaoguang; Zhao, Guozhong; Jiang, Lu; Li, Ming; Zeng, Bin

2016-03-01

To characterize biochemically the lipid metabolism-regulating acyl-CoA binding protein (ACBP) from the industrially-important fungus Aspergillus oryzae. A full-length cDNA encoding a candidate ACBP from A. oryzae (AoACBP) was cloned and expressed in Escherichia coli as a maltose-binding protein (MBP) fusion protein. The MBP-AoACBP protein was purified by an amylose resin chromatography column. SDS-PAGE showed that MBP-AoACBP has an estimated molecular weight of 82 kDa. Microscale thermophoresis binding assay showed that the recombinant AoACBP displayed much greater affinity for palmitoyl-CoA (K d = 80 nM) than for myristoyl-CoA (K d = 510 nM), thus demonstrating the preference of AoACBP for long-chain acyl-CoA. The data support the identification of AoACBP as a long-chain ACBP in A. oryzae.

Secretory expression of the non-secretory-type Lentinula edodes laccase by Aspergillus oryzae.

Science.gov (United States)

Yano, Akira; Kikuchi, Sayaka; Nakagawa, Yuko; Sakamoto, Yuichi; Sato, Toshitsugu

2009-01-01

The shiitake mushroom, Lentinula edodes, has an extracelluar secretory-type laccase, Lcc1, and a fruiting-body-accumulation-type laccase, Lcc4. We previously reported the production of Lcc1 by plant cells, but had difficulty producing Lcc4. Here, we report the production of Lcc1 and Lcc4 by Aspergillus oryzae and the extracellular secretory production of Lcc4 using a modified secretion signal peptide (SP) from Lcc1. Sp-Lcc4 produced by A. oryzae had biochemical activities similar to Lcc4 produced by L. edodes. Lcc1 did not react with beta-(3,4-dihydroxyphenol) alanine (DOPA), but Lcc4 from L. edodes and A. oryzae could oxidize DOPA. K(M) values for the substrates 2,2'-azino-di-(3-ethylbenzthiazolinsulfonate), 2,6-dimethoxyphenol, guaiacol, pyrogallol, and catechol were similar for Lcc4 and Sp-Lcc4. In conclusion, a non-secretory-type fungal laccase is secreted into the culture media with its original enzymatic properties by exploiting modified secretory signal peptide. 2008 Elsevier GmbH.

Effect of low oxygen concentrations on growth and alpha-amylase production of Aspergillus oryzae in model solid-state fermentation systems

NARCIS (Netherlands)

Rahardjo, Y.S.P.; Sie, S.; Weber, F.J.; Tramper, J.; Rinzema, A.

2005-01-01

Oxygen transfer in the fungal mat is a major concern in solid-state fermentation (SSF). Oxygen supply into the mycelial layers is hampered by diffusion limitation. For aerobic fungi, like Aspergillus oryzae, this oxygen depletion can be a severely limiting factor for growth and metabolite

Subtractive cloning of cDNA from Aspergillus oryzae differentially regulated between solid-state culture and liquid (submerged) culture.

Science.gov (United States)

Akao, Takeshi; Gomi, Katsuya; Goto, Kuniyasu; Okazaki, Naoto; Akita, Osamu

2002-07-01

In solid-state cultures (SC), Aspergillus oryzae shows characteristics such as high-level production and secretion of enzymes and hyphal differentiation with asexual development which are absent in liquid (submerged) culture (LC). It was predicted that many of the genes involved in the characteristics of A. oryzae in SC are differentially expressed between SC and LC. We generated two subtracted cDNA libraries with bi-directional cDNA subtractive hybridizations to isolate and identify such genes. Among them, we identified genes upregulated in or specific to SC, such as the AOS ( A. oryzae SC-specific gene) series, and those downregulated or not expressed in SC, such as the AOL ( A. oryzae LC-specific) series. Sequencing analyses revealed that the AOS series and the AOL series contain genes encoding extra- and intracellular enzymes and transport proteins. However, half were functionally unclassified by nucleotide sequences. Also, by expression profile, the AOS series comprised two groups. These gene products' molecular functions and physiological roles in SC await further investigation.

Effect of selenium application on arsenic uptake in rice (Oryza sativa L.).

Science.gov (United States)

Kaur, Sumandeep; Singh, Dhanwinder; Singh, Kuldip

2017-09-01

Alluvial aquifers of the agrarian state of Punjab of southwestern arid zone used for irrigation of rice crops are rich in arsenic concentration. In the present study, rice (Oryza sativa L.) crops were raised in pots in a greenhouse with a purpose to study whether selenium (Se) application was effective in ameliorating As uptake. The rice crop was irrigated with arsenic laced water (0, 2.5, 5.0, 10.0 μM As L -1 ) throughout the growing period, without and with selenium (0.05 and 0.10 mg kg -1 ) added through mustard biomass, grown ex situ in seleniferous soil. Arsenic uptake and dry matter yield in different parts of the rice crop were assayed after application of As alone and simultaneous supplementations (As + Se). An antagonistic interaction between Se and As was observed. Addition of As through irrigation water significantly reduced yield of rice grain, straw and root. However, subsequent addition of Se helped in mitigating the harmful effect of As and countered the yield reduction caused due to As toxicity. The effect of Se on dry matter yield was more pronounced at its higher dose (0.10 mg kg -1 ) as compared to its lower dose (0.05 mg kg -1 ). The presence of Se either alone or added along with As significantly reduced the As concentration and its uptake by different parts of rice and higher reduction in As concentration was observed with addition of the highest level of applied Se (0.10 mg kg -1 ). Our observations indicated that Se supplementation might be favourable to reduce As accumulation and toxicity in rice crops.

Crystallization and diffraction analysis of β-N-acetylhexosaminidase from Aspergillus oryzae

International Nuclear Information System (INIS)

Vaněk, Ondřej; Brynda, Jiří; Hofbauerová, Kateřina; Kukačka, Zdeněk; Pachl, Petr; Bezouška, Karel; Řezáčová, Pavlína

2011-01-01

The fungal β-N-acetylhexosaminidase from A. oryzae was crystallized and diffraction data were collected from two crystal forms to 3.2 and 2.4 Å resolution, respectively. Fungal β-N-acetylhexosaminidases are enzymes that are used in the chemoenzymatic synthesis of biologically interesting oligosaccharides. The enzyme from Aspergillus oryzae was produced and purified from its natural source and crystallized using the hanging-drop vapour-diffusion method. Diffraction data from two crystal forms (primitive monoclinic and primitive tetragonal) were collected to resolutions of 3.2 and 2.4 Å, respectively. Electrophoretic and quantitative N-terminal protein-sequencing analyses confirmed that the crystals are formed by a complete biologically active enzyme consisting of a glycosylated catalytic unit and a noncovalently attached propeptide

KB425796-A, a novel antifungal antibiotic produced by Paenibacillus sp. 530603.

Science.gov (United States)

Kai, Hirohito; Yamashita, Midori; Takase, Shigehiro; Hashimoto, Michizane; Muramatsu, Hideyuki; Nakamura, Ikuko; Yoshikawa, Koji; Ezaki, Masami; Nitta, Kumiko; Watanabe, Masato; Inamura, Noriaki; Fujie, Akihiko

2013-08-01

The novel antifungal macrocyclic lipopeptidolactone, KB425796-A (1), was isolated from the fermentation broth of bacterial strain 530603, which was identified as a new Paenibacillus species based on morphological and physiological characteristics, and 16S rRNA sequences. KB425796-A (1) was isolated as white powder by solvent extraction, HP-20 and ODS-B column chromatography, and lyophilization, and was determined to have the molecular formula C79H115N19O18. KB425796-A (1) showed antifungal activities against Aspergillus fumigatus and the micafungin-resistant infectious fungi Trichosporon asahii, Rhizopus oryzae, Pseudallescheria boydii and Cryptococcus neoformans.

Protection of Grain Products from Sitophilus oryzae (L.) Contamination by Anti-Insect Pest Repellent Sachet Containing Allyl Mercaptan Microcapsule.

Science.gov (United States)

Chang, Yoonjee; Lee, Soo-Hyun; Na, Ja Hyun; Chang, Pahn-Shick; Han, Jaejoon

2017-11-01

The purpose of this study was to develop an anti-insect pest repellent sachet to prevent Sitophilus oryzae (L.) (Coleoptera: Curculionidae) contamination in grain packaging. The anti-insect pest activities of essential oils (EOs) from garlic (Allium Sativum), ginger (Zingiber Officinalis), black pepper (Piper nigrum), onion (Allium cepa), and fennel (Foeniculum vulgare) as well as major compounds (allyl disulfide, AD; allyl mercaptan, AM) isolated from of garlic and onion (AD and AM) were measured against S. oryzae. The results revealed that garlic EO, onion EO, AD, and AM showed strong fumigant insecticidal activities. Among these, AM showed the highest acetylcholinesterase (AChE) inhibition rate, indicating that the fumigation insecticidal efficacy of AM is related with its AChE inhibition ability. Subsequently, the microcapsules were produced with a high efficiency (80.02%) by using AM as a core material and rice flour as a wall material. Finally, sachet composed of rice flour microcapsule containing 2% AM (RAM) was produced. Repellent assay was performed to measure anti-insect pest ability of the RAM sachet, showed remarkable repelling effect within 48 h both in the presence or absence of attractant. In a release profile of RAM sachet, it was expected to last over 20 mo during the distribution period of brown rice. Moreover, RAM sachet showed no undesirable changes to the sensory properties of the rice both before and after cooking. Taken together, these results suggest that the newly developed RAM sachet could be used as a packaging material to protect grain products from S. oryzae contamination. The rice weevil, Sitophilus oryzae (L.) (Coleoptera: Curculionidae), causes damages to stored products and its contamination in grain products has become a major problem in cereal market. To preserve brown rice, an anti-insect pest repellent sachet containing 2% allyl mercaptan was newly developed and it showed remarkable repellent abilities against S. oryzae. It

Rim 2/Hipa CACTA transposon display ; A new genetic marker technique in Oryza species

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Lee Ju

2005-03-01

Full Text Available Abstract Background Transposons constitute the major fractions of repetitive sequences in eukaryotes, and have been crucial in the shaping of current genomes. Transposons are generally divided into two classes according to the mechanism underlying their transposition: RNA intermediate class 1 and DNA intermediate class 2. CACTA is a class 2 transposon superfamily, which is found exclusively in plants. As some transposons, including the CACTA superfamily, are highly abundant in plant species, and their nucleotide sequences are highly conserved within a family, they can be utilized as genetic markers, using a slightly modified version of the conventional AFLP protocol. Rim2 /Hipa is a CACTA transposon family having 16 bp consensus TIR sequences to be present in high copy numbers in rice genome. This research was carried out in order to develop a Rim2/Hipa CACTA-AFLP or Rim2/Hipa CACTA-TD (transposon display, hereafter Rim2/Hipa-TD protocol for the study of genetic markers in map construction and the study of genetic diversity in rice. Results Rim2/Hipa-TD generated ample polymorphic profiles among the different rice accessions, and the amplification profiles were highly reproducible between different thermocyclers and Taq polymerases. These amplification profiles allowed for clear distinction between two different ecotypes, Japonica and Indica, of Oryza sativa. In the analysis of RIL populations, the Rim2/Hipa-TD markers were found to be segregated largely in a dominant manner, although in a few cases, non-parental bands were observed in the segregating populations. Upon linkage analysis, the Rim2/Hipa-TD markers were found to be distributed in the regions proximal to the centromeres of the chromosomes. The distribution of the Rim2/Hipa CACTA elements was surveyed in 15 different Oryza species via Rim2/Hipa-TD. While Rim2/Hipa-TD yielded ample amplification profiles between 100 to 700 bp in the AA diploid Oryza species, other species having BB, CC

Analysis of nucleotide diversity among alleles of the major bacterial blight resistance gene Xa27 in cultivars of rice (Oryza sativa) and its wild relatives.

Science.gov (United States)

Bimolata, Waikhom; Kumar, Anirudh; Sundaram, Raman Meenakshi; Laha, Gouri Shankar; Qureshi, Insaf Ahmed; Reddy, Gajjala Ashok; Ghazi, Irfan Ahmad

2013-08-01

Xa27 is one of the important R-genes, effective against bacterial blight disease of rice caused by Xanthomonas oryzae pv. oryzae (Xoo). Using natural population of Oryza, we analyzed the sequence variation in the functionally important domains of Xa27 across the Oryza species. DNA sequences of Xa27 alleles from 27 rice accessions revealed higher nucleotide diversity among the reported R-genes of rice. Sequence polymorphism analysis revealed synonymous and non-synonymous mutations in addition to a number of InDels in non-coding regions of the gene. High sequence variation was observed in the promoter region including the 5'UTR with 'π' value 0.00916 and 'θ w ' = 0.01785. Comparative analysis of the identified Xa27 alleles with that of IRBB27 and IR24 indicated the operation of both positive selection (Ka/Ks > 1) and neutral selection (Ka/Ks ≈ 0). The genetic distances of alleles of the gene from Oryza nivara were nearer to IRBB27 as compared to IR24. We also found the presence of conserved and null UPT (upregulated by transcriptional activator) box in the isolated alleles. Considerable amino acid polymorphism was localized in the trans-membrane domain for which the functional significance is yet to be elucidated. However, the absence of functional UPT box in all the alleles except IRBB27 suggests the maintenance of single resistant allele throughout the natural population.

Co-Production of Fungal Biomass Derived Constituents and Ethanol from Citrus Wastes Free Sugars without Auxiliary Nutrients in Airlift Bioreactor.

Science.gov (United States)

Satari, Behzad; Karimi, Keikhosro; Taherzadeh, Mohammad J; Zamani, Akram

2016-02-26

The potential of two zygomycetes fungi, Mucor indicus and Rhizopus oryzae, in assimilating citrus waste free sugars (CWFS) and producing fungal chitosan, oil, and protein as well as ethanol was investigated. Extraction of free sugars from citrus waste can reduce its environmental impact by decreasing the possibility of wild microorganisms growth and formation of bad odors, a typical problem facing the citrus industries. A total sugar concentration of 25.1 g/L was obtained by water extraction of citrus waste at room temperature, used for fungal cultivation in shake flasks and airlift bioreactor with no additional nutrients. In shake flasks cultivations, the fungi were only able to assimilate glucose, while fructose remained almost intact. In contrast, the cultivation of M. indicus and R. oryzae in the four-liter airlift bioreactor resulted in the consumption of almost all sugars and production of 250 and 280 g fungal biomass per kg of consumed sugar, respectively. These biomasses correspondingly contained 40% and 51% protein and 9.8% and 4.4% oil. Furthermore, the fungal cell walls, obtained after removing the alkali soluble fraction of the fungi, contained 0.61 and 0.69 g chitin and chitosan per g of cell wall for M. indicus and R. oryzae, respectively. Moreover, the maximum ethanol yield of 36% and 18% was obtained from M. indicus and R. oryzae, respectively. Furthermore, that M. indicus grew as clump mycelia in the airlift bioreactor, while R. oryzae formed spherical suspended pellets, is a promising feature towards industrialization of the process.

Co-Production of Fungal Biomass Derived Constituents and Ethanol from Citrus Wastes Free Sugars without Auxiliary Nutrients in Airlift Bioreactor

Directory of Open Access Journals (Sweden)

Behzad Satari

2016-02-01

Full Text Available The potential of two zygomycetes fungi, Mucor indicus and Rhizopus oryzae, in assimilating citrus waste free sugars (CWFS and producing fungal chitosan, oil, and protein as well as ethanol was investigated. Extraction of free sugars from citrus waste can reduce its environmental impact by decreasing the possibility of wild microorganisms growth and formation of bad odors, a typical problem facing the citrus industries. A total sugar concentration of 25.1 g/L was obtained by water extraction of citrus waste at room temperature, used for fungal cultivation in shake flasks and airlift bioreactor with no additional nutrients. In shake flasks cultivations, the fungi were only able to assimilate glucose, while fructose remained almost intact. In contrast, the cultivation of M. indicus and R. oryzae in the four-liter airlift bioreactor resulted in the consumption of almost all sugars and production of 250 and 280 g fungal biomass per kg of consumed sugar, respectively. These biomasses correspondingly contained 40% and 51% protein and 9.8% and 4.4% oil. Furthermore, the fungal cell walls, obtained after removing the alkali soluble fraction of the fungi, contained 0.61 and 0.69 g chitin and chitosan per g of cell wall for M. indicus and R. oryzae, respectively. Moreover, the maximum ethanol yield of 36% and 18% was obtained from M. indicus and R. oryzae, respectively. Furthermore, that M. indicus grew as clump mycelia in the airlift bioreactor, while R. oryzae formed spherical suspended pellets, is a promising feature towards industrialization of the process.

Extent of Microbial Contamination of Refined and Unrefined Vegetable oils sold in South-west Nigeria

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Oluwafemi Flora

2018-04-01

Full Text Available Oils constitute a major source of plant-based protein. A major limitation to optimal oil consumption in sub-tropical region is fungal infestation and consequent mycotoxin contamination. Ten refined and eight unrefined vegetable oils were randomly purchase from open markets and screened for microbial contamination using standard microbial procedures. Twenty six fungi isolates were obtained from the vegetable oil samples, the isolates were identified as Aspergillus fumigatus (43.0%, Mucor (17.9%, Saccharomyces cerevisiae (10.7%, Aspergillus niger (7.1%, Aspergillus flavus (7.1%, Penicillium spp (7.1%, Aspergillus oryzae (3.6%, Mucor (17.9% and Rhizopus spp (3.6%. Five out of the ten refined vegetable oil samples had no fungal contamination. A. flavus and A. oryzae were absent in all the refined oil samples while A. niger was absent in all the unrefined oil samples. Isolation of mycotoxigenic fungi such as Aspergillus spp. is of vital importance in the food industry. Education and training of processors and consumers is recommended.

Tanggap Pertumbuhan dan Produksi Padi (Oryza sativa L.) Varietas Ciherang Terhadap Pemberian Pupuk Organik

OpenAIRE

Sari, Vira Irma

2011-01-01

VIRA IRMA SARI. Response of Growth and Production of Rice (Oryza sativa L.) Variety Ciherang to The Application of Organic Fertilizer. Scarcity of inorganic fertilizer and the negative effect caused by inorganic fertilizer continuing to make organic fertilizer as a solution to overcome these problems. The availability of organic fertilizers is expected that the farmers can get the optimal production and profit, because the organic fertilizer is cheaper and easy ...

Cyclopiazonic Acid Biosynthesis of Aspergillus flavus and Aspergillus oryzae

Science.gov (United States)

Cyclopiazonic acid (CPA) is an indole-tetramic acid neurotoxin produced by some of the same strains of A. flavus that produce aflatoxins and by some Aspergillus oryzae strains. Despite its discovery 40 years ago, few reviews of its toxicity and biosynthesis have been reported. This review examines w...

Endocytosis of a maltose permease is induced when amylolytic enzyme production is repressed in Aspergillus oryzae.

Science.gov (United States)

Hiramoto, Tetsuya; Tanaka, Mizuki; Ichikawa, Takanori; Matsuura, Yuka; Hasegawa-Shiro, Sachiko; Shintani, Takahiro; Gomi, Katsuya

2015-09-01

In the filamentous fungus Aspergillus oryzae, amylolytic enzyme production is induced by the presence of maltose. Previously, we identified a putative maltose permease (MalP) gene in the maltose-utilizing cluster of A. oryzae. malP disruption causes a significant decrease in α-amylase activity and maltose consumption, indicating that MalP is a maltose transporter required for amylolytic enzyme production in A. oryzae. Although the expression of amylase genes and malP is repressed by the presence of glucose, the effect of glucose on the abundance of functional MalP is unknown. In this study, we examined the effect of glucose and other carbon sources on the subcellular localization of green fluorescence protein (GFP)-tagged MalP. After glucose addition, GFP-MalP at the plasma membrane was internalized and delivered to the vacuole. This glucose-induced internalization of GFP-MalP was inhibited by treatment with latrunculin B, an inhibitor of actin polymerization. Furthermore, GFP-MalP internalization was inhibited by repressing the HECT ubiquitin ligase HulA (ortholog of yeast Rsp5). These results suggest that MalP is transported to the vacuole by endocytosis in the presence of glucose. Besides glucose, mannose and 2-deoxyglucose also induced the endocytosis of GFP-MalP and amylolytic enzyme production was inhibited by the addition of these sugars. However, neither the subcellular localization of GFP-MalP nor amylolytic enzyme production was influenced by the addition of xylose or 3-O-methylglucose. These results imply that MalP endocytosis is induced when amylolytic enzyme production is repressed. Copyright © 2015 Elsevier Inc. All rights reserved.

Biological Activity of Essential Oils from Leaves and Fruits of Pepper Tree (Schinus molle L. to Control Rice Weevil (Sitophilus oryzae L. Bioactividad de aceites esenciales de hojas y frutos del aguaribay (Schinus molle L. en el gorgojo del arroz (Sitophilus oryzae L.

Directory of Open Access Journals (Sweden)

Verónica Benzi

2009-06-01

Full Text Available Rice weevil (Sitophilus oryzae L. is a primary insect pest of stored grain. The development of resistance resulted in the application of synthetic insecticides. In recent years many plant essential oils have provided potential alternatives to currently used insect control agents. The Brazilian pepper tree (Schinus molle L. var. areira (L. DC. (Anacardiaceae has different biological properties such as insecticidal activity. In this study, repellent, fumigant activity, nutritional indices, and feeding deterrent action were evaluated on S. oryzae adults. Filter paper impregnation was used to test fumigant toxicity, whereas treated whole wheat was used to evaluate repellent activity and a flour disk bioassay was done to evaluate feeding deterrent action and nutritional index alteration. Leaf essential oils showed repellent effects at both concentrations (0.04 and 0.4% w/w, while fruit essential oils lacked repellent activity. Both plant oils altered nutritional indices. Fruit essential oils had a strong feeding deterrent action (62% while leaves had a slight effect (40.6%. With respect to fumigant activity, neither of the essential oils was found to be toxic.El gorgojo del arroz (Sitophilus oryzae. L. es un insecto-plaga de infestación primaria de granos. El uso de insecticidas sintéticos ha desarrollado fenómenos de resistencia. En los últimos años los aceites esenciales se presentan como una alternativa en el control de insectos-plaga. El aguaribay (Schinus molle L. var. areira (L. DC. (Anacardiaceae es una planta con diferentes propiedades biológicas entre las que se destacan el uso como insecticida. El objetivo de este estudio fue evaluar la actividad fumigante, repelente, los índices nutricionales y la actividad antialimentaria de los aceites esenciales de hojas y frutos de S. molle var. areira en adultos de S. oryzae. Para la actividad fumigante se utilizó la técnica de impregnación de papeles de filtro; para la actividad repelente

Transfer of bacterial blight resistance from Oryza meyeriana to O.Sativa L.by asymmetric somatic hybridization

Institute of Scientific and Technical Information of China (English)

ZHU Yongsheng; CHEN Baotang; YU Shunwu; ZHANG Duanpin; ZHANG Xueqin; YAN Qiusheng

2004-01-01

Asymmetric somatic hybrid plants were produced between cultivated rice (Oryza sativa L.) and wild species [O. Meyeriana (Zoll. etMor, exSteud.)] with high resistance to rice bacterial blight. X-ray-irradiated protoplasts of the wild species were used as donor and chemically fused with iodoacetamide-inactivated protoplasts of rice cv. 02428to produce hybrids. Seventy-two plants were regenerated from 623 calli based on metabolic complementation. The morphological characters of the plants closely resembled that of the rice. Simple sequence repeats were employed to identify their hybridity. Cytological analysis of root-tips revealed that their chromosome number varied in the range of 27-38. The somatic hybrids were inoculated with strains of Xanthamonas oryzae pv. Oryzae at adult growth stage and demonstrated the resistance to bacterial blight introgression from the O. Meyeriana.

The Population Structure of African Cultivated Rice Oryza glaberrima (Steud.)

DEFF Research Database (Denmark)

Semon, Mande; Nielsen, Rasmus; Jones, Monty P.

2005-01-01

Genome-wide linkage disequilibrium (LD) was investigated for 198 accessions of Oryza glaberrima using 93 nuclear microsatellite markers. Significantly elevated levels of LD were detected, even among distantly located markers. Free recombination among loci at the population genetic level was shown...

A reductive aminase from Aspergillus oryzae

OpenAIRE

Grogan, Gideon James; Aleku, Godwin; France, Scott; Man, Henry Wing-Hong; Mangas-Sanchez, Juan; Sharma, Mahima; Montgomery, Sarah L; Leipold, Friedemann; Hussain, Shahed; Turner, Nicholas

2017-01-01

Reductive amination is one of the most important methods for the synthesis of chiral amines. Here we report the discovery of an NADP(H)-dependent reductive aminase from Aspergillus oryzae (AspRedAm, Uniprot code Q2TW47) which can catalyse the reductive coupling of a broad set of carbonyl compounds with a variety of primary and secondary amines with up to >98% conversion and with up to >98% enantiomeric excess. In cases where both carbonyl and amine show high reactivity, it is possible to empl...

Effects of aqueous extract of Cinnamomum verum on growth of bread spoilage fungi

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Monir Doudi

2016-01-01

Full Text Available Food waste has been identified as a considerable problem and bread is the most wasted food. This study aimed to evaluate In-vitro anti-fungal activity of cinnamon extract on bread spoilage fungi and to determine its anti-fungal effect in the bread slices. At first, the MIC and MFC values of the extract were determined against Aspergillus niger, Aspergillus flavus, Aspergillus fumigatus, Penicillium chrysogenum, Penicillium notatum and Rhizopus oryzae. Then, Aspergillus sp was selected to assess antifungal activities of different doses of cinnamon extract in bread slices. Cinnamon extract at a dose of 64 mg/ml completely inhibited all standard and bread isolated fungi. This concentration of extract also inhibited Aspergillus growth on bread slices and delayed colony formation but adversely affected the sensory characteristics of bread. Cinnamon extract at 32 mg/ml not only delayed fungal growth, but also improved bread shelf life and delayed its staling. Moreover, 32mg/ml of extract did not adversely affect bread aroma, flavor and texture. However, sodium acetate inhibited the growth of Aspergillus sp but is not recommended for fungal control because it is considered as chemical. Therefore 32 mg/ml of extract is recommended for increasing the shelf-life of flat bread.

Expression of Aspergillus hemoglobin domain activities in Aspergillus oryzae grown on solid substrates improves growth rate and enzyme production

NARCIS (Netherlands)

Biesebeke, te R.; Boussier, A.; Biezen van, N.; Braaksma, M.; Hondel, van den C.A.M.J.J.; Vos, de W.M.; Punt, P.J.

2006-01-01

DNA fragments coding for hemoglobin domains (HBD) were isolated from Aspergillus oryzae and Aspergillus niger. The HBD activities were expressed in A. oryzae by introduction of HBD gene fragments under the control of the promoter of the constitutively expressed gpdA gene. In the transformants,

Lead Biosorption by Self-Immobilized Rhizopus nigricans Pellets in a Laboratory Scale Packed Bed Column: Mathematical Model and Experiment

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Adela Kogej

2010-01-01

Full Text Available The biosorption of lead ions from aqueous solution on a self-immobilized Rhizopus nigricans biomass has been studied. Experiments were performed in a laboratory scale packed bed column at different liquid flow rates and biosorbent bed heights. Recorded experimental breakthrough curves were compared to those predicted by a mathematical model, which was developed to simulate a packed bed biosorption process by a soft, self-immobilized fungal biosorbent. In the range of examined experimental conditions, the biomass characteristics such as pellet porosity and biosorption capacity substantially affected the predicted response curve. General correlations for the estimation of the intra-pellet effective diffusivity, the external mass transfer coefficient, as well as axial dispersion were successfully applied in this biological system with specific mechanical properties. Under the experimental conditions, mass transfer is controlled by the external film resistance, while the intra-pellet mass transfer resistance, as well as the effect of axial dispersion, can be neglected. A new parameter α, the fraction of active biomass, with an average value of α=0.7, was introduced to take into account the specific biomass characteristics, and consequently the observed non-ideal liquid flow through the bed of fungal pellets.

Purification, characterization and antiproliferative activity of l-asparaginase from Aspergillus oryzae CCT 3940 with no glutaminase activity

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Fernanda Furlan Gonçalves Dias

2016-09-01

Conclusions: The sensitivity of the cells lines to purified l-asparaginase from A. oryzae CCT 3940 appeared to be concentration dependent affording a more significant decrease in cell growth than that observed for the commercial l-asparaginase from Escherichia coli. The l-asparaginase from A. oryzae CCT 3940 has a high potential for pharmaceutical exploitation in the treatment of leukemia.

Identification of Pectin Degrading Enzymes Secreted by Xanthomonas oryzae pv. oryzae and Determination of Their Role in Virulence on Rice.

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Lavanya Tayi

Full Text Available Xanthomonas oryzae pv.oryzae (Xoo causes the serious bacterial blight disease of rice. Xoo secretes a repertoire of plant cell wall degrading enzymes (CWDEs like cellulases, xylanases, esterases etc., which act on various components of the rice cell wall. The major cellulases and xylanases secreted by Xoo have been identified and their role in virulence has been determined. In this study, we have identified some of the pectin degrading enzymes of Xoo and assessed their role in virulence. Bioinformatics analysis indicated the presence of four pectin homogalacturonan (HG degrading genes in the genome of Xoo. The four HG degrading genes include one polygalacturonase (pglA, one pectin methyl esterase (pmt and two pectate lyases (pel and pelL. There was no difference in the expression of pglA, pmt and pel genes by laboratory wild type Xoo strain (BXO43 grown in either nutrient rich PS medium or in plant mimic XOM2 medium whereas the expression of pelL gene was induced in XOM2 medium as indicated by qRT-PCR experiments. Gene disruption mutations were generated in each of these four genes. The polygalacturonase mutant pglA- was completely deficient in degrading the substrate Na-polygalacturonicacid (PGA. Strains carrying mutations in the pmt, pel and pelL genes were as efficient as wild type Xoo (BXO43 in cleaving PGA. These observations clearly indicate that PglA is the major pectin degrading enzyme produced by Xoo. The pectin methyl esterase, Pmt, is the pectin de-esterifying enzyme secreted by Xoo as evident from the enzymatic activity assay performed using pectin as the substrate. Mutations in the pglA, pmt, pel and pelL genes have minimal effects on virulence. This suggests that, as compared to cellulases and xylanases, the HG degrading enzymes may not have a major role in the pathogenicity of Xoo.

Identification of Pectin Degrading Enzymes Secreted by Xanthomonas oryzae pv. oryzae and Determination of Their Role in Virulence on Rice.

Science.gov (United States)

Tayi, Lavanya; Maku, Roshan V; Patel, Hitendra Kumar; Sonti, Ramesh V

2016-01-01

Xanthomonas oryzae pv.oryzae (Xoo) causes the serious bacterial blight disease of rice. Xoo secretes a repertoire of plant cell wall degrading enzymes (CWDEs) like cellulases, xylanases, esterases etc., which act on various components of the rice cell wall. The major cellulases and xylanases secreted by Xoo have been identified and their role in virulence has been determined. In this study, we have identified some of the pectin degrading enzymes of Xoo and assessed their role in virulence. Bioinformatics analysis indicated the presence of four pectin homogalacturonan (HG) degrading genes in the genome of Xoo. The four HG degrading genes include one polygalacturonase (pglA), one pectin methyl esterase (pmt) and two pectate lyases (pel and pelL). There was no difference in the expression of pglA, pmt and pel genes by laboratory wild type Xoo strain (BXO43) grown in either nutrient rich PS medium or in plant mimic XOM2 medium whereas the expression of pelL gene was induced in XOM2 medium as indicated by qRT-PCR experiments. Gene disruption mutations were generated in each of these four genes. The polygalacturonase mutant pglA- was completely deficient in degrading the substrate Na-polygalacturonicacid (PGA). Strains carrying mutations in the pmt, pel and pelL genes were as efficient as wild type Xoo (BXO43) in cleaving PGA. These observations clearly indicate that PglA is the major pectin degrading enzyme produced by Xoo. The pectin methyl esterase, Pmt, is the pectin de-esterifying enzyme secreted by Xoo as evident from the enzymatic activity assay performed using pectin as the substrate. Mutations in the pglA, pmt, pel and pelL genes have minimal effects on virulence. This suggests that, as compared to cellulases and xylanases, the HG degrading enzymes may not have a major role in the pathogenicity of Xoo.

Comparative Transcriptome Profiling of Rice Near-Isogenic Line Carrying Xa23 under Infection of Xanthomonas oryzae pv. oryzae.

Science.gov (United States)

Tariq, Rezwan; Wang, Chunlian; Qin, Tengfei; Xu, Feifei; Tang, Yongchao; Gao, Ying; Ji, Zhiyuan; Zhao, Kaijun

2018-03-02

Bacterial blight, caused by Xanthomonas oryzae pv. oryzae ( Xoo ), is an overwhelming disease in rice-growing regions worldwide. Our previous studies revealed that the executor R gene Xa23 confers broad-spectrum disease resistance to all naturally occurring biotypes of Xoo . In this study, comparative transcriptomic profiling of two near-isogenic lines (NILs), CBB23 (harboring Xa23 ) and JG30 (without Xa23 ), before and after infection of the Xoo strain, PXO99 A , was done by RNA sequencing, to identify genes associated with the resistance. After high throughput sequencing, 1645 differentially expressed genes (DEGs) were identified between CBB23 and JG30 at different time points. Gene Ontlogy (GO) analysis categorized the DEGs into biological process, molecular function, and cellular component. KEGG analysis categorized the DEGs into different pathways, and phenylpropanoid biosynthesis was the most prominent pathway, followed by biosynthesis of plant hormones, flavonoid biosynthesis, and glycolysis/gluconeogenesis. Further analysis led to the identification of differentially expressed transcription factors (TFs) and different kinase responsive genes in CBB23, than that in JG30. Besides TFs and kinase responsive genes, DEGs related to ethylene, jasmonic acid, and secondary metabolites were also identified in both genotypes after PXO99 A infection. The data of DEGs are a precious resource for further clarifying the network of Xa23 -mediated resistance.

Growth responses of NaCl stressed rice (Oryza sativa L.) plants ...

African Journals Online (AJOL)

GREGORY

2010-09-27

Sep 27, 2010 ... 3Department of Statistics, University of Sindh Jamshoro, Pakistan. 4Mityari Sugar Mills ... Key words: Oryza sativa L., seedling biomass, epidermal cells, proline content. ... Attempts to reduce the soil salinity, using mechanical.

Molecular and genetic characterization of OSH6 ( Oryza sativa ...

African Journals Online (AJOL)

Genetic studies of dissociation (Ds) insertion mutant rice plants indicated that ectopic expression of truncated OSH6 (Oryza sativa Homeobox 6) mRNA may be responsible for the mutant phenotype of knotted leaf formation at the peduncle. Additionally, ectopic expression of truncated OSH6 mRNA in the OSH6-Ds mutant ...

Analysis of the temporal variation of radiation balance components in arid rice (Oryza sativa L.) culture

International Nuclear Information System (INIS)

Prates, J.E.; Coelho, D.T.; Steinmetz, S.

1988-01-01

The time variation of measured radiation balance components in a cultived rice area (Oryza sativa L.) under arid conditions in the Brazil central-west region was analysed. The relation between global solar radiation, radiation balance, reflected radiation and terrestrial effective radiation in three different stages of the culture development: vegetative stage; blooming and maturation, was determined. (M.C.K.) [pt

Nuclear and original DNA application in Oryza taxonomy and phylogeny

International Nuclear Information System (INIS)

Romero, Gabriel O.

1998-01-01

Conventional taxonomy and phylogeny of germplasm are based on the tedious characterization of morphological variation. The ability to assay DNA variation that underlies morphological variation offers great promise as a convenient alternative for the genetic characterization of germplasm. Restriction fragment length polymorphism (RFLP) was used to survey DNA variation in 22 species of the genus Oryza. At the ribosomal DNA (rDNA) multigene family, 15 rDNA spacer length (sl) variants were identified using restriction enzyme Sst1 and wheatrDNA unit as probe. Particular sl variants predominated in certain isozyme groups of O. sativa, indicating a potential of sl ploymorphism in varietal classification. The distribution of sl variants supports the origin of O. sativa and O. nivara from O. rufipogon, and that O. spontanea arose from introgressions among O. sativa, O. nivara, and O. rufipogon. The distribution also suggests that the CCgenome, of all the genomes in the Officinalis complex, may be closest to the Sativa complex genomes, and it affirms the genetic position of the Officinalis complex intermediate between the Sativa and Ridleyi complexes. Variation at the Oryza organelle genomes was probed with a maize mitochondrial gene, atpA, a wheat chloroplast inverted repeat segment, p6. Results indicated that the complexes can be differentiated by their mitochondrial genome, but not their chloroplast genome when digested by Sst1 or BamH1. Therefore, the natural DNA variation in the nuclear and mitochondrial genomes has demonstrated great potential in complementing the conventional basis of taxa classification and phylogeny in the genus Oryza. (Author)

Jewelry boxes contaminated by Aspergillus oryzae: an occupational health risk?

Science.gov (United States)

Bellanger, Anne-Pauline; Roussel, Anaïs; Millon, Laurence; Delaforge, Marcel; Reboux, Gabriel

2012-01-01

In 2009, 100,000 jewelry boxes, manufactured in China, were delivered to a jewelry manufacturer in Besançon, France. All the boxes were contaminated by mold. Because the workers refused to handle these jewelry boxes, the company contacted our laboratory to determine how to deal with the problem. Three choices were available: (1) decontaminate the boxes, (2) return the boxes to the Chinese manufacturer, or (3) destroy the entire shipment. Based on microscopic identification, the culture analysis was positive for A. oryzae. This could not be confirmed by molecular techniques because of the genetic proximity of A. oryzae and A. flavus. Because A. flavus can produce aflatoxins, we tested for them using mass spectrometry. Aflatoxins B1, B2, G1, G2, and M1 were not detected; however, given the specifics of this situation, we could not discard the possibility of the presence of other aflatoxins, such as P1, B3, GM2, and ethoxyaflatoxin B2. We concluded that the contamination by A. oryzae was probably due to food products. However, because of the possible presence of aflatoxins, occupational health risks could not be entirely ruled out. The decision was therefore taken to destroy all the jewelry boxes by incineration. To avoid a similar situation we propose: (1) to maintain conditions limiting mold contamination during production (not eating on the work site, efficient ventilation systems); (2) to desiccate the products before sending them; and (3) to closely control the levels of dampness during storage and transport.

PENENTUAN PATOTIPE DAN KERAGAMAN GENETIK Xanthomonas oryzae pv. oryzae PADA TANAMAN PADI DI WILAYAH KARESIDENAN BANYUMAS

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Heru Adi Djatmiko

2011-11-01

Full Text Available One of the major diseases of rice paddy fields in Indonesia and the Asian countries is bacterial leaf blight or kresek caused by X. oryzae pv. oryzae (Xoo. Losses caused by the disease in Indonesia reached 70–80%, in India reached 74–81%, and Japan reached 20–50%, thus causing great losses in the economy. The objectives of that research were: 1 Characterize Xoo from Karesidenan Banyumas; 2 To study of the amount of damage and AUDPC (the area under disease progress curve of bacterial leaf blight disease at Karesidenan Banyumas; 3 To obtain of Xoo pathotype by using the test varieties; 4 To obtaining genetic diversity of Xoo that found in Banjarnegara, Purbalingga, Banyumas, Cilacap dan Kebumen region. Research was carried out in several stages: isolation and characterization of Xoo from Barlingmascakeb region, testing of Xoo with five varieties testing, assesment of disease intensity of bacterial leaf blight and AUDPC in the field, and testing the genetic diversity of Xoo. The results showed that pathogen of bacterial blight on rice is Xoo characterized yellow colour of colonies on SPA medium, negative gram reaction, catalase positive, oxidase negative, negative growth at 0.1% TZC, negative starch hydrolisis , and resistance to 0.001% Cu(NO32 positive. Xoo pathotype isolats found in Banjarnegara was pathotype X, Cilacap were pathotype I and II and Purbalingga was pathotype II. Eighteen of Xoo from Karesidenan Banyumas (Banjarnegara, Purbalingga, Banyumas, Cilacap and Kebumen of RAPD differ one from the others.

Discovery and analysis of an active long terminal repeat-retrotransposable element in Aspergillus oryzae.

Science.gov (United States)

Jie Jin, Feng; Hara, Seiichi; Sato, Atsushi; Koyama, Yasuji

2014-01-01

Wild-type Aspergillus oryzae RIB40 contains two copies of the AO090005001597 gene. We previously constructed A. oryzae RIB40 strain, RKuAF8B, with multiple chromosomal deletions, in which the AO090005001597 copy number was found to be increased significantly. Sequence analysis indicated that AO090005001597 is part of a putative 6,000-bp retrotransposable element, flanked by two long terminal repeats (LTRs) of 669 bp, with characteristics of retroviruses and retrotransposons, and thus designated AoLTR (A. oryzae LTR-retrotransposable element). AoLTR comprised putative reverse transcriptase, RNase H, and integrase domains. The deduced amino acid sequence alignment of AoLTR showed 94% overall identity with AFLAV, an A. flavus Tf1/sushi retrotransposon. Quantitative real-time RT-PCR showed that AoLTR gene expression was significantly increased in the RKuAF8B, in accordance with the increased copy number. Inverse PCR indicated that the full-length retrotransposable element was randomly integrated into multiple genomic locations. However, no obvious phenotypic changes were associated with the increased AoLTR gene copy number.

Detection of Alien Oryza punctata Kotschy Chromosomes in Rice, Oryza sativa L., by Genomic in situ Hybridization

OpenAIRE

Yasui, Hideshi; Nonomura, Ken-ichi; Iwata, Nobuo; 安井, 秀; 野々村, 賢一; 岩田, 伸夫

1997-01-01

Genomic in situ hybridization (GIS H) using total Oryza punctata Kotschy genomic DNA as a probe was applied to detect alien chromosomes transferred from O. punctata (W1514: 2n=2x=24: BB) to O. sativa Japonica cultivar, Nipponbare (2n=2x=24: AA). Only 12 chromosomes in the interspecific hybrids (2n=3x=36: AAB) between autotetraploid of O. sativa cultivar Nipponbare and a diploid strain of O. punctata (W1514) showed intense staining by FITC in mitotic metaphase spreads. Only one homologous pair...

Identification and characterization of filamentous fungi isolated from fermentation starters for Hong Qu glutinous rice wine brewing.

Science.gov (United States)

Lv, Xu-Cong; Huang, Zhi-Qing; Zhang, Wen; Rao, Ping-Fan; Ni, Li

2012-01-01

Hong Qu glutinous rice wine is one of the most popular traditional rice wines in China. Traditionally, this wine is brewed from glutinous rice with the addition of wine fermentation starters (Hong Qu (also called red yeast rice) and White Qu). The objective of this study was to investigate the variability of filamentous fungi associated with traditional fermentation starters through a traditional culture-dependent method and a molecular identification approach. In this study, forty-three filamentous fungi were separated by traditional culture-dependent means (macro- and microscopic characteristics) from 10 fermentation starters and classified into 16 different species based on morphological examination and the internal transcribed spacer (ITS) sequences analysis. It was observed that the genus Aspergillus had the highest number (14 isolates) of isolates followed by Rhizopus (11 isolates), Monascus (5 isolates) and Penicillium (4 isolates). The species R. oryzae, A. niger, A. flavus and M. purpureus were frequently found in wine starter samples, among which R. oryzae was the most frequent species. The enzyme-producing properties (glucoamylase, α-amylase and protease) of all fungal isolates from different starters were also evaluated. A. flavus, R. oryzae and M. purpureus were found to be better glucoamylase producers. A. flavus, R. oryzae and A.oryzae exhibited higher activity of α-amylase. A. flavus and A. oryzae had higher protease activity. However, some fungal isolates of the same species exhibited a significant variability in the production levels for all determined enzyme activity. This study is the first to identify filamentous fungi associated with the starter of Hong Qu glutinous rice wine using both traditional and molecular methods. The results enrich our knowledge of liquor-related micro-organisms, and can be used to promote the development of the traditional fermentation technology.

Caracterização agromorfológica interpopulacional em Oryza glumaepatula Interpopulational agro-morphological haracterization of Oryza glumaepatula

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Mariana Silva Rosa

2006-01-01

Full Text Available O gênero Oryza apresenta duas espécies cultivadas e 21 espécies silvestres, sendo quatro originárias da América do Sul e Central. Dentre essas, a única espécie diplóide é Oryza glumaepatula Steud., compatível em cruzamentos com a espécie cultivada O. sativa L. O objetivo deste trabalho foi caracterizar, por meio de caracteres agromorfológicos, oito populações de O. glumaepatula, coletadas em diferentes bacias hidrográficas brasileiras. O experimento foi realizado em casa de vegetação utilizando-se o delineamento em blocos ao acaso com oito tratamentos e seis repetições. Cada parcela foi constituída de quatro plantas, obtendo-se o total de 24 plantas por população. Foram avaliadas três características agronômicas e 18 características morfológicas. Os dados foram analisados utilizando-se estatísticas univariadas e multivariadas. Os resultados mostraram diferenças significativas entre populações para todos os caracteres avaliados, o que indica a grande variabilidade genética observada para todas as populações. Os resultados da análise de componentes principais foram similares aos da análise de agrupamento que classificou as populações em quatro grupos, sendo um grupo para cinco populações da Amazônia, um grupo para uma população do Rio Negro, e dois grupos, um para cada população originária do Rio Xingu e do Rio Paraguai, no Pantanal. A população com características agronômicas mais desejáveis, maior produção de sementes e maior número de perfilhos foi a PG-4, originária do Pantanal Matogrossense, seguida da população JA-4, do Rio Japurá.The genus Oryza contains two cultivated and 21 wild species, and four of these wild species originated from South and Central America. Among these, the only diploid species, compatible in crossing experiments with the cultivated O. sativa L. is O. glumaepatula Steud. The objective of this research was to characterize with morpho-agronomic traits eight

Rice diversity panels available through the genetic stocks oryza collection

Science.gov (United States)

The Genetic Stocks Oryza (GSOR) Collection was established in 2004 at the USDA-ARS, Dale Bumpers National Rice Research Center (DBNRRC) located in Stuttgart, AR. The mission of GSOR is to provide unique genetic resources to the rice research community for genetic and genomics related research. GSOR ...

Effect of gamma-irradiated sludge on the growth and yield of rice (Oryza sativa L. var. GR-3)

International Nuclear Information System (INIS)

Pandya, G.A.; Prakash, L.; Devasia, Preston; Modi, V.V.

1988-01-01

The effects of gamma-irradiated sludge on the growth and yield of rice (Oryza sativa L. var. GR-3) in pot cultures have been studied. Compared to plants grown only in soil, shoot length, root length, fresh weight, dry weight, total proteins, total soluble sugars, starch and chlorophyll content of plants grown in soil supplemented with unirradiated or gamma-irradiated sludge were found to be significantly increased. Irradiation of sludge significantly stimulated the linear growth of shoot and root systems as well as fresh and dry weights of plants, compared to those grown in soil containing unirradiated sludge. There was also an improvement in the grain yield (weight of seed) when plants were grown in soil supplemented with irradiated sludge. The results obtained suggest that the gamma-irradiated sewage sludge can be beneficially recycled for agricultural uses. (author)

In Vitro Activity of Isavuconazole and Comparators against Clinical Isolates of the Mucorales Order.

Science.gov (United States)

Arendrup, Maiken Cavling; Jensen, Rasmus Hare; Meletiadis, Joseph

2015-12-01

The in vitro activity of isavuconazole against Mucorales isolates measured by EUCAST E.Def 9.2 and CLSI M38-A2 methodologies was investigated in comparison with those of amphotericin B, posaconazole, and voriconazole. Seventy-two isolates were included: 12 of Lichtheimia corymbifera, 5 of Lichtheimia ramosa, 5 of group I and 9 of group II of Mucor circinelloides, 9 of Rhizomucor pusillus, 26 of Rhizopus microsporus, and 6 of Rhizopus oryzae. Species identification was confirmed by internal transcribed spacer (ITS) sequencing. EUCAST MICs were read on day 1 (EUCAST-d1) and day 2 (EUCAST-d2), and CLSI MICs were read on day 2 (CLSI-d2). Isavuconazole MIC50s (range) (mg/liter) by EUCAST-d1, CLSI-d2, and EUCAST-d2 were 1 (0.125 to 16), 1 (0.125 to 2), and 4 (0.5 to >16), respectively, across all isolates. The similar values for comparator drugs were as follows: posaconazole, 0.25 (≤ 0.03 to >16), 0.25 (0.06 to >16), and 1 (0.06 to >16); amphotericin, 0.06 (≤ 0.03 to 0.5), 0.06 (≤ 0.03 to 0.25), and 0.125 (≤ 0.03 to 1); voriconazole, 16 (2 to >16), 8 (1 to >16), and >16 (8 to >16), respectively. Isavuconazole activity varied by species: Lichtheimia corymbifera, 1 (0.5 to 2), 1 (1 to 2), and 2 (1 to 4); Lichtheimia ramosa, 0.25 (0.125 to 0.5), 1 (0.5 to 2), and 2 (0.5 to 4); Rhizomucor pusillus, 0.5 (0.5 to 1), 1 (0.125 to 1), and 2 (1 to 2); Rhizopus microsporus, 1 (0.5 to 4), 0.5 (0.125 to 1), and 4 (1 to 8); and Rhizopus oryzae, 1 (0.5 to 4), 1 (0.125 to 2), and 4 (0.5 to 8), respectively, were more susceptible than Mucor circinelloides: group I, 8 (4 to 8), 4 (2 to 4), and 16 (2 to 16), respectively, and group II, 8 (1 to 16), 8 (1 to 8), and 16 (4 to >16), respectively. This was also observed for posaconazole. The essential agreement was best between EUCAST-d1 and CLSI-d2 (75% to 83%). Isavuconazole displayed in vitro activity against Mucorales isolates with the exception of Mucor circinelloides. The MICs were in general 1 to 3 steps higher than those for

Feeding Aspergillus oryzae Fermentation Culture (AOFC to Growing Sheep: 1. The Effect of AOFC on Rumen Fermentation

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Darwinsyah Lubis

2002-10-01

Full Text Available Cultures of fungi, especially Aspergillus oryzae, have been of interest to animal nutritionists to increase feed efficiency. Many experiments have been done and showed positive results on rumen fermentation and productivity of ruminants. This paper reports the results of an in vivo study on feeding Aspergillus oryzae, fermentation culture (AOFC to growing sheep. ‘Onggok’ (tapioca processing waste was used as media for AO cultivation after being enriched with a mineral mixture. Commercial concentrate (GT-03 was fed to 15 growing sheep supplemented with 0% (C0, 5% (C1, and 10% (C2 AOFC (w/w. Chopped fresh King grass was used as a basal diet. The 3 treatments were randomly allotted to the sheep according to randomized block design with 5 replications. The study was carried out for 14 weeks. Digestion trial was conducted in the last 10 days of experiment. All feed and fecal samples were analyzed for nutrients. Rumen fluid was sampled at the mid experimental period. Analyses were done on rumen pH, ammonia content, (VFA volatile fatly acids concentration, and also total digestive tract digestibility of dry and organic matter, crude protein, and total fiber (NDF. Differences in treatment means were analyzed by Duncan’s MRT. Feeding AOFC resulted in increased (P<0.05 digestibility of crude protein from 59.6% in control sheep to 65.5% in sheep fed concentrate with 10% AOFC supplementation. The same pattern also occurred for NDF, but no effect was found on dry and organic matter. Higher fiber digestibility with AOFC supplementation was in line with an increase (P<0.05 in cellulolytic bacteria population in the rumen. VFA produced also increased (P<0.05, as well as individual acids content, primarily acetate and propionate. No differences (P<0.05 were detected in rumen pH and ammonia content. It appears that AOFC is more suitable for the purpose of meat production.

Purification, characterization and antimicrobial activity of chitinase from marine-derived Aspergillus terreus

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Aida M. Farag

2016-06-01

Full Text Available Chitinase (EC 3.2.1.14 was produced from the culture filtrate of marine-derived Aspergillus terreus and purified by 65% ammonium sulphate precipitation, followed by gel filtration on Sephadex G-100 and DEAE-Sephadex A-50 ion exchange chromatography, with 5.16-fold of purification and specific activity of 182.08 U/mg protein. The molecular weight of the purified chitinase was 60 kDa, determined by a sodium dodecyl sulphate polyacrylamide gel electrophoresis. The optimum pH and temperature of purified chitinase were 5.6 and 50 °C, respectively. The chitinase enzyme was stable from pH 5 to 7.5 and stable up to 70 °C. The effect of activators and inhibitors was studied, Hg+, pb, EDTA, ethanol, methanol and acetone strongly inhibited the enzyme activity, while, metal ions such as Ca2+, Mn2+ and Na2+ highly increased chitinase activity. The purified chitinase produced by A. terreus inhibited the growth of Aspergillus niger, Aspergillus oryzae, Penicillum oxysporium, Rhizocotonia solani, Candida albicans and Fusarium solani, while did not inhibit the growth of Rhizopus oryzae. Moreover, the purified enzyme had antibacterial effects against some pathogenic bacteria such as; Staphylococcus aureus, Salmonella typhi and Pseudomonas aeruginosa, while, it had not any activity against Escherichia coli, Aeromonas hydrophila and Photobacterium damsela.

AoS28D, a proline-Xaa carboxypeptidase secreted by Aspergillus oryzae.

Science.gov (United States)

Salamin, Karine; Eugster, Philippe J; Jousson, Olivier; Waridel, Patrice; Grouzmann, Eric; Monod, Michel

2017-05-01

Prolyl peptidases of the MEROPS S28 family are of particular interest because they are key enzymes in the digestion of proline-rich peptides. A BLAST analysis of the Aspergillus oryzae genome revealed sequences coding for four proteases of the S28 family. Three of these proteases, AoS28A, AoS28B, and AoS28C, were previously characterized as acidic prolyl endopeptidases. The fourth protease, AoS28D, showed high sequence divergence with other S28 proteases and belongs to a phylogenetically distinct cluster together with orthologous proteases from other Aspergillus species. The objective of the present paper was to characterize AoS28D protease in terms of substrate specificity and activity. AoS28D produced by gene overexpression in A. oryzae and in Pichia pastoris was a 70-kDa glycoprotein with a 10-kDa sugar moiety. In contrast with other S28 proteases, AoS28D did not hydrolyze internal Pro-Xaa bonds of several tested peptides. Similarly, to human lysosomal Pro-Xaa carboxypeptidase, AoS28D demonstrated selectivity for cleaving C-terminal Pro-Xaa bonds which are resistant to carboxypeptidases of the S10 family concomitantly secreted by A. oryzae. Therefore, AoS28D could act in synergy with these enzymes during sequential degradation of a peptide from its C-terminus.

Growth responses of NaCl stressed rice (Oryza sativa L.) plants ...

African Journals Online (AJOL)

GREGORY

2010-09-27

Sep 27, 2010 ... Growth responses of NaCl stressed rice (Oryza sativa. L.) plants ... 2008), which is a real threat to human's food security. Existed situation may ..... content and composition of essential oil and minerals in black cumin. (Nigella ...

Purification and properties of amylolytic enzyme from Aspergillus oryzae MIBA316

OpenAIRE

仮屋, 麻紀子; 矢野, めぐむ; 瀧井, 幸男; Makiko, Kariya; Megumu, Yano; Yukio, Takii

2003-01-01

Amylolytic enzyme was purified to electrophoretically homogeneous state from culture broth of Aspergillus oryzae MIBA316. This enzyme hydrolyzed preferentially amylopectin, starch and glycogen. Approaches to complete breakdown of starch to its components and their utilization in food processing were discussed.

Antimicrobial efficacy of secondary metabolites from Glomerella cingulata Eficiência antimicrobiana do extrato bruto de Glomerella cingulata

Directory of Open Access Journals (Sweden)

K. Hara Kishore

2007-03-01

Full Text Available Fungi are known to produce a vast array of secondary metabolites that are gaining importance for their biotechnological applications. Early reports suggest that G. cingulata has the capability to transform many compounds by various enzymatic actions. Therefore, the focus of this study was to determine the antibacterial and antifungal activity of crude ethyl acetate extract of G. cingulata using agar cup bioassay method. Crude extract of G. cingulata exhibited remarkable antifungal activity against Rhizopus oryzae, Chrysoporium tropicum and Beauveria bassiana but no antifungal activity was found against Alternaria tenuissima and Aspergillus niger at any concentrations. The crude extract presented no antibacterial activity against Gram positive and Gram negative bacteria at any concentration.Fungos são conhecidos produtores de uma vasta coleção de metabólitos secundários que vem mostrando importância crescente na sua aplicação biotecnológica. Publicações anteriores sugerem que G. cingulata tem a capacidade de transformar vários componentes por diferentes ações enzimáticas. Logo, o foco deste estudo foi determinar a atividade antibacteriana e antifúngica do extrato bruto de G. cingulata obtido por acetato de etila utilizando-se um método envolvendo bloco de agar. O extrato bruto de G. cingulata demonstrou marcante atividade antifungica contra Rhizopus oryzae, Chrysoporium tropicum e Beauveria bassiana entretanto, não foi possível detectar, em nenhuma concentração, atividade antifungica contra Alternaria tenuissima e Aspergillus niger. O mesmo extrato não apresentou atividade antibacteriana, em nenhuma concentração, contra bactérias Gram negativa e positiva.

Weissella oryzae DC6-facilitated green synthesis of silver nanoparticles and their antimicrobial potential.

Science.gov (United States)

Singh, Priyanka; Kim, Yeon J; Wang, Chao; Mathiyalagan, Ramya; Yang, Deok C

2016-09-01

Nanoparticles and nanomaterials are at the prominent edge of the rapidly developing field of nanotechnology. Recently, nanoparticle synthesis using biological resources has been found to be a new area with considerable prospects for development. Biological systems are the masters of ambient condition chemistry and are able to synthesize nanoparticles by utilizing metal salts. In the perspective of the current initiative to develop green technologies for the synthesis of nanoparticles, microorganisms are of considerable interest. Thus, the present study describes a bacterial strain-Weissella oryzae DC6-isolated from mountain ginseng, for the green and facile synthesis of silver nanoparticles. The particles were synthesized effectively without the need for any supplementary modification to maintain stability. The synthesized nanoparticles were evaluated by several instrumental techniques, comprising ultraviolet-visible spectrophotometry, field emission transmission electron microscopy, energy dispersive X-ray spectroscopy, elemental mapping, X-ray diffraction, and dynamic light scattering. In addition, the biosynthesized silver nanoparticles were explored for their antimicrobial activity against clinical pathogens including Vibrio parahaemolyticus, Bacillus cereus, Bacillus anthracis, Staphylococcus aureus, Escherichia coli, and Candida albicans. Furthermore, the potential of nanoparticles has been observed for biofilm inhibition against Staphylococcus aureus and Pseudomonas aeruginosa. Thus, the synthesis of silver nanoparticles by the strain W. oryzae DC6 may serve as a simple, green, cost-effective, consistent, and harmless method to produce antimicrobial silver nanoparticles.

Ectopic expression of a vesicle trafficking gene, OsRab7, from Oryza ...

African Journals Online (AJOL)

Jane

2011-07-18

Jul 18, 2011 ... OsRab7, from Oryza sativa, confers tolerance to several ... Key Laboratory of Molecular Biology and Gene Engineering, College of Life Science, Nanchang University, ... The reverse transcription was performed using MMLV.

Transposable elements as stress adaptive capacitors induce genomic instability in fungal pathogen Magnaporthe oryzae.

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Sonia Chadha

Full Text Available A fundamental problem in fungal pathogenesis is to elucidate the evolutionary forces responsible for genomic rearrangements leading to races with fitter genotypes. Understanding the adaptive evolutionary mechanisms requires identification of genomic components and environmental factors reshaping the genome of fungal pathogens to adapt. Herein, Magnaporthe oryzae, a model fungal plant pathogen is used to demonstrate the impact of environmental cues on transposable elements (TE based genome dynamics. For heat shock and copper stress exposed samples, eight TEs belonging to class I and II family were employed to obtain DNA profiles. Stress induced mutant bands showed a positive correlation with dose/duration of stress and provided evidences of TEs role in stress adaptiveness. Further, we demonstrate that genome dynamics differ for the type/family of TEs upon stress exposition and previous reports of stress induced MAGGY transposition has underestimated the role of TEs in M. oryzae. Here, we identified Pyret, MAGGY, Pot3, MINE, Mg-SINE, Grasshopper and MGLR3 as contributors of high genomic instability in M. oryzae in respective order. Sequencing of mutated bands led to the identification of LTR-retrotransposon sequences within regulatory regions of psuedogenes. DNA transposon Pot3 was identified in the coding regions of chromatin remodelling protein containing tyrosinase copper-binding and PWWP domains. LTR-retrotransposons Pyret and MAGGY are identified as key components responsible for the high genomic instability and perhaps these TEs are utilized by M. oryzae for its acclimatization to adverse environmental conditions. Our results demonstrate how common field stresses change genome dynamics of pathogen and provide perspective to explore the role of TEs in genome adaptability, signalling network and its impact on the virulence of fungal pathogens.

Enzymatic synthesis of novel oligosaccharides from N-acetylsucrosamine using β-fructofuranosidase from Aspergillus oryzae.

Science.gov (United States)

Nishio, Toshiyuki; Juami, Mai; Wada, Toru; Sugimoto, Yuta; Senou, Hiroki; Komori, Wataru; Sakuma, Chiseko; Hirano, Takako; Hakamata, Wataru; Tashiro, Mitsuru

2013-12-15

Mycelia of Aspergillus oryzae NBRC100959 contain 2 types of β-fructofuranosidases, transfructosylation-catalyzing enzyme (βFFaseI), and hydrolysis-catalyzing enzyme (βFFaseII). Using βFFaseI extracted from the mycelia of strain NBRC100959, two novel oligosaccharides consisting of GlcNAc and fructose, β-d-fructofuranosyl-(2→1)-β-d-fructofuranosyl-(2↔1)-2-acetamido-2-deoxy-α-d-glucopyranoside (N-acetyl-1-kestosamine, 1-KesNAc) and β-d-fructofuranosyl-(2→1)-β-d-fructofuranosyl-(2→1)-β-d-fructofuranosyl-(2↔1)-2-acetamido-2-deoxy-α-d-glucopyranoside (N-acetylnystosamine, NysNAc), were synthesized from β-d-fructofuranosyl-(2↔1)-2-acetamido-2-deoxy-α-d-glucopyranoside (N-acetylsucrosamine, SucNAc). We next planned to synthesize 1-KesNAc and NysNAc using A. oryzae mycelia. However, it was thought that the presence of βFFaseII is disadvantageous for the production of these oligosaccharides by βFFaseI, because βFFaseII hydrolyzed 1-KesNAc and NysNAc. We succeeded to produce A. oryzae mycelia containing βFFaseI as the major β-fructofuranosidase, by increasing sucrose concentration in the culture medium. Then, using a dried sample of these A. oryzae mycelia, reaction for the oligosaccharide production was performed. As the results, 190mg of 1-KesNAc and 60mg of NysNAc were obtained from 0.6g of SucNAc. This whole-cell catalysis method facilitates the synthesis of 1-KesNAc and NysNAc because extraction and purification of βFFaseI from mycelia are unnecessary. Copyright © 2013 Elsevier Ltd. All rights reserved.

Can rice (Oryza sativa) mitigate pesticides and nutrients in agricultural runoff?

Science.gov (United States)

Phytoremediation of nutrients and pesticides in runoff is a growing conservation effort, particularly in agriculturally intensive areas such as the lower Mississippi River Valley. In the current study, rice (Oryza sativa) was examined for its mitigation capacity of nitrogen, phosphorus, diazinon, a...

HrcQ is necessary for Xanthomonas oryzae pv. oryzae HR-induction in non-host tobacco and pathogenicity in host rice

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Xiaoping Zhang

2013-12-01

Full Text Available Bacterial blight, caused by Xanthomonas oryzae pv. oryzae (Xoo, is one of the most destructive diseases of rice (Oryza sativa L. worldwide. The type III secretion system (T3SS of Xoo, encoded by the hrp (hypersensitive response and pathogenicity genes, plays critical roles in conferring pathogenicity in host rice and triggering a hypersensitive response (HR in non-host plants. To investigate the major genes conferring the pathogenicity and avirulence of Xoo, we previously constructed a random Tn5-insertion mutant library of Xoo strain PXO99A. We report here the isolation and characterization of a Tn5-insertion mutant PXM69. Tn5-insertion mutants were screened on indica rice JG30, which is highly susceptible to PXO99A, by leaf-cutting inoculation. Four mutants with reduced virulence were obtained after two rounds of screening. Among them, the mutant PXM69 had completely lost virulence to the rice host and ability to elicit HR in non-host tobacco. Southern blotting analysis showed a single copy of a Tn5-insertion in the genome of PXM69. PCR walking and sequencing analysis revealed that the Tn5 transposon was inserted at nucleotide position 70,192–70,201 in the genome of PXO99A, disrupting the type III hrc (hrp-conserved gene hrcQ, the first gene in the D operon of the hrp cluster in Xoo. To confirm the relationship between the Tn5-insertion and the avirulence phenotype of PXM69, we used the marker exchange mutagenesis to create a PXO99A mutant, ΔhrcQ::KAN, in which the hrcQ was disrupted by a kanamycin-encoding gene cassette at the same site as that of the Tn5-insertion. ΔhrcQ::KAN showed the same phenotype as mutant PXM69. Reintroduction of the wild-type hrcQ gene partially complemented the pathogenic function of PXM69. RT-PCR and cellulase secretion assays showed that the Tn5-disruption of hrcQ did not affect transcription of downstream genes in the D operon and function of the type II secretion system. Our results provide new insights into

HrcQ is necessary for Xanthomonas oryzae pv. oryzae HR-induction in non-host tobacco and pathogenicity in host rice

Institute of Scientific and Technical Information of China (English)

Xiaoping; Zhang; Chunlian; Wang; Chongke; Zheng; Jinying; Che; Yanqiang; Li; Kaijun; Zhao

2013-01-01

Bacterial blight, caused by Xanthomonas oryzae pv. oryzae(Xoo), is one of the most destructive diseases of rice(Oryza sativa L.) worldwide. The type III secretion system(T3SS) of Xoo, encoded by the hrp(hypersensitive response and pathogenicity) genes, plays critical roles in conferring pathogenicity in host rice and triggering a hypersensitive response(HR) in non-host plants. To investigate the major genes conferring the pathogenicity and avirulence of Xoo, we previously constructed a random Tn5-insertion mutant library of Xoo strain PXO99A. We report here the isolation and characterization of a Tn5-insertion mutant PXM69. Tn5-insertion mutants were screened on indica rice JG30, which is highly susceptible to PXO99A, by leaf-cutting inoculation.Four mutants with reduced virulence were obtained after two rounds of screening. Among them, the mutant PXM69 had completely lost virulence to the rice host and ability to elicit HR in non-host tobacco. Southern blotting analysis showed a single copy of a Tn5-insertion in the genome of PXM69. PCR walking and sequencing analysis revealed that the Tn5 transposon was inserted at nucleotide position 70,192–70,201 in the genome of PXO99A, disrupting the type III hrc(hrp-conserved) gene hrcQ, the first gene in the D operon of the hrp cluster in Xoo. To confirm the relationship between the Tn5-insertion and the avirulence phenotype of PXM69, we used the marker exchange mutagenesis to create a PXO99Amutant, ΔhrcQ::KAN, in which the hrcQ was disrupted by a kanamycin-encoding gene cassette at the same site as that of the Tn5-insertion. ΔhrcQ::KAN showed the same phenotype as mutant PXM69. Reintroduction of the wild-type hrcQ gene partially complemented the pathogenic function of PXM69. RT-PCR and cellulase secretion assays showed that the Tn5-disruption of hrcQ did not affect transcription of downstream genes in the D operon and function of the type II secretion system. Our results provide new insights into the pathogenic

In silico Analysis of 3′-End-Processing Signals in Aspergillus oryzae Using Expressed Sequence Tags and Genomic Sequencing Data

Science.gov (United States)

Tanaka, Mizuki; Sakai, Yoshifumi; Yamada, Osamu; Shintani, Takahiro; Gomi, Katsuya

2011-01-01

To investigate 3′-end-processing signals in Aspergillus oryzae, we created a nucleotide sequence data set of the 3′-untranslated region (3′ UTR) plus 100 nucleotides (nt) sequence downstream of the poly(A) site using A. oryzae expressed sequence tags and genomic sequencing data. This data set comprised 1065 sequences derived from 1042 unique genes. The average 3′ UTR length in A. oryzae was 241 nt, which is greater than that in yeast but similar to that in plants. The 3′ UTR and 100 nt sequence downstream of the poly(A) site is notably U-rich, while the region located 15–30 nt upstream of the poly(A) site is markedly A-rich. The most frequently found hexanucleotide in this A-rich region is AAUGAA, although this sequence accounts for only 6% of all transcripts. These data suggested that A. oryzae has no highly conserved sequence element equivalent to AAUAAA, a mammalian polyadenylation signal. We identified that putative 3′-end-processing signals in A. oryzae, while less well conserved than those in mammals, comprised four sequence elements: the furthest upstream U-rich element, A-rich sequence, cleavage site, and downstream U-rich element flanking the cleavage site. Although these putative 3′-end-processing signals are similar to those in yeast and plants, some notable differences exist between them. PMID:21586533

Development and validation of cross-transferable and polymorphic DNA markers for detecting alien genome introgression in Oryza sativa from Oryza brachyantha.

Science.gov (United States)

Ray, Soham; Bose, Lotan K; Ray, Joshitha; Ngangkham, Umakanta; Katara, Jawahar L; Samantaray, Sanghamitra; Behera, Lambodar; Anumalla, Mahender; Singh, Onkar N; Chen, Meingsheng; Wing, Rod A; Mohapatra, Trilochan

2016-08-01

African wild rice Oryza brachyantha (FF), a distant relative of cultivated rice Oryza sativa (AA), carries genes for pests and disease resistance. Molecular marker assisted alien gene introgression from this wild species to its domesticated counterpart is largely impeded due to the scarce availability of cross-transferable and polymorphic molecular markers that can clearly distinguish these two species. Availability of the whole genome sequence (WGS) of both the species provides a unique opportunity to develop markers, which are cross-transferable. We observed poor cross-transferability (~0.75 %) of O. sativa specific sequence tagged microsatellite (STMS) markers to O. brachyantha. By utilizing the genome sequence information, we developed a set of 45 low cost PCR based co-dominant polymorphic markers (STS and CAPS). These markers were found cross-transferrable (84.78 %) between the two species and could distinguish them from each other and thus allowed tracing alien genome introgression. Finally, we validated a Monosomic Alien Addition Line (MAAL) carrying chromosome 1 of O. brachyantha in O. sativa background using these markers, as a proof of concept. Hence, in this study, we have identified a set molecular marker (comprising of STMS, STS and CAPS) that are capable of detecting alien genome introgression from O. brachyantha to O. sativa.

Mutation induced enhanced biosynthesis of lipases by Rhizopus oligosporus var. microsporus

International Nuclear Information System (INIS)

Iftikhar, T.; Ikram-ul-Haq; Niaz, M.; Abbas, S.Q.; Zia, M.A.; Ashraf, I.; Lee, K.J.

2010-01-01

The present study describes the isolation, identification and screening of fugal strain Rhizopus oligosporus (var. microsporus) for the production of extracellular lipases. One hundred and sixty seven cultures of fungi were isolated from different environments such as soil, air, milk, pickle, oily bread, decayed fruits and vegetables by serial dilution method. The strains were initially selected qualitatively on Tween 80-Agar plates and were shifted to the slants of PDA for maintenance and storage at 4 deg. C. Quantitative screening for extracellular lipase production by isolated strains was carried out in shake flasks and the most potent strain producing 3.20 +- 0.003 U mL/sup -1/ of enzyme was selected. The strain was then identified on the basis of standard morphological measurements and was assigned the code IIB-63. The selected strain was then subjected to physical (UV and Gamma radiations) and chemical mutagenic (MNNG/NTG, NA, EtBr) treatments in order to improve its lipolytic potential. During the treatment, mutants were qualitatively and quantitatively selected and IIB-63 NTG-7 was found to be the mutant showing highest lipases production (10.37 +- 0.06a U mL/sup -1/) with a zone size of 12.3 mm on Luria-Bertani-tributyrin agar plates. This mutant showed an overall 325% increase in activity over its parent strain for the production of extracellular lipase. (author)

Cloning and heterologous expression of the antibiotic peptide (ABP) genes from Rhizopus oligosporus NBRC 8631.

Science.gov (United States)

Yamada, Osamu; Sakamoto, Kazutoshi; Tominaga, Mihoko; Nakayama, Tasuku; Koseki, Takuya; Fujita, Akiko; Akita, Osamu

2005-03-01

We carried out protein sequencing of purified Antibiotic Peptide (ABP), and cloned two genes encoding this peptide as abp1 and abp2, from Rhizopus oligosporus NBRC 8631. Both genes contain an almost identical 231-bp segment, with only 3 nucleotide substitutions, encoding a 77 amino acid peptide. The abp gene product comprises a 28 amino acid signal sequence and a 49 amino acid mature peptide. Northern blot analysis showed that at least one of the abp genes is transcribed in R. oligosporus NBRC 8631. A truncated form of abp1 encoding only the mature peptide was fused with the alpha-factor signal peptide and engineered for expression in Pichia pastoris SMD1168H. Culture broth of the recombinant Pichia displayed ABP activity against Bacillus subtilis NBRC 3335 after induction of heterologous gene expression. This result indicates that mature ABP formed the active structure without the aid of other factors from R. oligosporus, and was secreted.

Metabolomic Profiles of Aspergillus oryzae and Bacillus amyloliquefaciens During Rice Koji Fermentation

Directory of Open Access Journals (Sweden)

Da Eun Lee

2016-06-01

Full Text Available Rice koji, used early in the manufacturing process for many fermented foods, produces diverse metabolites and enzymes during fermentation. Using gas chromatography time-of-flight mass spectrometry (GC-TOF-MS, ultrahigh-performance liquid chromatography linear trap quadrupole ion trap tandem mass spectrometry (UHPLC-LTQ-IT-MS/MS, and multivariate analysis we generated the metabolite profiles of rice koji produced by fermentation with Aspergillus oryzae (RK_AO or Bacillus amyloliquefaciens (RK_BA for different durations. Two principal components of the metabolomic data distinguished the rice koji samples according to their fermenter species and fermentation time. Several enzymes secreted by the fermenter species, including α-amylase, protease, and β-glucosidase, were assayed to identify differences in expression levels. This approach revealed that carbohydrate metabolism, serine-derived amino acids, and fatty acids were associated with rice koji fermentation by A. oryzae, whereas aromatic and branched chain amino acids, flavonoids, and lysophospholipids were more typical in rice koji fermentation by B. amyloliquefaciens. Antioxidant activity was significantly higher for RK_BA than for RK_AO, as were the abundances of flavonoids, including tricin, tricin glycosides, apigenin glycosides, and chrysoeriol glycosides. In summary, we have used MS-based metabolomics and enzyme activity assays to evaluate the effects of using different microbial species and fermentation times on the nutritional profile of rice koji.

ALOGAMIA EM ARROZ (Oryza sativa L. E RELAÇÃO COM CARACTERÍSTICAS AGRONÔMICAS RICE (Oryza sativa L. ALLOGAMY AND RELATIONSHIP WITH AGRONOMIC TRAITS

Directory of Open Access Journals (Sweden)

Péricles de Carvalho Ferreira Neves

2007-09-01

Full Text Available

A produção de sementes híbridas de arroz, utilizando a técnica chinesa, requer uma grande quantidade de mão-de-obra e é cara. Alternativas para aumentar a taxa de polinização cruzada podem ajudar a reduzir tal custo. A Embrapa Arroz e Feijão desenvolve um projeto de produção de híbridos através da transferência de caracteres alogâmicos (comprimento de estigma e antera da Oryza longistaminata A. Chev. para a cultivada Oryza sativa L. O objetivo deste estudo foi determinar as correlações genética e fenotípica entre esses caracteres e os agronômicos. O. longistaminata foi cruzada com a O. sativa e retrocruzada duas vezes para a cultivada. Foram obtidas 25 linhagens F3:6 e estudadas as correlações entre os caracteres alogâmicos (comprimento de estigma, antera e espigueta e os agronômicos (comprimento de estigma, antera, espigueta, arista e panícula, esterilidade, degrane, altura de planta, perfilhos por planta, e exserção de panícula. O desenho experimental utilizado foi o de blocos ao acaso com quatro repetições. Os ensaios foram conduzidos em dois locais dentro da área experimental da Embrapa Arroz e Feijão. Em geral, foram observadas pequenas correlações genética e fenotípica entre caracteres alogâmicos e agronômicos. Elevada e significativa associação foi observada entre o comprimento de estigma e o de anteras, o comprimento de estigma e o de arista, o comprimento de antera e o de arista, e o comprimento de panícula e a altura de planta.

PALAVRAS-CHAVE: Oryza longistaminata; taxa de autopolinização; arroz híbrido; produção de sementes.

Variation of photosynthetic tolerance of rice cultivars (Oryza sativa L ...

African Journals Online (AJOL)

STORAGESEVER

2010-03-01

Mar 1, 2010 ... (Oryza sativa L.) to chilling temperature in the light. Xia Li*, Kun Cao, Chao .... 2 mol) formed red-brown trimethine that can be detected quantitatively with spectrophoto- ..... ses through the generation of appropriate signals (H2O2) and the balance ..... was under weak light intensity (Murata, 1989). Light.

Occurrence of fungus Rhizopus sp in bioassays with Allium cepa germinated in the presence of uranium to study the effect of natural radioactivity

Energy Technology Data Exchange (ETDEWEB)

Xavier, Magno N.; Oliveira, Andressa L.; Maffei, Eliane M.D.; Campos, Simara S., E-mail: simaracampos@gmail.com [Universidade Estadual do Sudoeste da Bahia (UESB), Vitoria da Conquista, BA (Brazil). Departamento de Ciencias Exatas e Tecnologicas; Gennari, Roseli F. [Universidade de Sao Paulo (USP), SP (Brazil). Instituto de Fisica. Departamento de Fisica Nuclear

2015-07-01

The demographic and global economic growth has promoted increasing power consumption. In this context, several studies point to nuclear energy as being promising to meet such demand. Although Brazil composes the seventh position in the world ranking of uranium reserves, the ore is still little explored in Brazil, considering its vast existing arsenal. On the one hand, despite nuclear energy has brought great benefits, technological and socio-economic development, it generates controversy about environmental contamination and risks to public health. Studies on this subject indicate that areas where uranium ore concentration is high, natural environmental radiation exposure levels are already higher than in other regions. The aim of this study is to observe the simple germination of the bio-indicator (Allium cepa), typically used to assess potential chromosomal aberrations, suffer any adverse effect caused by natural radiation uranium. The choice of this bio-indicator is based on its potential for evaluating the mutation caused by countless chemical compounds. Four treatments with three replicates were designed. In each treatment, 10 seeds of onion A. cepa without any pesticides were packed in the Petri dish lined with germination paper and room temperature (25°C) was kept until the root reaches approximately 1cm long. As a result, growth of the fungus Rhizopus sp was observed, in the experiments where uranium ore was added. It is important to mention according to the literature, this fungus can cause serious infections (and often fatal) in humans and animals, due to its high growth rate and also to their ability on surviving in relatively high temperatures. (author)

Occurrence of fungus Rhizopus sp in bioassays with Allium cepa germinated in the presence of uranium to study the effect of natural radioactivity

International Nuclear Information System (INIS)

Xavier, Magno N.; Oliveira, Andressa L.; Maffei, Eliane M.D.; Campos, Simara S.; Gennari, Roseli F.

2015-01-01

The demographic and global economic growth has promoted increasing power consumption. In this context, several studies point to nuclear energy as being promising to meet such demand. Although Brazil composes the seventh position in the world ranking of uranium reserves, the ore is still little explored in Brazil, considering its vast existing arsenal. On the one hand, despite nuclear energy has brought great benefits, technological and socio-economic development, it generates controversy about environmental contamination and risks to public health. Studies on this subject indicate that areas where uranium ore concentration is high, natural environmental radiation exposure levels are already higher than in other regions. The aim of this study is to observe the simple germination of the bio-indicator (Allium cepa), typically used to assess potential chromosomal aberrations, suffer any adverse effect caused by natural radiation uranium. The choice of this bio-indicator is based on its potential for evaluating the mutation caused by countless chemical compounds. Four treatments with three replicates were designed. In each treatment, 10 seeds of onion A. cepa without any pesticides were packed in the Petri dish lined with germination paper and room temperature (25°C) was kept until the root reaches approximately 1cm long. As a result, growth of the fungus Rhizopus sp was observed, in the experiments where uranium ore was added. It is important to mention according to the literature, this fungus can cause serious infections (and often fatal) in humans and animals, due to its high growth rate and also to their ability on surviving in relatively high temperatures. (author)

Genome-scale analysis of the high-efficient protein secretion system of Aspergillus oryzae

DEFF Research Database (Denmark)

Liu, Lifang; Feizi, Amir; Osterlund, Tobias

2014-01-01

related fungal species such as Aspergillus nidulans and Aspergillus niger. To evaluate the defined component list, we performed transcriptome analysis on three a-amylase over-producing strains with varying levels of secretion capacities. Specifically, secretory components involved in the ER......Background: The koji mold, Aspergillus oryzae is widely used for the production of industrial enzymes due to its particularly high protein secretion capacity and ability to perform post-translational modifications. However, systemic analysis of its secretion system is lacking, generally due...... to the poorly annotated proteome. Results: Here we defined a functional protein secretory component list of A. oryzae using a previously reported secretory model of S. cerevisiae as scaffold. Additional secretory components were obtained by blast search with the functional components reported in other closely...

Genomics reveals traces of fungal phenylpropanoid-flavonoid metabolic pathway in the f ilamentous fungus Aspergillus oryzae.

Science.gov (United States)

Juvvadi, Praveen Rao; Seshime, Yasuyo; Kitamoto, Katsuhiko

2005-12-01

Fungal secondary metabolites constitute a wide variety of compounds which either play a vital role in agricultural, pharmaceutical and industrial contexts, or have devastating effects on agriculture, animal and human affairs by virtue of their toxigenicity. Owing to their beneficial and deleterious characteristics, these complex compounds and the genes responsible for their synthesis have been the subjects of extensive investigation by microbiologists and pharmacologists. A majority of the fungal secondary metabolic genes are classified as type I polyketide synthases (PKS) which are often clustered with other secondary metabolism related genes. In this review we discuss on the significance of our recent discovery of chalcone synthase (CHS) genes belonging to the type III PKS superfamily in an industrially important fungus, Aspergillus oryzae. CHS genes are known to play a vital role in the biosynthesis of flavonoids in plants. A comparative genome analyses revealed the unique character of A. oryzae with four CHS-like genes (csyA, csyB, csyC and csyD) amongst other Aspergilli (Aspergillus nidulans and Aspergillus fumigatus) which contained none of the CHS-like genes. Some other fungi such as Neurospora crassa, Fusarium graminearum, Magnaporthe grisea, Podospora anserina and Phanerochaete chrysosporium also contained putative type III PKSs, with a phylogenic distinction from bacteria and plants. The enzymatically active nature of these newly discovered homologues is expected owing to the conservation in the catalytic residues across the different species of plants and fungi, and also by the fact that a majority of these genes (csyA, csyB and csyD) were expressed in A. oryzae. While this finding brings filamentous fungi closer to plants and bacteria which until recently were the only ones considered to possess the type III PKSs, the presence of putative genes encoding other principal enzymes involved in the phenylpropanoid and flavonoid biosynthesis (viz

Differences in the Sensitivity to UVB Radiation of Two Cultivars of Rice (Oryza sativa L.)

OpenAIRE

Jun, Hidema; Hye-Sook, Kang; Tadashi, Kumagai; Institute of Genetic Ecology, Tohoku University; Institute of Genetic Ecology, Tohoku University; Institute of Genetic Ecology, Tohoku University

1996-01-01

The effects of UVB radiation on the growth of two cultivars of Japanese lowland rice (Oryza sativa L.), Sasanishiki and Norin 1, were examined in a phytotron. Supplementation of visible radiation with UVB radiation reduced plant length, tiller number, the fresh and dry weights of the aboveground parts of plants, and the amounts of total leaf nitrogen, chlorophyll, soluble protein and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) in the eighth leaf, the youngest fully expanded leaf...

Effect of coated urea on cadmium accumulation in Oryza sativa L. grown in contaminated soil.

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Xu, Chao; Wu, Zisong; Zhu, Qihong; Zhu, Hanhua; Zhang, Yangzhu; Huang, Daoyou

2015-11-01

Experiments were conducted to determine the effects of three types of coated urea on the accumulation of cadmium (Cd) in rice (Oryza sativa L.) grown in contaminated soil. Pot-culture experiments were conducted in a greenhouse from July to November 2012 on the rice cultivar "Hua Hang Si Miao" in Guangzhou (China). The experimental design was completely randomized with four treatments and three replications. The treatments were control (CK) (N 0 mg/kg), prilled urea (PU) (N 200 mg/kg), polymer-coated urea (PCU) (N 200 mg/kg), and sulfur-coated urea (SCU) (N 200 mg/kg). Our results indicated that applications of PCU and SCU slightly increased the dry weight of rice grains. The application of SCU significantly decreased the CaCl2 and toxicity characteristic leaching procedure (TCLP)-extractable Cd concentrations by 15.4 and 56.1%, respectively. Sequential extractions showed that PCU and SCU applications led to a significant decrease in Cd in the exchangeable fraction and an increase in the bound iron (Fe) and manganese (Mn) oxides fractions. Cd concentrations in grains treated with PCU were reduced by 11.7%, whereas SCU significantly reduced Cd concentrations by 29.1%. SCU reduced Cd transfer from the straws to the grain. Our results demonstrated that PCU and SCU may be effective in mitigating Cd accumulation in rice grown in acidic Cd-contaminated soil, especially in plants receiving SCU.

Comprehensive Secondary Metabolite Profiling Toward Delineating the Solid and Submerged-State Fermentation of Aspergillus oryzae KCCM 12698

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Su Y. Son

2018-05-01

Full Text Available Aspergillus oryzae has been commonly used to make koji, meju, and soy sauce in traditional food fermentation industries. However, the metabolic behaviors of A. oryzae during fermentation in various culture environments are largely uncharacterized. Thus, we performed time resolved (0, 4, 8, 12, 16 day secondary metabolite profiling for A. oryzae KCCM 12698 cultivated on malt extract agar and broth (MEA and MEB under solid-state fermentation (SSF and submerged fermentation (SmF conditions using the ultrahigh performance liquid chromatography-linear trap quadrupole-ion trap-mass spectrometry (UHPLC-LTQ-IT-MS/MS followed by multivariate analyses. We observed the relatively higher proportions of coumarins and oxylipins in SSF, whereas the terpenoids were abundant in SmF. Moreover, we investigated the antimicrobial efficacy of metabolites that were extracted from SSF and SmF. The SSF extracts showed higher antimicrobial activities as compared to SmF, with higher production rates of bioactive secondary metabolites viz., ketone-citreoisocoumarin, pentahydroxy-anthraquinone, hexylitaconic acid, oxylipins, and saturated fatty acids. The current study provides the underpinnings of a metabolomic framework regarding the growth and bioactive compound production for A. oryzae under the primarily employed industrial cultivation states. Furthermore, the study holds the potentials for rapid screening and MS-characterization of metabolites helpful in determining the consumer safety implications of fermented foods involving Koji mold.

Effectiveness of Aspergillus oryzae fermentation culture to improve digestion of fibrous feeds

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Darwinsyah Lubis

2002-06-01

Full Text Available Studies regarding the use of living microorganisms as supplement have been done in the last two decades to improve digestion process in the rumen. Many factors affect the use of the supplement, such as ration composition and physiological status of the animals. Materials used in this experiment are Aspergillus oryzae (AO derived from ‘tauco’ (salty fermented soybean and soybean sauce processing factories located in Bogor, Cianjur, and Sukabumi districts (15 sources, and also AO culture collections of the Indonesian Res. Inst. for Anim. Prod. (2 strains and one strain from Indonesian Res. Inst. for Vet. Sci. in Bogor. The fungus was first isolated in potato dextrose agar (PDA, incubated at room temp. (26-300C for 5 days for purification. The AO from PDA then inoculated into cooked rice enriched with mineral mix, oven-dried (40-450C, then ground and kept in refrigerator as a stock culture for further use. To produce more AO (scaling-up, the culture is inoculated in two basal media, i.e. soybean meal and ‘onggok’ (tapioca processing waste enriched with mineral mix, dried and ground, then kept in refrigerator as A. oryzae fermentation culture (AOFC. Ground-dried King grass was used for in vitro digestion trials using sheep rumen fluid, which was not supplemented (control or supplemented with the AOFC (10% w/w from various sources. Three best AOFC (by origin were chosen and used for further digestion study. The study was run using 2 (media x 3 (AOFC origin factorial experiment based on a completely randomized design and Duncan’s MRT was applied to test differences among treatment means. Preliminary results indicated of the AOFC-SP66, -F172, and -CT4 used, the best AOFC was SP66, as it increased (P<0.05 fiber (NDF digestion (10.5% better than the control. Total VFA productions were similar, but acetate content in the rumen fluid was lowered (P<0.05, while propionate and butyrate levels were alleviated (P<0.05 by the AOFC-SP66. Ammonia

A rare sugar, d-allose, confers resistance to rice bacterial blight with upregulation of defense-related genes in Oryza sativa.

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Kano, Akihito; Gomi, Kenji; Yamasaki-Kokudo, Yumiko; Satoh, Masaru; Fukumoto, Takeshi; Ohtani, Kouhei; Tajima, Shigeyuki; Izumori, Ken; Tanaka, Keiji; Ishida, Yutaka; Tada, Yasuomi; Nishizawa, Yoko; Akimitsu, Kazuya

2010-01-01

We investigated responses of rice plant to three rare sugars, d-altrose, d-sorbose, and d-allose, due to establishment of mass production methods for these rare sugars. Root growth and shoot growth were significantly inhibited by d-allose but not by the other rare sugars. A large-scale gene expression analysis using a rice microarray revealed that d-allose treatment causes a high upregulation of many defense-related, pathogenesis-related (PR) protein genes in rice. The PR protein genes were not upregulated by other rare sugars. Furthermore, d-allose treatment of rice plants conferred limited resistance of the rice against the pathogen Xanthomonas oryzae pv. oryzae but the other tested sugars did not. These results indicate that d-allose has a growth inhibitory effect but might prove to be a candidate elicitor for reducing disease development in rice.

Rewiring the reductive tricarboxylic acid pathway and L-malate transport pathway of Aspergillus oryzae for overproduction of L-malate.

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Liu, Jingjing; Xie, Zhipeng; Shin, Hyun-Dong; Li, Jianghua; Du, Guocheng; Chen, Jian; Liu, Long

2017-07-10

Aspergillus oryzae finds wide application in the food, feed, and wine industries, and is an excellent cell factory platform for production of organic acids. In this work, we achieved the overproduction of L-malate by rewiring the reductive tricarboxylic acid (rTCA) pathway and L-malate transport pathway of A. oryzae NRRL 3488. First, overexpression of native pyruvate carboxylase and malate dehydrogenase in the rTCA pathway improved the L-malate titer from 26.1gL -1 to 42.3gL -1 in shake flask culture. Then, the oxaloacetate anaplerotic reaction was constructed by heterologous expression of phosphoenolpyruvate carboxykinase and phosphoenolpyruvate carboxylase from Escherichia coli, increasing the L-malate titer to 58.5gL -1 . Next, the export of L-malate from the cytoplasm to the external medium was strengthened by overexpression of a C4-dicarboxylate transporter gene from A. oryzae and an L-malate permease gene from Schizosaccharomyces pombe, improving the L-malate titer from 58.5gL -1 to 89.5gL -1 . Lastly, guided by transcription analysis of the expression profile of key genes related to L-malate synthesis, the 6-phosphofructokinase encoded by the pfk gene was identified as a potential limiting step for L-malate synthesis. Overexpression of pfk with the strong sodM promoter increased the L-malate titer to 93.2gL -1 . The final engineered A. oryzae strain produced 165gL -1 L-malate with a productivity of 1.38gL -1 h -1 in 3-L fed-batch culture. Overall, we constructed an efficient L-malate producer by rewiring the rTCA pathway and L-malate transport pathway of A. oryzae NRRL 3488, and the engineering strategy adopted here may be useful for the construction of A. oryzae cell factories to produce other organic acids. Copyright © 2017 Elsevier B.V. All rights reserved.

Effects of silicon on Oryza sativa L. seedling roots under simulated acid rain stress.

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Ju, Shuming; Yin, Ningning; Wang, Liping; Zhang, Cuiying; Wang, Yukun

2017-01-01

Silicon (Si) has an important function in reducing the damage of environmental stress on plants. Acid rain is a serious abiotic stress factor, and Si can alleviate the stress induced by acid rain on plants. Based on these assumptions, we investigated the effects of silicon on the growth, root phenotype, mineral element contents, hydrogen peroxide (H2O2) and antioxidative enzymes of rice (Oryza sativa L.) seedling roots under simulated acid rain (SAR) stress. The results showed that the combined or single effects of Si and/or SAR on rice roots depend on the concentration of Si and the pH of the SAR. The combined or single effects of a low or moderate concentration of Si (1.0 or 2.0 mM) and light SAR (pH 4.0) enhanced the growth of rice roots, and the combined effects were stronger than those of the single treatment. A high concentration of Si (4.0 mM) or severe SAR (pH 2.0) exerted deleterious effects. The incorporation of Si (1.0, 2.0 or 4.0 mM) into SAR with pH 3.0 or 2.0 promoted the rice root growth, decreased the H2O2 content, increased the Si concentration and the superoxide dismutase (SOD), catalase (CAT), peroxidase (POD) and ascorbate peroxidase (APX) activities, maintained the balance of mineral element (K, Ca, Mg, Fe, Zn, and Cu) concentrations in the roots of rice seedlings compared with SAR alone. The alleviatory effects observed with a moderate concentration of Si (2.0 mM) were better than the effects obtained with a low or high concentration of Si (1.0 or 4.0 mM). The observed effects were due to disruptions in the absorption and utilization of mineral nutrients and impacts on the activity of antioxidant enzymes in roots, and this conclusion suggests that the degree of rice root damage caused by acid rain might be attributed to not only acid rain but also the level of Si in the soil.

Effects of silicon on Oryza sativa L. seedling roots under simulated acid rain stress

Science.gov (United States)

Wang, Liping; Zhang, Cuiying; Wang, Yukun

2017-01-01

Silicon (Si) has an important function in reducing the damage of environmental stress on plants. Acid rain is a serious abiotic stress factor, and Si can alleviate the stress induced by acid rain on plants. Based on these assumptions, we investigated the effects of silicon on the growth, root phenotype, mineral element contents, hydrogen peroxide (H2O2) and antioxidative enzymes of rice (Oryza sativa L.) seedling roots under simulated acid rain (SAR) stress. The results showed that the combined or single effects of Si and/or SAR on rice roots depend on the concentration of Si and the pH of the SAR. The combined or single effects of a low or moderate concentration of Si (1.0 or 2.0 mM) and light SAR (pH 4.0) enhanced the growth of rice roots, and the combined effects were stronger than those of the single treatment. A high concentration of Si (4.0 mM) or severe SAR (pH 2.0) exerted deleterious effects. The incorporation of Si (1.0, 2.0 or 4.0 mM) into SAR with pH 3.0 or 2.0 promoted the rice root growth, decreased the H2O2 content, increased the Si concentration and the superoxide dismutase (SOD), catalase (CAT), peroxidase (POD) and ascorbate peroxidase (APX) activities, maintained the balance of mineral element (K, Ca, Mg, Fe, Zn, and Cu) concentrations in the roots of rice seedlings compared with SAR alone. The alleviatory effects observed with a moderate concentration of Si (2.0 mM) were better than the effects obtained with a low or high concentration of Si (1.0 or 4.0 mM). The observed effects were due to disruptions in the absorption and utilization of mineral nutrients and impacts on the activity of antioxidant enzymes in roots, and this conclusion suggests that the degree of rice root damage caused by acid rain might be attributed to not only acid rain but also the level of Si in the soil. PMID:28291806

Usaha Pemasaran Produsen Tempe

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R. Nana Kusdiana

2013-05-01

Full Text Available Tempe is a food made from the fermentation of soybeans or some other materials using somekind of fungus Rhizopus, such as Rhizopus oligosporus, Rh. oryzae, Rh. stolonifer or also known as bread mold, or Rh. arrhizus. Fermentation preparations are commonly known as "yeast tempe". Tempe has been known since ancient times, particularly in Java as supplement staple food which is high protein and low cholesterol. Tempe and tofu selling price is still low compared to other protein food sources, such as fish and chicken. According to market research, the majority of tempe and tofu products is still the lower class’ consumption, since the quality and processing methods are traditional distribution, including in sanitation and hygiene. It is necessary to stimulate and generate interest in the bank to develop relationship with the Indonesian Tofu Tempe Producers Cooperative (KOPTI and its members to tempe producers can increase production and performance. Tempe and Tofu Development Centers Manufacturers through Integrated Partnership Program by KOPTI as a company with a core group of tempe and tofu manufacturers, which are members of the KOPTI, is the modernization of tempe and tofu small business activity to be an independent food business industry.

Non-aflatoxigenicity of commercial Aspergillus oryzae strains due to genetic defects compared to aflatoxigenic Aspergillus flavus.

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Tao, Lin; Chung, Soo Hyun

2014-08-01

Aspergillus oryzae is generally recognized as safe, but it is closely related to A. flavus in morphology and genetic characteristics. In this study, we tested the aflatoxigenicity and genetic analysis of nine commercial A. oryzae strains that were used in Korean soybean fermented products. Cultural and HPLC analyses showed that none of the commercial strains produced detectable amount of aflatoxins. According to the molecular analysis of 17 genes in the aflatoxin (AF) biosynthetic pathway, the commercial strains could be classified into three groups. The group I strains contained all the 17 AF biosynthetic genes tested in this study; the group II strains deleted nine AF biosynthetic genes and possessed eight genes, including aflG, aflI, aflK, aflL, aflM, aflO, aflP, and aflQ; the group III strains only had six AF biosynthetic genes, including aflG, aflI, aflK, aflO, aflP, and aflQ. With the reverse transcription polymerase chain reaction, the group I A. oryzae strains showed no expression of aflG, aflQ and/or aflM genes, which resulted in the lack of AF-producing ability. Group II and group III strains could not produce AF owing to the deletion of more than half of the AF biosynthetic genes. In addition, the sequence data of polyketide synthase A (pksA) of group I strains of A. oryzae showed that there were three point mutations (two silent mutations and one missense mutation) compared with aflatoxigenic A. flavus used as the positive control in this study.

Ortholog Alleles at Xa3/Xa26 Locus Confer Conserved Race-Specific Resistance against Xanthomonas oryzae in Rice

Institute of Scientific and Technical Information of China (English)

Hong-Jing Li; Xiang-Hua Li; Jing-Hua Xiao; Rod A. Wing; Shi-Ping Wang

2012-01-01

The rice disease resistance (R) gene Xa3/Xa26 (having also been named Xa3 and Xa26) against Xanthomonas oryzae pv.oryzae (Xoo),which causes bacterial blight disease,belongs to a multiple gene family clustered in chromosome 11 and is from an AA genome rice cultivar (Oryza sativa L.).This family encodes leucine-rich repeat (LRR) receptor kinasetype proteins.Here,we show that the orthologs (alleles) of Xa3/Xa26,Xa3/Xa26-2,and Xa3/Xa26-3,from wild Oryza species O.officinalis (CC genome) and O.minuta (BBCC genome),respectively,were also R genes against Xoo.Xa3/Xa26-2 and Xa3/Xa26-3 conferred resistance to 16 of the 18 Xoo strains examined.Comparative sequence analysis of the Xa3/Xa26 families in the two wild Oryza species showed that Xa3/Xa26-3 appeared to have originated from the CC genome of O.minuta.The predicted proteins encoded by Xa3/Xa26,Xa3/Xa26-2,and Xa3/Xa26-3 share 91-99％ sequence identity and 94-99％ sequence similarity.Transgenic plants carrying a single copy of Xa3/Xa26,Xa3/Xa26-2,or Xa3/Xa26-3,in the same genetic background,showed a similar resistance spectrum to a set of Xoo strains,although plants carrying Xa3/Xa26-2 or Xa3/Xa26-3 showed lower resistance levels than the plants carrying Xa3/Xa26.These results suggest that the Xa3/Xa26 locus predates the speciation of A and C genome,which is approximately 7.5 million years ago.Thus,the resistance specificity of this locus has been conserved for a long time.

What Does Genetic Diversity of Aspergillus flavus Tell Us About Aspergillus oryzae?

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Aspergillus flavus and Aspergillus oryzae belong to Aspergillus section Flavi. They are closely related and are of significant economic importance. The former species has the ability to produce harmful aflatoxins while the latter is widely used in food fermentation and industrial enzyme production. ...

Caracterização físico-química e tecnológica da farinha de soja integral fermentada com Aspergillus oryzae Physicochemical and technological characterization of whole soybean flour fermented by Aspergillus oryzae

OpenAIRE

Leomar Hackbart da Silva; Paula Fernanda Pinto da Costa; Gabriela Wakayama Nomiyama; Ingrid Paula de Souza; Yoon Kil Chang

2012-01-01

Neste trabalho, estudou-se o efeito das condições de fermentação da farinha de soja integral, utilizando-se o fungo Aspergillus oryzae, sobre as características físico-químicas e as propriedades tecnológicas da farinha de soja integral autoclavada fermentada. A farinha de soja integral autoclavada foi fermentada com o fungo Aspergillus oryzae CCT 4359, incubada às temperaturas de 30 °C e 40 °C por 24 horas e 48 horas, e seca em estufa a vácuo, a 60 °C, até 10% de umidade final, obtendo-se a f...

Improved production of kojic acid by mutagenesis of Aspergillus flavus HAk1 and Aspergillus oryzae HAk2 and their potential antioxidant activity.

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Ammar, Hala A M; Ezzat, Saeid M; Houseny, Asmaa M

2017-10-01

Two wild-type (WT) Aspergillus strains, A. flavus HAk1 and A. oryzae HAk2, were selected for kojic acid (KA) biosynthesis. Malt extract sucrose culture medium (MES) was the best culture medium for maximum production of KA. The maximum production of KA has been estimated at pH 4 after 7 days of incubation at 30 °C. Overproduction of KA was attained by mutagenesis of both A. flavus HAk1 and A. oryzae HAk2 through their exposer to different doses of gamma irradiation. The mutant strains (MT) A. flavus HAk1-M2 and A. oryzae HAk2-M26 were the most stable mutants for maximum production of KA through four generations. Yield of KA by A. oryzae HAk2-M26 and A. flavus HAk1-M2 has been 2.03-fold and 1.9-fold, respectively, higher than their wild-type strains. All WT and MT strains were used for KA production from different agricultural raw materials. Apple peel was the best waste for KA production by WT strains of A. flavus and A. oryzae, while orange peel and rice stalk are best material for KA production by MT strains, A. flavus HAk1-M2 and A. oryzae HAk2-M26, respectively. All experimental strains have the ability to produce considerable amounts of KA from sugarcane molasse (SCM) and sugar-beet molasse (SBM). SBM was better than SCM for KA production by all strains. The antioxidant activity of biosynthesizing KA was strongly affected with production conditions, where the highest antioxidant activity of all strains was recorded at the optimum environmental and nutritional conditions for KA production.

Sequence analysis and overexpression of a pectin lyase gene (pel1) from Aspergillus oryzae KBN616.

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Kitamoto, N; Yoshino-Yasuda, S; Ohmiya, K; Tsukagoshi, N

2001-01-01

A gene (pel1) encoding pectin lyase (Pel1) was isolated from a shoyu koji mold, Aspergillus oryzae KBN616, and characterized. The structural gene comprised 1,196 bp with a single intron. The ORF encoded 381 amino acids with a signal peptide of 20 amino acids. The deduced amino acid sequence showed high similarity to those of Aspergillus niger pectin lyases and Glomerella cingulata PnlA. The pel1 gene was successfully overexpressed under the promoter of the A. oryzae TEF1 gene. The molecular mass of the recombinant pectin lyase substantially coincided with that calculated based on nucleotide sequence.

Evaluation of the ORYZA2000 Rice Growth Model under Nitrogen-Limited Conditions in an Irrigated Mediterranean Environment Evaluación del Modelo de Crecimiento de Arroz ORYZA2000 bajo Condiciones Limitadas de Nitrógeno en un Ambiente Mediterráneo Regado

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Pamela Artacho

2011-03-01

Full Text Available ORYZA2000 is a growth model for tropical lowland rice ( Oryza sativa L. developed by the International Rice Research Institute and Wageningen University. This model has been evaluated extensively in a wide range of environments. However, reports examining japonica cultivars growing in temperate climates are scarce. In this study, ORYZA2000 was calibrated and evaluated using data from experiments carried out in the South-Central area of Chile. These experiments were performed on a japonica rice cultivar growing under an irrigated Mediterranean environment at various N rates. ORYZA2000 was then applied to explore potential yield and grain yield response to N fertilization under likely weather conditions in the major rice-producing area in Chile. ORYZA2000 was sufficiently accurate to simulate grain yield and crop N uptake at the end of the season. Final crop N uptake was simulated with a root mean squared error (RMSE of 20 kg ha-1 (15% and grain yield with a RMSE of 1666 kg ha-1 (19%. However, the prediction of biomass and N uptake of individual organs throughout the season was poor. A long-term simulation study confirmed a potential yield as high as 12 000 kg ha-1 in the Parral area, as well as the existence of a scope for yield increase. The yield response to N fertilization was predicted even at rates of 300 kg ha-1, although a significant probability of low yields was also observed. This trend supports the need to incorporate dynamic N management in Chilean rice production.ORYZA2000 es un modelo para arroz tropical regado desarrollado por el Instituto Internacional de Investigación en Arroz y Universidad de Wageningen, que ha sido evaluado en un amplio rango de ambientes. Sin embargo, los reportes para cultivares japónica creciendo en ambientes templados son escasos. En este estudio, ORYZA2000 fue calibrado y evaluado usando datos de experimentos realizados en el centro-sur de Chile, que incluyeron una variedad japónica cultivada bajo un

Construction and application of a novel genetically engineered Aspergillus oryzae for expressing proteases

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Xiao-Chun Yu

2017-09-01

Conclusions: These findings suggest that cloning of microbial protease genes with good physicochemical properties and expressing them in an ideal host such as A. oryzae is a novel strategy to enhance the value of soybean meal.

Three extracellular dipeptidyl peptidases found in Aspergillus oryzae show varying substrate specificities.

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Maeda, Hiroshi; Sakai, Daisuke; Kobayashi, Takuji; Morita, Hiroto; Okamoto, Ayako; Takeuchi, Michio; Kusumoto, Ken-Ichi; Amano, Hitoshi; Ishida, Hiroki; Yamagata, Youhei

2016-06-01

Three extracellular dipeptidyl peptidase genes, dppB, dppE, and dppF, were unveiled by sequence analysis of the Aspergillus oryzae genome. We investigated their differential enzymatic profiles, in order to gain an understanding of the diversity of these genes. The three dipeptidyl peptidases were expressed using Aspergillus nidulans as the host. Each recombinant enzyme was purified and subsequently characterized. The enzymes displayed similar optimum pH values, but optimum temperatures, pH stabilities, and substrate specificities varied. DppB was identified as a Xaa-Prolyl dipeptidyl peptidase, while DppE scissile substrates were similar to the substrates for Aspergillus fumigatus DPPV (AfDPPV). DppF was found to be a novel enzyme that could digest both substrates for A. fumigatus DPPIV and AfDPPV. Semi-quantitative PCR revealed that the transcription of dppB in A. oryzae was induced by protein substrates and repressed by the addition of an inorganic nitrogen source, despite the presence of protein substrates. The transcription of dppE depended on its growth time, while the transcription of dppF was not affected by the type of the nitrogen source in the medium, and it started during the early stage of the fungal growth. Based on these results, we conclude that these enzymes may represent the nutrition acquisition enzymes. Additionally, DppF may be one of the sensor peptidases responsible for the detection of the protein substrates in A. oryzae environment. DppB may be involved in nitrogen assimilation control, since the transcription of dppB was repressed by NaNO3, despite the presence of protein substrates.

Cooperative role of calnexin and TigA in Aspergillus oryzae glycoprotein folding.

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Wang, Ning; Seko, Akira; Takeda, Yoichi; Kikuma, Takashi; Ito, Yukishige

2015-10-01

Calnexin (CNX), known as a lectin chaperone located in the endoplasmic reticulum (ER), specifically recognizes G1M9GN2-proteins and facilitates their proper folding with the assistance of ERp57 in mammalian cells. However, it has been left unidentified how CNX works in Aspergillus oryzae, which is a filamentous fungus widely exploited in biotechnology. In this study, we found that a protein disulfide isomerase homolog TigA can bind with A. oryzae CNX (AoCNX), which was revealed to specifically recognize monoglucosylated glycans, similarly to CNX derived from other species, and accelerate the folding of G1M9GN2-ribonuclease (RNase) in vitro. For refolding experiments, a homogeneous monoglucosylated high-mannose-type glycoprotein G1M9GN2-RNase was chemoenzymatically synthesized from G1M9GN-oxazoline and GN-RNase. Denatured G1M9GN2-RNase was refolded with highest efficiency in the presence of both soluble form of AoCNX and TigA. TigA contains two thioredoxin domains with CGHC motif, mutation analysis of which revealed that the one in N-terminal regions is involved in binding to AoCNX, while the other in catalyzing protein refolding. The results suggested that in glycoprotein folding process of A. oryzae, TigA plays a similar role as ERp57 in mammalian cells, as a partner protein of AoCNX. © The Author 2015. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Comparative analysis of solid-state bioprocessing and enzymatic treatment of finger millet for mobilization of bound phenolics.

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Yadav, Geetanjali; Singh, Anshu; Bhattacharya, Patrali; Yuvraj, Jude; Banerjee, Rintu

2013-11-01

The present work investigates the probable bioprocessing technique to mobilize the bound phenolics naturally found in finger millet cell wall for enriching it with dietary antioxidants. Comparative study was performed between the exogenous enzymatic treatment and solid-state fermentation of grain (SSF) with a food grade organism Rhizopus oryzae. SSF results indicated that at the 6th day of incubation, total phenolic content (18.64 mg gallic acid equivalent/gds) and antioxidant property (DPPH radical scavenging activity of 39.03 %, metal chelating ability of 54 % and better reducing power) of finger millet were drastically enhanced when fermented with GRAS filamentous fungi. During the enzymatic bioprocessing, most of the phenolics released during the hydrolysis, leached out into the liquid portion rather than retaining them within the millet grain, resulting in overall loss of dietary antioxidant. The present study establishes the most effective strategy to enrich the finger millet with phenolic antioxidants.

Gene expansion shapes genome architecture in the human pathogen Lichtheimia corymbifera: an evolutionary genomics analysis in the ancient terrestrial mucorales (Mucoromycotina).

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Schwartze, Volker U; Winter, Sascha; Shelest, Ekaterina; Marcet-Houben, Marina; Horn, Fabian; Wehner, Stefanie; Linde, Jörg; Valiante, Vito; Sammeth, Michael; Riege, Konstantin; Nowrousian, Minou; Kaerger, Kerstin; Jacobsen, Ilse D; Marz, Manja; Brakhage, Axel A; Gabaldón, Toni; Böcker, Sebastian; Voigt, Kerstin

2014-08-01

Lichtheimia species are the second most important cause of mucormycosis in Europe. To provide broader insights into the molecular basis of the pathogenicity-associated traits of the basal Mucorales, we report the full genome sequence of L. corymbifera and compared it to the genome of Rhizopus oryzae, the most common cause of mucormycosis worldwide. The genome assembly encompasses 33.6 MB and 12,379 protein-coding genes. This study reveals four major differences of the L. corymbifera genome to R. oryzae: (i) the presence of an highly elevated number of gene duplications which are unlike R. oryzae not due to whole genome duplication (WGD), (ii) despite the relatively high incidence of introns, alternative splicing (AS) is not frequently observed for the generation of paralogs and in response to stress, (iii) the content of repetitive elements is strikingly low (<5%), (iv) L. corymbifera is typically haploid. Novel virulence factors were identified which may be involved in the regulation of the adaptation to iron-limitation, e.g. LCor01340.1 encoding a putative siderophore transporter and LCor00410.1 involved in the siderophore metabolism. Genes encoding the transcription factors LCor08192.1 and LCor01236.1, which are similar to GATA type regulators and to calcineurin regulated CRZ1, respectively, indicating an involvement of the calcineurin pathway in the adaption to iron limitation. Genes encoding MADS-box transcription factors are elevated up to 11 copies compared to the 1-4 copies usually found in other fungi. More findings are: (i) lower content of tRNAs, but unique codons in L. corymbifera, (ii) Over 25% of the proteins are apparently specific for L. corymbifera. (iii) L. corymbifera contains only 2/3 of the proteases (known to be essential virulence factors) in comparison to R. oryzae. On the other hand, the number of secreted proteases, however, is roughly twice as high as in R. oryzae.

Structural and Functional Studies of A. oryzae Cutinase: Enhanced Thermostability and Hydrolytic Activity of Synthetic Ester and Polyester Degradation

OpenAIRE

Liu, Zhiqiang; Gosser, Yuying; Baker, Peter James; Ravee, Yaniv; Lu, Ziying; Alemu, Girum; Li, Huiguang; Butterfoss, Glenn L.; Kong, Xiang-Peng; Gross, Richard; Montclare, Jin Kim

2009-01-01

Cutinases are responsible for hydrolysis of the protective cutin lipid polyester matrix in plants and thus have been exploited for hydrolysis of small molecule esters and polyesters. Here we explore the reactivity, stability, and structure of Aspergillus oryzae cutinase and compare it to the well-studied enzyme from Fusarium solani. Two critical differences are highlighted in the crystallographic analysis of the A. oryzae structure: (i) an additional disulfide bond and (ii) a topologically fa...

BASAL MEDIA FORMULATION USING CANAVALIA ENSIFORMIS AS CARBON AND NITROGEN SOURCE FOR THE GROWTH OF SOME FUNGI SPECIES

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B.J. Akinyele2

2012-02-01

Full Text Available The possibility of developing alternative media to commercial potato dextrose agar was assessed using, Canavalia ensiformis (Linn (jack beans as carbon and nitrogen source. Six leguminous meal media were used as substitute for either carbon or nitrogen or both, while potato dextrose broth (PDB was used as a positive control and basal medium as a negative control. Six species of fungi Aspergillus flavus, A. niger, Meria coniospora, Mucor sp, Neurospora crassa and Rhizopus oryzae were aseptically inoculated into the formulated media and allowed to grow. Their mycelia dry weights were taken after 24, 48, 72, 96 and 120 hours. Growth of all fungal species was observed to be slightly lower, about the same or better in the formulated media relative to the control. Aspergillus flavus had its highest biomass of 1.70g in the media formulated with Canavalia ensiformis as the carbon source relative to 1.42g as the standard at the 120 hour. A. niger had a growth of 0.62g relative to 0.61g at 120 hours of the control. Meria coniospora had a growth of 0.27g relative to 0.38g at 120 hours. Mucor sp had a growth of 0.54g relative to 0.44g at 120 hours. Neurospora crassa had a growth of 1.05g relative to 0.24g at 120 hours. Rhizopus oryzae had a growth of 0.14g relative to 0.25g at 120 hours. The study revealed that Canavalia ensiformis contains minerals and nutrients that is able to provide the nutritional requirements of these fungi. Thus, it can be used as an alternative material in the preparation of culture media for in vitro cultivation of these fungi for teaching and research purposes.

Ehd4 encodes a novel and Oryza-genus-specific regulator of photoperiodic flowering in rice.

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He Gao

Full Text Available Land plants have evolved increasingly complex regulatory modes of their flowering time (or heading date in crops. Rice (Oryza sativa L. is a short-day plant that flowers more rapidly in short-day but delays under long-day conditions. Previous studies have shown that the CO-FT module initially identified in long-day plants (Arabidopsis is evolutionary conserved in short-day plants (Hd1-Hd3a in rice. However, in rice, there is a unique Ehd1-dependent flowering pathway that is Hd1-independent. Here, we report isolation and characterization of a positive regulator of Ehd1, Early heading date 4 (Ehd4. ehd4 mutants showed a never flowering phenotype under natural long-day conditions. Map-based cloning revealed that Ehd4 encodes a novel CCCH-type zinc finger protein, which is localized to the nucleus and is able to bind to nucleic acids in vitro and transactivate transcription in yeast, suggesting that it likely functions as a transcriptional regulator. Ehd4 expression is most active in young leaves with a diurnal expression pattern similar to that of Ehd1 under both short-day and long-day conditions. We show that Ehd4 up-regulates the expression of the "florigen" genes Hd3a and RFT1 through Ehd1, but it acts independently of other known Ehd1 regulators. Strikingly, Ehd4 is highly conserved in the Oryza genus including wild and cultivated rice, but has no homologs in other species, suggesting that Ehd4 is originated along with the diversification of the Oryza genus from the grass family during evolution. We conclude that Ehd4 is a novel Oryza-genus-specific regulator of Ehd1, and it plays an essential role in photoperiodic control of flowering time in rice.

Presence and Functionality of Mating Type Genes in the Supposedly Asexual Filamentous Fungus Aspergillus oryzae

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Wada, Ryuta; Maruyama, Jun-ichi; Yamaguchi, Haruka; Yamamoto, Nanase; Wagu, Yutaka; Paoletti, Mathieu; Archer, David B.; Dyer, Paul S.

2012-01-01

The potential for sexual reproduction in Aspergillus oryzae was assessed by investigating the presence and functionality of MAT genes. Previous genome studies had identified a MAT1-1 gene in the reference strain RIB40. We now report the existence of a complementary MAT1-2 gene and the sequencing of an idiomorphic region from A. oryzae strain AO6. This allowed the development of a PCR diagnostic assay, which detected isolates of the MAT1-1 and MAT1-2 genotypes among 180 strains assayed, including industrial tane-koji isolates. Strains used for sake and miso production showed a near-1:1 ratio of the MAT1-1 and MAT1-2 mating types, whereas strains used for soy sauce production showed a significant bias toward the MAT1-2 mating type. MAT1-1 and MAT1-2 isogenic strains were then created by genetic manipulation of the resident idiomorph, and gene expression was compared by DNA microarray and quantitative real-time PCR (qRT-PCR) methodologies under conditions in which MAT genes were expressed. Thirty-three genes were found to be upregulated more than 10-fold in either the MAT1-1 host strain or the MAT1-2 gene replacement strain relative to each other, showing that both the MAT1-1 and MAT1-2 genes functionally regulate gene expression in A. oryzae in a mating type-dependent manner, the first such report for a supposedly asexual fungus. MAT1-1 expression specifically upregulated an α-pheromone precursor gene, but the functions of most of the genes affected were unknown. The results are consistent with a heterothallic breeding system in A. oryzae, and prospects for the discovery of a sexual cycle are discussed. PMID:22327593

Metabolomics analysis of TiO2 nanoparticles induced toxicological effects on rice (Oryza sativa L.).

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Wu, Biying; Zhu, Lizhong; Le, X Chris

2017-11-01

The wide occurrence and high environmental concentration of titanium dioxide nanoparticles (nano-TiO 2 ) have raised concerns about their potential toxic effects on crops. In this study, we employed a GC-MS-based metabolomic approach to investigate the potential toxicity of nano-TiO 2 on hydroponically-cultured rice (Oryza sativa L.) after exposed to 0, 100, 250 or 500 mg/L of nano-TiO 2 for fourteen days. Results showed that the biomass of rice was significantly decreased and the antioxidant defense system was significantly disturbed after exposure to nano-TiO 2 . One hundred and five identified metabolites showed significant difference compared to the control, among which the concentrations of glucose-6-phosphate, glucose-1-phosphate, succinic and isocitric acid were increased most, while the concentrations of sucrose, isomaltulose, and glyoxylic acid were decreased most. Basic energy-generating ways including tricarboxylic acid cycle and the pentose phosphate pathway, were elevated significantly while the carbohydrate synthesis metabolism including starch and sucrose metabolism, and glyoxylate and dicarboxylate metabolism were inhibited. However, the biosynthetic formation of most of the identified fatty acids, amino acids and secondary metabolites which correlated to crop quality, were increased. The results suggest that the metabolism of rice plants is distinctly disturbed after exposure to nano-TiO 2 , and nano-TiO 2 would have a mixed effect on the yield and quality of rice. Copyright © 2017 Elsevier Ltd. All rights reserved.

High-level expression and efficient purification of bioactive swollenin in Aspergillus oryzae.

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Wang, Meihua; Cai, Jin; Huang, Lei; Lv, Zhengbin; Zhang, Yaozhou; Xu, Zhinan

2010-11-01

The bioactivity of swollenin is beneficial to cellulose decomposition by cellulase despite the lack of hydrolytic activity itself. In order to improve the productivity of swollenin, the effects of culture conditions on the expression level in recombinant Aspergillus oryzae were investigated systematically. With regard to the bioactivity of swollenin, glycerin and peanut meal were the optimal carbon or nitrogen source, respectively. The highest level production of swollenin (50 mg L(-1)) was attained after 88 h cultivation with the initial pH of 5.6 in the culture medium. Then the soluble swollenin was effectively purified from the cultural supernatant by ammonium sulfate precipitation and cationic exchange chromatography with recovery yield of 53.2%. The purified swollenin was fully bioactive due to its strong synergistic activity with cellulose.

Effect of external mass transfer on activation energy of butyl oleate ester synthesis using a whole cell bio catalyst

International Nuclear Information System (INIS)

Shahhoseini, Sh.; Nasernejad, B.; Vahabzadeh, F.

2016-01-01

In the present research, synthesis of butyl oleate ester from oleic acid and butanol using loofa-immobilized Rhizopus oryzae as a whole cell biocatalyst (LIC) was studied in which hexane was used as the hydrophobic solvent. Decrease of mass transfer limitations as result of the interface formation between the two immiscible substrates, positively affected on the reaction progress (87% as the ester product yielded within 10 h). By applying Arrhenius equation, the activation energy of the ester synthesis was determined as Ea=18.2 kJ/mol within temperature range of 15-45°C. It was notable to test appearance of the nonlinearity in Arrhenius plot which was indicative of presence of two sections. The reaction limited region was 15-35°C; Ea=27 kJ/mol and diffusion limited region was >35°C; Ea=6.8 kJ/mol. Eventually, in this research, influence of external mass transfer on activation energy with reference to the catalytic role of the LIC in the ester synthesis was discussed.

Effect of Exogenous _D-Alanine on _D-Alanyl-_D-alanine Content in Leaf Blades of Oryza australiensis Domin

OpenAIRE

Hisashi, Manabe; Aizu Junior College of Fukushima Prefecture

1986-01-01

In seedlings of Oryza australiensis Domin (W0008), most of the D-alanyl-D-alanine was distributed in the leaf blades. In excised leaf blades of W0008, exogenous D-alanine was incorporated into D-alanyl-D-alanine irrespective of the light condition as in Sasanishiki. With cultivation in D-alanine medium for several days, the D-alanyl-D-alanine content in W0008 leaf blades was found to increase, but no other D-alanine-containing dipeptides such as D-alanylglycine or D-alanyl-L-alanine were dete...

Crystallization and preliminary X-ray analysis of Aspergillus oryzae catechol oxidase

International Nuclear Information System (INIS)

Kaljunen, Heidi; Gasparetti, Chiara; Kruus, Kristiina; Rouvinen, Juha; Hakulinen, Nina

2011-01-01

Catechol oxidase from A. oryzae was crystallized by the hanging-drop vapour-diffusion method. Catechol oxidase is an enzyme that catalyzes the oxidation of o-diphenols to the corresponding o-quinones. It is a copper-containing enzyme with a binuclear copper active site. Here, the crystallization and multiple-wavelength anomalous dispersion data collection of catechol oxidase from the mould fungus Aspergillus oryzae are described. During the purification, three forms of the enzyme (39.3, 40.5 and 44.3 kDa) were obtained. A mixture of these three forms was initially crystallized and gave crystals that diffracted to 2.5 Å resolution and belonged to space group P3 2 21, with unit-cell parameters a = b = 118.9, c = 84.5 Å, α = β = 90, γ = 120°. A preparation containing only the shorter form (39.3 kDa) produced crystals that diffracted to 2.9 Å resolution and belonged to space group P2 1 2 1 2 1 , with unit-cell parameters a = 51.8, b = 95.3, c = 139.5 Å, α = β = γ = 90°

Enzyme activity of β-galactosidase from Kluyveromyces lactis and Aspergillus oryzae on simulated conditions of human gastrointestinal system

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Alessandra Bosso

2015-09-01

Full Text Available An alternative to relieve the symptoms of lactose intolerance is the intake of the enzyme β-galactosidase in pharmaceutical dosage forms. The ability of β-galactosidase produced by Kluyveromyces lactis and Aspergillus oryzae to hydrolyze lactose in simulated conditions of the human gastrointestinal tract was investigated. The experiment was carried out in the optimum temperature for each enzyme activity, 40 and 55°C, respectively, and at the normal human body temperature (37°C at concentrations of 1.5, 3.0, and 5.0 g/L (enzyme from A. oryzae or mL/L (enzyme from K. lactis. Both enzymes were completely inactivated under simulated gastric conditions (pH 2. When the enzymes were subjected to simulated small intestine conditions (pH 7.4, lactose hydrolysis has occurred, but at 37°C the percentage was lower than that under the optimal temperatures. At concentrations of 1.5, 3.0, and 5.0 mL/L the enzyme from K. lactis hydrolyzed 76.63%, 88.91% and 94.80% of lactose at 40°C, and 55.99%, 80.91% and 81.53% at 37°C, respectively. In contrast, the enzyme from A. oryzae hydrolyzed 7.11%, 16.18% and 21.29% at 55°C, and 8.4%, 11.85% and 16.43% at 37°C. It was observed that under simulated intestinal conditions, the enzyme from K. lactis was more effective on lactose hydrolysis as compared to the enzyme from A. oryzae. Considering the findings of this study, it is extremely necessary to use an enteric coating on β-galactosidase capsules so that this enzyme is released only in the small intestine, which is its site of action, thus not suffering the action of the stomach pH.Keywords: Lactase. Hydrolysis. Lactose intolerance. Gastrointestinal tract. RESUMOAtividade de β-galactosidase de Kluyveromyces lactis e Aspergillus oryzae, em condições simuladas do sistema gastrintestinal humanoUma das alternativas para amenizar os sintomas da intolerância à lactose é a ingestão de β-galactosidase em formas farmacêuticas. Neste trabalho avaliou-se a

Overexpression of Rice Auxilin-Like Protein, XB21, Induces Necrotic Lesions, up-Regulates Endocytosis-Related Genes, and Confers Enhanced Resistance to Xanthomonas oryzae pv. oryzae.

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Park, Chang-Jin; Wei, Tong; Sharma, Rita; Ronald, Pamela C

2017-12-01

The rice immune receptor XA21 confers resistance to the bacterial pathogen, Xanthomonas oryzae pv. oryzae (Xoo). To elucidate the mechanism of XA21-mediated immunity, we previously performed a yeast two-hybrid screening for XA21 interactors and identified XA21 binding protein 21 (XB21). Here, we report that XB21 is an auxilin-like protein predicted to function in clathrin-mediated endocytosis. We demonstrate an XA21/XB21 in vivo interaction using co-immunoprecipitation in rice. Overexpression of XB21 in rice variety Kitaake and a Kitaake transgenic line expressing XA21 confers a necrotic lesion phenotype and enhances resistance to Xoo. RNA sequencing reveals that XB21 overexpression results in the differential expression of 8735 genes (4939 genes up- and 3846 genes down-regulated) (≥2-folds, FDR ≤0.01). The up-regulated genes include those predicted to be involved in 'cell death' and 'vesicle-mediated transport'. These results indicate that XB21 plays a role in the plant immune response and in regulation of cell death. The up-regulation of genes controlling 'vesicle-mediated transport' in XB21 overexpression lines is consistent with a functional role for XB21 as an auxilin.

Purification and Characterisation of a Fibrinolytic Enzyme from Rhizopus micro sporus var. tuberosus

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Shuli Zhang

2015-01-01

Full Text Available Extracellular fibrinolytic enzyme from Rhizopus microsporus var. tuberosus was purified and characterised. The microorganism was isolated in a distillery from daqu, a fermentative agent used in the production of Chinese liquor and vinegar at diff erent temperatures. The fibrinolytic enzyme was partially purifi ed by ammonium sulphate precipitation, dialysis, DEAE Sepharose® Fast Flow ion exchange chromatography and Sephadex G-75 gel filtration chromatography. The molecular mass of the fi brinolytic enzyme was estimated to be 24.5 kDa by SDS-PAGE. The purified enzyme showed optimal activity at pH=7.0 and 37 °C by fibrin plate method. It showed stronger resistance to the inhibition by trypsin and was stable at 37 °C retaining 96.1 % residual activity aft er 4 h of incubation. The fibrinolytic activity of the enzyme was enhanced by Na+, Ca2+, Mg2+ and Mn2+. Conversely, Zn2+ and Cu2+ partly inhibited enzymatic activity. Using fibrin plate method, we found that the enzyme not only degrades fibrin directly, but also activates plasminogen into plasmin to degrade fibrin. The results indicate that the pure enzyme has a potential in dissolving blood clot, and the possibility for application in the treatment of thrombosis.

Comparative proteome analysis of Aspergillus oryzae 3.042 and A. oryzae 100-8 strains: Towards the production of different soy sauce flavors.

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Zhao, Guozhong; Hou, Lihua; Yao, Yunping; Wang, Chunling; Cao, Xiaohong

2012-07-16

Aspergillus oryzae plays a central role in soybean fermentation, particularly in its contribution to the flavor of soy sauce. We present a comparative assessment of the intracellular differences between wild-type strain 3.042 and mutant strain A100-8, at the proteome level. 522 different protein spots were identified by MALDI-TOF MS, with 134 spots being confirmed by MALDI-TOF MS/MS. Of these, 451 were differentially expressed proteins (DEPs). There was at least a two-fold increase for 288 spots, and at least a two-fold decrease for 163 spots, in strain A100-8 when compared to 3.042. Further analysis showed that 63 of the more abundant proteins were involved in glycolysis and the citrate cycle; 43 more abundant proteins and 10 less abundant proteins were related to amino acid biosynthesis and metabolism; two of the more abundant proteins were involved in vitamin biosynthesis; and five of the more abundant proteins and four of the less abundant proteins were related to secondary metabolites. Moreover, quantitative real time PCR showed that the mRNA expression levels of six typical genes we selected were consistent with changes in protein expression. We postulate that there may be a relationship between DEPs and the flavor formation mechanism in A. oryzae. Crown Copyright © 2012. Published by Elsevier B.V. All rights reserved.

Combining regio- and enantioselectivity of lipases for the preparation of (R)-4-chloro-2-butanol.

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Méndez, Jonh J; Oromi, Mireia; Cervero, Maria; Balcells, Mercè; Torres, Mercè; Canela, Ramon

2007-01-01

Preparation of 98% ee (R)-4-chloro-2-butanol was carried out by the enzymatic hydrolysis of chlorohydrin esters, using fungal resting cells and commercial enzymes. Hydrolyzes were carried out using lipases from Candida antarctica (Novozym 435), C. rugosa, Rhizomucor miehei (Lipozyme IM), Burkolia cepacia, and resting cells of Rhizopus oryzae and Aspergillus flavus. The influence of the enzyme, the solvent, the temperature, and the alkyl chain length on the selectivity of hydrolyzes of isomeric mixtures of chlorohydrin esters is described. Regioselectivity was higher than 95% for some of the tested lipases. Novozym 435 allowed preparation of the (R)-4-chloro-2-butanol after 15 min of reaction at 30-40 degrees C. (c) 2006 Wiley-Liss, Inc.

Preparation of (S)-1-Halo-2-octanols Using Ionic Liquids and Biocatalysts.

Science.gov (United States)

Oromí-Farrús, Mireia; Eras, Jordi; Sala, Núria; Torres, Mercè; Canela, Ramon