Reduction of hexavalent chromium by Rhizopus Oryzae

African Journals Online (AJOL)

EJIRO

reduction data and the specific growth rate constant value was calculated as 0.082 and the ... Key words: Hexavalent chromium, Rhizopus Oryzae, leather tanning, Monod and Haldane models. ... composition; Glucose 1 g; K2HPO4 0.5 g; NaCl 0.5 g; MgCl2 1.0 g; ... ficantly, because of the inhibitor role of high concentration.

Secretory expression of Rhizopus oryzae α-amylase in ...

African Journals Online (AJOL)

Kluyveromyces lactis is a non-conventional yeast species extensively used in the expression of heterologous genes. In this study, a genetically modified K. lactis with high-level expression of α- amylase from Rhizopus oryzae was obtained, which could successfully hydrolyze and use starch for growth very well. Shake flask ...

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.

Comparative study on the antifungal activity of hydroalcoholic extract of Iranian Propolis and Royal jelly against Rhizopus oryzae

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Moghim Hassan

2015-07-01

Full Text Available Introduction: Mucormycosis is an opportunistic fungal infection. Rhizopus oryzae is major cause of mucormycosis in humans. This disease is the most common form of the acute fungal infection with rapid progress. Iranian Propolis extract and Royal jelly are honey bee products which have been used by human over the past centuries in traditional medicine. In this study the effects of Iranian Propolis and Royal jelly were investigated against Rhizopus oryzae and Candida albicans. Methods: The used method in this study was microdilution. To perform it, the prepared dilutions of Royal jelly and alcoholic extract of Iranian Propolis were added to tubes containing Sabouraud dextrose broth culture media except to control group and then Rhizopus oryzae suspension was added to all microtubes. In the next stage, microtubes were maintained in incubator at 25oC for 48 hours and then 10 ml of the content of each microtube was transferred to Sabouraud dextrose agar media. The minimum inhibitory concentration (MIC and minimum fungicidal concentration (MFC of the agents were calculated. Results: In this study, the MIC and MFC of Iranian Propolis alcoholic extract on Rhizopus oryzae were respectively 0.1 and 0.25 mg/ml and the MFC of Royal jelly on Rhizopus oryzae were respectively 100 ± 34 and 133 ± 46 mg/ml. Conclusion: The results indicate that Propolis might be used as an ideal combination for the treatment of fungal infections like Rhizopus oryzae. However, clinical studies are needed to confirm the effects of these drugs.

Evaluasi Perlakuan Pendahuluan Menggunakan Kalsium Hidroksida untuk Biokonversi Jerami Padi Menjadi L-Asam Laktat oleh Rhizopus oryzae AT3 (Evaluation of Lime Pretreatment for Bioconversion of Rice Straw to L-Lactic Acid by Rhizopus Oryzae AT3

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Dhina Aprilia Nurani Widyahapsari

2016-12-01

Full Text Available L-lactic acid can be used as a precursor of polylactic acid (PLA. PLA is a biodegradable biomaterial commonly used for biodegradable plastics. Lactic acid can be produced from lignocelluloses materials such as rice straw. Rice straw is composed of cellulose and hemicellulose that can be hydrolyzed to fermentable sugar by cellulolytic and hemicellulolytic enzymes then converted to L-lactic acid by Rhizopus oryzae. As most cellulose and hemicellulose present in lignocellulose biomass are not readily accessible for these enzyme, pretreatment is required to alter the structure of lignocellulose substrates. This research aimed to investigate the effect of lime pretreatment on rice straw bioconversion to L-lactic acid by Rhizopus oryzae AT3. Rice straw was pretreated with lime (Ca(OH2 at 85 °C for 16 hours. Unpretreated and pretreated rice straw were hydrolyzed using crude enzyme that produced by Trichoderma reesei Pk1J2. Enzyme production was carried out by solid state fermentation using rice straw and rice brand as substrate. Enzymatic hydrolysis was carried out in flasks. Each flask was added with unpretreated or pretreated rice straw, buffer citrate solution and crude enzyme then hydrolyzed for 0-96 hours. Hydrolysate was fermented by Rhizopus oryzae AT3 for 0-6 days by using adsorbed carrier solid-state fermentation method with polyurethane foam as inert support material. Lime pretreatment at 85 °C for 16 hour led to significant solubilisation of lignin and hemicellulose. It involved lignocellulose structure modified that enhance enzymatic hydrolysis and resulted higher reducing sugars than unpretreated rice straw. The high reducing sugars was not related to high lactic acid yields. Fermentation of pretreated rice straw hydrolysate by Rhizopus oryzae AT3 did not only produce L-lactic acid but also other compound. On the other hand, fermentation of unpretreated rice straw hydrolysate only produced L-lactic acid.   ABSTRAK Polimerisasi asam

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

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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...

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 ...

Biosynthesis of tannase and gallic acid from tannin rich substrates by Rhizopus oryzae and Aspergillus foetidus.

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Mukherjee, Gargi; Banerjee, Rintu

2004-01-01

Modified solid-state fermentation (MSSF) of tannin-rich substrates for production of tannase and gallic acid was carried out using two fungal cultures, Rhizopus oryzae (RO IIT RB-13, NRRL 21498) and Aspergillus foetidus (GMRB013 MTCC 3557). The tannin rich substrates included powdered fruits of Terminalia chebula and Caesalpinia digyna pod cover powder. The different environmental parameters for the maximum production of tannase and gallic acid were optimized through media engineering. The highest yield of tannase and gallic acid was obtained after 60 h in case of Rhizopus oryzae and after 72 h by Aspergillus foetidus with 3 ml of induced inoculum. The optimum initial pH of the fermentation was found to be 4.5 in case of Rhizopus oryzae and 5.0 for Aspergillus foetidus. MSSF was carried out at the optimum conditions of 30 degrees C and 80% relative humidity. Collectively, the data reveal the potential of the modified solid-state fermentation process for the production of tannase and gallic acid from tannin-rich substrates with R. oryzae and A. foetidus.

Immobilization of Isolated Lipase From Moldy Copra (Aspergillus Oryzae

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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.

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

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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.

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.

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

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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.

Screening of allyl alcohol resistant mutant of Rhizopus oryzae and ...

African Journals Online (AJOL)

Ethanol is a main by-product in the fermentation broth of Rhizopus oryzae during the production of high-optical purity L-lactic acid. By screening the lower activity of alcohol dehydrogenase (ADH) mutant, thus decreasing the flux of pyruvic acid to ethanol may be a virtual method for increasing the conversion rate of glucose ...

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...

Pyogenic liver abscess and peritonitis due to Rhizopus oryzae in a child with Papillon-Lefevre syndrome.

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Dalgic, Buket; Bukulmez, Aysegul; Sari, Sinan

2011-06-01

Papillon-Lefevre syndrome (PLS) is an autosomal recessive disease that is characterized by symmetric palmoplantar keratodermatitis and severe periodontal destruction. Mutations in the cathepsin C gene (CTSC) have recently been detected in PLS. Immune dysregulation, due to a mutation in CTSC, increases the risk of pyogenic infections in PLS patients. A child with PLS is presented here with liver abscesses and peritonitis caused by Rhizopus oryzae. His liver abscess and peritonitis were cured with amphotericin B without surgical care. This is the first case in the literature liver abscess due to Rhizopus oryzae in a child with PLS.

Effect of Rhizopus oryzae Fermentation on Kenaf-Based Polylactic Acid’s Monomer

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Nur Aimi Mohd Nasir

2011-12-01

Full Text Available Kenaf biomass is the potential as raw materials used to produce polylactic acid's monomer which is lactic acid via fermentation by Rhizopus oryzae. Kenaf biomass' structure is complex due to its lignin and cellulose content. This matter had encouraged it to undergo pre- treatment process as the initial step before fermentation process can be done. In this paper, kenaf biomass was treated with dilute sulphuric acid (H2SO4 to hydrolyze the cellulose content in it as well as to convert the cellulose into glucose- a carbon source for Rhizopus to grow. Then, the fermentation process was carried out in shake flask for 3 days at pH 6. Several conditions for fermentation process had been chosen which were 25oC at 150 rpm, 25 oC at 200 rpm, 37 oC at 150 rpm and 37oC at 200 rpm. In this fermentation process, 0.471 g/L, 0.428 g/L, 0.444 g/L and 0.38 g/L of lactic acid was produced respectively. Sample at 25oC at 200 rpm produced maximum amount of lactic acid compared to others.ABSTRAK: Biojisim kenaf berpotensi sebagai bahan mentah dalam penghasilan monomer asid polylactic (poliester alifatik termoplastik diterbitkan daripada sumber boleh diperbaharu seperti kanji jagung yang merupakan asid laktik menerusi penapaian oleh Rhizopus oryzae (sejenis fungus yang hidup dalam jirim organik yang telah mati. Struktur biojisim kenaf adalah kompleks disebabkan kandungan lignin dan selulosanya. Hal ini menyebabkan ia perlu melalui proses pra-rawatan sebagai langkah awal sebelum proses penapaian dijalankan. Dalam kertas ini, biojirim kenaf dirawat dengan asid sulfurik (H2SO4 yang dicairkan untuk menghidrolisis kandungan selulosa di dalamnya di samping menukar selulosa menjadi glukosa - sumber karbon bagi tumbesaran Rhizopus. Kemudian, proses penapaian dijalankan di dalam kelalang goncang selama 3 hari pada pH 6. Beberapa ciri proses penapaian telah dipilih iaitu 25 oC pada 150 rpm, 25 oC pada 200 rpm, 37 oC pada 150 rpm dan 37 oC pada 200 rpm. Dalam proses penapaian

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)

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...

An unusual ulcer: A case of cutaneous mucormycosis caused by Rhizopus oryzae

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Bradley J. Gardiner

2015-03-01

Full Text Available Mucormycoses are high-mortality infections feared by clinicians worldwide. They predominantly affect immunocompromised hosts and are associated with a spectrum of disease. We describe a case of cutaneous mucormycosis caused by Rhizopus oryzae in a patient with multiple risk factors cured with complete surgical excision and a short course of antifungal therapy.

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.

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.

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.

Production of extracellular lipases by Rhizopus oligosporus in a stirred fermentor

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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.

Promoter sequence of 3-phosphoglycerate kinase gene 1 of lactic acid-producing fungus rhizopus oryzae and a method of expressing a gene of interest in fungal species

Energy Technology Data Exchange (ETDEWEB)

Gao, Johnway [Richland, WA; Skeen, Rodney S [Pendleton, OR

2002-10-15

The present invention provides the promoter clone discovery of phosphoglycerate kinase gene 1 of a lactic acid-producing filamentous fungal strain, Rhizopus oryzae. The isolated promoter can constitutively regulate gene expression under various carbohydrate conditions. In addition, the present invention also provides a design of an integration vector for the transformation of a foreign gene in Rhizopus oryzae.

Promoter sequence of 3-phosphoglycerate kinase gene 2 of lactic acid-producing fungus rhizopus oryzae and a method of expressing a gene of interest in fungal species

Energy Technology Data Exchange (ETDEWEB)

Gao, Johnway [Richland, WA; Skeen, Rodney S [Pendleton, OR

2003-03-04

The present invention provides the promoter clone discovery of phosphoglycerate kinase gene 2 of a lactic acid-producing filamentous fungal strain, Rhizopus oryzae. The isolated promoter can constitutively regulate gene expression under various carbohydrate conditions. In addition, the present invention also provides a design of an integration vector for the transformation of a foreign gene in Rhizopus oryzae.

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

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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.

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.

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)

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.

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)

Effect of Rhizopus oryzae Fermentation on Kenaf-Based Polylactic Acid’s Monomer

OpenAIRE

Nur Aimi Mohd Nasir; Mohd Adlan Mustafa Kamalbahrin; Nurhafizah Mohamad; Hazleen Anuar; Maizirwan Mel; and Rashidi Othman

2011-01-01

Kenaf biomass is the potential as raw materials used to produce polylactic acid's monomer which is lactic acid via fermentation by Rhizopus oryzae. Kenaf biomass' structure is complex due to its lignin and cellulose content. This matter had encouraged it to undergo pre- treatment process as the initial step before fermentation process can be done. In this paper, kenaf biomass was treated with dilute sulphuric acid (H2SO4) to hydrolyze the cellulose content in it as well as to convert the cell...

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...

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.

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

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...

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.

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.

Combining regio- and enantioselectivity of lipases for the preparation of (R)-4-chloro-2-butanol.

Science.gov (United States)

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.

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.

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. 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.

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.

Highly efficient biodiesel production by a whole-cell biocatalyst employing a system with high lipase expression in Aspergillus oryzae

Energy Technology Data Exchange (ETDEWEB)

Takaya, Tomohiro; Koda, Risa; Adachi, Daisuke; Ogino, Chiaki; Kondo, Akihiko [Kobe Univ. (Japan). Dept. of Chemical Science and Engineering; Nakashima, Kazunori [Kobe Univ. (Japan). Organization of Advanced Science and Technology; Wada, Junpei; Bogaki, Takayuki [Ozeki Co., Nishinomiya-shi, Hyogo (Japan)

2011-05-15

In the present study, a system with high lipase expression in Aspergillus oryzae was developed using an improved enolase promoter (P-enoA124) and the 5{sup '} untranslated region of a heat-shock protein (Hsp-UTR). P-enoA142 enhanced the transcriptional level of a heterologous lipase gene and Hsp-UTR improved its translational efficiency. Fusarium heterosporum lipase (FHL) was inserted into a pSENSU-FHL expression vector harboring P-enoA142 and Hsp-UTR and was transformed into an A. oryzae NS4 strain. Transformants possessing pSENSU-FHL in single (pSENSU-FHL1) and double copies (pSENSU-FHL2) were selected to evaluate the lipase activity of the whole-cell biocatalyst. The two strains, pSENSU-FHL1 and 2, showed excellent lipase activity in hydrolysis compared with the strain transformed with conventional expression vector pNAN8142-FHL. Furthermore, by using pSENSU-FHL2, methanolysis could proceed much more effectively without deactivation, which allowed a swift addition of methanol to the reaction mixture, thereby reducing reaction time. (orig.)

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.

Scientific Opinion on Lipase from a Genetically Modified Strain of Aspergillus oryzae (strain NZYM-LH)

OpenAIRE

EFSA Panel on Food Contact Materials, Enzymes, Flavourings and Processing Aids (CEF)

2014-01-01

The food enzyme considered in this opinion is a lipase (triacylglycerol lipase; EC 3.1.1.3) produced with a genetically modified strain of Aspergillus oryzae. The genetic modifications do not raise safety concern. The food enzyme contains neither the production organism nor recombinant DNA. The lipase is intended to be used in a number of food manufacturing processes, such as in baking and other cereal-based processes. The dietary exposure was assessed on the basis of data retrieved from the ...

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

Directory of Open Access Journals (Sweden)

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.

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.

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.

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.

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.

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. 

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

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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.

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%.

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.

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

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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.

Mechanism of acetaldehyde-induced deactivation of microbial lipases

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Jaeger Karl E

2011-02-01

Full Text Available Abstract Background Microbial lipases represent the most important class of biocatalysts used for a wealth of applications in organic synthesis. An often applied reaction is the lipase-catalyzed transesterification of vinyl esters and alcohols resulting in the formation of acetaldehyde which is known to deactivate microbial lipases, presumably by structural changes caused by initial Schiff-base formation at solvent accessible lysine residues. Previous studies showed that several lipases were sensitive toward acetaldehyde deactivation whereas others were insensitive; however, a general explanation of the acetaldehyde-induced inactivation mechanism is missing. Results Based on five microbial lipases from Candida rugosa, Rhizopus oryzae, Pseudomonas fluorescens and Bacillus subtilis we demonstrate that the protonation state of lysine ε-amino groups is decisive for their sensitivity toward acetaldehyde. Analysis of the diverse modification products of Bacillus subtilis lipases in the presence of acetaldehyde revealed several stable products such as α,β-unsaturated polyenals, which result from base and/or amino acid catalyzed aldol condensation of acetaldehyde. Our studies indicate that these products induce the formation of stable Michael-adducts at solvent-accessible amino acids and thus lead to enzyme deactivation. Further, our results indicate Schiff-base formation with acetaldehyde to be involved in crosslinking of lipase molecules. Conclusions Differences in stability observed with various commercially available microbial lipases most probably result from different purification procedures carried out by the respective manufacturers. We observed that the pH of the buffer used prior to lyophilization of the enzyme sample is of utmost importance. The mechanism of acetaldehyde-induced deactivation of microbial lipases involves the generation of α,β-unsaturated polyenals from acetaldehyde which subsequently form stable Michael-adducts with the

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.

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...

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.

Lipase - Catalyzed glycerolysis of sunflower oil to produce partial glycerides.

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Zaher, F. A.

1998-12-01

Full Text Available Partial glycerides were prepared by glycerolysis of sunflower oil in presence of lipase enzyme as catalyst. Six lipases of different origins were used and compared for their catalytic activity. These include Chromobacterium lipase, pancreatic lipase, Rhizopus arrhizus lipase, lyophilized lipase (plant lipase in addition to two lipase preparations derived from Rhizopus japonicas; Lilipase A-10 and Lilipase B-2. Chromobacterium lipase was found to be the most active as glycerolysis catalyst whereas lyophilized lipase; a plant preparation from wheat germ was the least active. The results have also shown that the lipase type affects also the product polarity and hence its field of application as a food emulsifier. Less polar products can be obtained using Chromobacterium lipase whereas the more polar ones using a fungal lipase preparation «Lipase A-10». The product polarity is also influenced by the process temperature but the mode of its effect is different for different lipases.

Se prepararon glicéridos parciales mediante glicerolisis de aceite de girasol en presencia de lipasa como catalizador. Seis lipasas de orígenes diferentes se utilizaron y compararon en función de su actividad catalítica. Estas incluyeron lipasa de Chromobacterium, lipasa pancreática, lipasa de Rhizopus arrhizus, lipasa liofilizada (lipasa vegetal además de dos preparaciones de lipasa derivadas de Rhizopus japonicus: lilipase A-10 y lilipase B-2. Se encontró que la lipasa de Chromobacterium fue la más activa como catalizador en la glicerolisis mientras que la lipasa liofilizada, preparación vegetal a partir de germen de trigo, fue la menos activa. Los resultados mostraron que los tipos de lipasa afectan también a la polaridad de los productos y por tanto a los rendimientos en su aplicación como emulsificantes alimentarios. Los productos menos polares pueden obtenerse usando lipasa de

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.

Mutation induced enhanced biosynthesis of lipase | Bapiraju ...

African Journals Online (AJOL)

The purpose of the present investigation is to enhance production of biomedically important enzyme lipase by subjecting the indigenous lipase producing strain Rhizopus sp. BTS-24 to improvement by natural selection and random mutagenesis (UV and N-methyl-N'-nitro-N-nitroso guanidine, NTG). The isolation of mutants ...

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.

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 ...

Extraction of microalgal lipids and the influence of polar lipids on biodiesel production by lipase-catalyzed transesterification.

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

2016-09-01

In order to obtain microalgal saponifiable lipids (SLs) fractions containing different polar lipid (glycolipids and phospholipids) contents, SLs were extracted from wet Nannochloropsis gaditana microalgal biomass using seven extraction systems, and the polar lipid contents of some fractions were reduced by low temperature acetone crystallization. We observed that the polar lipid content in the extracted lipids depended on the polarity of the first solvent used in the extraction system. Lipid fractions with polar lipid contents between 75.1% and 15.3% were obtained. Some of these fractions were transformed into fatty acid methyl esters (FAMEs, biodiesel) by methanolysis, catalyzed by the lipases Novozym 435 and Rhizopus oryzae in tert-butanol medium. We observed that the reaction velocity was higher the lower the polar lipid content, and that the final FAME conversions achieved after using the same lipase batch to catalyze consecutive reactions decreased in relation to an increase in the polar lipid content. Copyright © 2016 Elsevier Ltd. All rights reserved.

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

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%.

[State of Fungal Lipases of Rhizopus microsporus, Penicillium sp. and Oospora lactis in Border Layers Water-Solid Phase and Factors Affecting Catalytic Properties of Enzymes].

Science.gov (United States)

Khasanov, Kh T; Davranov, K; Rakhimov, M M

2015-01-01

We demonstrated that a change in the catalytic activity of fungal lipases synthesized by Rhizopus microsporus, Penicillium sp. and Oospora lactis and their ability to absorb on different sorbents depended on the nature of groups on the solid phase surface in the model systems water: lipid and water: solid phase. Thus, the stability of Penicillium sp. lipases increased 85% in the presence ofsorsilen or DEAE-cellulose, and 55% of their initial activity respectively was preserved. In the presence of silica gel and CM-cellulose, a decreased rate of lipid hydrolysis by Pseudomonas sp. enzymes was observed in water medium, and the hydrolysis rate increased by 2.4 and 1.5 times respectively in the presence of aminoaerosil and polykefamid. In an aqueous-alcohol medium, aminoaerosil and polykefamid decreased the rate of substrate hydrolysis by more than 30 times. The addition of aerosil to aqueous and aqueous-alcohol media resulted in an increase in the hydrolysis rate by 1.2-1.3 times. Sorsilen stabilized Penicillium sp. lipase activity at 40, 45, 50 and 55 degrees C. Either stabilization or inactivation of lipases was observed depending on the pH of the medium and the nature of chemical groups localized on the surface of solid phase. The synthetizing activity of lipases also changed depending on the conditions.

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 ...

A robust whole-cell biocatalyst that introduces a thermo- and solvent-tolerant lipase into Aspergillus oryzae cells: characterization and application to enzymatic biodiesel production.

Science.gov (United States)

Adachi, Daisuke; Koh, FookHee; Hama, Shinji; Ogino, Chiaki; Kondo, Akihiko

2013-05-10

To develop a robust whole-cell biocatalyst that works well at moderately high temperature (40-50°C) with organic solvents, a thermostable lipase from Geobacillus thermocatenulatus (BTL2) was introduced into an Aspergillus oryzae whole-cell biocatalyst. The lipase-hydrolytic activity of the immobilized A. oryzae (r-BTL) was highest at 50°C and was maintained even after an incubation of 24-h at 60°C. In addition, r-BTL was highly tolerant to 30% (v/v) organic solvents (dimethyl carbonate, ethanol, methanol, 2-propanol or acetone). The attractive characteristics of r-BTL also worked efficiently on palm oil methanolysis, resulting in a nearly 100% conversion at elevated temperature from 40 to 50°C. Moreover, r-BTL catalyzed methanolysis at a high methanol concentration without a significant loss of lipase activity. In particular, when 2 molar equivalents of methanol were added 2 times, a methyl ester content of more than 90% was achieved; the yield was higher than those of conventional whole-cell biocatalyst and commercial Candida antarctica lipase (Novozym 435). On the basis of the results regarding the excellent lipase characteristics and efficient biodiesel production, the developed whole-cell biocatalyst would be a promising biocatalyst in a broad range of applications including biodiesel production. Copyright © 2013 Elsevier Inc. All rights reserved.

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.

EFSA CEF Panel (EFSA Panel on Food Contact Materials, Enzymes, Flavourings and Processing Aids), 2014. Scientific Opinion on lipase from a genetically modified strain of Aspergillus oryzae (strain NZYM-FL)

DEFF Research Database (Denmark)

Poulsen, Morten; Hallas-Møller, Torben; Binderup, Mona-Lise

The food enzyme considered in this opinion is a lipase (triacylglycerol lipase; EC 3.1.1.3) produced with a genetically modified strain of Aspergillus oryzae. The genetic modifications do not raise safety concern. The food enzyme contains neither the production organism nor recombinant DNA...

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.

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.

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

Science.gov (United States)

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.

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

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)

Lipase production by recombinant strains of Aspergillus niger expressing a lipase-encoding gene from Thermomyces lanuginosus

DEFF Research Database (Denmark)

Prathumpai, Wai; Flitter, S.J.; Mcintyre, Mhairi

2004-01-01

Two recombinant strains of Aspergillus niger (NW 297-14 and NW297-24) producing a heterologous lipase from Thermomyces lanuginosus were constructed. The heterologous lipase was expressed using the TAKA amylase promoter from Aspergillus oryzae. The production kinetics of the two strains on different...... shows that it is possible to obtain high productivities of heterologous fungal enzymes in A. niger. However, SDS-PAGE analysis showed that most of the produced lipase was bound to the cell wall....

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

Science.gov (United States)

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.

Maximization of Intracellular Lipase Production in a Lipase-Overproducing Mutant Derivative of Rhizopus oligosporus DGM 31: A Kinetic Study

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Tehreema Iftikhar

2008-01-01

Full Text Available Regulation and maximization of lipase production in a mutant derivative of R. oligosporus has been investigated using different substrates, inoculum sizes, pH of the medium, temperature, and nitrogen sources in shake flask experiments and batch fermentation in a fermentor. The production of intracellular lipase was improved 3 times following medium optimization involving one-at-a-time approach and aeration in the fermentor. Interestingly, intracellular lipase was poorly induced by oils, instead its production was induced by sugars, mainly starch, lactose, sucrose, xylose, glucose and glycerol. Dependent variables studied were cell mass, lipase activity, lipase yield, lipase specific and volumetric rate of formation. It was confirmed that lipase production in the derepressed mutant is sufficiently uncoupled from catabolite repression. The results of average specific productivities at various temperatures worked out according to the Arrhenius equation revealed that mutation decreased the magnitude of enthalpy and entropy demand in the inactivation equilibrium during product formation, suggesting that mutation made the metabolic network of the organism thermally more stable. The highest magnitudes of volumetric productivity (QP=490 IU/(L·h and other product attributes of lipase formation occurring on optimized medium in the fermentor are greater than the values reported by other workers. The purified enzyme is monomeric in nature and exhibits stability up to 80 °C and pH=6.0–8.0. Activation energy, enthalpy and entropy of catalysis at 50 °C, and magnitudes of Gibbs free energy for substrate binding, transition state stabilization and melting point indicated that this lipase is highly thermostable.

Stereoselectivity of Mucorales lipases toward triradylglycerols--a simple solution to a complex problem.

Science.gov (United States)

Scheib, H.; Pleiss, J.; Kovac, A.; Paltauf, F.; Schmid, R. D.

1999-01-01

The lipases from Rhizopus and Rhizomucor are members of the family of Mucorales lipases. Although they display high sequence homology, their stereoselectivity toward triradylglycerols (sn-2 substituted triacylglycerols) varies. Four different triradylglycerols were investigated, which were classified into two groups: flexible substrates with rotatable O'-C1' ether or ester bonds adjacent to C2 of glycerol and rigid substrates with a rigid N'-C1' amide bond or a phenyl ring in sn-2. Although Rhizopus lipase shows opposite stereopreference for flexible and rigid substrates (hydrolysis in sn-1 and sn-3, respectively), Rhizomucor lipase hydrolyzes both groups of triradylglycerols preferably in sn-1. To explain these experimental observations, computer-aided molecular modeling was applied to study the molecular basis of stereoselectivity. A generalized model for both lipases of the Mucorales family highlights the residues mediating stereoselectivity: (1) L258, the C-terminal neighbor of the catalytic histidine, and (2) G266, which is located in a loop contacting the glycerol backbone of a bound substrate. Interactions with triradylglycerol substrates are dominated by van der Waals contacts. Stereoselectivity can be predicted by analyzing the value of a single substrate torsion angle that discriminates between sn-1 and sn-3 stereopreference for all substrates and lipases investigated here. This simple model can be easily applied in enzyme and substrate engineering to predict Mucorales lipase variants and synthetic substrates with desired stereoselectivity. PMID:10210199

Encapsulation of protease from Aspergillus oryzae and lipase from Thermomyces lanuginoseus using alginate and different copolymer types

Directory of Open Access Journals (Sweden)

Truong Thi Mong Thu

2016-05-01

Full Text Available Although the application of enzymes in food as a food processing aid and enzyme supplement is of interest and widely used, the enzymes can be easily deactivated or lose their activity due to many causes such as pH and moisture as well as through the introduction of incompatible ingredients during food processing and storage. These problems can be solved by the encapsulation technique, especially in a gel matrix. The influences were studied of the alginate concentration, types of copolymer and their concentrations on the bead size, encapsulation yield (EY, encapsulation efficiency (EE, leakage and the retention of enzyme activity during storage period of encapsulated protease from Aspergillus oryzae and lipase from Thermomyces lanuginosus beads. A solution of purified protease or lipase was encapsulated in calcium alginate-chitosan beads (CACB, calcium alginate-xanthan gum beads (CAXB and calcium alginate-maltodextrin beads (CAMB using the extrusion method. Increasing the alginate and copolymer concentrations in the solution increased the bead size, EY, EE and the retention of enzyme activity during the storage period and reduced leakage of both the encapsulated protease and lipase. In addition, different types of copolymer significantly (p ≤ 0.05 affected these properties of both encapsulated enzymes. Furthermore, protease encapsulated using 2.0% alginate and 0.2% chitosan provided the highest EY (81.7% and EE (77.2% with a bead size of 1.85 mm and 8.1% leakage. The retention of encapsulated protease activity and the shelf-life of encapsulated enzyme which was expressed as half-life, the time required for the enzyme activity to decrease by half (thalf life were 75.8% and 27.2 wk, respectively after storage at 4 °C for 10 wk. For lipase, encapsulation using 2.0% alginate and 0.4% xanthan gum provided the highest EY (42.5% and EE (43.9% and the bead size and leakage were 1.81 mm and 6.2%, respectively. The retention of encapsulated

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

Science.gov (United States)

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

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.

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,...

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

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.)

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.

Screening for Extracellular Lipase Enzymes with Transesterification Capacity in Mucoromycotina Strains

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Alexandra Kotogán

2014-01-01

Full Text Available In this study, 169 zygomycetes fungal strains including some cold-tolerant isolates were screened for their extracellular lipolytic activity towards tributyrin. Nineteen of them were outstanding in their enzyme production as they developed the largest lipolytic halo around the colonies in plate tests. Mortierella alpina, M. echinosphaera, Mucor corticolus, Rhizomucor miehei, Rhizopus oryzae, Rh. stolonifer, Umbelopsis autotrophica, U. isabellina, U. ramanniana var. angulispora and U. versiformis were selected for further studies to characterise their lipolytic enzyme production in detail. In these assays, effect of Tween 80 and palm, soybean, sunflower, olive, extra virgin olive, wheat germ, corn germ, sesame seed, pumpkin seed and cottonseed oils on the enzyme activities was investigated, and wheat bran-based submerged and solid-state fermentations were also tested. Tween 80 and olive oil proved to be efficient inductors for lipolytic enzyme production, which was also enhanced when wheat bran was used as support. Addition of mineral salts and olive oil to the solid fermentation medium resulted in at least 1.5-fold increment in the enzyme activities of the crude extracts. Organic synthesis was also assayed by the selected lipases, in which enzymes from the fungi R. miehei, Rh. stolonifer and M. echinosphaera gave the best yields during transesterification reactions between p-nitrophenyl palmitate and ethanol.

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)

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

Science.gov (United States)

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.

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.

Immobilization of lipases in PSS/PEO blends and applications in esters synthesis

International Nuclear Information System (INIS)

Vecchia, Roberto D.; Nascimento, Maria G.; Soldi, Valdir

2001-01-01

Various lipases were immobilized in PSS/PEO blends and used as bio catalysts in the esterification reaction of lauric acid with n-pentanol, in hexane as a solvent for 24 h at 35 deg C. The best results in the ester conversion, were obtained by using lipase from Rhryzopus oryzae immobilized in PSS/PEO 80:20 blend. The data are in agreement with DSC and TGA values, which showed that these systems (blend/lipase) were very stable with low mass loss. No product was obtained by using lipase FAP-15 immobilized in PSS film , showing the strong influence of the polymer on enzyme activity. (author)

Enzymatic resolution of (R,S-ibuprofen and (R,S-ketoprofen by microbial lipases from native and commercial sources Resolução enzimática do (R,S-ibuprofeno e (R,S-cetoprofeno por lipases microbianas de fontes nativas e comerciais

Directory of Open Access Journals (Sweden)

Patrícia de Oliveira Carvalho

2006-09-01

Full Text Available The enantioselectivity (E of native lipases from Aspergillus niger, Aspergillus terreus, Fusarium oxysporum, Mucor javanicus, Penicillium solitum and Rhizopus javanicus in the resolution of (R,S-ibuprofen and (R,S-ketoprofenenantiomers by esterification reaction with 1-propanol in isooctane was compared with known commercial Candida rugosa (Sigma and Candida antarctica (Novozym®435 lipases. In the resolution of (R,S-ibuprofen, C. rugosa lipase showed good selectivity (E = 12 while Novozym®435 (E = 6.7 and A. niger (E = 4.8 lipases had intermediate selectivities. Other enzymes were much less selective (E around 2.3 and 1.5, under tested conditions. After preliminary optimization of reaction conditions (water content, enzyme concentration and presence of additives the enantioselectivity of native A. niger lipase could be enhanced substantially (E = 15. All tested lipases showed low selectivity in the resolution of (R,S-ketoprofen because poor ester yields and low enantiomeric excess of the acid remaining were achieved.A enantioseletividade (E das lipases nativas de Aspergillus niger, Aspergillus terreus, Fusarium oxysporum, Mucor javanicus, Penicillium solitum e Rhizopus javanicus na resolução dos enantiômeros do (R,S-ibuprofeno e (R,S-cetoprofeno na reação de esterificação com 1-propanol em isoctano foi comparada com as lipases comerciais de Candida rugosa (Sigma e Candida antarctica (Novozym®435. A lipase de C. rugosa mostrou boa enantioseletividade (E = 12 comparada com as da Novozym®435 (E = 6.7, de A. niger (E=4.8 e com as outras lipases que foram muito menos seletivas (E por volta de 2.3 e 1.5 na resolução do (R,S-ibuprofeno, dentro das condições testadas. Após uma otimização preliminar das condições da reação (conteúdo de água, concentração da enzima e presença de aditivos a enantioseletividade da lipase de A. niger pôde ser substancialmente aumentada (E = 15. Todas as lipases testadas mostraram baixa

OPTIMASI PRODUKSI ENZIMATIS DIASILGLISEROL MELALUI GLISEROLISIS KONTINU [Optimization of Enzymatic Diacylglycerol Production through Continuous Glycerolysis

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Tri-Panji*

2014-06-01

Full Text Available Diacylglycerol (DAG produced from crude palm oil (CPO is one of the healthy oils that can be consumed for daily human diet. DAG production in Indonesia is constrained by the high cost of the mostly imported lipase. To overcome this problem, research of DAG production has been carried out using crude extracts of lipase produced by local species of fungi Rhizopus oryzae. This study aims to develop a continuous process of enzymatic glycerolysis of CPO for DAG production; to establish optimum conditions of DAG production which includes flow rate of CPO and glycerolysis time; and to test the performance of lipase from the local mold R. oryzae in catalyzing continuous process of glycerolysis for the production of DAG. Lipase isolation was carried out by acetone precipitation and lipase was used as a catalyst in the continuous glycerolysis process. The glycerolysis was conducted by reacting CPO with glycerol continuously at various time periods. The optimum condition of automatic continuous glycerolysis process was achieved at a CPO flow rate of 3 mL/min with a glycerolysis time at the 18 cycles (9 hours. The conversion of DAG was 29%. The performance of lipase was proven to remain stable up to 3 times changes of CPO substrate for 9 hours of glycerolysis process with the best condition at the 3 cycles and can improved conversion of DAG until 37%.

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.

Ecological screening of lipolytic cultures and process optimization for extracellular lipase production from fungal hyperproducer

International Nuclear Information System (INIS)

Iftikhar, T.; Niaz, M.; Anwer, M.; Abbas, S.Q.; Saleem, M.; Jabeen, R.

2011-01-01

Present investigation describes the biosynthesis of extracellular lipases by various local fungal strains isolated from various lipid rich habitats of Faisalabad. The isolated cultures of Aspergillus niger, Penicillium chrysogenum, Rhizopus microsporus, Mucor mucedo, Alternaria alternata, Trichophyton sp., Fusarium semitectum, E (un-identified), Curvularia sp., Aspergillus flavus, G (un-identified), F (Mucor sp.) and H (Synnematous) were identified and screened for the extracelluler lipases production. Different environmental parameters such as pH, temperature, inoculum size, amount of substrate and incubation time were optimized for the selected hyper producer. It was found that maximum production of lipases by Trichophyton sp., was obtained after 48 h of batch fermentation. Similarly, the diluent pH of 7.0 and incubation temperature of 30 deg. C were found optimum for enzyme production by the microorganism. The maximum production of lipases during the course of present studies was 65.20 +- 1.13a U/g. (author)

Lipase immobilized on the hydrophobic polytetrafluoroethene membrane with nonwoven fabric and its application in intensifying synthesis of butyl oleate.

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Wang, Shu-Guang; Zhang, Wei-Dong; Li, Zheng; Ren, Zhong-Qi; Liu, Hong-Xia

2010-11-01

The synthesis of butyl oleate was studied in this paper with immobilized lipase. Five types of membrane were used as support to immobilize Rhizopus arrhizus lipase by following a procedure combining filtration and protein cross-linking. Results showed that hydrophobic polytetrafluoroethene membrane with nonwoven fabric (HO-PTFE-NF) was the favorite choice in terms of higher protein loading, activity, and specific activity of immobilized lipase. The factors including solvent polarity, lipase dosage, concentration, and molar ratio of substrate and temperature were found to have significant influence on conversion. Results showed that hexane (logP = 3.53) was a favorable solvent for the biosynthesis of butyl oleate in our studies. The optimal conditions were experimentally determined of 50 U immobilized lipase, molar ratio of oleic acid to butanol of 1.0, substrate concentration of 0.12 mol/L, temperature of 37 °C, and reaction time of 2 h. The conversion was beyond 91% and decreased slightly after 18 cycles. Lipase immobilization can improve the conversion and the repeated use of immobilized lipase relative to free lipase.

Multifunctionality and diversity of GDSL esterase/lipase gene family in rice (Oryza sativa L. japonica genome: new insights from bioinformatics analysis

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Chepyshko Hanna

2012-07-01

Full Text Available Abstract Background GDSL esterases/lipases are a newly discovered subclass of lipolytic enzymes that are very important and attractive research subjects because of their multifunctional properties, such as broad substrate specificity and regiospecificity. Compared with the current knowledge regarding these enzymes in bacteria, our understanding of the plant GDSL enzymes is very limited, although the GDSL gene family in plant species include numerous members in many fully sequenced plant genomes. Only two genes from a large rice GDSL esterase/lipase gene family were previously characterised, and the majority of the members remain unknown. In the present study, we describe the rice OsGELP (Oryza sativa GDSL esterase/lipase protein gene family at the genomic and proteomic levels, and use this knowledge to provide insights into the multifunctionality of the rice OsGELP enzymes. Results In this study, an extensive bioinformatics analysis identified 114 genes in the rice OsGELP gene family. A complete overview of this family in rice is presented, including the chromosome locations, gene structures, phylogeny, and protein motifs. Among the OsGELPs and the plant GDSL esterase/lipase proteins of known functions, 41 motifs were found that represent the core secondary structure elements or appear specifically in different phylogenetic subclades. The specification and distribution of identified putative conserved clade-common and -specific peptide motifs, and their location on the predicted protein three dimensional structure may possibly signify their functional roles. Potentially important regions for substrate specificity are highlighted, in accordance with protein three-dimensional model and location of the phylogenetic specific conserved motifs. The differential expression of some representative genes were confirmed by quantitative real-time PCR. The phylogenetic analysis, together with protein motif architectures, and the expression profiling were

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

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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.

Lipase immobilization and production of fatty acid methyl esters from canola oil using immobilized lipase

International Nuclear Information System (INIS)

Yuecel, Yasin; Demir, Cevdet; Dizge, Nadir; Keskinler, Buelent

2011-01-01

Lipase enzyme from Aspergillus oryzae (EC 3.1.1.3) was immobilized onto a micro porous polymeric matrix which contains aldehyde functional groups and methyl esters of long chain fatty acids (biodiesel) were synthesized by transesterification of crude canola oil using immobilized lipase. Micro porous polymeric matrix was synthesized from styrene-divinylbenzene (STY-DVB) copolymers by using high internal phase emulsion technique and two different lipases, Lipozyme TL-100L ® and Novozym 388 ® , were used for immobilization by both physical adsorption and covalent attachment. Biodiesel production was carried out with semi-continuous operation. Methanol was added into the reactor by three successive additions of 1:4 M equivalent of methanol to avoid enzyme inhibition. The transesterification reaction conditions were as follows: oil/alcohol molar ratio 1:4; temperature 40 o C and total reaction time 6 h. Lipozyme TL-100L ® lipase provided the highest yield of fatty acid methyl esters as 92%. Operational stability was determined with immobilized lipase and it indicated that a small enzyme deactivation occurred after used repeatedly for 10 consecutive batches with each of 24 h. Since the process is yet effective and enzyme does not leak out from the polymer, the method can be proposed for industrial applications. -- Research highlights: → Lipozyme TL-100L and Novozym 388 were immobilized onto micro porous polymeric matrix by both physical adsorption and covalent linking. → Immobilized enzymes were used for synthesis of fatty acid methyl esters by transesterification of canola oil and methanol using semi-continuous operation system. → According to chromatographic analysis, Lipase Lipozyme TL-100L resulted in the highest yield of methyl ester as 92%.

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...

Production of oleic acid ethyl ester catalyzed by crude rice bran (Oryza sativa lipase in a modified fed-batch system: problem and its solution

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Indro Prastowo

2015-01-01

Full Text Available A fed-batch system was modified for the enzymatic production of Oleic Acid Ethyl Ester (OAEE using rice bran (Oryza sativa lipase by retaining the substrate molar ratio (ethanol/oleic acid at 2.05: 1 during the reaction. It resulted in an increase in the ester conversion up to 76.8% in the first 6 h of the reaction, and then followed by a decrease from 76.8% to 22.9% in 6 h later. Meanwhile, the production of water in the reaction system also showed a similar trend to the trend of ester production. The water was hypothesized to lead lipase to reverse the reaction which resulted in a decrease in both (water and esters in the last 6 h of the reaction. In order to overcome the problem, zeolite powders (25 and 50 mg/ml were added into the reaction system at 5 h of the reaction. As the result, final ester conversions increased drastically up to 90 - 95.7% (1.17 – 1.24 times. The addition also proved a hypothesis that the water was involved in reducing the ester conversion in the last 6 h of the reaction. Thus, the combination was effective to produce the high final ester conversion.

The effect of olive cake types on lipase production by isolated Rhizopus sp. and process statistical optimization

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Gholam Khayati

2013-01-01

Full Text Available The aim of this work was to study the production of extracellular lipase by solid-state fermentation with different olive cakes varieties including Mary, Shenghe and Yellow from isolated fungi using agro-industries waste such as rice straw, rice barn and wheat straw. The highest yields of enzyme were obtained in solid-state fermentation using rice straw as solid substrate in combination with 40% Mary olive cakes as inducer. The initial screening by using Plackett-Burman's design demonstrated that among the tested factors, lactose and ammonium sulfate of the medium significantly (p < 0.05 enhanced the lipase production. Further optimization of lipase production by isolated fungi in solid-state fermentation by applying response surface methodology was achieved, which revealed these as follows: 0.42 (% w/v for lactose and 0.09 (% w/v for ammonium sulfate. Also the enzyme kinetics parameters, biochemical properties, thermodynamic of thermal deactivation and deactivation rate constant of enzyme were determined.

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.

Biochemical Properties of a New Cold-Active Mono- and Diacylglycerol Lipase from Marine Member Janibacter sp. Strain HTCC2649

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Dongjuan Yuan

2014-06-01

Full Text Available Mono- and di-acylglycerol lipase has been applied to industrial usage in oil modification for its special substrate selectivity. Until now, the reported mono- and di-acylglycerol lipases from microorganism are limited, and there is no report on the mono- and di-acylglycerol lipase from bacteria. A predicted lipase (named MAJ1 from marine Janibacter sp. strain HTCC2649 was purified and biochemical characterized. MAJ1 was clustered in the family I.7 of esterase/lipase. The optimum activity of the purified MAJ1 occurred at pH 7.0 and 30 °C. The enzyme retained 50% of the optimum activity at 5 °C, indicating that MAJ1 is a cold-active lipase. The enzyme activity was stable in the presence of various metal ions, and inhibited in EDTA. MAJ1 was resistant to detergents. MAJ1 preferentially hydrolyzed mono- and di-acylglycerols, but did not show activity to triacylglycerols of camellia oil substrates. Further, MAJ1 is low homologous to that of the reported fungal diacylglycerol lipases, including Malassezia globosa lipase 1 (SMG1, Penicillium camembertii lipase U-150 (PCL, and Aspergillus oryzae lipase (AOL. Thus, we identified a novel cold-active bacterial lipase with a sn-1/3 preference towards mono- and di-acylglycerides for the first time. Moreover, it has the potential, in oil modification, for special substrate selectivity.

Preparation of (S)-1-Halo-2-octanols Using Ionic Liquids and Biocatalysts.

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Oromí-Farrús, Mireia; Eras, Jordi; Sala, Núria; Torres, Mercè; Canela, Ramon

2009-10-23

Preparation of (S)-1-chloro-2-octanol and (S)-1-bromo-2-octanol was carried out by the enzymatic hydrolysis of halohydrin palmitates using biocatalysts. Halohydrin palmitates were prepared by various methods from palmitic acid and 1,2-octanediol. A tandem hydrolysis was carried out using lipases from Candida antarctica (Novozym 435), Rhizomucor miehei (Lipozyme IM), and "resting cells" from a Rhizopus oryzae strain that was not mycotoxigenic. The influence of the enzyme and the reaction medium on the selective hydrolysis of isomeric mixtures of halohydrin esters is described. Novozym 435 allowed preparation of (S)-1-chloro-2-octanol and (S)-1-bromo-2-octanol after 1-3 h of reaction at 40 degrees C in [BMIM][PF(6)].

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

Engineering a disulfide bond in the lid hinge region of Rhizopus chinensis lipase: increased thermostability and altered acyl chain length specificity.

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Xiao-Wei Yu

Full Text Available The key to enzyme function is the maintenance of an appropriate balance between molecular stability and structural flexibility. The lid domain which is very important for "interfacial activation" is the most flexible part in the lipase structure. In this work, rational design was applied to explore the relationship between lid rigidity and lipase activity by introducing a disulfide bond in the hinge region of the lid, in the hope of improving the thermostability of R. chinensis lipase through stabilization of the lid domain without interfering with its catalytic performance. A disulfide bridge between F95C and F214C was introduced into the lipase from R. chinensis in the hinge region of the lid according to the prediction of the "Disulfide by Design" algorithm. The disulfide variant showed substantially improved thermostability with an eleven-fold increase in the t(1/2 value at 60°C and a 7°C increase of T(m compared with the parent enzyme, probably contributed by the stabilization of the geometric structure of the lid region. The additional disulfide bond did not interfere with the catalytic rate (k(cat and the catalytic efficiency towards the short-chain fatty acid substrate, however, the catalytic efficiency of the disulfide variant towards pNPP decreased by 1.5-fold probably due to the block of the hydrophobic substrate channel by the disulfide bond. Furthermore, in the synthesis of fatty acid methyl esters, the maximum conversion rate by RCLCYS reached 95% which was 9% higher than that by RCL. This is the first report on improving the thermostability of the lipase from R. chinensis by introduction of a disulfide bond in the lid hinge region without compromising the catalytic rate.

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.

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

Directory of Open Access Journals (Sweden)

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

Comparative analyses of lipoprotein lipase, hepatic lipase, and endothelial lipase, and their binding properties with known inhibitors.

Directory of Open Access Journals (Sweden)

Ziyun Wang

Full Text Available The triglyceride lipase gene subfamily plays a central role in lipid and lipoprotein metabolism. There are three members of this subfamily: lipoprotein lipase, hepatic lipase, and endothelial lipase. Although these lipases are implicated in the pathophysiology of hyperlipidemia and atherosclerosis, their structures have not been fully solved. In the current study, we established homology models of these three lipases, and carried out analysis of their activity sites. In addition, we investigated the kinetic characteristics for the catalytic residues using a molecular dynamics simulation strategy. To elucidate the molecular interactions and determine potential key residues involved in the binding to lipase inhibitors, we analyzed the binding pockets and binding poses of known inhibitors of the three lipases. We identified the spatial consensus catalytic triad "Ser-Asp-His", a characteristic motif in all three lipases. Furthermore, we found that the spatial characteristics of the binding pockets of the lipase molecules play a key role in ligand recognition, binding poses, and affinities. To the best of our knowledge, this is the first report that systematically builds homology models of all the triglyceride lipase gene subfamily members. Our data provide novel insights into the molecular structures of lipases and their structure-function relationship, and thus provides groundwork for functional probe design towards lipase-based therapeutic inhibitors for the treatment of hyperlipidemia and atherosclerosis.

Change in hyphal morphology of Aspergillus Oryzae during fed-batch cultivation

DEFF Research Database (Denmark)

Haack, Martin Brian; Olsson, Lisbeth; Hansen, K

2006-01-01

the batch phase from 2.8-2.9 up to 4.0-4.4 mu m. The diameter of the hyphal elements remained constant, around 4 mu m, after the feed was started. However, the diameter of the immediate hyphal tip, where the enzyme secretion is thought to take place, increased dramatically with up to a factor 2.5 during......Industrial enzymes are often produced by filamentous fungi in fed-batch cultivations. During cultivation, the different morphological forms displayed by the fungi have an impact on the overall production. The morphology of a recombinant lipase producing Aspergillus oryzae strain was investigated...

Synthesis of 2-monoacylglycerols and structured triacylglycerols rich in polyunsaturated fatty acids by enzyme catalyzed reactions.

Science.gov (United States)

Rodríguez, Alicia; Esteban, Luis; Martín, Lorena; Jiménez, María José; Hita, Estrella; Castillo, Beatriz; González, Pedro A; Robles, Alfonso

2012-08-10

This paper studies the synthesis of structured triacylglycerols (STAGs) by a four-step process: (i) obtaining 2-monoacylglycerols (2-MAGs) by alcoholysis of cod liver oil with several alcohols, catalyzed by lipases Novozym 435, from Candida antartica and DF, from Rhizopus oryzae, (ii) purification of 2-MAGs, (iii) formation of STAGs by esterification of 2-MAGs with caprylic acid catalyzed by lipase DF, from R. oryzae, and (iv) purification of these STAGs. For the alcoholysis of cod liver oil, absolute ethanol, ethanol 96% (v/v) and 1-butanol were compared; the conditions with ethanol 96% were then optimized and 2-MAG yields of around 54-57% were attained using Novozym 435. In these 2-MAGs, DHA accounted for 24-31% of total fatty acids. In the operational conditions this lipase maintained a stable level of activity over at least 11 uses. These results were compared with those obtained with lipase DF, which deactivated after only three uses. The alcoholysis of cod liver oil and ethanol 96% catalyzed by Novozym 435 was scaled up by multiplying the reactant amounts 100-fold and maintaining the intensity of treatment constant (IOT=3g lipase h/g oil). In these conditions, the 2-MAG yield attained was about 67%; these 2-MAGs contained 36.6% DHA. The synthesized 2-MAGs were separated and purified from the alcoholysis reaction products by solvent extraction using solvents of low toxicity (ethanol and hexane); 2-MAG recovery yield and purity of the target product were approximately 96.4% and 83.9%, respectively. These 2-MAGs were transformed to STAGs using the optimal conditions obtained in a previous work. After synthesis and purification, 93% pure STAGs were obtained, containing 38% DHA at sn-2 position and 60% caprylic acid (CA) at sn-1,3 positions (of total fatty acids at these positions), i.e. the major TAG is the STAG with the structure CA-DHA-CA. Copyright © 2012 Elsevier Inc. All rights reserved.

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

A novel biocatalytic approach to acetylation of 1-β-D-arabinofuranosylcytosine by Aspergillus oryzae whole cell in organic solvents.

Science.gov (United States)

Li, Xiao-Feng; Zhu, Zhen; Zhao, Guang-Lei; Yu, Yi-Gang; Lai, Fu-Rao; Wu, Hui

2012-01-01

Biocatalytic acylation of 1-β-D-arabinofuranosylcytosine (ara-C) was developed using whole cell of Aspergillus oryzae as a novel catalyst. (13)C nuclear magnetic resonance (NMR) analysis indicated that the whole-cell biocatalyst had more specific activity toward the 3'-hydroxyl group than 5'-hydroxyl group among the available hydroxyl groups in sugar moiety of ara-C. Except for glucose and maltose, 11 carbon sources supplemented to basal media, including Spans, Tweens, olive oil and oleic acid, exhibited notable enhancement effects on both the cell growth and the acylation reactions. It was suggested that the carbon sources containing controlled-release oleic acid were the important substrates for the production of fungal cell-bound lipase with specific activity, partially due to a gradual induction effect of their released oleic acid on the cell-bound lipase production. Despite the low initial reaction rate and substrate conversion, the addition of 2.0 g/l Span 80 resulted in a higher 3'-regioselectivity of the cells than 81%. By using Tween 85 at its optimum concentration of 5.0 g/l, however, the highest initial rates (3.2 mmol/l h) and substrate conversion (76%) of the whole-cell catalyzed acylation of ara-C can be achieved. It was also found that the 3'-regioselectivity of the cells showed observable increase by extending the culture time. And the activity of cell-bound lipase drastically increased in the early stage of cell growth and then declined in the late culture stage, whatever the culture media used. Our results thus indicated that A. oryzae whole cell was a promising green tool for biosynthesis of nucleoside esters with potential bioactivities.

Improvement of selected strains through gamma irradiation for enhanced lipolytic potential

International Nuclear Information System (INIS)

Iftikhar, T.; Mubashir, N.; Hussain, Y.; Abbas, S.Q.; Ashraf, I.

2010-01-01

The purpose of the present investigation was to enhance the production of industrially important enzyme lipase by subjecting the wild lipase producing fungal strains i.e. Aspergillus niger, Rhizopus microsporus and Penicillium atrovenetum to various doses of gamma irradiation (20, 40, 60, 80, 100, 120, 140 and 160 Gy). The isolation and lipolytic activity of selected mutant derived strains is described in this paper. Among all the mutants tested, MBL-5 obtained at 140Gy of Aspergillus niger strain showed highest extracellular lipase activity (13.75 +- 0.15 U mL/sup -1/) while MBL-1 Rhizopus microsporus at the rate 20Gy showed the lowest activity i.e., 1.06 +- 0.11 U mL/sup -1/. A range of pH 3, 5, 7, 9 and 11 was used to check the lipolytic potential of various mutants along with their wild type. It was observed that MBL-5 (Aspergillus niger) and MBL-2 (Rhizopus microsporus) showed enhanced extracellular lipase activity at pH 11 while MBL-3 (Penicillium atrovenetum) showed the highest extracellular lipase activity 22.53 +- 0.21 U mL/sup -1/ at pH 9. It indicates a possible role for the MBL-2, MBL-3 and MBL-5 mutant strains in the detergent industry for the development of eco-friendly technologies. (author)

Evaluation of cellulose-binding domain fused to a lipase for the lipase immobilization.

Science.gov (United States)

Hwang, Sangpill; Ahn, Jungoh; Lee, Sumin; Lee, Tai Gyu; Haam, Seungjoo; Lee, Kangtaek; Ahn, Ik-Sung; Jung, Joon-Ki

2004-04-01

A cellulose-binding domain (CBD) fragment of a cellulase gene of Trichoderma hazianum was fused to a lipase gene of Bacillus stearothermophilus L1 to make a gene cluster for CBD-BSL lipase. The specific activity of CBD-BSL lipase for oil hydrolysis increased by 33% after being immobilized on Avicel (microcrystalline cellulose), whereas those of CBD-BSL lipase and BSL lipase decreased by 16% and 54%, respectively, after being immobilized on silica gel. Although the loss of activity of an enzyme immobilized by adsorption has been reported previously, the loss of activity of the CBD-BSL lipase immobilized on Avicel was less than 3% after 12 h due to the irreversible binding of CBD to Avicel.

Bacterial lipases

NARCIS (Netherlands)

Jaeger, Karl-Erich; Ransac, Stéphane; Dijkstra, Bauke W.; Colson, Charles; Heuvel, Margreet van; Misset, Onno

Many different bacterial species produce lipases which hydrolyze esters of glycerol with preferably long-chain fatty acids. They act at the interface generated by a hydrophobic lipid substrate in a hydrophilic aqueous medium. A characteristic property of lipases is called interfacial activation,

Bacterial lipases for biotechnological applications

NARCIS (Netherlands)

Jaeger, Karl-Erich; Schneidinger, Bernd; Rosenau, Frank; Werner, Michael; Lang, Dietmar; Dijkstra, Bauke W.; Schimossek, Klaus; Zonta, Albin; Reetz, Manfred T.

1997-01-01

Lipase genes originating from the Gram-negative bacteria Serrutiu marcescens and Pseudomonus urruginosa were cloned. S. marcescens lipase was overexpressed in Escherichia coli yielding inclusion bodies which were purified and finally refolded to give enzymatically active lipase. The lipase operon of

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...

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

Directory of Open Access Journals (Sweden)

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

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/.

High-level extracellular production and characterization of Candida antarctica lipase B in Pichia pastoris.

Science.gov (United States)

Eom, Gyeong Tae; Lee, Seung Hwan; Song, Bong Keun; Chung, Keun-Wo; Kim, Young-Wun; Song, Jae Kwang

2013-08-01

The gene encoding lipase B from Candida antarctica (CalB) was expressed in Pichia pastoris after it was synthesized by the recursive PCR and cloned into the Pichia expression plasmid, pPICZαA. The CalB was successfully secreted in the recombinant P. pastoris strain X-33 with an apparent molecular weight of 34 kDa. For 140 h flask culture, the dry cell weight and the extracellular lipase activity reached at 5.4 g/l and 57.9 U/l toward p-nitrophenyl palmitate, respectively. When we performed the fed-batch fermentation using a methanol feeding strategy for 110 h, the dry cell weight and the extracellular lipase activity were increased to 135.7 g/l and 11,900 U/l; the CalB protein concentration was 1.18 g/l of culture supernatant. The characteristics of CalB recovered from the P. pastoris culture were compared with the commercial form of CalB produced in Aspergillus oryzae. The kinetic constants and specific activity, the effects of activity and stability on temperature and pH, the glycosylation extent, the degree of immobilization on macroporous resin and the yield of esterification reaction between oleic acid and n-butanol were almost identical to each other. Therefore, we successfully proved that the Pichia-based expression system for CalB in this study was industrially promising compared with one of the most efficient production systems. Copyright © 2013 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

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.

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.

Acid Lipase Disease

Science.gov (United States)

... of Neurological Disorders and Stroke conducts and supports research to understand lipid storage diseases such as acid lipase deficiency and ... of Neurological Disorders and Stroke conducts and supports research to understand lipid storage diseases such as acid lipase deficiency and ...

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.

A cell wall-degrading esterase of Xanthomonas oryzae requires a unique substrate recognition module for pathogenesis on rice.

Science.gov (United States)

Aparna, Gudlur; Chatterjee, Avradip; Sonti, Ramesh V; Sankaranarayanan, Rajan

2009-06-01

Xanthomonas oryzae pv oryzae (Xoo) causes bacterial blight, a serious disease of rice (Oryza sativa). LipA is a secretory virulence factor of Xoo, implicated in degradation of rice cell walls and the concomitant elicitation of innate immune responses, such as callose deposition and programmed cell death. Here, we present the high-resolution structural characterization of LipA that reveals an all-helical ligand binding module as a distinct functional attachment to the canonical hydrolase catalytic domain. We demonstrate that the enzyme binds to a glycoside ligand through a rigid pocket comprising distinct carbohydrate-specific and acyl chain recognition sites where the catalytic triad is situated 15 A from the anchored carbohydrate. Point mutations disrupting the carbohydrate anchor site or blocking the pocket, even at a considerable distance from the enzyme active site, can abrogate in planta LipA function, exemplified by loss of both virulence and the ability to elicit host defense responses. A high conservation of the module across genus Xanthomonas emphasizes the significance of this unique plant cell wall-degrading function for this important group of plant pathogenic bacteria. A comparison with the related structural families illustrates how a typical lipase is recruited to act on plant cell walls to promote virulence, thus providing a remarkable example of the emergence of novel functions around existing scaffolds for increased proficiency of pathogenesis during pathogen-plant coevolution.

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

Quantitative Metabolomics and Instationary 13C-Metabolic Flux Analysis Reveals Impact of Recombinant Protein Production on Trehalose and Energy Metabolism in Pichia pastoris

Directory of Open Access Journals (Sweden)

Joel Jordà

2014-05-01

Full Text Available Pichia pastoris has been recognized as an effective host for recombinant protein production. In this work, we combine metabolomics and instationary 13C metabolic flux analysis (INST 13C-MFA using GC-MS and LC-MS/MS to evaluate the potential impact of the production of a Rhizopus oryzae lipase (Rol on P. pastoris central carbon metabolism. Higher oxygen uptake and CO2 production rates and slightly reduced biomass yield suggest an increased energy demand for the producing strain. This observation is further confirmed by 13C-based metabolic flux analysis. In particular, the flux through the methanol oxidation pathway and the TCA cycle was increased in the Rol-producing strain compared to the reference strain. Next to changes in the flux distribution, significant variations in intracellular metabolite concentrations were observed. Most notably, the pools of trehalose, which is related to cellular stress response, and xylose, which is linked to methanol assimilation, were significantly increased in the recombinant strain.

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 Tallow As Lipase Inducer on Total of Aspergillus Niger, Lipolitic Activity and Lipase Yield

Directory of Open Access Journals (Sweden)

Manik Eirry Sawitri

2012-02-01

Full Text Available The objectives of this research was to determined of tallow addition with different concentration as lipase Aspergillus niger inducer to total of A. niger, lipolitic activity and lipase yield. The result showed that tallow addition as inducer in the lipase A. niger production gave no significant effect on total of A. niger (5.3 x 107 – 1.7 x 108 cfu/gram in the medium. Tallow addition gave a highly significant effect on lipolytic activity and yield of lipase A. niger. Lipolytic activity ranged between 32.0354 – 53.1197 U/mg protein, while the yield of lipase was 6.6418–7.8941 µg/ml. The conclusion of this research was the addition of tallow for 8% as the lipase inducer of A. niger on lipase production was  more effective to obtain the optimal result. Keywords : Tallow, lipase, inducer, Aspergillus niger

Lipase H, a new member of the triglyceride lipase family synthesized by the intestine

NARCIS (Netherlands)

Jin, Weijun; Broedl, Uli C.; Monajemi, Houshang; Glick, Jane M.; Rader, Daniel J.

2002-01-01

We report here the molecular cloning of a novel member of the triglyceride lipase family, a 2.4-kb cDNA encoding human lipase H (LIPH) and the mouse ortholog (Liph). The human LIPH cDNA encodes a 451-amino-acid protein with a lipase domain. Mouse Liph shows 85% amino acid identity and 75% nucleotide

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.

Activity and stability of immobilized lipases in lipase-catalyzed modification of peanut oil

OpenAIRE

Soumanou Mohamed M.; Edorh Aleodjrodo P.; Bornscheuer Uwe T.

2004-01-01

Fatty acid release during lipolysis of peanut oil using microbial free and immobilized lipases in aqueous media was developed. Immobilized lipase from Rhizomucor miehei (RML) gave the best result from its ability to clive different fatty acids from peanut oil in such media. In organic solvent, interesterification of peanut oil with tricaprylin using immobilized lipases from RML, Chromobacterium viscosum (CVL) and Candida rugosa (CRL) was performed. The best substrate molar ratio of tricapryli...

Biodiesel production with immobilized lipase: A review.

Science.gov (United States)

Tan, Tianwei; Lu, Jike; Nie, Kaili; Deng, Li; Wang, Fang

2010-01-01

Fatty acid alkyl esters, also called biodiesel, are environmentally friendly and show great potential as an alternative liquid fuel. Biodiesel is produced by transesterification of oils or fats with chemical catalysts or lipase. Immobilized lipase as the biocatalyst draws high attention because that process is "greener". This article reviews the current status of biodiesel production with immobilized lipase, including various lipases, immobilization methods, various feedstocks, lipase inactivation caused by short chain alcohols and large scale industrialization. Adsorption is still the most widely employed method for lipase immobilization. There are two kinds of lipase used most frequently especially for large scale industrialization. One is Candida antartica lipase immobilized on acrylic resin, and the other is Candida sp. 99-125 lipase immobilized on inexpensive textile membranes. However, to further reduce the cost of biodiesel production, new immobilization techniques with higher activity and stability still need to be explored. Copyright 2010 Elsevier Inc. All rights reserved.

Organic Solvent Tolerant Lipases and Applications

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Shivika Sharma

2014-01-01

Full Text Available Lipases are a group of enzymes naturally endowed with the property of performing reactions in aqueous as well as organic solvents. The esterification reactions using lipase(s could be performed in water-restricted organic media as organic solvent(s not only improve(s the solubility of substrate and reactant in reaction mixture but also permit(s the reaction in the reverse direction, and often it is easy to recover the product in organic phase in two-phase equilibrium systems. The use of organic solvent tolerant lipase in organic media has exhibited many advantages: increased activity and stability, regiospecificity and stereoselectivity, higher solubility of substrate, ease of products recovery, and ability to shift the reaction equilibrium toward synthetic direction. Therefore the search for organic solvent tolerant enzymes has been an extensive area of research. A variety of fatty acid esters are now being produced commercially using immobilized lipase in nonaqueous solvents. This review describes the organic tolerance and industrial application of lipases. The main emphasis is to study the nature of organic solvent tolerant lipases. Also, the potential industrial applications that make lipases the biocatalysts of choice for the present and future have been presented.

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 ...

FRAKSINASI ENZIM LIPASE DARI ENDOSPERM KELAPA DENGAN METODE SALTING OUT (Lipase fractionation of Coconut Endosperm by Salting out Method

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Moh. Su'i

2014-02-01

Full Text Available This research learns about fractionation of lipases activity from coconut endosperm by using ammonium sulphate of 0–15%; 15-30 %, 30–45 %, 45–60 %, 60–75 % and 75–90 %. The results showed that the fractions of 0–15% ; 30–45 %, 45–60 % and 60–75 % have lipase activity. Meanwhile, the highest activity was fractions of 60-75%. fractions of 15-30% and 75-90%  have no lipase enzym activity. Molecule weigh of lipase enzyme was 72 kDa. Keywords: Lipases, endosperm, coconut, fractionation, ammonium sulphate   ABSTRAK Penelitian ini mempelajari fraksinasi enzim lipase dari endosperm kelapa menggunakan ammonium sulfat. fraksinasi dilakukan dengan variasi konsentrasi ammonium sulfat 0–15% ; 15-30%; 30–45 %, 45–60 %, 60–75 % dan 75–90 %. Hasil penelitian menunjukkan bahwa enzim lipase terdapat pada fraksi 0–15% ; 30–45 %, 45–60 % dan fraksi 60–75 % dengan aktivitas enzim tertinggi pada fraksi 60-75%. Sedangkan fraksi 15-30% dan 75-90% tidak ada enzim lipase. Berat molekul enzim lipase pada semua fraksi 72 kDa. Kata kunci: Lipase, endosperm, fraksinasi, ammonium sulfat

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

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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.

Lipase kinetics: hydrolysis of triacetin by lipase from Candida cylindracea in a hollow-fiber membrane reactor

NARCIS (Netherlands)

Guit, R.P.M.; Kloosterman, M.; Meindersma, G.W.; Mayer, M.; Meijer, E.M.

1991-01-01

The aptitude of a hollow-fiber membrane reactor to det. lipase kinetics was investigated using the hydrolysis of triacetin catalyzed by lipase from C. cylindracea as a model system. The binding of the lipase to the membrane appears not to be very specific (surface adsorption), and probably its

OPTIMASI ISOLASI LIPASE INDIGENOUS BIJI KAKAO (Theobroma cacao L. The Optimizing of Isolation of Cocoa Bean Indogenous Lipase (Theobroma cacao L.

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I D. G. Mayun Permana

2012-05-01

Full Text Available The aim of the research is to optimize the isolation method of cocoa bean lipase. The research is held by determining the position of lipase on cocoa bean, varying extraction medium and isolation process. The result shows that the lipase of cocoa bean is   cytosolic enzyme. The defatting process do not increase the lipase activity. Polyphenols inhibit the lipase activity, so that removal of the polyphenol will increase the activity. Blocking the polyphenol with polyvinilpolypirrolidone (PVPP will also increase the activity.The optimum consentration of PVPP is 8 %. The lipase activity will reach the highest when homogenized for 10 menit at 10,000 rpm. The best medium extraction for lipase isolation is 0.15 M phosphate buffer pH 7.5 containing sucrose 0.6 M and CaCl  1.0 mM.   ABSTRAK Penelitian ini bertujuan untuk mengoptimasi isolasi lipase indigenous biji kakao. Optimasi diawali dengan menentukan keberadaan lipase kemudian optimasi medium ekstraksi dan proses ekstraksi. Hasil penelitian menunjukkan bahwa lipase berada dalam sitosol. Penghilangan lemak tidak meningkatkan aktivitas lipase. Senyawa polifenol menghambat aktivitas lipase dan penghilangan polifenol dapat meningkatkan aktivitas lipase. Polyvinilpolypirrolidone (PVPP dapat menghambat polifenol sehingga dapat meningkatkan aktivitas lipase. Konsentrasi PVPP optimum adalah 8 % dari berat biji kakao. Proses homogenisasi optimum diperoleh dalam waktu 10 menit pada kecepatan 10.000 rpm. Medium ekstraksi untuk isolasi lipase biji kakao terbaik adalah bufer fosfat 0,15 M  dan pH 7,5 yang mengandung sukrosa 0,6 M dan 1,0 mM CaCl .

Familial lipoprotein lipase deficiency

Science.gov (United States)

... lack an enzyme called lipoprotein lipase. Without this enzyme, the body cannot break down fat from digested food. Fat particles called chylomicrons build up in the blood. Risk factors include a family history of lipoprotein lipase deficiency. The condition is usually ...

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

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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.

Potencial de biocatálise enantiosseletiva de lipases microbianas Potential of enantioselective biocatalysis by microbial lipases

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Patrícia de O. Carvalho

2005-08-01

Full Text Available Microbial lipases have a great potential for commercial applications due to their stability, selectivity and broad substrate specificity because many non-natural acids, alcohols or amines can be used as the substrate. Three microbial lipases isolated from Brazilian soil samples (Aspergillus niger; Geotrichum candidum; Penicillium solitum were compared in terms of their stability and as biocatalysts in the enantioselective esterification using racemic substrates in organic medium. The lipase from Aspergillus niger showed the highest activity (18.2 U/mL and was highly thermostable, retaining 90% and 60% activity at 50 ºC and 60 ºC after 1 hour, respectively. In organic medium, this lipase provided the best results in terms of enantiomeric excess of the (S-active acid (ee = 6.1% and conversion value (c = 20% in the esterification of (R,S-ibuprofen with 1-propanol in isooctane. The esterification reaction of the racemic mixture of (R,S-2-octanol with decanoic acid proceeded with high enantioselectivity when lipase from Aspergillus niger (E = 13.2 and commercial lipase from Candida antarctica (E = 20 were employed.

Lipase Catalyzed Kinetic Resolution of rac-2-(3-Methoxy-4-methylphenyl) propan-1-ol and rac-2-(3-Hydroxy-4-methylphenyl)propyl propanoate for S-(+)-Xanthorrhizol

International Nuclear Information System (INIS)

Shafioul, Azam Sharif Mohammed; Cheong, Chan Seong

2012-01-01

Xanthorrhizol is a bisabolane type of natural sesquiterpene, the major component of essential oils of Curcuma xanthorrhiza. 2-(3-Methoxy-4-methylphenyl)propan-1-ol and 2-(3-hydroxy-4-methyl phenyl)propan-1-ol could be essential building block for enantioselective synthesis of xanthorrhizol. Enantioselective (c = 53%, E = 80 ± 3) for R-(+)-2-(3-hydroxy-4-methylphenyl) propan-1-ol and (c = 58%, E = 27 ± 1) for R-(+)-2-(3- methoxy-4-methylphenyl) propan-1-ol resolution processes were developed via lipase-catalyzed reaction. We found lipase Aspergillus oryzae (AOL) and Porcine pancreas (PPL) are selective to transesterification and hydrolysis in organic and aqueous phase. Modified demethylated substrate is appropriate for enantioselective hydrolysis reaction without any additives. Enantiopure chiral alcohol was crystallized from ethyl acetate/ n-hexane co-solvent system. Gram scale resolved chiral intermediate will facilitate the synthesis of the unnatural S-(+)-xanthorrhizol, the corresponding isomer of the natural one

Lipase Catalyzed Kinetic Resolution of rac-2-(3-Methoxy-4-methylphenyl) propan-1-ol and rac-2-(3-Hydroxy-4-methylphenyl)propyl propanoate for S-(+)-Xanthorrhizol

Energy Technology Data Exchange (ETDEWEB)

Shafioul, Azam Sharif Mohammed [University of Science and Technology, Daejeon (Korea, Republic of); Cheong, Chan Seong [Korea Institute of Science and Technology, Seoul (Korea, Republic of)

2012-02-15

Xanthorrhizol is a bisabolane type of natural sesquiterpene, the major component of essential oils of Curcuma xanthorrhiza. 2-(3-Methoxy-4-methylphenyl)propan-1-ol and 2-(3-hydroxy-4-methyl phenyl)propan-1-ol could be essential building block for enantioselective synthesis of xanthorrhizol. Enantioselective (c = 53%, E = 80 ± 3) for R-(+)-2-(3-hydroxy-4-methylphenyl) propan-1-ol and (c = 58%, E = 27 ± 1) for R-(+)-2-(3- methoxy-4-methylphenyl) propan-1-ol resolution processes were developed via lipase-catalyzed reaction. We found lipase Aspergillus oryzae (AOL) and Porcine pancreas (PPL) are selective to transesterification and hydrolysis in organic and aqueous phase. Modified demethylated substrate is appropriate for enantioselective hydrolysis reaction without any additives. Enantiopure chiral alcohol was crystallized from ethyl acetate/ n-hexane co-solvent system. Gram scale resolved chiral intermediate will facilitate the synthesis of the unnatural S-(+)-xanthorrhizol, the corresponding isomer of the natural one.

Application of alkaline thermo-stable lipase(s) enzyme produced from irradiated microbial isolate in the field of detergent technology

International Nuclear Information System (INIS)

Ahmed, O.E.A.M.S

2010-01-01

Due to continuous demand for manufacture of high quality, low coast industrial detergents containing lipolytic enzymes and due to continuous accumulation of enviro-agro-industrial wastes which are good and suitable conditions for growth and reproduction of pathogenic microorganisms, our study aims at isolating thermoalkalophilic lipase producer microorganisms from enviro-agro-industrial wastes and selection of the most potent isolate for studying physiological conditions controlling enzyme formation also purification characterization and some applications on purified and crude enzyme as bio-detergent. Some environmental and industrial wastes were collected from different places. The industrial wastes include, cotton seed, soyabean, sun flower, lin seed and olive oil wastes. Environmental wastes include poultry and fish wastes, all these wastes were dried at 70 degree C, grounded and used for isolation of microorganisms and lipase(s) production.Nine thermoalkalophilic bacterial isolates were isolated from enviro-agro-industrial wastes at ph 11.5 and 70 degree C. They were purified and screening for their ability of thermoalkalo-stable lipase(s) formation, this is followed by examining the effect of different nutritional media and exposure of bacterial isolates to different doses of gamma irradiation and the influence of these radiation on lipase(s) productivity by these isolates. From the results it was found that.1- The most potent lipase(s) forming bacterial isolates were isolates number B 2 and B 3 which cultivated on medium A amended with fish-wastes as being the best nutritional medium for enzyme formation. 2-Bacterial isolate B 2 finally was selected as being the most potent lipase(s) forming bacterial isolate cultivated on fish-wastes and yeast extract (in tap water) and identified according to key's of Bergey Manual of Systematic Bacteriology (1984) as being Bacillus brevis B 2 .The optimum culture conditions for maximum biosynthesis of extracellular lipase(s

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

Directory of Open Access Journals (Sweden)

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

Assessing the reaction conditions to mediate the milkfat-soybean oil enzymatic interesterification

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Ariela Veloso de Paula

Full Text Available Summary A food grade lipase from Rhizopus oryzae immobilized on a hybrid polysiloxane-polyvinyl alcohol matrix (SiO2-PVA was used as the biocatalyst to mediate the interesterification reactions of a blend containing 65% milkfat and 35% soybean oil. All the reactions occurred in an inert nitrogen atmosphere in cylindrical glass reactors (80 mL with 40 g of the milkfat-soybean oil blend. The influence of the following variables was evaluated: biocatalyst loading (250-1500 activity units per gram of blend, biocatalyst moisture content (5-20%, temperature (45-60 °C and incubation time (2-48 h. The reactions were monitored by determining the free fatty acid content, triacylglycerol (TAGs composition in carbon species, and the consistency of the interesterified (IE products. The reaction conditions were set based on the parameters that provided a high interesterification yield and good consistency of the final product within the ideal range (200 to 800 gf cm-2. Hence the best results were obtained using a biocatalyst loading of 500 U g-1 of blend with 10% moisture content at 45 °C for 4 h. Under these conditions the consistency of the interesterified product was 539.7 ± 38 gf cm-2. The results demonstrated the potential of the immobilized lipase to alter the TAGs profile of the milkfat-soybean oil blend, allowing for the production of structured lipids.

Activity and stability of immobilized lipases in lipase-catalyzed modification of peanut oil

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Soumanou Mohamed M.

2004-11-01

Full Text Available Fatty acid release during lipolysis of peanut oil using microbial free and immobilized lipases in aqueous media was developed. Immobilized lipase from Rhizomucor miehei (RML gave the best result from its ability to clive different fatty acids from peanut oil in such media. In organic solvent, interesterification of peanut oil with tricaprylin using immobilized lipases from RML, Chromobacterium viscosum (CVL and Candida rugosa (CRL was performed. The best substrate molar ratio of tricaprylin to peanut oil found was in the range 0.7 to 0.8. Using substrate molar ratio 0.7, high amount of structured triglyceride ST (about 35% MLM, 44% LML triglyceride fractions was obtained with lipase from RML in n-hexane. The results found in solvent free system were in some cases quite similar to that obtained in organic solvent. In nine successive batch interesterification in solvent free medium using immobilized RML and CRL, no significant loss of amount of both produced triacylglycerol fractions until batch 7 was observed with RML.

Lipase Test: MedlinePlus Lab Test Information

Science.gov (United States)

... page: https://medlineplus.gov/labtests/lipasetest.html Lipase Test To use the sharing features on this page, please enable JavaScript. What is a lipase test? Lipase is a type of protein made by ...

21 CFR 184.1415 - Animal lipase.

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Animal lipase. 184.1415 Section 184.1415 Food and... Substances Affirmed as GRAS § 184.1415 Animal lipase. (a) Animal lipase (CAS Reg. No. 9001-62-1) is an enzyme preparation obtained from edible forestomach tissue of calves, kids, or lambs, or from animal pancreatic...

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

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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.

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%.

Biochemical Characterization of Lipases Obtained from Acinetobacter psychrotolerans Strains

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Şule SEREN

2017-12-01

Full Text Available In this study, extracellular lipases obtained from Acinetobacter psychrotolerans strains (Xg1 and Xg2 were characterized. The effects of varying pH values (3.0-10.0 and various temperatures (10-90 °C on lipase activities were examined. Also the effects of different metal ions, organic solvents and detergents on lipases were studied. The extracellular crude lipases were concentrated using ultrafiltration. Zymogram analysis of these lipases was performed. Lipases exhibited maximum activity at pH 8 and 30 °C.  While lipase obtained from the Xg1 strain exhibited the highest stability in the presence of various organic solvents, including hexane, ethyl acetate, chloroform and N,N dietil formamide, lipase obtained from the Xg2 strain was sensitive in the presence of isopropanol, acetonitrile, and butan-1-ol. The lipases of the Xg1 and Xg2 strains were inhibited in the presence of Cu2+ and Zn2+. Also, the lipase of the Xg1 strain was inhibited in the presence of Fe3+. In the presence of EDTA, the lipase activities of the Xg1 and Xg2 strains were partially inhibited. In presence of SDS, they were exactly inhibited. According to the zymogram results, the molecular weights of the lipases obtained from the Acinetobacter psychrotolerans Xg1 and Xg2 strains have been found approximately 37 and 30 kDa, respectively.

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

Role of Hepatic Lipase and Endothelial Lipase in High-Density Lipoprotein-Mediated Reverse Cholesterol Transport

NARCIS (Netherlands)

Annema, Wijtske; Tietge, Uwe J. F.

Reverse cholesterol transport (RCT) constitutes a key part of the atheroprotective properties of high-density lipoproteins (HDL). Hepatic lipase (HL) and endothelial lipase (EL) are negative regulators of plasma HDL cholesterol levels. Although overexpression of EL decreases overall

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.

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

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 ...

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.

Biodegradable products by lipase biocatalysis.

Science.gov (United States)

Linko, Y Y; Lämsä, M; Wu, X; Uosukainen, E; Seppälä, J; Linko, P

1998-11-18

The interest in the applications of biocatalysis in organic syntheses has rapidly increased. In this context, lipases have recently become one of the most studied groups of enzymes. We have demonstrated that lipases can be used as biocatalyst in the production of useful biodegradable compounds. A number of examples are given. 1-Butyl oleate was produced by direct esterification of butanol and oleic acid to decrease the viscosity of biodiesel in winter use. Enzymic alcoholysis of vegetable oils without additional organic solvent has been little investigated. We have shown that a mixture of 2-ethyl-1-hexyl esters can be obtained in a good yield by enzymic transesterification from rapeseed oil fatty acids for use as a solvent. Trimethylolpropane esters were also similarly synthesized as lubricants. Finally, the discovery that lipases can also catalyze ester syntheses and transesterification reactions in organic solvent systems has opened up the possibility of enzyme catalyzed production of biodegradable polyesters. In direct polyesterification of 1,4-butanediol and sebacic acid, polyesters with a mass average molar mass of the order of 56,000 g mol-1 or higher, and a maximum molar mass of about 130,000 g mol-1 were also obtained by using lipase as biocatalyst. Finally, we have demonstrated that also aromatic polyesters can be synthesized by lipase biocatalysis, a higher than 50,000 g mol-1 mass average molar mass of poly(1,6-hexanediyl isophthalate) as an example.

Lipase entrapment in PVA/Chitosan biodegradable film for reactor coatings

Energy Technology Data Exchange (ETDEWEB)

Batista, Karla A. [Departamento de Bioquímica e Biologia Molecular, Laboratório de Química de Proteínas, Universidade Federal de Goiás, Cx. Postal 131, 74001-970, Goiânia, GO (Brazil); Lopes, Flavio Marques [Departamento de Bioquímica e Biologia Molecular, Laboratório de Química de Proteínas, Universidade Federal de Goiás, Cx. Postal 131, 74001-970, Goiânia, GO (Brazil); Unidade Universitária de Ciências Exatas e Tecnológicas, Universidade Estadual de Goiás, Anápolis, GO (Brazil); Yamashita, Fabio [Departamento de Tecnologia de Alimentos e Medicamentos, Laboratório de Tecnologia, Universidade Estadual de Londrina, Cx. Postal 6001, CEP 86051-990, Londrina, PR (Brazil); Fernandes, Kátia Flávia, E-mail: katia@icb.ufg.br [Departamento de Bioquímica e Biologia Molecular, Laboratório de Química de Proteínas, Universidade Federal de Goiás, Cx. Postal 131, 74001-970, Goiânia, GO (Brazil)

2013-04-01

This study reports the development and characterization of novel biodegradable film, based on chitosan and polyvinyl alcohol containing lipase entrapped. The films showed a thickness of 70.4 and 79 μm to PVA/Chitosan and PVA/Chitosan/Lipase, respectively. The entrapment of lipase in PVA/Chitosan film resulted in increasing of 69.4% tensile strength (TS), and 52.4% of elongation. SEM images showed the formation of a continuous film, without pores or cracks. The lipase entrapment efficiency was estimated in 92% and the films were repeatedly used for 25 hydrolytic cycles, maintaining 62% of initial activity. The PVA/Chitosan/Lipase film was used for olive oil hydrolysis of high performance. These results indicate that PVA/Chitosan/Lipase is a promising material for biotechnology applications such as triacylglycerol hydrolysis and biodiesel production. - Highlights: ► Development and characterization of PVA/Chitosan biodegradable film ► Lipase immobilization onto PVA/Chitosan film ► PVA/Chitosan/Lipase film for reactor coating ► Olive oil hydrolysis using PVA/Chitosan/Lipase film.

Lipase entrapment in PVA/Chitosan biodegradable film for reactor coatings

International Nuclear Information System (INIS)

Batista, Karla A.; Lopes, Flavio Marques; Yamashita, Fabio; Fernandes, Kátia Flávia

2013-01-01

This study reports the development and characterization of novel biodegradable film, based on chitosan and polyvinyl alcohol containing lipase entrapped. The films showed a thickness of 70.4 and 79 μm to PVA/Chitosan and PVA/Chitosan/Lipase, respectively. The entrapment of lipase in PVA/Chitosan film resulted in increasing of 69.4% tensile strength (TS), and 52.4% of elongation. SEM images showed the formation of a continuous film, without pores or cracks. The lipase entrapment efficiency was estimated in 92% and the films were repeatedly used for 25 hydrolytic cycles, maintaining 62% of initial activity. The PVA/Chitosan/Lipase film was used for olive oil hydrolysis of high performance. These results indicate that PVA/Chitosan/Lipase is a promising material for biotechnology applications such as triacylglycerol hydrolysis and biodiesel production. - Highlights: ► Development and characterization of PVA/Chitosan biodegradable film ► Lipase immobilization onto PVA/Chitosan film ► PVA/Chitosan/Lipase film for reactor coating ► Olive oil hydrolysis using PVA/Chitosan/Lipase film

Rat lingual lipase: partial purification, hydrolytic properties, and comparison with pancreatic lipase.

Science.gov (United States)

Roberts, I M; Montgomery, R K; Carey, M C

1984-10-01

We have partially purified lingual lipase from the serous glands of rat tongue. With a combination of Triton X-100 extraction or Triton X-114 phase-separation techniques, Bio-Bead SM-2 treatment, dialysis, and gel filtration on Sephadex G-200 or Sephacryl S-300, we obtained a sparingly soluble lipid-free protein demonstrating hydrolytic activity against triglycerides and negligible phospholipase or cholesteryl esterase activities. Compared with homogenate, specific activities of the enzyme were enriched 3- to 5-fold prior to gel filtration and 10-fold after gel filtration. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration under denaturing conditions (6 M guanidine X HCl or 0.1% sodium dodecyl sulfate) revealed one major glycoprotein band with Mr approximately 50,000. Gel filtration of the active enzyme in 0.1% Triton X-100 gave an Mr approximately 270,000-300,000, suggesting extensive self-aggregation. With both tributyrin and triolein, the pH optimum of the purified enzyme was 4.0 and activity extended from pH 2.0 to 8.0. In contrast to purified human pancreatic lipase, lingual lipase hydrolyzed triglyceride emulsions and mixed micelles stabilized with both short-chain (dihexanoyl) and long-chain (egg) lecithin and were inhibited only slightly (18-25%) by micellar concentrations of two common bile salts, taurodeoxycholate and taurocholate. Our results suggest that the hydrolysis of dietary fat by lingual lipase may extend from the pharynx through the esophagus and stomach and into the upper small intestine.

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.

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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.

Endothelial lipase is a major determinant of HDL level

Energy Technology Data Exchange (ETDEWEB)

Ishida, Tatsuro; Choi, Sungshin; Kundu, Ramendra K.; Hirata, Ken-Ichi; Rubin, Edward M.; Cooper, Allen D.; Quertermous, Thomas

2003-01-30

For the past three decades, epidemiologic studies have consistently demonstrated an inverse relationship between plasma HDL cholesterol (HDL-C) concentrations and coronary heart disease (CHD). Population-based studies have provided compelling evidence that low HDL-C levels are a risk factor for CHD, and several clinical interventions that increased plasma levels of HDL-C were associated with a reduction in CHD risk. These findings have stimulated extensive investigation into the determinants of plasma HDL-C levels. Turnover studies using radiolabeled apolipoprotein A-I, the major protein component of HDL, suggest that plasma HDL-C concentrations are highly correlated with the rate of clearance of apolipoprotein AI. However, the metabolic mechanisms by which HDL are catabolized have not been fully defined. Previous studies in humans with genetic deficiency of cholesteryl ester transfer protein, and in mice lacking the scavenger receptor BI (SR-BI), have demonstrated that these proteins participate in the removal of cholesterol from HDL, while observations in individuals with mutations in hepatic lipase indicate that this enzyme hydrolyzes HDL triglycerides. In this issue of the JCI, reports from laboratories of Tom Quertermous and Dan Rader now indicate that endothelial lipase (LIPG), a newly identified member of the lipase family, catalyzes the hydrolysis of HDL phospholipids and facilitates the clearance of HDL from the circulation. Endothelial lipase was initially cloned by both of these laboratories using entirely different strategies. Quertermous and his colleagues identified endothelial lipase as a transcript that was upregulated in cultured human umbilical vein endothelial cells undergoing tube formation, whereas the Rader group cloned endothelial lipase as a transcript that was upregulated in the human macrophage-like cell line THP-1 exposed to oxidized LDL. Database searches revealed that endothelial lipase shows strong sequence similarity to lipoprotein

Catalytic Properties of Lipase Extracts from Aspergillus niger

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Cintia M. Romero

2006-01-01

Full Text Available Screening of lipolytic strains using Rhodamine-B/olive oil plate technique allowed the selection of Aspergillus niger MYA 135. Lipase production in submerged culture containing 2 % olive oil was enhanced by more than 50 % compared to basal cultural conditions. Optimal catalytic conditions for olive oil-induced lipase were pH=6.5 and 30–35 °C. These values were shifted to the acid region (4.0–6.5 and 35–37 °C when lipase extract was produced under basal conditions. Slight changes of the residual lipase activity against the pH were found. However, preincubation at either 37 or 40 °C caused an increase in the olive oil-inducible lipolytic activity. On the contrary, lipase residual activity decreases in the 30–55 °C range when it was produced in basal medium. Lipolytic extracts were almost not deactivated in presence of 50 % water-miscible organic solvents. However, water-immiscible aliphatic solvents reduced the lipase activity between 20 and 80 %.

Isolation and analysis of lipase-overproducing mutants of Serratia marcescens.

Science.gov (United States)

Kawai, E; Akatsuka, H; Sakurai, N; Idei, A; Matsumae, H; Shibatani, T; Komatsubara, S; Omori, K

2001-01-01

We have isolated a lipase-overproducing mutant, GE14, from Serratia marcescens 8000 after three rounds of N-methyl-N'-nitro-N-nitrosoguanidine mutagenesis. The mutant GE14 produced 95 kU/ml of extracellular lipase in the lipase medium, which was about threefold higher than that of produced by the original strain 8000. Enzymatic characteristics including specific activity of purified lipases from culture supernatants of GE14 and 8000 were almost same. The lipase gene (lipA) of GE14 contained two base substitutions; one in the promoter region and another in the N-terminal region of the lipA gene without an amino acid substitution. Promoter analysis using lipA-lacZ fusion plasmids revealed that these substitutions were responsible for the increase in the lipA expression level, independently. In contrast, no base substitution was found in the genes encoding the lipase secretion device, the Lip system. In addition, the genes coding for metalloprotease and the cell surface layer protein which are both secreted through the Lip system and associated with extracellular lipase production, also contained no base substitution. The strain GE14 carrying a high-copy-number lipA plasmid produced a larger amount of the extracellular lipase than the recombinant strains of 8000 and other mutants also did, indicating that GE14 was not only a lipase-overproducing strain, but also an advantageous host strain for overproducing the lipase by a recombinant DNA technique. These results suggest that the lipase-overproducing mutant GE14 and its recombinant strains are promising candidates for the industrial production of the S. marcescens lipase.

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%.

Production of Fungal Biomass for Feed, Fatty Acids, and Glycerol by Aspergillus oryzae from Fat-Rich Dairy Substrates

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Amir Mahboubi

2017-09-01

Full Text Available Dairy waste is a complex mixture of nutrients requiring an integrated strategy for valorization into various products. The present work adds insights into the conversion of fat-rich dairy products into biomass, glycerol, and fatty acids via submerged cultivation with edible filamentous fungi. The pH influenced fat degradation, where Aspergillus oryzae lipase was more active at neutral than acidic pH (17 g/L vs. 0.5 g/L of released glycerol; the same trend was found during cultivation in crème fraiche (12 g/L vs. 1.7 g/L of released glycerol. In addition to glycerol, as a result of fat degradation, up to 3.6 and 4.5 g/L of myristic and palmitic acid, respectively, were released during A. oryzae growth in cream. The fungus was also able to grow in media containing 16 g/L of lactic acid, a common contaminant of dairy waste, being beneficial to naturally increase the initial acidic pH and trigger fat degradation. Considering that lactose consumption is suppressed in fat-rich media, a two-stage cultivation for conversion of dairy waste is also proposed in this work. Such an approach would provide biomass for possibly feed or human consumption, fatty acids, and an effluent of low organic matter tackling environmental and social problems associated with the dairy sector.

Stability of immobilized candida sp. 99-125 Lipase for biodiesel production

Energy Technology Data Exchange (ETDEWEB)

Lu, J. [Beijing Bioprocess Key Laboratory, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing (China); Bioengineering Department, Zhengzhou University, Zhengzhou (China); Deng, L.; Nie, K.; Wang, F.; Tan, T. [Beijing Bioprocess Key Laboratory, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing (China)

2012-12-15

The stability of the immobilized lipase from Candida sp. 99-125 during biodiesel production was investigated. The lipase was separately incubated in the presence of various reaction components such as soybean oil, oleic acid methyl ester, n-hexane, water, methanol, and glycerol, or the lipase was stored at 60, 80, 100 and 120 C. Thereafter the residual lipase activity was determined by methanolysis reaction. The results showed that the lipase was rather stable in the reaction media, except for methanol and glycerol. The stability study performed in a reciprocal shaker indicated that enzyme desorption from the immobilized lipase mainly contributed to the lipase inactivation in the water system. So the methanol and glycerol contents should be controlled more precisely to avoid lipase inactivation, and the immobilization method should be improved with regard to lipase desorption. (Copyright copyright 2012 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

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...

Molecular characterization of a proteolysis-resistant lipase from Bacillus pumilus SG2

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R. Sangeetha

2014-06-01

Full Text Available Proteolysis-resistant lipases can be well exploited by industrial processes which employ both lipase and protease as biocatalysts. A proteolysis resistant lipase from Bacillus pumilus SG2 was isolated, purified and characterized earlier. The lipase was resistant to native and commercial proteases. In the present work, we have characterized the lip gene which encodes the proteolysis-resistant lipase from Bacillus pumilus SG2. The parameters and structural details of lipase were analysed. The lip gene consisted of 650 bp. The experimental molecular weight of SG2 lipase was nearly double that of its theoretical molecular weight, thus suggesting the existence of the functional lipase as a covalent dimer. The proteolytic cleavage sites of the lipase would have been made inaccessible by dimerisation, thus rendering the lipase resistant to protease.

Applications of immobilized Thermomyces lanuginosa lipase in interesterification

DEFF Research Database (Denmark)

Yang, Tiankui; Fruekilde, Maj-Britt; Xu, Xuebing

2003-01-01

(RSM). Thermomyces lanuginosa lipase had an activity similar to that of immobilized Rhizomucor miehei lipase (Lipozyme RM IM) in the glycerolysis of sunflower oil, but the former had higher activity at a low reaction temperature (5degreesC). Thermomyces lanuginosa lipase was found to have much lower...... catalytic activity than Lipozyme RM IM in the acidolysis of sunflower oil with caprylic acid. However, the activity of T. lanuginosa lipase was only slightly lower than that of Lipozyme RM IM in the ester-ester exchange between tripalmitin (PPP) and the ethyl esters of EPA and DHA (EE). For this reason...

Altering the activation mechanism in Thermomyces lanuginosus lipase

DEFF Research Database (Denmark)

Skjold-Jørgensen, Jakob; Vind, Jesper; Svendsen, Allan

2014-01-01

It is shown by rational site-directed mutagenesis of the lid region in Thermomyces lanuginosus lipase that it is possible to generate lipase variants with attractive features, e.g., high lipase activity, fast activation at the lipid interface, ability to act on water-soluble substrates......, and enhanced calcium independence. The rational design was based on the lid residue composition in Aspergillus niger ferulic acid esterase (FAEA). Five constructs included lipase variants containing the full FAEA lid, a FAEA-like lid, an intermediate lid of FAEA and TlL character, and the entire lid region...... from Aspergillus terreus lipase (AtL). To investigate an altered activation mechanism for each variant compared to that of TlL, a combination of activity- and spectroscopic-based measurements were applied. The engineered variant with a lid from AtL displayed interfacial activation comparable...

Butyl acetate synthesis using immobilized lipase in calcium alginate beads

International Nuclear Information System (INIS)

Mohd Zulkhairi Abdul Rahim; Lee, Pat M.; Lee, Kong H.

2008-01-01

The esterification reaction of acetic acid and n-butanol using immobilized lipase encapsulated in calcium alginate beads (Lipase - CAB) and in chitosan coated calcium alginate beads (Lipase-CCAB) in n-hexane under mild reaction conditions were studied. Effects of temperature and substrate concentration (acetic acid and n-butanol) using Lipase - CAB, Lipase - CCAB and free lipase on the esterification reaction and their thermal stability towards esterification reaction were investigated. Results of temperature studies showed that the butyl acetate conversion increased with increase of temperature and reached the highest yield of about 70% around 50 degree Celsius for both immobilized systems but the yield of product catalyzed by free enzyme decreased as temperature was increased. Thermal stabilities studies showed that the Lipase-CCAB and Lipase-CAB were stable throughout the temperature range of 30-60 degree Celsius. However, free lipase became less stable at temperatures higher than 50 degree Celsius. The substrates, n-butanol and acetic acid exerted different effects on the esterification reaction and the reaction was favoured by higher acetic acid concentration than butanol. Kinetics parameters, Km and Vmax values for both substrates and the specific activities of the three enzyme system were also determined. The beads morphology was examined using SEM. Batch-wise operational stability studies for both immobilized systems demonstrated that the immobilized lipase performed better in the batch wise reactor system than the continuous bioreactor system and that the immobilized lipase remained active for at least 5 cycles of batch wise esterification reactions. (author)

Gastric lipase: localization of the enzyme in the stomach

International Nuclear Information System (INIS)

DeNigris, S.J.; Hamosh, M.; Hamosh, P.; Kasbekar, D.K.

1986-01-01

Isolated gastric glands prepared from human and rabbit stomach secrete lipase in response to secretagogues. They have investigated the localization of this enzyme in three species (rabbit, baboon, guinea pig). Gastric mucosa was sampled from the cardia (C), fundus-smooth (FS), fundus-ruggae (FR) and the antral area (A). Lipase activity was measured in mucosal homogenates using 3 H-triolein as substrate and is expressed in units (U) = nmols free fatty acid released/min/mg wet weight. The localization of lipase is compared with that of pepsin (measured by hydrolysis of 2% hemoglobin at pH 1.8 and expressed in I.U.). Lipase is localized in a well defined area in the rabbit and is diffusely distributed in both guinea pig and baboon. The distribution of lipase and pepsin containing cells differs in all three species. The cellular origin of gastric lipase remains to be determined

Differential effect of combined lipase deficiency (cld/cld) on human hepatic lipase and lipoprotein lipase secretion.

Science.gov (United States)

Boedeker, J C; Doolittle, M H; White, A L

2001-11-01

Combined lipase deficiency (cld) is a recessively inherited disorder in mice associated with a deficiency of LPL and hepatic lipase (HL) activity. LPL is synthesized in cld tissues but is retained in the endoplasmic reticulum (ER), whereas mouse HL (mHL) is secreted but inactive. In this study we investigated the effect of cld on the secretion of human HL (hHL) protein mass and activity. Differentiated liver cell lines were derived from cld mice and their normal heterozygous (het) littermates by transformation of hepatocytes with SV40 large T antigen. After transient transfection with lipase expression constructs, secretion of hLPL activity from cld cells was only 12% of that from het cells. In contrast, the rate of secretion of hHL activity and protein mass per unit of expressed hHL mRNA was identical for the two cell lines. An intermediate effect was observed for mHL, with a 46% reduction in secretion of activity from cld cells. The ER glucosidase inhibitor, castanospermine, decreased secretion of both hLPL and hHL from het cells by approximately 70%, but by only approximately 45% from cld cells. This is consistent with data suggesting that cld may result from a reduced concentration of the ER chaperone calnexin. In conclusion, our results demonstrate a differential effect of cld on hLPL, mHL, and hHL secretion, suggesting differential requirements for activation and exit of the enzymes from the ER.

Lipase in milk, curd and cheese

NARCIS (Netherlands)

Geurts, T.J.; Lettink, F.J.; Wouters, J.T.M.

2003-01-01

Presence of lipase in milk, curd, whey and cheese was studied. A small amount of the product was added to a large volume of lipase-free whole milk that had been made sensitive to lipolysis by homogenization. Increase of the acidity of the fat in the mixture, determined after incubation, was

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...

Seed lipases: sources, applications and properties - a review

Directory of Open Access Journals (Sweden)

M. Barros

2010-03-01

Full Text Available This paper provides an overview regarding the main aspects of seed lipases, such as the reactions catalyzed, physiological functions, specificities, sources and applications. Lipases are ubiquitous in nature and are produced by several plants, animals and microorganisms. These enzymes exhibit several very interesting features, such as low cost and easy purification, which make their commercial exploitation as industrial enzymes a potentially attractive alternative. The applications of lipases in food, detergents, oils and fats, medicines and fine chemistry, effluent treatment, biodiesel production and in the cellulose pulp industry, as well as the main sources of oilseed and cereal seed lipases, are reviewed.

Hormone-sensitive lipase (HSL) expression and regulation in skeletal muscle

DEFF Research Database (Denmark)

Langfort, J; Ploug, T; Ihlemann, J

1998-01-01

Because the enzymatic regulation of muscle triglyceride metabolism is poorly understood we explored the character and activation of neutral lipase in muscle. Western blotting of isolated rat muscle fibers demonstrated expression of hormone-sensitive lipase (HSL). In incubated soleus muscle...... epinephrine increased neutral lipase activity by beta-adrenergic mechanisms involving cyclic AMP-dependent protein kinase (PKA). The increase was paralleled by an increase in glycogen phosphorylase activity and could be abolished by antiserum against HSL. Electrical stimulation caused a transient increase...... in activity of both neutral lipase and glycogen phosphorylase. The increase in lipase activity during contractions was not influenced by sympathectomy or propranolol. Training diminished the epinephrine induced lipase activation in muscle but enhanced the activation as well as the overall concentration...

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...

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.

Structural characterization of MAPLE deposited lipase biofilm

Energy Technology Data Exchange (ETDEWEB)

Aronne, Antonio [Department of Chemical Engineering, Materials and Industrial Production, Università degli Studi di Napoli Federico II, Piazzale V. Tecchio 80, 80125 Napoli (Italy); Ausanio, Giovanni; Bloisi, Francesco [CNR-SPIN and Department of Physics, Università degli Studi di Napoli Federico II, Piazzale V. Tecchio 80, 80125 Napoli (Italy); Calabria, Raffaela [Istituto Motori-CNR, via G. Marconi 8, 80125 Napoli (Italy); Califano, Valeria, E-mail: v.califano@im.cnr.it [Istituto Motori-CNR, via G. Marconi 8, 80125 Napoli (Italy); Fanelli, Esther [Department of Chemical Engineering, Materials and Industrial Production, Università degli Studi di Napoli Federico II, Piazzale V. Tecchio 80, 80125 Napoli (Italy); Massoli, Patrizio [Istituto Motori-CNR, via G. Marconi 8, 80125 Napoli (Italy); Vicari, Luciano R.M. [CNR-SPIN and Department of Physics, Università degli Studi di Napoli Federico II, Piazzale V. Tecchio 80, 80125 Napoli (Italy)

2014-11-30

Highlights: • Lipase from Candida Rugosa was deposited by Matrix Assisted Pulsed Laser Evaporation (MAPLE) on KBr pellets, mica and glass substrate. • The deposited film was characterized morphologically and structurally by optical microscopy, SEM and FTIR analysis. • Results of characterization underlined a phenomenon of aggregation taking place. • The aggregation phenomenon was reversible since lipase showed activity in the transesterification reaction between soybean oil and isopropyl alcohol once detached from the substrate. - Abstract: Lipases (triacylglycerol ester hydrolases) are enzymes used in several industrial applications. Enzymes immobilization can be used to address key issues limiting widespread application at industrial level. Immobilization efficiency is related to the ability to preserve the native conformation of the enzyme. MAPLE (Matrix Assisted Pulsed Laser Evaporation) technique, a laser deposition procedure for treating organic/polymeric/biomaterials, was applied for the deposition of lipase enzyme in an ice matrix, using near infrared laser radiation. Microscopy analysis showed that the deposition occurred in micrometric and submicrometric clusters with a wide size distribution. AFM imaging showed that inter-cluster regions are uniformly covered with smaller aggregates of nanometric size. Fourier transform infrared spectroscopy was used for both recognizing the deposited material and analyzing its secondary structure. Results showed that the protein underwent reversible self-association during the deposition process. Actually, preliminary tests of MAPLE deposited lipase used for soybean oil transesterification with isopropyl alcohol followed by gas chromatography–mass spectrometry gave results consistent with undamaged deposition of lipase.

Lipase or amylase for the diagnosis of acute pancreatitis?

Science.gov (United States)

Ismail, Ola Z; Bhayana, Vipin

2017-12-01

Acute pancreatitis is a rapid onset of inflammation of the pancreas causing mild to severe life threatening conditions [1, 2]. In Canada, acute pancreatitis is the 5th most expensive digestive disease in Canada with a considerable economic burden on the health care system [3]. The diagnosis of acute pancreatitis is usually based on the presence of abdominal pain and elevated levels of serum amylase and/or lipase. Many health care centers use either serum amylase, lipase or both to diagnose acute pancreatitis without considering which one could provide a better diagnostic accuracy. The aim of this review is to investigate whether serum lipase alone is a sufficient biomarker for the diagnosis of acute pancreatitis. We have examined various studies looking at the utilization, sensitivity, specificity and cost associated savings of lipase and amylase in the diagnosis of acute pancreatitis. When comparing different studies, serum lipase offers a higher sensitivity than serum amylase in diagnosing acute pancreatitis. Lipase also offers a larger diagnostic window than amylase since it is elevated for a longer time, thus allowing it to be a useful diagnostic biomarker in early and late stages of acute pancreatitis. Several recent evidence-based guidelines recommend the use of lipase over amylase. Nevertheless, both lipase and amylase alone lack the ability to determine the severity and etiology of acute pancreatitis. The co-ordering of both tests has shown little to no increase in the diagnostic sensitivity and specificity. Thus, unnecessary testing and laboratory expenditures can be reduced by testing lipase alone. Copyright © 2017 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

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.

Production of lipase free of citrinin by Penicillium citrinum.

Science.gov (United States)

Pimentel, M C; Melo, E H; Lima Filho, J L; Durán, N

1996-02-01

Lipase (Glycerol ester hydrolase E.G. 3.1.1.3) from a Brazilian strain of Penicillium citrinum free of the mycotoxin citrinin has been investigated. Citrinin production was inhibited by using culture medium containing olive oil, soybean oil and corn oil as carbon sources. Potassium concentration and pH play an important role in citrinin production. Potassium concentration lower than 30 mM and pH below 4.5 inhibited the mycotoxin production. P. citrinum produced lipase free of extraneous proteins and citrinin when cultured using, as nitrogen source, ammonium sulphate (lipase activity of 7.88 U/mg) and yeast extract (lipase activity of 4.95 U/mg) with olive oil as carbon source. This data is relevant to the larger scale production of lipases for food technology applications, from Penicillium citrinum.

Fiscal 1999 achievement report on regional consortium research and development project. Regional consortium research on energy (Research and development of biofuel production using highly functional bioreactor - 2nd year); 1999 nendo kokino bio reactor ni yoru bio nenryo seisan ni kansuru kenkyu kaihatsu seika hokokusho (dai 2 nendo)

Energy Technology Data Exchange (ETDEWEB)

NONE

2000-03-01

Microbes capable of a high biodiesel fuel (BDF) yield from moisture-containing oils assumedly waste oil are investigated. Lipase attributed to Rhizopus oryzae exhibits a high reaction rate of not lower than 90%. The process functions even when microbes are immobilized by BSPs. BDF does not affect driving performance, and black smoke is reduced. A process basic to industrial production is developed by use of a fixed bed reactor. In the production of ethanol from starch thanks to plural kinds of glucoamylase producing yeast, ethanol is produced at a rate of 7-8% under microaerophilic conditions in both proliferation and fermentation periods, which means a success achieved in growing arming yeast equipped with enhanced functions. A 20-liter class bench plant is installed and immobilization by BSPs is tested, when no problem is detected. In a reaction involving these immobilized microbes, a reaction rate near 16% is achieved. In the production of ethanol by yeast immobilized by BSPs, use of a fuzzy control system is studied, and it is found that prolonged stability is available when glucose concentration is sustained at 10-20g/liter. (NEDO)

A simple model-based control for Pichia pastoris allows a more efficient heterologous protein production bioprocess.

Science.gov (United States)

Cos, Oriol; Ramon, Ramon; Montesinos, José Luis; Valero, Francisco

2006-09-05

A predictive control algorithm coupled with a PI feedback controller has been satisfactorily implemented in the heterologous Rhizopus oryzae lipase production by Pichia pastoris methanol utilization slow (Mut(s)) phenotype. This control algorithm has allowed the study of the effect of methanol concentration, ranging from 0.5 to 1.75 g/L, on heterologous protein production. The maximal lipolytic activity (490 UA/mL), specific yield (11,236 UA/g(biomass)), productivity (4,901 UA/L . h), and specific productivity (112 UA/g(biomass)h were reached for a methanol concentration of 1 g/L. These parameters are almost double than those obtained with a manual control at a similar methanol set-point. The study of the specific growth, consumption, and production rates showed different patterns for these rates depending on the methanol concentration set-point. Results obtained have shown the need of implementing a robust control scheme when reproducible quality and productivity are sought. It has been demonstrated that the model-based control proposed here is a very efficient, robust, and easy-to-implement strategy from an industrial application point of view. (c) 2006 Wiley Periodicals, Inc.

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

Directory of Open Access Journals (Sweden)

André Luiz Cabral Monteiro Azevedo Santiago

2006-09-01

Full Text Available A presença de metais pesados no solo proporciona impacto sobre os microrganismos, vegetação e os processos funcionais do ecossistema. Visando comprovar que Mucorales são afetados pela extração de cobre na Mineradora Caraíba, Jaguarari, BA, espécies de Mucorales foram isoladas de amostras de solo coletadas nas épocas seca e chuvosa. Os Mucorales foram identificados e caracterizados quanto à capacidade de degradar amido e inulina. Utilizando os métodos de diluição e placa de solo, foram obtidos 46 isolados de Mucorales pertencentes a sete diferentes espécies. Absidia blakesleeana Lendner, A. cylindrospora Hagem, A. hialospora (Saito Lendn., Cunninghamella elegans Lendner, Rhizopus microsporus V. Thieghen, R. oryzae Went. & Prinsen Geerl. e Syncephalastrum racemosum (Cohn. Schroet. foram isolados na época seca. A. blakesleeana e A. hialospora foram isoladas apenas na época chuvosa. Rhizopus oryzae apresentou maior número de isolados (30,43%, seguido por A. blakesleeana (26,09% e C. elegans (21,74%. O gênero que apresentou maior número de espécies foi Absidia seguido por Rhizopus. Entre as áreas, houve diferenças significativas no número de isolados de Mucorales, comprovando os efeitos negativos do impacto ambiental na abundância de Mucorales nos solos impactados. Todas as espécies degradaram o amido e apenas C. elegans degradou também inulina. Culturas de mesma espécie isoladas de áreas diferentes apresentaram o mesmo comportamento quanto à degradação de amido e inulina.The presence of heavy metals in soil causes impact on microorganisms, vegetation and functional processes in ecosystems. Aiming to prove that Mucorales are affected by copper extraction at the Caraíba mining site, Jaguarari, Bahia, species of Mucorales were isolated from samples collected in the dry and wet seasons. Mucorales were identified and characterized according to their capacity to degrade starch and inulin. Forty six isolates from seven

Engineering Lipases: walking the fine line between activity and stability

Science.gov (United States)

Dasetty, Siva; Blenner, Mark A.; Sarupria, Sapna

2017-11-01

Lipases are enzymes that hydrolyze lipids and have several industrial applications. There is a tremendous effort in engineering the activity, specificity and stability of lipases to render them functional in a variety of environmental conditions. In this review, we discuss the recent experimental and simulation studies focused on engineering lipases. Experimentally, mutagenesis studies have demonstrated that the activity, stability, and specificity of lipases can be modulated by mutations. It has been particularly challenging however, to elucidate the underlying mechanisms through which these mutations affect the lipase properties. We summarize results from experiments and molecular simulations highlighting the emerging picture to this end. We end the review with suggestions for future research which underscores the delicate balance of various facets in the lipase that affect their activity and stability necessitating the consideration of the enzyme as a network of interactions.

Study of enzymatic reactors with microencapsulated lipase. Doctoral thesis. Estudo de reactores enzimaticos com lipase microencapsulada

Energy Technology Data Exchange (ETDEWEB)

de Franca Teixeira dos Prazeres, D.M.

1992-10-01

The work reports the development of a membrane reactor for the hydrolysis of triglycerides catalyzed by lipase B from Chromobacterium viscosum in AOT/isooctane reversed miceller system. In a preliminary phase the potential of the organic system was evaluated with comparative studies on the activity and stability of lipase B in aqueous media (emulsion) and in the alternative miceller media. A tubular ceramic membrane reactor with recirculation was selected for the olive oil hydrolysis in a reversed miceller system. The influence of the hydration degree, recirculation rate, AOT, olive oil and lipase concentration in the operation of the reactor were investigated in a batch mode. The hydration degree was identified as a critical parameter due to its influence in the separation process and in the kinetics of the reaction.

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.

Genetics Home Reference: lysosomal acid lipase deficiency

Science.gov (United States)

... lipase deficiency develop multi-organ failure and severe malnutrition and generally do not survive past 1 year. In the later-onset form of lysosomal acid lipase deficiency , signs and symptoms vary and usually begin in mid-childhood, although they can appear anytime up to late ...

Structural studies on lipoprotein lipase

International Nuclear Information System (INIS)

Socorro, L.

1985-01-01

The structure of lipoprotein lipase is not known. The lack of information on its primary sequence has been due to the inability of preparing it in homogeneous and stable form. This research has focused on the structural characterization of lipoprotein lipase. The first approach taken was to develop a purification method using bovine milk and affinity chromatography on heparin-Sepharose. The protein obtained was a heterogeneous peak with the activity shifted towards the trailing edge fractions. These fractions only presented a 55 Kdalton band on polyacrylamide gel electrophoresis. Monoclonal antibodies against this band detected an endogenous, phenyl methane sulfonyl fluoride-sensitive protease responsible for the presence of lower molecular weight fragments. The second approach was to label the lipoprotein lipase with a radioactive, active site, directed probe. After incubation and affinity chromatography a complex [ 3 H]inhibitor enzyme was isolated with a stoichiometry of 1.00 +/- 0.2. The complex was digested with CNBr and the insoluble peptides at low ionic strength (>90% [ 3 H]dpm) were used for further purification. Differential extraction of the [ 3 H]-peptide, digestion with S. aureus V8 protease, and high performance liquid chromatography yielded a hexapeptide with a composition consistent with the consensus sequence of the active site peptides of many serine-esterase. This and the kinetic data imply this being the mechanism of action for lipoprotein lipase

Lipase from a Brazilian strain of Penicillium citrinum.

Science.gov (United States)

Pimentel, M C; Krieger, N; Coelho, L C; Fontana, J O; Melo, E H; Ledingham, W M; Lima Filho, J L

1994-10-01

A lipases (glycerol ester hydrolases E. C. 3.1.1.3) from a brazilian strain of Penicillium citrinum has been investigated. When the microorganism was cultured in the simple medium (1.0% olive oil and 0.5% yeast extract), using olive oil in as carbon source in the inocula, the enzyme extracted showed maximum activity (409 IU/mL). In addition, decrease of yeast extract concentration also reduces the lipase activity. Nevertheless, when yeast extract was replaced by ammonium sulfate, no activity was detected. Purification by precipitation with ammonium sulfate showed best activity in the 40-60% fraction. The optimum temperature for enzyme activity was found in the range of 34-37 degrees C. However, after 30 min at 60 degrees C, the enzyme was completely inactivated. The enzyme showed optimum at pH 8.0. The dried concentrated fraction (after dialysis and lyophilization) maintained its lipase activity at room temperature (28 degrees C) for 8 mo. This result in lipase stability suggests an application of lipases from P. citrinum in detergents and other products that require a high stability at room temperature.

Dependency of water concentration on ethanolysis of trioleoylglycerol by lipases

DEFF Research Database (Denmark)

Piyatheerawong, W.; Iwasaki, Y; Xu, Xuebing

2004-01-01

tested (Rhizomucor miehei lipase, Burkholderia cepacia lipase and Thermomyces lanuginosus lipase) required larger amounts of free water (ca. 7-9 wt.%) for their best performance and exhibited no ethanolysis reaction at low free water concentrations. The CALB's anomalous behavior was also observed...

Bioremediation of cooking oil waste using lipases from wastes.

Directory of Open Access Journals (Sweden)

Clarissa Hamaio Okino-Delgado

Full Text Available Cooking oil waste leads to well-known environmental impacts and its bioremediation by lipase-based enzymatic activity can minimize the high cytotoxic potential. In addition, they are among the biocatalysts most commercialized worldwide due to the versatility of reactions and substrates. However, although lipases are able to process cooking oil wastes, the products generated from this process do not necessarily become less toxic. Thus, the aim of the current study is to analyze the bioremediation of lipase-catalyzed cooking oil wastes, as well as their effect on the cytotoxicity of both the oil and its waste before and after enzymatic treatment. Thus, assessed the post-frying modification in soybean oil and in its waste, which was caused by hydrolysis reaction catalyzed by commercial and home-made lipases. The presence of lipases in the extracts obtained from orange wastes was identified by zymography. The profile of the fatty acid esters formed after these reactions was detected and quantified through gas chromatography and fatty acids profile compared through multivariate statistical analyses. Finally, the soybean oil and its waste, with and without enzymatic treatment, were assessed for toxicity in cytotoxicity assays conducted in vitro using fibroblast cell culture. The soybean oil wastes treated with core and frit lipases through transesterification reaction were less toxic than the untreated oils, thus confirming that cooking oil wastes can be bioremediated using orange lipases.

Screening of thermophilic neutral lipase-producing Pseudomonas ...

African Journals Online (AJOL)

From oil-contaminated soil, three lipase-producing microorganisms were selected as good lipase producers using rhodamine B-olive oil plate agar and they were identified as from Pseudomonas, Burkholderia and Klebsiella genera by morphology, biochemical characterization and 16S rRNA gene sequencing. Among the ...

Frozen Microemulsions for MAPLE Immobilization of Lipase

Directory of Open Access Journals (Sweden)

Valeria Califano

2017-12-01

Full Text Available Candida rugosa lipase (CRL was deposited by matrix assisted pulsed laser evaporation (MAPLE in order to immobilize the enzyme with a preserved native conformation, which ensures its catalytic functionality. For this purpose, the composition of the MAPLE target was optimized by adding the oil phase pentane to a water solution of the amino acid 3-(3,4-dihydroxyphenyl-2-methyl-l-alanine (m-DOPA, giving a target formed by a frozen water-lipase-pentane microemulsion. Fourier transform infrared (FTIR spectroscopy and atomic force microscopy (AFM were used to investigate the structure of MAPLE deposited lipase films. FTIR deconvolution of amide I band indicated a reduction of unfolding and aggregation, i.e., a better preserved lipase secondary structure in the sample deposited from the frozen microemulsion target. AFM images highlighted the absence of big aggregates on the surface of the sample. The functionality of the immobilized enzyme to promote transesterification was determined by thin layer chromatography, resulting in a modified specificity.

Microbial lipases: Production, properties and biotechnological applications

Directory of Open Access Journals (Sweden)

Josana Maria Messias

2011-09-01

Full Text Available Lipases belong to the group of hydrolases that catalyze the hydrolysis of triacylglycerol lipids to free fatty acids and glycerol. They have significant potential biotechnological applications in catalyzing organic synthesis reactions in non-aqueous solvents using simplified procedures resulting in conversions of high yields. Lipase production has conventionally been performed by submerged fermentation; however, solid-state fermentation processes have been prominent when residues are used as substrates because they serve as low-cost nutrient sources. Microbial lipases can be used as additives in foods to modify and enhance organoleptic properties, as well as in detergents to hydrolyse fats in the treatment of oily effluents, and also have value for pharmaceutical, cosmetic, agrochemical, and oil chemical industries. More recently, they are used in transesterification reactions to convert plant seed oils into biodiesel. The objective of this work was to review the published literature on the production, properties and applications of microbial lipases, and its biotechnological role in producing biodiesel.

Structure of product-bound SMG1 lipase: active site gating implications.

Science.gov (United States)

Guo, Shaohua; Xu, Jinxin; Pavlidis, Ioannis V; Lan, Dongming; Bornscheuer, Uwe T; Liu, Jinsong; Wang, Yonghua

2015-12-01

Monoacylglycerol and diacylglycerol lipases are industrially interesting enzymes, due to the health benefits that arise from the consumption of diglycerides compared to the traditional triglyceride oils. Most lipases possess an α-helix (lid) directly over the catalytic pocket which regulates the activity of the enzyme. Generally, lipases exist in active and inactive conformations, depending on the positioning of this lid subdomain. However, lipase SMG1, a monoacylglycerol and diacylglycerol specific lipase, has an atypical activation mechanism. In the present study we were able to prove by crystallography, in silico analysis and activity tests that only two positions, residues 102 and 278, are responsible for a gating mechanism that regulates the active and inactive states of the lipase, and that no significant structural changes take place during activation except for oxyanion hole formation. The elucidation of the gating effect provided data enabling the rational design of improved lipases with 6-fold increase in the hydrolytic activity toward diacylglycerols, just by providing additional substrate stabilization with a single mutation (F278N or F278T). Due to the conservation of F278 among the monoacylglycerol and diacylglycerol lipases in the Rhizomucor miehei lipase-like family, the gating mechanism described herein might represent a general mechanism applicable to other monoacylglycerol and diacylglycerol lipases as well. Database: Structural data are available in the Protein Data Bank under the accession numbers 4ZRE (F278D mutant) and 4ZRD (F278N mutant). © 2015 FEBS.

The specificity of Several Kinds Lipases on n-3 Polyunsaturated Fatty Acids

Directory of Open Access Journals (Sweden)

Jenny Elisabeth, T Yuliani, P M Tambunan, J M Purba

2001-04-01

Full Text Available Several lipases from microbial and plant, i.e Rhizomucor miehei, Pseudomonas sp., Candida antartica, rice bran, and Carica papaya latex (CPL were examined for synthesis of omega-3 (n-3 PUFA-rich glyceride by hydrolysis and acidolysis reaction. Tuna oil was used in hydrolysis reaction, whereas tuna and palm oils were used as source of triglyceride (TAG molecules and n-3 PUFA concentrate from tuna oil as source of EPA and DHA in acidolysis reaction.For hydrolysis reaction, the rice bran and CPL lipases showed the lowest hydrolytic activity of the tuna oil, whereas the R. miehei lipase showed the highest hydrolytic activity but was unable to hydrolyze EPA and DHA. On the contrary, the C. antartica and Pseudomonas sp. lipases acted stronger on hydrolysis of DHA ester bond than EPA.For acidolysis reaction, all the lipases showed ability to incorporate n-3 PUFA into tuna and palm oils. C. antartica lipase had the maximum DHA incorporation into tuna and palm oils, rice bran lipase had relatively similar ability with R. miehei lipase, and the CPL lipase had the lowest ability. This study proved that rice bran and CPL lipases also had transesterification activity and showed the feasibility of the rice bran lipase to be a biocatalyst for n-3 PUFA-rich glyceride production. Increasing the substrate ratio, of n-3 PUFA concentrate and tuna or palm oil, could increase the EPA and DHA incorporation. The R. miehei, rice bran, and CPL lipases unabled to incorporate DHA into DHA-containing glyceride molecule, whereas C. antartica lipase had the capability in high ratio of n-3 PUFA concentrate to oil. Therefore, the lipases were easier to incorporate n-3 PUFA into palm oil than tuna oil, since the TAG molecules of palm oil was not as complex as tuna oil. It could be suggested that the lipases did not only have acyl chain and positional specificity, but also the whole glyceride structure specificity.

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.

Lipase and esterase: to what extent can this classification be applied accurately?

Directory of Open Access Journals (Sweden)

Danielle Branta Lopes

2011-09-01

Full Text Available Enzyme technology is an ever-growing field of knowledge and, in recent years, this technology has raised renewed interest, due to the search for new paradigms in several productive processes. Lipases, esterases and cutinases are enzymes used in a wide range of processes involving synthesis and hydrolysis reactions. The objective of this work was to investigate and compare the specific lipase and esterase activities of five enzymes - four already classified as lipases and one classified as cutinase - in the presence of natural and synthetic substrates. All tested enzymes presented both esterase and lipase specific activities. The highest specific esterase activity was observed for Aspergillus 1068 lipase in natural substrate and for F. oxysporum cutinase in synthetic substrate, while the highest specific lipase activity was observed for Geotrichum sp. lipase in natural substrate and for F. oxysporum cutinase in synthetic substrate. These results display some interface-independent lipolytic activity for all lipases tested. This is in accordance with the rationale that a new and broader definition of lipases may be necessary.

Lipase inactivation in wheat germ by gamma irradiation

International Nuclear Information System (INIS)

Jha, Pankaj Kumar; Kudachikar, V.B.; Kumar, Sourav

2013-01-01

An attempt was made to improve the shelf life of wheat germ by optimizing processing conditions involving γ-irradiation. Studies were carried out to investigate the effect of γ-irradiation (0–30 kGy doses) on the chemical composition of wheat germ with respect to variation in moisture, total ash, crude fat, free fatty acid, protein and lipase activity. The results demonstrate that shelf stability of wheat germ was achieved by inactivation of lipase at doses of γ-irradiation greater than 12 kGy. - Highlights: Ø γ-irradiation was found to inactivate Lipase present in Wheat Germ. Ø The treatment did not result in significant changes in Total Ash, Moisture and Protein Content of Wheat Germ. Ø The irradiation at 30 kGy resulted in 31.2 % inactivation of Lipase in Wheat Germ

Production of Cold Active Lipase from Bacillus sp.

OpenAIRE

Yasemin, Sara; Arabacı, Nihan; Korkmaz Güvenmez, Hatice

2018-01-01

A cold active lipase producing Bacillus sp. strains were isolated from sewage of oil. Bacillus sp. strain SY-7 was determined as the best lipase producing isolate. The highest enzyme production was found at 20°C and pH 8.0 on tributyrin media. Analyses of molecular mass of the partial purified lipase was carried out by SDS-PAGE which revealed a single band as 110.5 kDa. The enzyme activity and stability were determined by spectrophotometric and titrimetric methods. The enzyme was active betwe...

Zymography Detection of a Bacterial Extracellular Thermoalkaline Esterase/Lipase Activity.

Science.gov (United States)

Tapizquent, María; Fernández, Marleny; Barreto, Georgina; Hernández, Zully; Contreras, Lellys M; Kurz, Liliana; Wilkesman, Jeff

2017-01-01

Lipases are esterases that occur widely in nature, yet those with commercial relevance are exclusively from microbial origin. Glycerol and long-chain fatty acids are the products after hydrolysis of esters bonds in saponifiable lipids catalyzed by lipases. In this work, we describe lipase/esterase activity contained in cell-free fractions from thermophilic bacteria, cultured in medium containing olive oil. Analysis of the cell-free fractions by electrotransference zymography, using tributyrin as substrate, revealed bands corresponding to lipase activity. The method is simple, fast, and inexpensive.

Purification and characterization of a new cold active lipase, EnL A ...

African Journals Online (AJOL)

Search of lipase engineering data base (LED) revealed that this protein belongs to a newly introduced super family of Candida antarctica lipase A like and to the homologous family of Aspergillus lipase like. Key words: Cold active lipase, Emericella nidulans, hydrophobic interaction chromatography, Candida antarctica ...

Realm of Thermoalkaline Lipases in Bioprocess Commodities

Directory of Open Access Journals (Sweden)

Ahmad Firdaus B. Lajis

2018-01-01

Full Text Available For decades, microbial lipases are notably used as biocatalysts and efficiently catalyze various processes in many important industries. Biocatalysts are less corrosive to industrial equipment and due to their substrate specificity and regioselectivity they produced less harmful waste which promotes environmental sustainability. At present, thermostable and alkaline tolerant lipases have gained enormous interest as biocatalyst due to their stability and robustness under high temperature and alkaline environment operation. Several characteristics of the thermostable and alkaline tolerant lipases are discussed. Their molecular weight and resistance towards a range of temperature, pH, metal, and surfactants are compared. Their industrial applications in biodiesel, biodetergents, biodegreasing, and other types of bioconversions are also described. This review also discusses the advance of fermentation process for thermostable and alkaline tolerant lipases production focusing on the process development in microorganism selection and strain improvement, culture medium optimization via several optimization techniques (i.e., one-factor-at-a-time, surface response methodology, and artificial neural network, and other fermentation parameters (i.e., inoculums size, temperature, pH, agitation rate, dissolved oxygen tension (DOT, and aeration rate. Two common fermentation techniques for thermostable and alkaline tolerant lipases production which are solid-state and submerged fermentation methods are compared and discussed. Recent optimization approaches using evolutionary algorithms (i.e., Genetic Algorithm, Differential Evolution, and Particle Swarm Optimization are also highlighted in this article.

Realm of Thermoalkaline Lipases in Bioprocess Commodities.

Science.gov (United States)

Lajis, Ahmad Firdaus B

2018-01-01

For decades, microbial lipases are notably used as biocatalysts and efficiently catalyze various processes in many important industries. Biocatalysts are less corrosive to industrial equipment and due to their substrate specificity and regioselectivity they produced less harmful waste which promotes environmental sustainability. At present, thermostable and alkaline tolerant lipases have gained enormous interest as biocatalyst due to their stability and robustness under high temperature and alkaline environment operation. Several characteristics of the thermostable and alkaline tolerant lipases are discussed. Their molecular weight and resistance towards a range of temperature, pH, metal, and surfactants are compared. Their industrial applications in biodiesel, biodetergents, biodegreasing, and other types of bioconversions are also described. This review also discusses the advance of fermentation process for thermostable and alkaline tolerant lipases production focusing on the process development in microorganism selection and strain improvement, culture medium optimization via several optimization techniques (i.e., one-factor-at-a-time, surface response methodology, and artificial neural network), and other fermentation parameters (i.e., inoculums size, temperature, pH, agitation rate, dissolved oxygen tension (DOT), and aeration rate). Two common fermentation techniques for thermostable and alkaline tolerant lipases production which are solid-state and submerged fermentation methods are compared and discussed. Recent optimization approaches using evolutionary algorithms (i.e., Genetic Algorithm, Differential Evolution, and Particle Swarm Optimization) are also highlighted in this article.

Liver lipase and high-density lipoprotein. Lipoprotein changes after incubation of human serum with rat liver lipase.

Science.gov (United States)

Groot, P H; Scheek, L M; Jansen, H

1983-05-16

Human sera were incubated with rat liver lipase after inactivation of lecithin:cholesterol acyltransferase, and the changes in serum lipoprotein composition were measured. In the presence of liver lipase serum triacylglycerol and phosphatidylcholine were hydrolyzed. The main changes in the concentrations of these lipids were found in the high-density lipoprotein fraction. Subfractionation of high-density lipoprotein by rate-zonal ultracentrifugation showed a prominent decrease in all constituents of high-density lipoprotein2, a smaller decrease in the 'light' high-density lipoprotein3 and an increase in the 'heavy' high-density lipoprotein3. These data support a concept in which liver lipase is involved in high-density lipoprotein2 phospholipid and triacylglycerol catabolism and suggest that as a result of this action high-density lipoprotein2 is converted into high-density lipoprotein3.

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

Directory of Open Access Journals (Sweden)

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

21 CFR 173.140 - Esterase-lipase derived from Mucor miehei.

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Esterase-lipase derived from Mucor miehei. 173.140... HUMAN CONSUMPTION Enzyme Preparations and Microorganisms § 173.140 Esterase-lipase derived from Mucor miehei. Esterase-lipase enzyme, consisting of enzyme derived from Mucor miehei var. Cooney et Emerson by...

Beneficial effects of adding lipase enzyme to broiler diet

International Nuclear Information System (INIS)

Elbarkouky, E.M.A.

2005-01-01

A total number of 300 Ross broiler chicks were obtained from commercial hatchery at one day of age. The chicks were divided into three groups (50 males and 50 females in each). The first and second groups were supplemented with 3000 and 2000 lU/kg diet of lipase enzyme, respectively, while the third group served as control and fed on basal diet. Birds fed on diets that supplemented with lipase enzyme showed significant increase in body weight and dry matter intake, as well as fats and protein content dry matters. The serum lipase activity showed significant increase in treated groups compared to the control. Non-significant changes were determined in serum total lipids, T3, T4 and ash content. Birds supplemented with lipase showed significant decrease in cholesterol concentration. It could be concluded that birds fed diets containing 2000 or 3000 lU/kg diet of lipase enzyme exhibited improvement in broiler performance

Anti- and pro-lipase activity of selected medicinal, herbal and aquatic plants, and structure elucidation of an anti-lipase compound.

Science.gov (United States)

Ado, Muhammad Abubakar; Abas, Faridah; Mohammed, Abdulkarim Sabo; Ghazali, Hasanah M

2013-11-26

Plants that help in slowing down the digestion of triacylglycerols (TAGs) in the pancreas and small intestine of humans play an important role in the reduction of obesity. On the other hand, there may be plants or plant parts that stimulate intestinal lipolytic activity, thus contributing to greater TAG assimilation. The aim of this study was to evaluate the aqueous methanolic extracts of ninety eight (98) medicinal, herbal and aquatic plant materials from Malaysia for their effect on porcine pancreatic lipase (PPL) activity and to identify the structure of an anti-lipase compound from one of the sources. The degree of inhibition was also quantified as relative to orlistat activity against PPL (orlistat equivalents). Results revealed that while 19.4% of the extracts were found to have anti-lipase activity ≥80%, 12% were actually found to promote PPL activity. Twenty two percent (22.4%) exhibited moderate inhibition (41%-80%) and 2% were neutral toward PPL activity. The ripe fruit of Averrhoa carambola and the leaves of Archidendron jiringa (Jack) I.C Nielsen L. (jering), Cynometra cauliflora (nam-nam) and Aleurites moluccana (L.) Willd (candle nut/buah keras) had the highest (100%) anti-lipase activity and are equivalent to 0.11 µg orlistat/mL. Plants that stimulated lipase activity included Pimpinella anisum L. (aniseed/jintan manis), activating the enzyme by 186.5%. Kaempferol 3-O-rhamnoside was isolated from the ethyl acetate fraction of C. cauliflora leaves and found to be an active lipase inhibitor. The structure was elucidated using 1H-NMR, 13C-NMR and 2D-NMR analyses.

ENZYMATIC BIODIESEL SYNTHESIS FROM ACID OIL USING A LIPASE MIXTURE

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Kelly C. N. R. Pedro

Full Text Available The conventional biodiesel production process has some disadvantages. It is necessary to use refined vegetable oils with low free fatty acids (FFAs content. An alternative route is to use low-cost acid oils in an enzymatic process. The use of lipases allows simultaneous esterification of FFAs and transesterification of triglycerides present in raw material forming alkyl esters. The aim of this work was to study the production of biodiesel using soybean oils with different acid contents (Acid Value of 8.5, 50, 90 and ethanol catalyzed by commercial immobilized lipases (Novozym 435, Lipozyme RM IM and Lipozyme TL IM. A significant decrease of acid value was observed mainly with Novozym 435 and Lipozyme RM IM. The use of a mixture of two immobilized lipases was also investigated to decrease catalyst cost and increase the amount of ester produced. The three commercial immobilized lipases were mixed in a dual system and tested for biodiesel synthesis from acid oil (AV of 8.5, 50 and 90. A positive synergistic effect occurred mainly for Lipozyme TL IM (1,3-specific lipase and Novozym 435 (non-specific lipase blend. The ester content doubled when this lipase mixture was used in ethanolysis of acid oil with AV of 90.

Biotechnological applications of halophilic lipases and thioesterases.

Science.gov (United States)

Schreck, Steven D; Grunden, Amy M

2014-02-01

Lipases and esterases are enzymes which hydrolyze ester bonds between a fatty acid moiety and an esterified conjugate, such as a glycerol or phosphate. These enzymes have a wide spectrum of use in industrial applications where their high activity, broad substrate specificity, and stability under harsh conditions have made them integral in biofuel production, textile processing, waste treatment, and as detergent additives. To date, these industrial applications have mainly leveraged enzymes from mesophilic and thermophilic organisms. However, increasingly, attention has turned to halophilic enzymes as catalysts in environments where high salt stability is desired. This review provides a brief overview of lipases and esterases and examines specific structural motifs and evolutionary adaptations of halophilic lipases. Finally, we examine the state of research involving these enzymes and provide an in-depth look at an exciting algal-based biofuel production system. This system uses a recombinant halophilic lipase to increase oil production efficiency by cleaving algal fatty acids from the acyl carrier protein, which eliminates feedback inhibition of fatty acid synthesis.

KARAKTERISASI SIFAT-SIFAT BIOKIMIA EKSTRAK KASAR LIPASE EKSTRASELULER BAKTERI Azospirillum sp.PRD1

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Santi Nur Handayani

2011-11-01

Full Text Available Enzim lipase mempunyai peranan penting dalam katalis berbagai reaksi industri satu diantaranya pembuatan flavor melalui reaksi esterifikasi. Lipase adalah biokatalis yang berperan besar dalam aplikasi bioteknologi, seperti dalam sintesis biopolimer, biodiesel, produksi obat, dan produksi flavor. Peningkatan penggunaan lipase untuk industri mendorong dilakukan penelitian untuk mendapatkan sumber-sumber lipase baru. Sumber lipase yang potensial salah satunya adalah bakteri Azospirillum sp.PRD1 dari isolat lokal Laboratorium Mikrobiologi, Fakultas Biologi Universitas Jenderal Soedirman. Tujuan penelitian adalah untuk mendapatkan ekstrak kasar lipase dan menentukan karakteristik sifat-sifat biokimiawinya. Metode yang digunakan antara lain peremajaan bakteri Azospirillum sp.PRD1, dan produksi inokulum, penentuan waktu produksi optimum dan fase pertumbuhan bakteri, ekstraksi dan produksi ekstrak kasar lipase dan penentuan karakteristik sifat-sifat biokimiawinya. Hasil penelitian diperoleh ekstrak kasar lipase dari inokulum berumur 7 jam dan medium produksi dengan induser minyak zaitun yang diinkubasi selama 3 jam memiliki aktivitas spesifik 7,0547 Unit/mg. Lipase ekstrak kasar optimum pada pH 7, suhu 40 oC dan waktu inkubasi selama 25 menit. Lipase merupakan metaloenzim dengan kofaktor Zn2+ , Mn2+, Hg2+, Ca2+, Co2+ and Mg2+.

Yeast Kluyveromyces lactis as host for expression of the bacterial lipase: cloning and adaptation of the new lipase gene from Serratia sp.

Science.gov (United States)

Šiekštelė, Rimantas; Veteikytė, Aušra; Tvaska, Bronius; Matijošytė, Inga

2015-10-01

Many microbial lipases have been successfully expressed in yeasts, but not in industrially attractive Kluyveromyces lactis, which among other benefits can be cultivated on a medium supplemented with whey--cheap and easily available industrial waste. A new bacterial lipase from Serratia sp. was isolated and for the first time expressed into the yeast Kluyveromyces lactis by heterologous protein expression system based on a strong promoter of Kluyveromyces marxianus triosephosphate isomerase gene and signal peptide of Kluyveromyces marxianus endopolygalacturonase gene. In addition, the bacterial lipase gene was synthesized de novo by taking into account a codon usage bias optimal for K. lactis and was expressed into the yeast K. lactis also. Both resulting strains were characterized by high output level of the target protein secreted extracellularly. Secreted lipases were characterized for activity and stability.

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,.

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.

Anti- and Pro-Lipase Activity of Selected Medicinal, Herbal and Aquatic Plants, and Structure Elucidation of an Anti-Lipase Compound

Directory of Open Access Journals (Sweden)

Muhammad Abubakar Ado

2013-11-01

Full Text Available Plants that help in slowing down the digestion of triacylglycerols (TAGs in the pancreas and small intestine of humans play an important role in the reduction of obesity. On the other hand, there may be plants or plant parts that stimulate intestinal lipolytic activity, thus contributing to greater TAG assimilation. The aim of this study was to evaluate the aqueous methanolic extracts of ninety eight (98 medicinal, herbal and aquatic plant materials from Malaysia for their effect on porcine pancreatic lipase (PPL activity and to identify the structure of an anti-lipase compound from one of the sources. The degree of inhibition was also quantified as relative to orlistat activity against PPL (orlistat equivalents. Results revealed that while 19.4% of the extracts were found to have anti-lipase activity ≥80%, 12% were actually found to promote PPL activity. Twenty two percent (22.4% exhibited moderate inhibition (41%–80% and 2% were neutral toward PPL activity. The ripe fruit of Averrhoa carambola and the leaves of Archidendron jiringa (Jack I.C Nielsen L. (jering, Cynometra cauliflora (nam-nam and Aleurites moluccana (L. Willd (candle nut/buah keras had the highest (100% anti-lipase activity and are equivalent to 0.11 µg orlistat/mL. Plants that stimulated lipase activity included Pimpinella anisum L. (aniseed/jintan manis, activating the enzyme by 186.5%. Kaempferol 3-O-rhamnoside was isolated from the ethyl acetate fraction of C. cauliflora leaves and found to be an active lipase inhibitor. The structure was elucidated using 1H-NMR, 13C-NMR and 2D-NMR analyses.

Serum Lipase as Clinical Laboratory Index for Chronic Renal Failure Diagnosis.

Science.gov (United States)

Zhu, Ying; Dong, Jing; Wang, Ping; Huang, Huifang; Jin, Xiaohua; Zhou, Jingou; Shi, Jingfang; Gu, Guohao; Chen, Jun; Xu, Jun; Song, Yanhui

2016-07-01

Measuring the level of serum lipase has been used for the clinical diagnosis of acute pancreatitis. Reports showed that the serum lipase level increased in patients of clinical renal failure. In this study, we aimed to measure the change of serum lipase levels in chronic kidney diseases and determine whether it could serve as a clinical laboratory index for clinical renal failure diagnosis. Materials: The OLYMPUS AU5400 automatic biochemical analyzer was used to determine the serum levels of lipase and creatinine. The study included 120 cases in the clinical renal failure group, 76 cases in the nephrotic syndrome group, 81 cases in the chronic nephritis group, and 80 healthy controls from our hospital volunteers in the same period. We then compared the lipase levels and conducted statistical analyses among these groups. The serum lipase levels were 15.3 U/L, 79.8 U/L, 45.1 U/L, and 51.0 U/L in the normal control, clinical renal failure, nephrotic syndrome, and chronic nephritis groups, respectively. The lipase levels in the groups with diseases were significantly different compared with that of the normal control group (p renal failure group was significantly higher than that of the nephrotic syndrome group and chronic nephritis group (p chronic nephritis group (p > 0.05) was observed. Moreover, an association of the serum lipase with disease progression was observed in the study. Serum lipase is an effective serological index which can reflect the clinical changes in the clinical renal failure and tends to increase through the progression of renal dysfunction.

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

Directory of Open Access Journals (Sweden)

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.

Enhancement of lipase catalyzed-fatty acid methyl esters production from waste activated bleaching earth by nullification of lipase inhibitors.

Science.gov (United States)

Dwiarti, Lies; Ali, Ehsan; Park, Enoch Y

2010-01-01

This study sought to identify inhibitory factors of lipase catalyzed-fatty acid methyl esters (FAME) production from waste activated bleaching earth (wABE). During the vegetable oil refinery process, activated bleaching earth (ABE) is used for removing the impure compounds, but adsorbs vegetable oil up to 35-40% as on a weight basis, and then the wABE is discarded as waste material. The impurities were extracted from the wABE with methanol and evaluated by infra-red (IR) spectroscopy, which revealed that some were chlorophyll-plant pigments. The chlorophylls inhibited the lipase during FAME conversion from wABE. The inhibition by a mixture of chlorophyll a and b was found to be competitive. The inhibition of the enzymatic hydrolysis of waste vegetable oil contained in wABE by chlorophyll a alone was competitive, while the inhibition by chlorophyll b alone was non-competitive. Furthermore, the addition of a small amount of alkali nullified this inhibitory effect and accelerated the FAME production rate. When 0.9% KOH (w/w wABE) was added to the transesterification reaction with only 0.05% lipase (w/w wABE), the maximum FAME production rate improved 120-fold, as compared to that without the addition of KOH. The alkali-combined lipase significantly enhanced the FAME production rate from wABE, in spite of the presence of the plant pigments, and even when a lower amount of lipase was used as a catalyst.

Lipase catalyzed ester synthesis for food processing industries

Directory of Open Access Journals (Sweden)

Aravindan Rajendran

2009-02-01

Full Text Available Lipases are one of the most important industrial biocatalyst which catalyzes the hydrolysis of lipids. It can also reverse the reaction at minimum water activity. Because of this pliable nature, it is widely exploited to catalyze the diverse bioconversion reactions, such as hydrolysis, esterification, interesterification, alcoholysis, acidolysis and aminolysis. The property to synthesize the esters from the fatty acids and glycerol promotes its use in various ester synthesis. The esters synthesized by lipase finds applications in numerous fields such as biodiesel production, resolution of the recemic drugs, fat and lipid modification, flavour synthesis, synthesis of enantiopure pharmaceuticals and nutraceuticals. It plays a crucial role in the food processing industries since the process is unaffected by the unwanted side products. Lipase modifications such as the surfactant coating, molecular imprinting to suit for the non-aqueous ester synthesis have also been reported. This review deals with lipase catalyzed ester synthesis, esterification strategies, optimum conditions and their applications in food processing industries.Lipases são catalizadores industriais dos mais importantes, os quais catalizam a hidrólise de lipídeos. Também podem reverter a reação a um mínimo de atividade de água. Devido sua natureza flexível, é amplamente explorada para catalizar uma diversidade de reações de bioconversão como hidrólise, esterificação, interesterificação, alcoólise, acidólise e aminólise. A propriedade de síntese de esteres a partir de ácidos graxos e glicerol promoveu seu uso em várias sínteses de esteres. Os esteres sintetizados por lipases encontram aplicação em numerosos campos como a produção de biodiesel, resolução de drogas racêmicas, modificação de gorduras e lipídios, sintese de aromas, síntese de produtos farmacêuticos enantiopuro e nutracêuticos. As lipases possuem um papel crucial nas indústrias de

Glycerol Monolaurate Inhibits Lipase Production by Clinical Ocular Isolates Without Affecting Bacterial Cell Viability.

Science.gov (United States)

Flanagan, Judith Louise; Khandekar, Neeta; Zhu, Hua; Watanabe, Keizo; Markoulli, Maria; Flanagan, John Terence; Papas, Eric

2016-02-01

We sought to determine the relative lipase production of a range of ocular bacterial isolates and to assess the efficacy of glycerol monolaurate (GML) in inhibiting this lipase production in high lipase-producing bacteria without affecting bacterial cell growth. Staphylococcus aureus,Staphylococcus epidermidis,Propionibacterium acnes, and Corynebacterium spp. were inoculated at a density of 10(6)/mL in varying concentrations of GML up to 25 μg/mL for 24 hours at 37 °C with constant shaking. Bacterial suspensions were centrifuged, bacterial cell density was determined, and production of bacterial lipase was quantified using a commercial lipase assay kit. Staphylococcus spp. produced high levels of lipase activity compared with P. acnes and Corynebacterium spp. GML inhibited lipase production by Staphylococcal spp. in a dose-dependent manner, with S. epidermidis lipase production consistently more sensitive to GML than S. aureus. Glycerol monolaurate showed significant (P < 0.05) lipase inhibition above concentrations of 15 μg/mL in S. aureus and was not cytotoxic up to 25 μg/mL. For S. epidermidis, GML showed significant (P < 0.05) lipase inhibition above 7.5 μg/mL. Lipase activity varied between species and between strains. Staphylococcal spp. produced higher lipase activity compared with P. acnes and Corynebacterium spp. Glycerol monolaurate inhibited lipase production by S. aureus and S. epidermidis at concentrations that did not adversely affect bacterial cell growth. GML can be used to inhibit ocular bacterial lipase production without proving detrimental to commensal bacteria viability.

Emulsifying triglycerides with dairy phospholipids instead of soy lecithin modulates gut lipase activity

DEFF Research Database (Denmark)

Mathiassen, Jakob Hovalt; Nejrup, Rikke Guldhammer; Frøkiær, Hanne

2015-01-01

in particular to limit fatty acid absorption in babies given infant formulas. Since interaction between the lipid droplet and the gastric and duodenal lipases occur through the hydrophobic/hydrophilic interface, the composition of the emulsifier may be crucial for efficient hydrolysis. We therefore determined...... hydrolytic rate of gastric lipase and pancreatic lipase, on their own or pancreatic lipase after gastric lipase on TAG droplets of similar size emulsified in either soy lecithin (SL) or in bovine milk phospholipids (MPL), more similar to human milk globule membrane lipids than soy lecithin. Gastric lipase...... for formulas for term-born infants....

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.

Study the effect of F17S mutation on the chimeric Bacillus thermocatenulatus lipase

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Seyed Hossein Khaleghinejad

2016-06-01

Full Text Available Lipases (triacylglycerol acylhydrolase, EC 3.1.1.3 are one of the highest value commercial enzymes as they have potential applications in biotechnology for detergents, food, pharmaceuticals, leather, textiles, cosmetics, and paper industries; and are currently receiving considerable attention because of their potential applications in biotechnology. Bacillus thermocatenulatus Lipase 2 (BTL2 is one of the most important research targets, because of its potential industrial applications. In this study, the effect of substitution Phe17 with Ser in mutated BTL2 lipase, which conserved pentapeptide (112Ala-His-Ser-Gln-Gly116 was replaced with similar sequences (207Gly-Glu-Ser-Ala-Gly211 of Candida rugosa lipase (CLR at the nucleophilic elbow region. Docking results confirmed the mutated lipase to be better than the chimeric lipase. So, cloning was conducted, and the mutated and chimeric btl2 genes were expressed in Escherichia coli, and then the enzymes were purified by anion exchange chromatography. The mutation increased lipase lipolytic activity against most of the applied substrates, with the exception of tributyrin when compared with chimeric lipase. Further, the mutated lipase exhibited higher activity than the chimeric lipase at all temperatures. Optimum pH of the mutated lipase was obtained at pH 9.5, which was more than the chimeric one. Enzyme activity of the mutated lipase in the presence of organic solvents, detergents, and metal ions was also improved than the chimeric lipase.

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

A lipase with broad temperature range from an alkaliphilic gamma-proteobacterium isolated in Greenland

DEFF Research Database (Denmark)

Schmidt, Mariane; Larsen, Dorte Møller; Stougaard, Peter

2010-01-01

A gamma-proteobacterium related to the genera Alteromonadales and Pseudomonadales , isolated from a cold and alkaline environment in Greenland, has been shown to produce a lipase active between 5 ° C and 80 ° C, with optimal activity at 55 ° C and pH 8. PCR-based screening of genomic DNA from...... the isolated bacterium, followed by genome walking, resulted in two complete open reading frames, which were predicted to encode a lipase and its helper protein, a lipase foldase. The amino acid sequence derived for the lipase showed resemblance to lipases from Pseudomonas , Rhodoferax, Aeromonas and Vibrio...... . The two genes were cloned into different expression systems in E. coli with or without a putative secretion sequence, but despite the fact that both recombinant lipase and lipase foldase were observed on SDS–PAGE, no recombinant lipase activity was detected. Attempts to refold the recombinant lipase...

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.

Desempenho de diferentes lipases imobilizadas na síntese de biodiesel de óleo de palma = Performance of different immobilized lipases in palm oil biodiesel synthesis

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Grazielle dos Santos Silva

2011-04-01

Full Text Available O presente trabalho teve como objetivo determinar as condicoes otimizadas da sintese enzimatica de biodiesel, a partir do oleo de palma e etanol, empregando diferentes lipases imobilizadas (lipase de Pseudomonas fluorescens imobilizada em SiO2-PVA e lipase de Candida antartica imobilizada em resina acrilica - Novozym„µ 435 em meio isento de solvente. Uma matriz de planejamento fatorial foi utilizada para avaliar a influencia da temperatura (42 ¡V 58„aC e a razao molar entre etanol e oleo de palma (6:1 ¡V 18:1 no rendimento detransesterificacao alcancado para cada preparacao de lipase. Os efeitos principais foram ajustados por analise de regressao multipla a modelos lineares e o rendimento maximo foi obtido quando o sistema operacional foi operado a 42„aC com substratos contendo etanol eoleo de palma na razao molar de 18:1. Os modelos matematicos que representam o rendimento global da reacao para cada lipase imobilizada foram considerados adequados para descrever os resultados experimentais.Optimized conditions for palm oil and ethanol enzymatic biodiesel synthesis were determined with different immobilized lipases SiO2-PVA-immobilized lipase from Pseudomonas fluorescens and acrylic resin-immobilized lipase, NovozymR435, from Candida antartica, in solvent-free medium. A full factorial design assessed the influence oftemperature (42 ¡V 58¢XC and ethanol: palm oil (6:1 ¡V 18:1 molar ratio on the transesterification yield. Main effects were adjusted by multiple regression analysis to linear models and the maximum transesterification yield was obtained at 42¢XC and 18:1 ethanol:palm oil molar ratio. Mathematical models featuring total yield for each immobilized lipase were suitable to describe the experimental results.

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.

AKTIVITAS HIDROLISIS ENZIM LIPASE DARI KENTOS KELAPA TERHADAP MINYAK KELAPA Hidrolysis Activity of Lipase Enzyme from Coconut Houstorium for Coconut Oil

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Mohammad Su’i

2012-05-01

Full Text Available This research was aimed to study hydrolysis conditions of houstorium lipases enzyme using coconut oil as substrate. Hydrolysis conditions studied were substrate (coconut oil concentration, enzyme substrate ratio, duration of hydro- lysis and effect of stirring to hydrolysis. The results show  that lipase of coconut houstorium may be optimally used at a coconut oil concentration of 10 %, enzyme to substrate ratio of 3 : 10 (v/v and hydrolysis for 60 minutes with stirring. ABSTRAK Penelitian ini mempelajari kondisi hidrolisis minyak kelapa yang optimum menggunakan enzim lipase dari kentos kelapa. Kondisi hidrolisis yang dipelajari meliputi konsentrasi substrat optium, perbandingan enzim : substrat dan lama hidrolisis yang optimum serta pengaruh pengadukan selama hidrolisis. Hasil penelitian menunjukkan bahwa, hidrolisis minyak kelapa menggunakan enzim lipase kentos kelapa menghasilkan asam lemak bebas paling banyak pada kon- sentrasi substrat (minyak kelapa 10 %, perbandingan enzim : substrat yaitu 3 : 10 (v/v, lama hidroloisa 60 menit dan dilakukan pengadukan selama hidrolisis.

Streptomyces rimosus GDS(L Lipase: Production, Heterologous Overexpression and Structure-Stability Relationship

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Marija Abramić

2003-01-01

Full Text Available Streptomyces rimosus lipase gene has been overexpressed in a heterologous host, S. lividans TK23. The maximal lipase activity was determined in the culture filtrates of the late stationary phase. Time course of lipase production was monitored by a modified plate assay. S. rimosus lipase gene has been located on the AseI B fragment approximately 2 Mb far from the left end of the S. rimosus linear chromosome. Out of eight examined streptomycetes, the presence of this rare type of bacterial lipase gene was detected in two belonging to the S. rimosus taxonomic cluster, and in one non-related species. Comparison of protein sequences of the Streptomyces lipolytic enzymes was performed. The result indicated the best structural stability of the putative S. coelicolor lipase-2.

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)

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.

A Quantitative Fluorescence-Based Lipase Assay

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Giovanna Lomolino

2012-01-01

Full Text Available An easy and fast gel diffusion assay for detecting and monitoring lipase activity by quantification of fluorescein is described. By measuring the intensity of fluorescein, it is possible to obtain a calibration curve with a regression coefficient better than by using the radius of fluorescent haloes. Through the quantification of fluorescence intensity of fluorescein released after the hydrolysis of a fluorescent ester, fluorescein dibutyrate, used as substrate in agar plates, commercial and skimmed milk lipase activity were studied. Moreover, with this method, lipase activity can be monitored in reaction medium that contains compounds which are affected by turbidity or cause measurement interference for UV-spectrophotometer and fluorimeter. In this experiment, boiled skimmed milk was dispersed in the agar gel with fluorescein dibutyrate, and it was used as a reaction medium to mimic natural conditions. The development of such an assay has a potential for applications in industries ranging from pharmaceuticals to food production and monitoring.

Gamma-irradiation sterilization of lipases for cheese making

Energy Technology Data Exchange (ETDEWEB)

Umanskij, M S; Borovkova, Yu A; Odegov, N I [Vsesoyuznyj Nauchno-Issledovatel' skij Inst. Maslodel' noj i Syrodel' noj Promyshlennosti, Uglich (USSR)

1979-03-01

The possibility of sterilizing the enzyme compounds of lipases from Oospora fragrans strains by gamma irradiation was studied. The enzyme compounds were exposed to gamma irradiation at the doses from 0.1 to 0.8 Mrad with the discreteness of 0.1 Mrad and at the dose of 2.0 Mrad. After the radiation treatment the lipases were investigated for bacterial invasion by the cultivation method and for the lipolytic activity by the titrometrical method. It is shown that the sterilization effect is achieved without losses of lipase activity and the radiation dose necessary for sterilization depends on initial invasion levels in the enzyme compounds.

Selection and optimization of extracellular lipase production using ...

African Journals Online (AJOL)

The aim of this study was to isolate and select lipase-producing microorganisms originated from different substrates, as well as to optimize the production of microbial lipase by submerged fermentation under different nutrient conditions. Of the 40 microorganisms isolated, 39 showed a halo around the colonies and 4 were ...

Optimization of lipase production by Staphylococcus sp. Lp12

African Journals Online (AJOL)

USER

2010-02-08

Feb 8, 2010 ... an enormous attention because of their biotechnological applications. Lipases remain ... selective transformations. The exponential increase in .... mass) lipase production and pH at regular intervals of time 24, 48 and 72 h on ...

New lipases by mining of Pleurotus ostreatus genome.

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Alessandra Piscitelli

Full Text Available The analysis of Pleurotus ostreatus genome reveals the presence of automatically annotated 53 lipase and 34 carboxylesterase putative coding-genes. Since no biochemical or physiological data are available so far, a functional approach was applied to identify lipases from P. ostreatus. In the tested growth conditions, four lipases were found expressed, with different patterns depending on the used C source. Two of the four identified proteins (PleoLip241 and PleoLip369, expressed in both analysed conditions, were chosen for further studies, such as an in silico analysis and their molecular characterization. To overcome limits linked to native production, a recombinant expression approach in the yeast Pichia pastoris was applied. Different expression levels were obtained: PleoLip241 reached a maximum activity of 4000 U/L, whereas PleoLip369 reached a maximum activity of 700 U/L. Despite their sequence similarity, these enzymes exhibited different substrate specificity and diverse stability at pH, temperature, and presence of metals, detergents and organic solvents. The obtained data allowed classifying PleoLip241 as belonging to the "true lipase" family. Indeed, by phylogenetic analysis the two proteins fall in different clusters. PleoLip241 was used to remove the hydrophobic layer from wool surface in order to improve its dyeability. The encouraging results obtained with lipase treated wool led to forecast PleoLip241 applicability in this field.

High cell density fed-batch fermentations for lipase production: feeding strategies and oxygen transfer.

Science.gov (United States)

Salehmin, M N I; Annuar, M S M; Chisti, Y

2013-11-01

This review is focused on the production of microbial lipases by high cell density fermentation. Lipases are among the most widely used of the enzyme catalysts. Although lipases are produced by animals and plants, industrial lipases are sourced almost exclusively from microorganisms. Many of the commercial lipases are produced using recombinant species. Microbial lipases are mostly produced by batch and fed-batch fermentation. Lipases are generally secreted by the cell into the extracellular environment. Thus, a crude preparation of lipases can be obtained by removing the microbial cells from the fermentation broth. This crude cell-free broth may be further concentrated and used as is, or lipases may be purified from it to various levels. For many large volume applications, lipases must be produced at extremely low cost. High cell density fermentation is a promising method for low-cost production: it allows a high concentration of the biomass and the enzyme to be attained rapidly and this eases the downstream recovery of the enzyme. High density fermentation enhances enzyme productivity compared with the traditional submerged culture batch fermentation. In production of enzymes, a high cell density is generally achieved through fed-batch operation, not through perfusion culture which is cumbersome. The feeding strategies used in fed-batch fermentations for producing lipases and the implications of these strategies are discussed. Most lipase-producing microbial fermentations require oxygen. Oxygen transfer in such fermentations is discussed.

Lipase polystyrene giant amphiphiles.

Science.gov (United States)

Velonia, Kelly; Rowan, Alan E; Nolte, Roeland J M

2002-04-24

A new type of giant amphiphilic molecule has been synthesized by covalently connecting a lipase enzyme headgroup to a maleimide-functionalized polystyrene tail (40 repeat units). The resulting biohybrid forms catalytic micellar rods in water.

Plant latex lipase as biocatalysts for biodiesel production | Mazou ...

African Journals Online (AJOL)

Plant latex lipase as biocatalysts for biodiesel production. ... This paper provides an overview regarding the main aspects of latex, such as the reactions catalyzed, physiological functions, specificities, sources and their industrial applications. Keywords: Plant latex, lipase, Transesterification, purification, biodiesel ...

Isolation and characterization of lipase-producing Bacillus strains ...

African Journals Online (AJOL)

Bacillus strains (B1 - B5) producing extra cellular lipase were isolated from the soil sample of coconut oil industry. The strains were identified by morphological and biochemical characters. Growth of the organisms and lipase production were measured with varying pH (4 - 9) temperature (27, 37 and 47ºC) and various ...

Production of extracellular lipase by the phytopathogenic fungus Fusarium solani FS1 Produção de lipase extracelular pelo fungo fitopatogênico Fusarium solani FS1

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Maria de Mascena Diniz Maia

1999-12-01

Full Text Available A Brazilian strain of Fusarium solani was tested for extracellular lipase production in peptone-olive oil medium. The fungus produced 10,500 U.l-1 of lipase after 72 hours of cultivation at 25oC in shake-flask at 120 rpm in a medium containing 3% (w/v peptone plus 0.5% (v/v olive oil. Glucose (1% w/v was found to inhibit the inductive effect of olive oil. Peptone concentrations below 3% (w/v resulted in a reduced lipase production while increased olive oil concentration (above 0.5% did not further stimulate lipase production. The optimum lipase activity was achieved at pH 8.6 and 30oC and a good enzyme stability (80% activity retention was observed at pH ranging from 7.6 to 8.6, and the activity rapidly dropped at temperatures above 50oC. Lipase activity was stimulated by the addition of n-hexane to the culture medium supernatants, in contrast to incubation with water-soluble solvents.

Rat liver contains a limited number of binding sites for hepatic lipase

NARCIS (Netherlands)

G.C. Schoonderwoerd (Kees); A.J.M. Verhoeven (Adrie); H. Jansen (Hans)

1994-01-01

textabstractThe binding of hepatic lipase to rat liver was studied in an ex vivo perfusion model. The livers were perfused with media containing partially purified rat hepatic lipase or bovine milk lipoprotein lipase. The activity of the enzymes was determined in the

Lipase inhibition and antiobesity effect of Atractylodes lancea.

Science.gov (United States)

Jiao, Ping; Tseng-Crank, Julie; Corneliusen, Brandon; Yimam, Mesfin; Hodges, Mandee; Hong, Mei; Maurseth, Catherine; Oh, Misun; Kim, Hyunjin; Chu, Min; Jia, Qi

2014-05-01

The ethanol extract of Atractylodes lancea rhizome displayed significant lipase inhibition with an IC50 value of 9.06 µg/mL in a human pancreatic lipase assay from high-throughput screening. Bioassay-guided isolation led to the identification of one new polyacetylene, syn-(5E,11E)-3-acetoxy-4-O-(3-methylbutanoyl)-1,5,11-tridecatriene-7,9-diyne-3,4-diol (7), along with six known compounds (1-6). The structure of compound 7 was determined based on the analysis of NMR and MS data. Among these seven lipase inhibitors, the major compound atractylodin (1) showed the highest lipase inhibitory activity (IC50 = 39.12 µM). The antiobesity effect of the ethanol extract of Atractylodes lancea rhizome was evaluated in a high-fat diet-induced obesity mice model at daily dosages of 250 mg/kg and 500 mg/kg body weight for 4 weeks, and treatment with this extract demonstrated a moderate efficacy at the 500 mg/kg dose level. Georg Thieme Verlag KG Stuttgart · New York.

Comparative performance of microbial lipases immobilized on magnetic polysiloxane polyvinyl alcohol particles

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Laura Maria Bruno

2008-10-01

Full Text Available Microbial lipase from Mucor miehei and Candida rugosa were immobilized by covalent binding onto magnetized polysiloxane polyvinyl alcohol particles (POS-PVA. The resulting immobilized derivatives were evaluated in aqueous solution (olive oil hydrolysis and organic solvent (butyl butyrate synthesis. Higher catalytic activities were found when the coupling procedure was made with M. miehei lipase. Immobilized M. miehei lipase also showed a better operational stability and a higher half-life than C. rugosa lipase after the successive batches of esterification. The performance of C. rugosa immobilized derivative was constrained by the low lipase loading used in the immobilizing step. Further information regarding the both immobilized derivatives was also obtained through chemical composition (FTIR.Lipases microbianas de Mucor miehei e Candida rugosa foram imobilizadas por ligação covalente em partículas magnetizadas de polisiloxano-álcool polivinílico (POS-PVA. Os derivados imobilizados resultantes foram avaliados em solução aquosa (hidrólise de azeite de oliva e em solvente orgânico (síntese de butirato de butila. As maiores atividades catalíticas foram encontradas quando o procedimento de ligação foi realizado com lipase de M. miehei. O derivado imobilizado de lipase de M. miehei também apresentou melhores resultados de estabilidade operacional e tempo de meia-vida do que o de lipase de C. rugosa, após sucessivas bateladas de esterificação. O desempenho do derivado imobilizado de C. rugosa foi restringido pelo baixo carregamento de lipase usado na etapa de imobilização. Informações adicionais a respeito de ambos derivados imobilizados também foram obtidas através da composição química (FTIR.

Post-heparin plasma lipoprotein lipase, but not hepatic lipase activity, is related to plasma adiponectin in type 2 diabetic patients and healthy subjects

NARCIS (Netherlands)

De Vries, R; Wolffenbuttel, BHR; Sluiter, WJ; Van Tol, A; Dullaart, RPF

2005-01-01

The aim of this study was to determine the relationships of plasma adiponectin with post-heparin plasma lipoprotein lipase (LPL) and hepatic lipase (HL) activities, and to evaluate whether plasma adiponectin contributes to diabetes-associated dyslipidaemia. Plasma adiponectin, post-heparin plasma

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

Lipases de látex vegetais: propriedades e aplicações industriais

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Paques Fernanda Wiermann

2006-01-01

Full Text Available Biocatalysts have innumerous advantages with respect to classical chemical processes, such as high specificity. Lipases (EC 3.1.1.3 are biocatalysts with large application in synthesis and hydrolysis reactions of triacylglycerols. The search for new sources of lipases has been intensified in the last years due to the high cost of microbial and animal lipases, wich restricts their use on an industrial scale. Lipases obtained from the latex of Carica papaya, Carica pentagona, Euphorbia characias, E. wulfenii, known for their proteolytic properties, are a good alternative source. In this review, we describe the well-known sources of vegetal lipases extracted from the latex and present some of their industrial applications.

Structural investigations of the regio- and enantioselectivity of lipases

NARCIS (Netherlands)

Lang, Dietmar A.; Dijkstra, Bauke W.

Although lipases are widely applied for the stereospecific resolution of racemic mixtures of esters, the atomic details of the factors that are responsible for their stereospecificity are largely obscure. We determined the X-ray structures of Pseudomonas cepacia lipase in complex with two

Endothelial and lipoprotein lipases in human and mouse placenta

DEFF Research Database (Denmark)

Lindegaard, Marie L S; Olivecrona, Gunilla; Christoffersen, Christina

2005-01-01

Placenta expresses various lipase activities. However, a detailed characterization of the involved genes and proteins is lacking. In this study, we compared the expression of endothelial lipase (EL) and LPL in human term placenta. When placental protein extracts were separated by heparin-Sepharos...

Purification and Characterization of Lipase from Aspergillus flavus ...

African Journals Online (AJOL)

USER

Abstract. Lipase from Aspergillus flavus was purified in a single step purification using MnFeO4 magnetic nano particles to achieve a 20.53- fold purification with specific activity of. 11.29 U/mg and a 59% recovery yield. SDS-PAGE of lipase showed a single pure band with corresponding molecular weight of 35 kDa.

Purification and Characterization of Lipase from Aspergillus flavus ...

African Journals Online (AJOL)

Lipase from Aspergillus flavus was purified in a single step purification using MnFeO4 magnetic nano particles to achieve a 20.53- fold purification with specific activity of 11.29 U/mg and a 59% recovery yield. SDS-PAGE of lipase showed a single pure band with corresponding molecular weight of 35 kDa. The optimal ...

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.

Lipase applications in oil hydrolysis with a case study on castor oil: a review.

Science.gov (United States)

Goswami, Debajyoti; Basu, Jayanta Kumar; De, Sirshendu

2013-03-01

Lipase (triacylglycerol acylhydrolase) is a unique enzyme which can catalyze various types of reactions such as hydrolysis, esterification, alcoholysis etc. In particular, hydrolysis of vegetable oil with lipase as a catalyst is widely studied. Free lipase, lipase immobilized on suitable support, lipase encapsulated in a reverse micelle and lipase immobilized on a suitable membrane to be used in membrane reactor are the most common ways of employing lipase in oil hydrolysis. Castor oil is a unique vegetable oil as it contains high amounts (90%) of a hydroxy monounsaturated fatty acid named ricinoleic acid. This industrially important acid can be obtained by hydrolysis of castor oil. Different conventional hydrolysis processes have certain disadvantages which can be avoided by a lipase-catalyzed process. The degree of hydrolysis varies widely for different lipases depending on the operating range of process variables such as temperature, pH and enzyme loading. Immobilization of lipase on a suitable support can enhance hydrolysis by suppressing thermal inactivation and estolide formation. The presence of metal ions also affects lipase-catalyzed hydrolysis of castor oil. Even a particular ion has different effects on the activity of different lipases. Hydrophobic organic solvents perform better than hydrophilic solvents during the reaction. Sonication considerably increases hydrolysis in case of lipolase. The effects of additives on the same lipase vary with their types. Nonionic surfactants enhance hydrolysis whereas cationic and anionic surfactants decrease it. A single variable optimization method is used to obtain optimum conditions. In order to eliminate its disadvantages, a statistical optimization method is used in recent studies. Statistical optimization shows that interactions between any two of the following pH, enzyme concentration and buffer concentration become significant in presence of a nonionic surfactant named Span 80.

Covalent immobilization of lipase from Candida rugosa on Eupergit®

Directory of Open Access Journals (Sweden)

Bezbradica Dejan I.

2005-01-01

Full Text Available An approach is presented for the stable covalent immobilization of Upase from Candida rugosa on Eupergit® with a high retention of hydrolytic activity. It comprises covalent bonding via lipase carbohydrate moiety previously modified by periodate oxidation, allowing a reduction in the involvement of the enzyme functional groups that are probably important in the catalytic mechanism. The hydrolytic activities of the lipase immobilized on Eupergif1 by two conventional methods (via oxirane group and via glutaralde-hyde and with periodate method were compared. Results of lipase assays suggest that periodate method is superior for lipase immobilization on Eupergit® among methods applied in this study with respect to both, yield of immobilization and hydrolytic activity of the immobilized enzyme.

Iodine-125-labeled lipoprotein lipase as a tool to detect and study spontaneous lipolysis in bovine milk

International Nuclear Information System (INIS)

Sundheim, G.; Bengtsson-Olivecrona, G.

1986-01-01

The distribution of lipoprotein lipase among cream, casein, and milk serum can be evaluated by addition of a trace amount of 125 I-labeled lipoprotein lipase to milk. Radioactive lipase was distributed in parallel to endogenous lipase under several conditions. In some milk samples, binding of lipase to cream increased when the milk was cooled. Correlation was good between bound labeled lipase and degree of cold-induced lipolysis in corresponding milk samples. Binding of lipase to cream or to casein was not saturable by addition of two-to threefold more lipase than is normally present in milk. In milk with a relatively high fraction of lipase bound to cream, a correspondingly lower fraction was associated with casein, whereas the fraction of lipase in milk serum was similar in all milk samples. Cold-induced binding of lipoprotein lipase to cream was not fully reversed when the milk was warmed again. Heparin released lipase from casein and increased the amount of lipase bound to cream after cooling

Effect of fermentation conditions on lipase production by Candida utilis

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SANJA Z. GRBAVCIC

2007-08-01

Full Text Available A wild yeast strain isolated from spoiled soybean oil and identified as Candida utilis initially presented rather low lipase activity (approximately 4 IU dm-3 in submerged culture in a universal yeast medium containing 2 % malt extract. Stu­dies were undertaken to improve the lipase production. The best yields of lipase were obtained with a medium supplemented with caprylic and oleic acids as indu­cers, but higher concentrations of the former (> 0.5 % had a negative effect on the lipase production and cell growth. The type of nitrogen source seemed also to be very important. The highest lipolytic activity of 284 IU dm-3 was achieved after 5 days of fermentation in a medium containing oleic acid and hydrolyzed casein as carbon and nitrogen sources, respectively, and supplemented with Tween 80®. It was shown that optimization of the fermentation conditions can lead to a significant improvement in the lipase production (more than 70-fold higher compared to the initial value obtained in the non-optimized medium.

Characteristics of lipase isolated from coconut (Cocos nucifera linn ...

African Journals Online (AJOL)

GREGO

2007-03-19

Mar 19, 2007 ... Lipase from coconut plant grown under complete nutrient conditions showed ... 35°C and had a broad optimum pH of 7.5 – 8.5. Key words: Lipase .... inhibited by the ex- cess of substrate concentration or change of physio-.

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

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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.

Covalent immobilization of lipases on monodisperse magnetic microspheres modified with PAMAM-dendrimer

Energy Technology Data Exchange (ETDEWEB)

Zhu, Weiwei [Lanzhou University, State Key Laboratory of Applied Organic Chemistry, Key Laboratory of Nonferrous Metal Chemistry and Resources Utilization of Gansu Province, College of Chemistry and Chemical Engineering, Institute of Biochemical Engineering and Environmental Technology (China); Zhang, Yimei [Suzhou Research Academy of North China Electric Power University (China); Hou, Chen; Pan, Duo; He, Jianjun; Zhu, Hao, E-mail: zhuhao07@lzu.edu.cn [Lanzhou University, State Key Laboratory of Applied Organic Chemistry, Key Laboratory of Nonferrous Metal Chemistry and Resources Utilization of Gansu Province, College of Chemistry and Chemical Engineering, Institute of Biochemical Engineering and Environmental Technology (China)

2016-02-15

This paper reported an immobilization of Candida rugosa lipase (CRL) onto PAMAM-dendrimer-grafted magnetic nanoparticles synthesized by a modified solvothermal reduction method. The dendritic magnetic nanoparticles were amply characterized by several instrumental measurements, and the CRL was covalently anchored on the three generation supports with glutaraldehyde as coupling reagent. The amount of immobilized enzyme was up to 150 mg/g support and the factors related with the enzyme activity were investigated. The immobilization of lipase improved their performance in wider ranges of pH and temperature. The immobilized lipase exhibited excellent thermal stability and reusability in comparison with free enzyme and can be reused 10 cycles with the enzymatic activity remained above 90 %. The properties of lipase improved obviously after being immobilized on the dendritic supports. The inactive immobilized lipase could be regenerated with glutaraldehyde and Cu{sup 2+}, respectively. This synthetic strategy was facile and eco-friendly for applications in lipase immobilization.

Síntese do butirato de n-butila empregando lipase microbiana imobilizada em copolímero de estireno-divinilbenzeno Synthesis of butyl butyrate by microbial lipase immobilized onto styrene-divinylbenzene copolymer

Directory of Open Access Journals (Sweden)

Pedro Carlos de Oliveira

2000-10-01

Full Text Available This work investigates the reaction parameters of an immobilized lipase in the esterification reaction of n-butanol and butyric acid. Microbial lipase from Candida rugosa was immobilized onto styrene-divinylbenzene copolymer (STY-DVB and subsequently introduced in an organic medium containing substrates in appropriate concentrations. Heptane was selected as solvent on the basis of its compatibility with the resin and the enzyme. The influence of molar ratio of acid to alcohol, amount of immobilized lipase and temperature on the butyl butyrate formation was determined. The results were compared with those achieved with free lipase and Lipozyme (commercially immobilized lipase under the same operational conditions.

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.

Influences of apolipoprotein E on soluble and heparin-immobilized hepatic lipase

International Nuclear Information System (INIS)

Landis, B.A.; Rotolo, F.S.; Meyers, W.C.; Clark, A.B.; Quarfordt, S.H.

1987-01-01

The effect of human apolipoprotein E (apoE), either alone or in combination with apoC, on the lipolysis of a radiolabeled triglyceride emulsion was studied with hepatic lipase in solution and immobilized on heparin-Sepharose. The soluble hepatic lipase was inhibited, whereas the heparin-immobilized lipase was stimulated by apoE. This stimulation was attenuated by combining apoE with either apoC-II or C-III. The heparin-immobilized lipase demonstrated much less lipolysis of the zwitterionic phosphatidylcholine-stabilized triglyceride emulsion than did the soluble enzyme. This difference was less when the emulsion was stabilized by a nonionic detergent. apoE inhibited lipase activity when assayed under conditions (0.4 M NaCl) of bound enzyme and unbound substrate. Increasing the emulsion apoE content beyond optimum inhibited lipolysis by the immobilized enzyme. Kinetic analysis of phosphatidylcholine-stabilized triglyceride emulsions revealed a significant decrease in immobilized enzyme K/sub m/ and an increase in V/sub max/ when the emulsion was supplemented with apoE. Distributing the immobilized lipase in clustered aggregates produced more lipolysis than when the same enzyme content was uniformly bound

Secoiridoids from the stem barks of Fraxinus rhynchophylla with pancreatic lipase inhibitory activity.

Science.gov (United States)

Ahn, Jong Hoon; Shin, Eunjin; Liu, Qing; Kim, Seon Beom; Choi, Kyeong-Mi; Yoo, Hwan-Soo; Hwang, Bang Yeon; Lee, Mi Kyeong

2013-01-01

Pancreatic lipase digests dietary fats by hydrolysis, which is a key enzyme for lipid absorption. Therefore, reduction of fat absorption by the inhibition of pancreatic lipase is suggested to be a therapeutic strategy for obesity. From the EtOAc-soluble fraction of the stem barks of Fraxinus rhynchophylla (Oleaceae), four secoiridoids such as ligstroside (1), oleuropein (2), 2"-hydroxyoleuropein (3) and hydroxyframoside B (4) were isolated. The inhibitory activity of these compounds on pancreatic lipase was assessed using porcine pancreatic lipase as an in vitro assay system. Compound 4 showed the strongest inhibition on pancreatic lipase, which followed by compounds 1-3. In addition, compound 4 exerted inhibitory effect on pancreatic lipase in a mixed mechanism of competitive and noncompetitive manner. Taken together, F. rhynchophylla and its constituents might be beneficial to obesity.

Improvement of lipase production at different stirring speeds and oxygen levels

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F.O.M. Alonso

2005-03-01

Full Text Available Lipase production by a Brazilian wild strain of Yarrowia lipolytica at different stirring speeds and air flow rates was studied. The relationship among lipid consumption, cell growth and lipase production by this microorganism is presented. The most pronounced effect of oxygen on lipase production was determined by stirring speed. Maximum lipase activity was detected in the late stationary phase at 200 rpm and an air flow rate of 1-2 dm³/min (0.8-1.7 vvm when the lipid source had been fully consumed. Higher stirring speeds resulted in mechanical and/or oxidative stress, while lower stirring speeds seemed to limit oxygen levels. An increase in the availability of oxygen at higher air flow rates led to faster lipid uptake and anticipation of enzyme release into the culture medium. The highest lipase production was obtained at 200 rpm and 1 dm³/min (0.8 vvm.

Statistical optimization for lipase production from solid waste of vegetable oil industry.

Science.gov (United States)

Sahoo, Rajesh Kumar; Kumar, Mohit; Mohanty, Swati; Sawyer, Matthew; Rahman, Pattanathu K S M; Sukla, Lala Behari; Subudhi, Enketeswara

2018-04-21

The production of biofuel using thermostable bacterial lipase from hot spring bacteria out of low-cost agricultural residue olive oil cake is reported in the present paper. Using a lipase enzyme from Bacillus licheniformis, a 66.5% yield of methyl esters was obtained. Optimum parameters were determined, with maximum production of lipase at a pH of 8.2, temperature 50.8°C, moisture content of 55.7%, and biosurfactant content of 1.693 mg. The contour plots and 3D surface responses depict the significant interaction of pH and moisture content with biosurfactant during lipase production. Chromatographic analysis of the lipase transesterification product was methyl esters, from kitchen waste oil under optimized conditions, generated methyl palmitate, methyl stearate, methyl oleate, and methyl linoleate.

Enzymes used in detergents: Lipases | Hasan | African Journal of ...

African Journals Online (AJOL)

This review describes the applications of microbial lipases in detergents. Enzymes can reduce the environmental load of detergent products as the chemicals used in conventional detergents are reduced; they are biodegradable, non-toxic and leave no harmful residues. Besides lipases, other enzymes are widely used in ...

Optimization of lipase production by Staphylococcus sp. Lp12

African Journals Online (AJOL)

USER

2010-02-08

Feb 8, 2010 ... of the genera Pseudomonas, Bacillus, Staphylococcus,. Achromobacter have been cloned and characterized. Bacterial lipases are mostly inducible enzymes and require some form of oil, fatty acid, fatty acid alcohol or fatty acid ester and surfactants for induction (Immanuel et al., 2008). Lipase biosynthesis ...

Fatty Acid Signaling: The New Function of Intracellular Lipases

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Zuzana Papackova

2015-02-01

Full Text Available Until recently, intracellular triacylglycerols (TAG stored in the form of cytoplasmic lipid droplets have been considered to be only passive “energy conserves”. Nevertheless, degradation of TAG gives rise to a pleiotropic spectrum of bioactive intermediates, which may function as potent co-factors of transcription factors or enzymes and contribute to the regulation of numerous cellular processes. From this point of view, the process of lipolysis not only provides energy-rich equivalents but also acquires a new regulatory function. In this review, we will concentrate on the role that fatty acids liberated from intracellular TAG stores play as signaling molecules. The first part provides an overview of the transcription factors, which are regulated by fatty acids derived from intracellular stores. The second part is devoted to the role of fatty acid signaling in different organs/tissues. The specific contribution of free fatty acids released by particular lipases, hormone-sensitive lipase, adipose triacylglycerol lipase and lysosomal lipase will also be discussed.

A simple and accurate two-step long DNA sequences synthesis strategy to improve heterologous gene expression in pichia.

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Jiang-Ke Yang

Full Text Available In vitro gene chemical synthesis is a powerful tool to improve the expression of gene in heterologous system. In this study, a two-step gene synthesis strategy that combines an assembly PCR and an overlap extension PCR (AOE was developed. In this strategy, the chemically synthesized oligonucleotides were assembled into several 200-500 bp fragments with 20-25 bp overlap at each end by assembly PCR, and then an overlap extension PCR was conducted to assemble all these fragments into a full length DNA sequence. Using this method, we de novo designed and optimized the codon of Rhizopus oryzae lipase gene ROL (810 bp and Aspergillus niger phytase gene phyA (1404 bp. Compared with the original ROL gene and phyA gene, the codon-optimized genes expressed at a significantly higher level in yeasts after methanol induction. We believe this AOE method to be of special interest as it is simple, accurate and has no limitation with respect to the size of the gene to be synthesized. Combined with de novo design, this method allows the rapid synthesis of a gene optimized for expression in the system of choice and production of sufficient biological material for molecular characterization and biotechnological application.

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

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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.

Characterization of biotechnologically relevant extracellular lipase produced by Aspergillus terreus NCFT 4269.10

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Bijay Kumar Sethi

2016-03-01

Full Text Available Abstract Enzyme production by Aspergillus terreus NCFT 4269.10 was studied under liquid static surface and solid-state fermentation using mustard oil cake as a substrate. The maximum lipase biosynthesis was observed after incubation at 30 °C for 96 h. Among the domestic oils tested, the maximum lipase biosynthesis was achieved using palm oil. The crude lipase was purified 2.56-fold to electrophoretic homogeneity, with a yield of 8.44%, and the protein had a molecular weight of 46.3 kDa as determined by SDS-PAGE. Enzyme characterization confirmed that the purified lipase was most active at pH 6.0, temperature of 50 °C, and substrate concentration of 1.5%. The enzyme was thermostable at 60 °C for 1 h, and the optimum enzyme–substrate reaction time was 30 min. Sodium dodecyl sulfate and commercial detergents did not significantly affect lipase activity during 30-min incubation at 30 °C. Among the metal ions tested, the maximum lipase activity was attained in the presence of Zn2+, followed by Mg2+ and Fe2+. Lipase activity was not significantly affected in the presence of ethylenediaminetetraacetic acid, sodium lauryl sulfate and Triton X-100. Phenylmethylsulfonyl fluoride (1 mM and the reducing, β-mercaptoethanol significantly inhibited lipase activity. The remarkable stability in the presence of detergents, additives, inhibitors and metal ions makes this lipase unique and a potential candidate for significant biotechnological exploitation.

Production of extracellular lipase by a new strain Staphylococcus ...

African Journals Online (AJOL)

Based on morphological, biochemical and 16S rRNA sequence analysis, the potent isolate was identified as Staphylococcus aureus. The lipase production of the isolate was increased by improving the conditions of production medium. Maximum lipase production (8.11 U/ml) was achieved when 2% punnakka oil was ...

Purification and Characterization of a Thermostable Lipase from Geobacillus thermodenitrificans IBRL-nra

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Anuradha Balan

2012-01-01

Full Text Available Thermostable lipase from Geobacillus thermodenitrificans IBRL-nra was purified and characterized. The production of thermostable lipase from Geobacillus thermodenitrificans IBRL-nra was carried out in a shake-flask system at 65°C in cultivation medium containing; glucose 1.0% (w/v; yeast extract 1.25% (w/v; NaCl 0.45% (w/v olive oil 0.1% (v/v with agitation of 200 rpm for 24 hours. The extracted extracellular crude thermostable lipase was purified to homogeneity by using ultrafiltration, Heparin-affinity chromatography, and Sephadex G-100 gel-filtration chromatography by 34 times with a final yield of 9%. The molecular weight of the purified enzyme was estimated to be 30 kDa after SDS-PAGE analysis. The optimal temperature for thermostable lipase was 65°C and it retained its initial activity for 3 hours. Thermostable lipase activity was highest at pH 7.0 and stable for 16 hours at this pH at 65°C. Thermostable lipase showed elevated activity when pretreated with BaCl2, CaCl2, and KCl with 112%, 108%, and 106%, respectively. Lipase hydrolyzed tripalmitin (C16 and olive oil with optimal activity (100% compared to other substrates.

IDENTIFIKASI POTENSI ENZIM LIPASE DAN SELULASE PADA SAMPAH KULIT BUAH HASIL FERMENTASI

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La Ode Sumarlin

2013-12-01

Full Text Available Fermentation is one of bioconversion to produce profitable anaerobic microbes and to produce various enzymes. Lipases and cellulases are widely used enzymes so far. Cellulases play an important role in bioconversion of organic waste cellulosic materials to glucose, single cell proteins, animal feed, and ethanol. Lipases can also degrade fatty ester bond. Therefore, both enzymes are potential to be used in industry as well as in households. Fermentation of fruit peel waste is an attempt to produce cellulase and lipase that can be carried out in a simple way. Cellulase as says was performed using DNS (3.5-dinitrosalicylic acid and acid-base titration for analysis of lipase using cooking oil as the substrate. The results showed that the highest cellulase activity was obtained from watermelon rind mixed with citrus fruit peel of 0.036 U/mL, and mixed of banana peel and citrus fruit, which was 0.035 U/mL. The optimum lipase activity was at 30 oC, pH 7, and reaction time of 60 minutes. The highest lipase activity (1.36 U/mL was obtained from mixture of watermelon and orange rind. Thus, the fruit peel waste is potential to produce cellulase and lipase by fermentation .

Adrenal and liver in normal and cld/cld mice synthesize and secrete hepatic lipase, but the lipase is inactive in cld/cld mice.

Science.gov (United States)

Schultz, C J; Blanchette-Mackie, E J; Scow, R O

2000-02-01

Combined lipase deficiency (cld) is a recessive mutation in mice that causes a severe lack of lipoprotein lipase (LPL) and hepatic lipase (HL) activities, hyperlipemia, and death within 3 days after birth. Earlier studies showed that inactive LPL and HL were synthesized by cld/cld tissues and that LPL synthesized by cld/cld brown adipocytes was retained in their ER. We report here a study of HL in liver, adrenal, and plasma of normal newborn and cld/cld mice. Immunofluorescence studies showed HL was present in extracellular space, but not in cells, in liver and adrenal of both normal and cld/cld mice. When protein secretion was blocked with monensin, HL was retained intracellularly in liver cell cultures and in incubated adrenal tissues of both groups of mice. These findings demonstrated that HL was synthesized and secreted by liver and adrenal cells in normal newborn and cld/cld mice. HL activities in liver, adrenal, and plasma in cld/cld mice were very low, cld/cld cells was inactive. Livers of both normal newborn and cld/cld mice synthesized LPL, but the level of LPL activity in cld/cld liver was very low, cld/cld mice, indicating that LPL was synthesized but not secreted by cld/cld liver cells. Immunofluorescent LPL was not found in normal newborn liver cells unless the cells were treated with monensin, thus demonstrating that normal liver cells synthesized and secreted LPL. Livers of both groups of mice contained an unidentified alkaline lipase activity which accounted for 34-54% of alkaline lipase activity in normal and 65% of that in cld/cld livers. Our findings indicate that liver and adrenal cells synthesized and secreted HL in both normal newborn and cld/cld mice, but the lipase was inactive in cld/cld mice. That cld/cld liver cells secreted inactive HL while retaining inactive LPL indicates that these closely related lipases were processed differently.

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.

Strategies to Characterize Fungal Lipases for Applications in Medicine and Dairy Industry

Science.gov (United States)

Gopinath, Subash C. B.; Anbu, Periasamy; Lakshmipriya, Thangavel; Hilda, Azariah

2013-01-01

Lipases are water-soluble enzymes that act on insoluble substrates and catalyze the hydrolysis of long-chain triglycerides. Lipases play a vital role in the food, detergent, chemical, and pharmaceutical industries. In the past, fungal lipases gained significant attention in the industries due to their substrate specificity and stability under varied chemical and physical conditions. Fungal enzymes are extracellular in nature, and they can be extracted easily, which significantly reduces the cost and makes this source preferable over bacteria. Soil contaminated with spillage from the products of oil and dairy harbors fungal species, which have the potential to secrete lipases to degrade fats and oils. Herein, the strategies involved in the characterization of fungal lipases, capable of degrading fatty substances, are narrated with a focus on further applications. PMID:23865040

MALDI imaging of enzymatic degradation of glycerides by lipase on textile surface

DEFF Research Database (Denmark)

Hall-Andersen, Jonatan; Kaasgaard, Svend G; Janfelt, Christian

2018-01-01

Most modern laundry detergents contain enzymes such as proteases, amylases, and lipases for more efficient removal of stains containing proteins, carbohydrates, and lipids during wash at low temperature. The function of the lipases is to hydrolyse the hydrophobic triglycerides from fats and oils...... stain and simulating washing cycles using well-defined detergents with lipase concentrations ranging between 0 and 0.5ppm. After washing, the textile swatches as well as cryo-sections of the swatches were imaged using MALDI imaging in positive ion mode at pixel sizes of 15-75μm. Similar samples were...... an inhomogeneous presence of diglycerides after lipase treatment both in planar images of the textile surface as well as in cross-sections suggesting a non-uniform enzyme effect or extraction of the lipase reaction products from the textile....

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...

Anaerobic biodegradability of dairy wastewater pretreated with porcine pancreas lipase

Directory of Open Access Journals (Sweden)

Adriano Aguiar Mendes

2010-12-01

Full Text Available Lipids-rich wastewater was partial hydrolyzed with porcine pancreas lipase and the efficiency of the enzymatic pretreatment was verified by the comparative biodegradability tests (crude and treated wastewater. Alternatively, simultaneous run was carried out in which hydrolysis and digestion was performed in the same reactor. Wastewater from dairy industries and low cost lipase preparation at two concentrations (0.05 and 0.5% w.v-1 were used. All the samples pretreated with enzyme showed a positive effect on organic matter removal (Chemical Oxygen Demand-COD and formation of methane. The best results were obtained when hydrolysis and biodegradation were performed simultaneously, attaining high COD and color removal independent of the lipase concentration. The enzymatic treatment considerably improved the anaerobic operational conditions and the effluent quality (lower content of suspended solids and less turbidity. Thus, the use of enzymes such as lipase seemed to be a very promising alternative for treating the wastewaters having high fat and grease contents, such as those from the dairy industry.O presente trabalho teve como objetivo o pré-tratamento de efluente da indústria de laticínios na hidrólise de lipídeos, empregando lipase de fonte de células animais de baixo custo disponível comercialmente (pâncreas de porco na formação de gás metano por biodegradabilidade anaeróbia empregando diferentes concentrações de lipase (0,05 e 0,5 % w.v-1. A utilização de lipase no pré-tratamento do efluente acelerou a hidrólise de lipídeos e, conseqüentemente, auxiliou o tratamento biológico resultando na redução da matéria orgânica em termos de Demanda Química de Oxigênio (DQO, cor e sólidos em suspensão como lipídeos. Os melhores resultados em termos de remoção de DQO e cor foram obtidos quando a hidrólise e biodigestão foram realizadas simultaneamente, independente da concentração de lipase empregada. Estes resultados

Lipase Activity in Fermented Oil Seeds of Africa Locust Bean ...

African Journals Online (AJOL)

acer

was determined. The peak lipase activity for fermented Africa locust bean, Castor seed, and African ..... Lipase by Penicillium restrictum in solid state ... sp. Rev. Microbiol. 28(2): 90-95. Martinek, G.H. (1969). Microbiology and amino acid ...

Enzymatic production of alkyl esters through alcoholysis: A critical evaluation of lipases and alcohols

DEFF Research Database (Denmark)

Li, Deng; Xu, Xuebing; Gudmundur G, Haraldsson

2005-01-01

This paper focuses on a detailed evaluation of commercially available immobilized lipases and simple monohydric alcohols for the production of alkyl esters from sunflower oil by enzymatic alcoholysis. Six lipases were tested with seven alcohols, including straight and branched-chain primary...... in an increased degree of conversion for all lipases except Novozym 435. The secondary alcohol 2-propanol significantly reduced the alcoholysis reaction with all lipases; however, the branch-chain isobutanol was more advantageous than linear 1-butanol for Novozym 435, Lipozyme RM IM, and Lipase PS-C. Many...

Lipase-nanoporous gold biocomposite modified electrode for reliable detection of triglycerides.

Science.gov (United States)

Wu, Chao; Liu, Xueying; Li, Yufei; Du, Xiaoyu; Wang, Xia; Xu, Ping

2014-03-15

For triglycerides biosensor design, protein immobilization is necessary to create the interface between the enzyme and the electrode. In this study, a glassy carbon electrode (GCE) was modified with lipase-nanoporous gold (NPG) biocomposite (denoted as lipase/NPG/GCE). Due to highly conductive, porous, and biocompatible three-dimensional structure, NPG is suitable for enzyme immobilization. In cyclic voltammetry experiments, the lipase/NPG/GCE bioelectrode displayed surface-confined reaction in a phosphate buffer solution. Linear responses were obtained for tributyrin concentrations ranging from 50 to 250 mg dl(-1) and olive oil concentrations ranging from 10 to 200 mg dl(-1). The value of apparent Michaelis-Menten constant for tributyrin was 10.67 mg dl(-1) and the detection limit was 2.68 mg dl(-1). Further, the lipase/NPG/GCE bioelectrode had strong anti-interference ability against urea, glucose, cholesterol, and uric acid as well as a long shelf-life. For the detection of triglycerides in human serum, the values given by the lipase/NPG/GCE bioelectrode were in good agreement with those of an automatic biochemical analyzer. These properties along with a long self-life make the lipase/NPG/GCE bioelectrode an excellent choice for the construction of triglycerides biosensor. © 2013 Elsevier B.V. All rights reserved.

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.

Biodiesel production by transesterification using immobilized lipase.

Science.gov (United States)

Narwal, Sunil Kumar; Gupta, Reena

2013-04-01

Biodiesel can be produced by transesterification of vegetable or waste oil catalysed by lipases. Biodiesel is an alternative energy source to conventional fuel. It combines environmental friendliness with biodegradability, low toxicity and renewability. Biodiesel transesterification reactions can be broadly classified into two categories: chemical and enzymatic. The production of biodiesel using the enzymatic route eliminates the reactions catalysed under acid or alkali conditions by yielding product of very high purity. The modification of lipases can improve their stability, activity and tolerance to alcohol. The cost of lipases and the relatively slower reaction rate remain the major obstacles for enzymatic production of biodiesel. However, this problem can be solved by immobilizing the enzyme on a suitable matrix or support, which increases the chances of re-usability. The main factors affecting biodiesel production are composition of fatty acids, catalyst, solvents, molar ratio of alcohol and oil, temperature, water content, type of alcohol and reactor configuration. Optimization of these parameters is necessary to reduce the cost of biodiesel production.

Recombinant Lipases and Phospholipases and Their Use as Biocatalysts for Industrial Applications

OpenAIRE

Borrelli, Grazia M.; Trono, Daniela

2015-01-01

Lipases and phospholipases are interfacial enzymes that hydrolyze hydrophobic ester linkages of triacylglycerols and phospholipids, respectively. In addition to their role as esterases, these enzymes catalyze a plethora of other reactions; indeed, lipases also catalyze esterification, transesterification and interesterification reactions, and phospholipases also show acyltransferase, transacylase and transphosphatidylation activities. Thus, lipases and phospholipases represent versatile bioc...

Enzymatic interesterification of palm stearin and coconut oil by a dual lipase system

DEFF Research Database (Denmark)

Ibrahim, Nuzul Amri Bin; Guo, Zheng; Xu, Xuebing

2008-01-01

greater than 100% over the theoretical value when the reaction proceeds for 2 h. The co-immobilization action of the carrier of the immobilized lipases towards the free lipase was proposed as being one of the reasons leading to the synergistic effect and this has been experimentally verified by a reaction......Enzymatic interesterification of palm stearin with coconut oil was conducted by applying a dual lipase system in comparison with individual lipase-catalyzed reactions. The results indicated that a synergistic effect occurred for many lipase combinations, but largely depending on the lipase species...... mixed and their ratios. The combination of Lipozyme TL IM and RM IM was found to generate a positive synergistic action at all test mixing ratios. Only equivalent amount mixtures of Lipozyme TL IM with Novozym 435 or Lipozyme RM IM with Novozym 435 produced a significant synergistic effect as well...

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.

Lipase biocatalysis for useful biodegradable products

Energy Technology Data Exchange (ETDEWEB)

Linko, Y Y; Wang, Zhuo Lin; Uosukainen, E; Seppaelae, J [Helsinki Univ. of Technology, Espoo (Finland); Laemsae, M [Raisio Group Oil Milling Industry, Raisio (Finland)

1997-12-31

It was shown that lipases can be used as biocatalysts in the production of useful biodegradable compounds such as 1-butyl oleate by direct esterification of butanol and oleic acid to decrease viscosity of biodiesel in winter use. By enzymic transesterification, a mixture of 2-ethyl-1-hexyl esters from rapeseed oil fatty acids can be obtained in good yields for use as a solvent, and of trimethylolpropane esters for use as a lubricant. Finally, it was demonstrated that polyesters with a mass average molar mass in excess of 75,000 g mol{sup -}1 can be obtained by esterification or transesterification by using lipase as biocatalyst. (author) (3 refs.)

Lipase biocatalysis for useful biodegradable products

Energy Technology Data Exchange (ETDEWEB)

Linko, Y.Y.; Wang, Zhuo Lin; Uosukainen, E.; Seppaelae, J. [Helsinki Univ. of Technology, Espoo (Finland); Laemsae, M. [Raisio Group Oil Milling Industry, Raisio (Finland)

1996-12-31

It was shown that lipases can be used as biocatalysts in the production of useful biodegradable compounds such as 1-butyl oleate by direct esterification of butanol and oleic acid to decrease viscosity of biodiesel in winter use. By enzymic transesterification, a mixture of 2-ethyl-1-hexyl esters from rapeseed oil fatty acids can be obtained in good yields for use as a solvent, and of trimethylolpropane esters for use as a lubricant. Finally, it was demonstrated that polyesters with a mass average molar mass in excess of 75,000 g mol{sup -}1 can be obtained by esterification or transesterification by using lipase as biocatalyst. (author) (3 refs.)

Structural basis of the chiral selectivity of Pseudomonas cepacia lipase

NARCIS (Netherlands)

Lang, Dietmar A.; Mannesse, Maurice L.M.; Haas, Gerard H. de; Verheij, Hubertus M.; Dijkstra, Bauke W.

1998-01-01

To investigate the enantioselectivity of Pseudomonas cepacia lipase, inhibition studies were performed with S(c)-and R(c)-(R(p),S(p))-1,2-dialkylcarbamoylglycero-3-O-p-nitrophenyl alkylphosphonates of different alkyl chain lengths. P. cepacia lipase was most rapidly inactivated by

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...

Metabolic fate of rat heart endothelial lipoprotein lipase

International Nuclear Information System (INIS)

Chajek-Shaul, T.; Bengtsson-Olivecrona, G.; Peterson, J.; Olivecrona, T.

1988-01-01

When isolated rat hearts were perfused with medium containing 125I-labeled bovine lipoprotein lipase (LPL), they bound both lipase activity and radioactivity. More than 80% of the bound lipase could be rapidly released by heparin. Low concentrations of bovine LPL displaced 50-60% of the endogeneous, endothelial-bound LPL. Higher concentrations caused additional binding. Both binding and exchange were rapid processes. The hearts continuously released endogenous LPL into the medium. An antiserum that inhibited bovine but not rat LPL was used to differentiate endogeneous and exogeneous LPL activity. When the pool of endothelial LPL was labeled with bovine 125I-labeled LPL and then chased with unlabeled bovine LPL, approximately 50% of the labeled lipase was rapidly displaced. During chase perfusion with medium only, catalytically active bovine LPL appeared in the perfusate. The rate of release was similar to that observed for endogeneous LPL activity and amounted to 10-13% of the heparin-releasable fraction in the first 5 min of perfusion. There was little or no degradation of bovine 125I-labeled LPL to fragments or acid-soluble products. These results indicate that endothelial LPL is accessible for exchange with exogeneous LPL and that detachment rather than degradation may be the pathway for catabolism of endothelial LPL

Comparison of immunoreactive serum trypsinogen and lipase in Cystic Fibrosis

International Nuclear Information System (INIS)

Lloyd-Still, J.D.; Weiss, S.; Wessel, H.; Fong, L.; Conway, J.J.

1984-01-01

The incidence of Cystic Fibrosis (CF) is 1 in 2,000. Early detection and treatment of CF may necessitate newborn screening with a reliable and cost-effective test. Serum immunoreactive trypsinogen (IRT) an enzyme produced by the pancreas, is detectable by radioimmunoassay (RIA) techniques. Recently, it has been shown that IRT is elevated in CF infants for the first few months of life and levels become subnormal as pancreatic insufficiency progresses. Other enzymes produced by the pancreas, such as lipase, are also elevated during this time. The author's earlier work confirmed previous reports of elevated IRT levels in CF infants. The development of a new RIA for lipase (nuclipase) has enabled comparison of these 2 pancreatic enzymes in C.F. Serum IRT and lipase determinations were performed on 2 groups of CF patients; infants under 1 year of age, and children between 1 and 18 years of age. Control populations of the same age groups were included. The results showed that both trypsin (161 +- 92 ng/ml, range 20 to 400) and lipase (167 +- 151 ng/ml, range 29 to 500) are elevated in CF in the majority of infants. Control infants had values of IRT ranging from 20 to 29.5 ng/ml and lipase values ranging from 23 to 34 ng/ml. IRT becomes subnormal in most CF patients by 8 years of age as pancreatic function insufficiency increases. Lipase levels and IRT levels correlate well in infancy, but IRT is a more sensitive indicator of pancreatic insufficiency in older patients with CF

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...

Structure and Function of Lipase

DEFF Research Database (Denmark)

Skjold-Jørgensen, Jakob

.e. the waterlipidinterface. For Thermomyces lanuginosus lipase (TlL) and related lipases, activation of the enzymeinvolves a rearrangement of a structural domain, called the “lid”, which covers the active site inhomogenous aqueous solution. At the water-lipid interface, the lid is displaced from the active site andmoves...... the water-lipid interface, structural movements occurring during activation have been difficult to probeexperimentally. In this work, novel variants of TlL were constructed based on rational design with amutated lid-region in order to elucidate the impact of the lid-residue composition and characteristics...... onthe activation mechanism. From characterization studies of these variants we have shown (Paper I) thatthe lid-region plays a crucial role in governing interfacial activation and enzymatic activity. Specifically,using a combination of spectroscopic and enzymatic activity-based methods we have...

The genotypic diversity and lipase production of some thermophilic bacilli from different genera

Directory of Open Access Journals (Sweden)

Melih Koc

2015-12-01

Full Text Available Abstract Thermophilic 32 isolates and 20 reference bacilli were subjected to Rep-PCR and ITS-PCR fingerprinting for determination of their genotypic diversity, before screening lipase activities. By these methods, all the isolates and references could easily be differentiated up to subspecies level from each other. In screening assay, 11 isolates and 7 references were found to be lipase producing. Their extracellular lipase activities were measured quantitatively by incubating in both tributyrin and olive oil broths at 60 °C and pH 7.0. During the 24, 48 and 72-h period of incubation, the changes in the lipase activities, culture absorbance, wet weight of biomass and pH were all measured. The activity was determined by using pNPB in 50 mM phosphate buffer at pH 7.0 at 60 °C. The lipase production of the isolates in olive oil broths varied between 0.008 and 0.052, whereas these values were found to be 0.002-0.019 (U/mL in the case of tyributyrin. For comparison, an index was established by dividing the lipase activities to cell biomass (U/mg. The maximum thermostable lipase production was achieved by the isolates F84a, F84b, and G. thermodenitrificans DSM 465T (0.009, 0.008 and 0.008 U/mg within olive oil broth, whereas G. stearothermophilus A113 displayed the highest lipase activity than its type strain in tyributyrin. Therefore, as some of these isolates displayed higher activities in comparison to references, new lipase producing bacilli were determined by presenting their genotypic diversity with DNA fingerprinting techniques.

Les lipases sont des hydrolases atypiques : principales caractéristiques et applications

Directory of Open Access Journals (Sweden)

Fickers P.

2008-01-01

Full Text Available ipases are atypical hydrolases: principal characteristics and applications. Due to their kinetic and substrate specificities, triacylglycerol acyl-hydrolases or lipases are atypical enzymes. In function of their microenvironment, lipases are able to act as hydrolases in aqueous solution or as biocatalysts in organic synthesis. As hydrolases, they are responsible of the triglycerids catabolism into fatty acids and glycerol. In many organisms, this reaction plays a major role in the fat and lipid metabolism. In addition, lipases are also able to hydrolyse phospholipids and cholesterol esters. In organic solvent, lipases could catalyse reactions such as esterifications, acidolysis or alcoolysis with enantio-, regio- and chimioselectivity. Lipases form a mixed class of enzyme due to their animal, vegetal or microbial origins. All those properties led to the development of many applications in the food and chemical industries but also in the medical and therapeutic field.

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.

Mycelium-Bound Lipase from a Locally Isolated Strain of Geotrichum candidum

Directory of Open Access Journals (Sweden)

Joo Ling Loo

2014-06-01

Full Text Available Mycelium-bound lipase (MBL, from a locally isolated Geotrichum candidum strain, was produced and characterized as a natural immobilized lipase. A time course study of its lipolytic activity in 1 L liquid broth revealed the maximum MBL activity at 4 h for mycelium cells harvested after 54 h. The yield and specific activity of MBL were 3.87 g/L dry weight and 508.33 U/g protein, respectively, while less than 0.2 U/mL lipase activity was detected in the culture supernatant. Prolonged incubation caused release of the bound lipase into the growth medium. The growth pattern of G. candidum, and production and properties of MBL were not affected by the scale. The stability of mycelia harboring lipase (MBL, harvested and lyophilized after 54 h, studied at 4 °C depicted a loss of 4.3% and 30% in MBL activity after 1 and 8 months, while the activity of free lipase was totally lost after 14 days of storage. The MBL from G. candidum displayed high substrate selectivity for unsaturated fatty acids containing a cis-9 double bond, even in crude form. This unique specificity of MBL could be a direct, simple and inexpensive way in the fats and oil industry for the selective hydrolysis or transesterification of cis-9 fatty acid residues in natural triacylglycerols.

Bioconjugation of lipase and cholesterol oxidase with graphene or graphene oxide

Energy Technology Data Exchange (ETDEWEB)

Silva, Rubens A.; Souza, Michele L.; Bloisi, Georgia D.; Corio, Paolo; Petri, Denise F. S., E-mail: dfsp@iq.usp.br [Universidade de São Paulo, Instituto de Química (Brazil)

2015-04-15

The catalytic behavior of lipase and cholesterol oxidase (ChOx) in the absence and in the presence of graphene (G) or graphene oxide (GO) was investigated at 24 ± 1 °C and pH 6.5. GO flat sheets (0.5–2 μm) were ∼2-nm thick, while G formed aggregates. The maximum reaction velocity (V{sub max}) values and turnover numbers (k{sub cat}) determined for reactions catalyzed by physical mixtures of lipase (at 0.01 g l{sup −1}) or ChOx (at 0.03 g l{sup −1}) and G (0.012 g l{sup −1}) increased six-fold or doubled, respectively, in comparison to neat enzymes. Circular dichroism (CD) and photoluminescence (PL) spectroscopic measurements revealed the preservation of native secondary structures of enzymes and bioconjugation driven by hydrophobic interaction and energy transfer (redshift) between lipase or ChOx and G, corroborating with the enhanced catalytic behavior. On the other hand, the interactions between GO, which has hydrophilic moieties on the basal plane, and ChOx caused enzyme deactivation, as evidenced by the absence of typical CD signal. At low GO concentration (<0.012 g l{sup −1}), bioconjugates of lipases with GO led to V{sub max} and k{sub cat} values four-fold lower than their counterparts with G, but the GO hydrophilic groups probably favored the affinity for the substrate, because the Michaelis constant (K{sub m}) values decreased in comparison to that of neat lipase. Upon increasing the GO concentration, lipases lost secondary structure and the typical lipase PL bands disappeared.

Bioconjugation of lipase and cholesterol oxidase with graphene or graphene oxide

International Nuclear Information System (INIS)

Silva, Rubens A.; Souza, Michele L.; Bloisi, Georgia D.; Corio, Paolo; Petri, Denise F. S.

2015-01-01

The catalytic behavior of lipase and cholesterol oxidase (ChOx) in the absence and in the presence of graphene (G) or graphene oxide (GO) was investigated at 24 ± 1 °C and pH 6.5. GO flat sheets (0.5–2 μm) were ∼2-nm thick, while G formed aggregates. The maximum reaction velocity (V max ) values and turnover numbers (k cat ) determined for reactions catalyzed by physical mixtures of lipase (at 0.01 g l −1 ) or ChOx (at 0.03 g l −1 ) and G (0.012 g l −1 ) increased six-fold or doubled, respectively, in comparison to neat enzymes. Circular dichroism (CD) and photoluminescence (PL) spectroscopic measurements revealed the preservation of native secondary structures of enzymes and bioconjugation driven by hydrophobic interaction and energy transfer (redshift) between lipase or ChOx and G, corroborating with the enhanced catalytic behavior. On the other hand, the interactions between GO, which has hydrophilic moieties on the basal plane, and ChOx caused enzyme deactivation, as evidenced by the absence of typical CD signal. At low GO concentration (<0.012 g l −1 ), bioconjugates of lipases with GO led to V max and k cat values four-fold lower than their counterparts with G, but the GO hydrophilic groups probably favored the affinity for the substrate, because the Michaelis constant (K m ) values decreased in comparison to that of neat lipase. Upon increasing the GO concentration, lipases lost secondary structure and the typical lipase PL bands disappeared

Characterization of an extracellular lipase by Pseudomonas koreensis BK-L07 isolated from soil.

Science.gov (United States)

Anbu, Periasamy

2014-01-01

Screening using spirit blue agar revealed that strain BK-L07 had the highest lipase activity. Furthermore, the isolated strain was identified as Pseudomonas sp. based on morphological, physiological, biochemical, and molecular analyses. The 16S rRNA gene sequence of strain BK-L07 shared a high similarity with that of Pseudomonas koreensis (99%). The nutritional conditions and physicochemical properties were influenced by P. koreensis BK-L07. The maximum lipase production was obtained in tryptic soy broth medium at pH 8.0 and a temperature of 25°C after 36 hr of incubation. In addition, the lipase activity was determined using different carbon sources and lipase inducers. The lipase production was greatest when 1% maltose was used as the carbon source and olive oil was used as the lipase inducer. The lipase production was significantly increased approximately threefold in the optimized medium when compared with the original medium. Further, the lipase was purified by ammonium sulfate precipitation and gel filtration chromatography with a purification yield of 10.8%. The molecular mass of lipase was 45 kDa. The optimum temperature and pH were 40°C and 8.0, respectively. The enzyme was stable up to 50°C and at pH from 7 to 9. In addition, the enzyme activity was stimulated by MgSO4 and completely inhibited by ethylenediamine tetraacetic acid (EDTA), indicating the metalloenzyme type. The lipase activity was toward medium to long chain length of fatty acids (C10 to C18). Supplemental materials are available for this article. Go to the publisher's online edition of Preparative Biochemistry and Biotechnology to view the supplemental file.

Pancreatic beta-cell lipotoxicity induced by overexpression of hormone-sensitive lipase

DEFF Research Database (Denmark)

Winzell, Maria Sörhede; Svensson, Håkan; Enerbäck, Sven

2003-01-01

Lipid perturbations associated with triglyceride overstorage in beta-cells impair insulin secretion, a process termed lipotoxicity. To assess the role of hormone-sensitive lipase, which is expressed and enzymatically active in beta-cells, in the development of lipotoxicity, we generated transgenic...... mice overexpressing hormone-sensitive lipase specifically in beta-cells. Transgenic mice developed glucose intolerance and severely blunted glucose-stimulated insulin secretion when challenged with a high-fat diet. As expected, both lipase activity and forskolin-stimulated lipolysis was increased...

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

Overexpression of Fusarium solani lipase in Pichia pastoris and its application in lipid degradation

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Jinaporn Wongwatanapaiboon

2016-09-01

Full Text Available Fusarium solani NAN103 lipase was successfully overexpressed in Pichia pastoris using inducible expression system and constitutive expression system under the control of alcohol oxidase 1 promoter (pAOX1 and glyceraldehyde-3-phosphate dehydrogenase promoter (pGAP, respectively. Lipase obtained using the constitutive promoter showed the highest activity of 18.8 U/mg in 3 days of cultivation time. Optimal lipase activity was observed at pH 7.0 and 35 °C using p-nitrophenyl laurate as the substrate. Lipase activity was enhanced by Mn2+, Ba2+, Li+, Ca2+, Ni2+, CHAPS and Triton X-100 but was inhibited by Hg2+, Ag+ and SDS. The addition of 10% v/v of octanol, p-xylene, hexane and isopropanol increased lipase activity. Cultivation of lipase-expressing P. pastoris under pGAP in synthetic wastewater containing 1% w/v palm oil resulted in degradation of 87% of the oil within 72 h. P. pastoris expressing F. solani lipase from constitutive expression system has the potential to be used as an alternative microorganism for lipid degradation.

The G-250A polymorphism in the hepatic lipase gene promoter is associated with changes in hepatic lipase activity and LDL cholesterol: The KANWU Study

DEFF Research Database (Denmark)

Lindi, Virpi; Schwab, Ursula; Louheranta, Anne

2007-01-01

BACKGROUND AND AIMS: Hepatic lipase (HL) catalyzes the hydrolysis of triglycerides and phospholipids from lipoproteins, and promotes the hepatic uptake of lipoproteins. A common G-250A polymorphism in the promoter of the hepatic lipase gene (LIPC) has been described. The aim was to study...

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.

Dual bioimprinting of Thermomyces lanuginosus lipase for synthesis of biodiesel

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Joyeeta Mukherjee

2016-06-01

Full Text Available Use of biodiesel as an alternative to non-renewable sources of energy has become an attractive option in recent years. The enzymatic synthesis of biodiesel by transesterification of fats/oils with an alcohol is a much more sustainable route than the chemical method. However, cost effectiveness of the enzymatic route is a major barrier in its commercialization. In this work, a high activity biocatalyst design of Thermomyces lanuginosus lipase is made by dually bioimprinting it with substrate and a surfactant (which is believed to open up the lid covering the active site of the lipase during precipitation of the lipase in organic solvent. When the lipase was bioimprinted with only the surfactants, 28 U of the enzyme/g of oil could yield 99% biodiesel from soybean oil in about 4 h. However, when dually bioimprinted even very low enzyme load 1.4 U/g of oil, yielded 99% biodiesel within 48 h.

Lipoprotein lipase and endothelial lipase in human testis and in germ cell neoplasms

DEFF Research Database (Denmark)

Nielsen, J E; Lindegaard, M L; Friis-Hansen, L

2009-01-01

. The results suggest that both EL and LPL participate in the supply of nutrients and steroidogenesis in the testes, and that especially EL may be important for the supply of cholesterol for testosterone production in the Leydig cells. The partial cellular separation of the expression of the two lipases...

Fat digestion by lingual lipase: mechanism of lipolysis in the stomach and upper small intestine.

Science.gov (United States)

Liao, T H; Hamosh, P; Hamosh, M

1984-05-01

Ten to 30% of dietary fat is hydrolyzed in the stomach by lingual lipase, an enzyme secreted from lingual serous glands. We investigated the substrate specificity of this enzyme as well as the potential of lingual lipase to act in the upper small intestine i.e., in the presence of bile salts and lecithin. The data presented show that partially purified preparations of rat lingual lipase and the lipase in gastric aspirates of newborn infants have identical substrate specificity: medium-chain triglycerides were hydrolyzed at rates 5-8-fold higher than long-chain triglycerides; the rat and human enzymes do not hydrolyze the ester bond of lecithin or cholesteryl-ester. In contrast to pancreatic lipase, the hydrolysis of triglycerides by lingual lipase is not inhibited by lecithin. But, similar to pancreatic lipase the activity of lingual lipase is inhibited by bile salts, the extent of inhibition varying with its nature and concentration. This inactivation is not prevented by colipase but is partially averted by lipids and protein, suggesting that lingual lipase can remain active in the duodenum. The pH optimum of the enzyme (2.2-6.5 in the rat and 3.5-6.0 in human gastric aspirates) is compatible with continued activity in the upper small intestine, especially during the neonatal period, when the luminal pH is under 6.5. The marked variation in lipase activity levels in gastric aspirates of newborn infants is probably due to individual variations in enzyme amounts. The characteristics of the lipase are however identical in infants with low, intermediate or high activity levels.(ABSTRACT TRUNCATED AT 250 WORDS)

Lipase genes in Mucor circinelloides: identification, sub-cellular location, phylogenetic analysis and expression profiling during growth and lipid accumulation.

Science.gov (United States)

Zan, Xinyi; Tang, Xin; Chu, Linfang; Zhao, Lina; Chen, Haiqin; Chen, Yong Q; Chen, Wei; Song, Yuanda

2016-10-01

Lipases or triacylglycerol hydrolases are widely spread in nature and are particularly common in the microbial world. The filamentous fungus Mucor circinelloides is a potential lipase producer, as it grows well in triacylglycerol-contained culture media. So far only one lipase from M. circinelloides has been characterized, while the majority of lipases remain unknown in this fungus. In the present study, 47 potential lipase genes in M. circinelloides WJ11 and 30 potential lipase genes in M. circinelloides CBS 277.49 were identified by extensive bioinformatics analysis. An overview of these lipases is presented, including several characteristics, sub-cellular location, phylogenetic analysis and expression profiling of the lipase genes during growth and lipid accumulation. All of these proteins contained the consensus sequence for a classical lipase (GXSXG motif) and were divided into four types including α/β-hydrolase_1, α/β-hydrolase_3, class_3 and GDSL lipase (GDSL) based on gene annotations. Phylogenetic analyses revealed that class_3 family and α/β-hydrolase_3 family were the conserved lipase family in M. circinelloides. Additionally, some lipases also contained a typical acyltransferase motif of H-(X) 4-D, and these lipases may play a dual role in lipid metabolism, catalyzing both lipid hydrolysis and transacylation reactions. The differential expression of all lipase genes were confirmed by quantitative real-time PCR, and the expression profiling were analyzed to predict the possible biological roles of these lipase genes in lipid metabolism in M. circinelloides. We preliminarily hypothesized that lipases may be involved in triacylglycerol degradation, phospholipid synthesis and beta-oxidation. Moreover, the results of sub-cellular localization, the presence of signal peptide and transcriptional analyses of lipase genes indicated that four lipase in WJ11 most likely belong to extracellular lipases with a signal peptide. These findings provide a platform

Chicken fat and inorganic nitrogen source for lipase production by ...

African Journals Online (AJOL)

SAM

2014-03-19

Mar 19, 2014 ... for lipase production, the production cost was $US 518.00/million Units of lipase. Key words: ... energetics, fine chemicals and pulp and paper industries. ... for enzyme production is extremely important in dictating ... fat is waste product of poultry processing industry ... Economic Research Service,” 2013).

In silico modeling of lipase H | Jabeen | African Journal of ...

African Journals Online (AJOL)

LAH 2 is a type of autosomal recessive hypotrichosis that affect hairs, eyebrows, scalp and eyelashes. Mutations in Lipase H gene result in LAH 2. Changes that result from mutation on physiochemical properties, post-translational modifications, functional sites, secondary structure and tertiary structure lipase H gene (LIPH) ...

Effect of culture conditions on lipase production by Fusarium solani in batch fermentation.

Science.gov (United States)

Maia, M M; Heasley, A; Camargo de Morais, M M; Melo, E H; Morais, M A; Ledingham, W M; Lima Filho, J L

2001-01-01

Lipase (Glycerol ester hydrolase EC 3.1.1.3.) from a Brazilian strain of Fusarium solani FSI has been investigated. The effect of different carbon sources and trace elements added to basal medium was observed with the aim of improving enzyme production. Lipase specific activity was highest (0.45 U mg(-1)) for sesame oil. When this medium was supplemented with trace elements using olive oil, corn oil and sesame oil the lipase specific activity increased to 0.86, 1.89 and 1.64 U mg(-1), respectively, after 96 h cultivation without any considerable biomass increase. The Km of this lipase using pNPP (p-nitrophenylpalmitate) as substrate, was 1.8 mM with a Vmax of 1.7 micromol min(-1) mg protein(-1). Lipase activity increased in the presence of increasing concentrations of hexane and toluene. In contrast, incubation of this enzyme with water-soluble solvents decreased its activity after 10% concentration (v/v) of the solvent. The lipase activity was stable below 35 degrees C but above this temperature activity losses were observed.

Lipase From Thermoalkalophilic Pseudomonas species as an Additive in Potential Laundry Detergent Formulations

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Ibrahim, C. O.

2009-01-01

Full Text Available Lipase isolated from a thermoalkalophilic Pseudomonas species was used as additive to improve the degree of olive oil removal from cotton fabric in the presence of surfactants. The lipase used in this study was found to be more effective with non ionic surfactants as compared to ionic surfactants. In terms of stability, there was no decrease in activity found in the presence of Tween 85, Span 80 and Span 20. Lipase from Pseudomonas species was most active in the presence of Tween 85, Span 80 and Span 20. The application of lipase from Pseudomonas species as an additive in the formulation containing Span 80 has improved oil removal by 36% using the washing system consisting 5 U/mL lipase, at 70 °C for 20 min and 0.8% of Span 80 as surfactant. Considering that lipase from Pseudomonas species is stable in high pH and temperatures in the presence of various surfactants, therefore it is suitable to be incorporated as additives in potential detergent formulations.

Characterization and spray drying of lipase produced by the endophytic fungus Cercospora kikuchii

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T. A. Costa-Silva

2014-12-01

Full Text Available A lipase from the endophytic fungus Cercospora kikuchii was purified, biochemically characterized and the effects of spray drying on stabilization of the purified enzyme were studied. The lipase was purified 9.31-fold with recovery of 26.6% and specific activity of 223.6 U/mg. The optimum pH and temperature were 4.6 and 35 ºC, respectively, while the Vmax was 10.28 µmol/min.mg-1 protein and Km 0.0324 mM. All the metal ions tested enhanced the enzyme activity. The lipase retained almost 100% activity in the presence of strong oxidants and was also resistant to Triton X, Tween 80 and 20 and SDS, as well as to proteases. The purified lipase was spray dried and kept until 85.2% of enzymatic activity. At least 70% of the enzymatic activity was maintained for spray dried purified lipase during the storage period. The lipase produced by Cercospora kikuchii has properties useful for industrial application and showed adequate stabilization and retention of its enzymatic activity after spray drying.

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 ...

Lipase-catalyzed biodiesel synthesis with different acyl acceptors

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Ognjanović Nevena D.

2008-01-01

Full Text Available Biodiesel is an alternative fuel for diesel engine that is environmentally acceptable. Conventionally, biodiesel is produced by transesterification of triglycerides and short alcohols in the presence of an acid or an alkaline catalyst. There are several problems associated with this kind of production that can be resolved by using lipase as the biocatalyst. The aim of the present work was to investigate novel acyl acceptors for biodiesel production. 2-Propanol and n-butanol have a less negative effect on lipase stability, and they also improve low temperature properties of the fuel. However, excess alcohol leads to inactivation of the enzyme, and glycerol, a major byproduct, can block the immobilized enzyme, resulting in low enzymatic activity. This problem was solved by using methyl acetate as acyl acceptor. Triacetylglycerol is produced instead of glycerol, and it has no negative effect on the activity of the lipase.

Dependence of PERT endpoint on endogenous lipase activity.

Science.gov (United States)

Gao, Wen-Yi; Mulberg, Andrew E

2014-11-01

To clarify and to understand the potential for misinterpretation of change in fecal fat quantitation during pancreatic enzyme replacement therapy (PERT) trials for treatment of exocrine pancreatic insufficiency. Analysis of clinical trials submitted to the U.S. Food and Drug Administration (FDA) for approval of PERT that enrolled 123 cystic fibrosis adult and pediatric patients treated with Creon, Pertzye, Ultresa, and Zenpep. The CFA% defines lipase activity as a percentage of converting substrate of "Total Daily Dietary Fat Intake." PERT trials performed to date have modified the definition to converting the "Shared Daily Fat Intake," generating "Partial CFA" for the exogenous lipase: the higher the activity of coexisting endogenous lipase, the lower the "Partial CFA" of exogenous measured. This review shows that "Partial CFA" is not CFA. Enrollment of patients with low HPLA during treatment may improve the interpretability of "Partial CFA" measured by PERT trials.

Novel magnetic cross-linked lipase aggregates for improving the resolution of (R, S)-2-octanol.

Science.gov (United States)

Liu, Ying; Guo, Chen; Liu, Chun-Zhao

2015-03-01

Novel magnetic cross-linked lipase aggregates were fabricated by immobilizing the cross-linked lipase aggregates onto magnetic particles with a high number of -NH2 terminal groups using p-benzoquinone as the cross-linking agent. At the optimal fabrication conditions, 100% of immobilization efficiency and 139% of activity recovery of the magnetic cross-linked lipase aggregates were achieved. The magnetic cross-linked lipase aggregates were able to efficiently resolve (R, S)-2-octanol, and retained 100% activity and 100% enantioselectivity after 10 cycles of reuse, whereas the cross-linked lipase aggregates only retained about 50% activity and 70% enantioselectivity due to insufficient cross-linking. These results provide a great potential for industrial applications of the magnetic cross-linked lipase aggregates. © 2014 Wiley Periodicals, Inc.

Novel One-Pot Green Synthesis of Indolizines Biocatalysed by Candida antarctica Lipases

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Simon Bonte

2013-02-01

Full Text Available Marine microorganisms are of considerable interest as a promising source of enzymes with unsuspected potentials as catalysts for chemical synthesis. We describe here an efficient method for one-pot indolizine synthesis that has been developed using lipase A and lipase B from Candida antarctica as biocatalysts. As showed by HPLC/MS analysis, the yield in indolizines was higher in the presence of the biocatalyst than in absence of enzyme. Lipase A, from Candida antarctica, showed high catalytic activity and selectivity for the cycloaddition reactions. When the reactions were performed under ultrasound irradiation, the Candida antarctica lipase catalyzed reactions yielded pure indolozines, in good yields and in very short time.

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

Science.gov (United States)

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.

Science.gov (United States)

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)

Synthesis activity-based zymography for detection of lipases and esterases.

Science.gov (United States)

Kwon, Min-A; Kim, Hyun Suk; Hahm, Dae-Hyun; Song, Jae Kwang

2011-04-01

A new zymography method for lipases and esterases was developed on the basis of the esterification reaction between fatty acids and alcohols. The enzymes were separated by SDS-PAGE and native PAGE. The gel was washed and then incubated in an aqueous solution containing fatty acids (oleic acid 18:1 or caprylic acid 8:0) and dodecanol. Synthesis was visualized by in situ precipitation of water-insoluble and non-diffusible fatty acid esters, such as dodecyl oleate and dodecyl octanoate. The synthesis activity-based zymography was confirmed with different enzyme samples, including commercial lipase preparations, purified recombinant lipase and cutinase, and crude culture supernatants of lipolytic enzyme-producing soil bacteria.

Transesterification Synthesis of Chloramphenicol Esters with the Lipase from Bacillus amyloliquefaciens

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Fengying Dong

2017-09-01

Full Text Available This work presents a synthetic route to produce chloramphenicol esters by taking advantage the high enantio- and regio-selectivity of lipases. A series of chloramphenicol esters were synthesized using chloramphenicol, acyl donors of different carbon chain length and lipase LipBA (lipase cloned from Bacillus amyloliquefaciens. Among acyl donors with different carbon chain lengths, vinyl propionate was found to be the best. The influences of different organic solvents, reaction temperature, reaction time, enzyme loading and water content on the synthesis of the chloramphenicol esters were studied. The synthesis of chloramphenicol propionate (0.25 M with 4.0 g L−1 of LipBA loading gave a conversion of ~98% and a purity of ~99% within 8 h at 50 °C in 1,4-dioxane as solvent. The optimum mole ratio of vinyl propionate to chloramphenicol was increased to 5:1. This is the first report of B. amyloliquefaciens lipase being used in chloramphenicol ester synthesis and a detailed study of the synthesis of chloramphenicol propionate using this reaction. The high enzyme activity and selectivity make lipase LipBA an attractive catalyst for green chemical synthesis of molecules with complex structures.

Influence of cosolvents on the hydrophobic surface immobilization topography of Candida antarctica lipase B

Science.gov (United States)

The presence of cosolvents and co-solutes during the immobilization of lipases on hydrophobic supports may influence the extent of lipase immobilization and the long-term catalytic stability of the biocatalyst. Candida antarctica B lipase immobilization was examined on a hydrophobic surface, i.e., ...

Structure of the human hepatic triglyceride lipase gene

International Nuclear Information System (INIS)

Cai, Shengjian; Wong, D.M.; Chen, Sanhwan; Chan, L.

1989-01-01

The structure of the human hepatic triglyceride lipase gene was determined from multiple cosmid clones. All the exons, exon-intron junctions, and 845 bp of the 5' and 254 bp of the 3' flanking DNA were sequenced. Comparison of the exon sequences to three previously published cDNA sequences revealed differences in the sequence of the codons for residue 133, 193, 202, and 234 that may represent sequence polymorphisms. By primer extension, hepatic lipase mRNA initiates at an adenine 77 bases upstream of the translation initiation site. The hepatic lipase gene spans over 60 kb containing 9 exons and 8 introns, the latter being all located within the region encoding the mature protein. The exons are all of average size (118-234 bp). Exon 1 encodes the signal peptide, exon 4, a region that binds to the lipoprotein substrate, and exon 5, an evolutionarily highly conserved region of potential catalytic function, and exons 6 and 9 encode sequences rich in basic amino acids thought to be important in anchoring the enzyme to the endothelial surface by interacting with acidic domains of the surface glycosaminoglycans. The human lipoprotein lipase gene has been recently reported to have an identical exon-intron organization containing the analogous structural domains. The observations strongly support the common evolutionary origin of these two lipolytic enzymes

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.

Cloning and characterization of newly isolated lipase from Enterobacter sp. Bn12.

Science.gov (United States)

Farrokh, Parisa; Yakhchali, Bagher; Karkhane, Ali Asghar

2014-01-01

A mesophilic Enterobacter sp. Bn12 producing an alkaline thermostable lipase was isolated from soil in Tehran, Iran. The lipase gene (ELBn12) was identified from a genomic library. Sequence analysis of the DNA fragment revealed an open reading frame of 879 bp encoding a lipase with a molecular mass of 31.3 kDa. The deduced amino acid sequence showed 96% identity with a lipase of Enterobacter sp. Ag1 and the identity of their DNA sequences was 88.9%. ELBn12 belongs to the lipase subfamily I.1 and its catalytic triad consists of Ser82, Asp237 and His259. The lipase was expressed in Escherichia coli (BL21) pLysS and partially purified by anion exchange chromatography. The maximum activity of ELBn12 was obtained at temperature of 60 °C and pH 8.0 towards tricaprylin (C8) and its specific activity was around 2900 U/mg. ELBn12 was stable within a broad pH range from 6.0 to 11.0. The enzyme showed high stability in both polar and nonpolar organic solvents at 50% (v/v). The lipase activity was enhanced in the presence of 10 mM of Ca(2+), Mg(2+) and K(+), while heavy metals (Fe(3+) and Zn(2+)) had strong inhibitory effect. ELBn12 showed high activity in the presence of 1% (w/v) nonionic surfactants, however ionic surfactants inhibited the lipolytic activity. ELBn12 characteristics show that it has a potential to be used in various industrial processes.

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

A newly high alkaline lipase: an ideal choice for application in detergent formulations

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Cherif Slim

2011-11-01

Full Text Available Abstract Background Bacterial lipases received much attention for their substrate specificity and their ability to function in extreme environments (pH, temperature.... Many staphylococci produced lipases which were released into the culture medium. Reports of thermostable lipases from Staphylococcus sp. and active in alkaline conditions are not previously described. Results A newly soil-isolated Staphylococcus sp. strain ESW secretes an induced lipase in the culture medium. The effects of temperature, pH and various components in a detergent on the activity and stability of Staphylococcus sp. lipase (SL1 were studied in a preliminary evaluation for use in detergent formulation solutions. The enzyme was highly active over a wide range of pH from 9.0 to 13.0, with an optimum at pH 12.0. The relative activity at pH 13.0 was about 60% of that obtained at pH 12.0. It exhibited maximal activity at 60°C. This novel lipase, showed extreme stability towards non-ionic and anionic surfactants after pre-incubation for 1 h at 40°C, and relative stability towards oxidizing agents. Additionally, the crude enzyme showed excellent stability and compatibility with various commercial solid and liquid detergents. Conclusions These properties added to the high activity in high alkaline pH make this novel lipase an ideal choice for application in detergent formulations.

Improved Performance of Pseudomonas fluorescens lipase by covalent immobilization onto Amberzyme

NARCIS (Netherlands)

Aslan, Yakup; Handayani, Nurrahmi; Stavila, Erythrina; Loos, Katja

2013-01-01

Objective: In this study, the conditions of covalent immobilization of Pseudomonas fluorescens lipase onto an oxirane-activated support (Amberzyme) were optimized to obtain a high activity yield. Furthermore, the operational and storage stabilities of immobilized lipase were tested. Methods: Optimum

Production of extracellular lipase by the phytopathogenic fungus Fusarium solani FS1 Produção de lipase extracelular pelo fungo fitopatogênico Fusarium solani FS1

OpenAIRE

Maria de Mascena Diniz Maia; Marcia Maria Camargo de Morais; Marcos Antonio de Morais Jr.; Eduardo Henrique Magalhães Melo; José Luiz de Lima Filho

1999-01-01

A Brazilian strain of Fusarium solani was tested for extracellular lipase production in peptone-olive oil medium. The fungus produced 10,500 U.l-1 of lipase after 72 hours of cultivation at 25oC in shake-flask at 120 rpm in a medium containing 3% (w/v) peptone plus 0.5% (v/v) olive oil. Glucose (1% w/v) was found to inhibit the inductive effect of olive oil. Peptone concentrations below 3% (w/v) resulted in a reduced lipase production while increased olive oil concentration (above 0.5%) did n...

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.

Direct solid phase radioimmunoassay for chicken lipoprotein lipase

International Nuclear Information System (INIS)

Cheung, A.H.; Bensadoun, A.; Cheng, C.

1979-01-01

A direct, noncompetitive immunoassay for chicken lipoprotein lipase (LPL) was developed. Antibodies to LPL were purified by immunoadsorption chromatography of goat antisera on an LPL-Sepharose column. Purified anti-LPL immunoglobulins were coupled covalently to hydrophilic polyacrylamide beads by a carbodiimide reagent. An excess amount of these beads was incubated with the sample on the standard to be assayed. The amount of LPL immobilized by the heads was then detected by an excess amount of 125 I-labeled anti-LPL immunoglobulin. A linear relationship was obtained between the radioactivity bound and the amount of highly purified LPL used as a standard. The range of the assay was from 0.1 to 1.1 ng PLP. The assay was specific for chicken LPL and showed no cross-reactivity with liver lipase. It does not distinguish heat-inactivated from catalytically active enzyme species. This assay should be useful in studies of lipoprotein lipase where both catalytic activity and enzyme mass need to be quantitated

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.

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.

Effect of Ascorbic Acid on Lipoprotein Lipase Activity | Kotze | South ...

African Journals Online (AJOL)

Baboons kept on hypovitaminotic C diets, but without clinical signs of scurvy, had significantly higher heart muscle lipoprotein lipase activity than baboons on vitamin C 34 mg/kg body mass/day. When the serum vitamin C levels were above 0,35 mg/100 ml the heart muscle lipoprotein lipase was repressed. Serum vitamin C ...

Oral lipase activities and fat-taste receptors for fat-taste sensing in chickens.

Science.gov (United States)

Kawabata, Yuko; Kawabata, Fuminori; Nishimura, Shotaro; Tabata, Shoji

2018-01-01

It has been reported that a functional fat-taste receptor, GPR120, is present in chicken oral tissues, and that chickens can detect fat taste in a behavioral test. However, although triglycerides need to be digested to free fatty acids to be recognized by fat-taste receptors such as GPR120, it remains unknown whether lipase activities exist in chicken oral tissues. To examine this question, we first cloned another fat-taste receptor candidate gene, CD36, from the chicken palate. Then, using RT-PCR, we determined that GPR120 and CD36 were broadly expressed in chicken oral and gastrointestinal tissues. Also by RT-PCR, we confirmed that several lipase genes were expressed in both oral and gastrointestinal tissues. Finally, we analyzed the lipase activities of oral tissues by using a fluorogenic triglyceride analog as a lipase substrate. We found there are functional lipases in oral tissues as well as in the stomach and pancreas. These results suggested that chickens have a basic fat-taste reception system that incorporates a triglycerides/oral-lipases/free fatty acids/GPR120 axis and CD36 axis. Copyright © 2017 Elsevier Inc. All rights reserved.

Improvement of lipase production from Geotrichum sp. in shaken flasks

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Maldonadoa Resende Rafael

2012-01-01

Full Text Available This work is focused on the study of different variables on inoculum build-up aiming to improve the lipase production by Geotrichum sp. by means a sequential strategy of experimental design. The effects of inoculum size, corn steep liquor concentration, volume of inoculum, pH of medium, age of inoculum and soybean oil concentration on lipase activity were assessed by means of two factorial experimental designs. A maximum lipase activity of 35.20±0.8 U/mL was obtained with a inoculum composed of one circular area of 0.78cm2 containing spores, 50 mL of inoculum volume medium, 12 hours of inoculum age, 15% w/v of corn steep liquor concentration, 1.0%w/v of soybean oil concentration and initial pH 5.0 at 30°C and 150 rpm in flasks. This work showed that an enhancement of lipase activity can be obtained using a sequential statistical factorial approach to define the variables for inoculum build-up.

Lipase-Secreting Bacillus Species in an Oil-Contaminated Habitat: Promising Strains to Alleviate Oil Pollution

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Li Pin Lee

2015-01-01

Full Text Available Lipases are of great interest for different industrial applications due to their diversity and versatility. Among different lipases, microbial lipases are preferable due to their broad substrate specificity, and higher stability with lower production costs compared to the lipases from plants and animals. In the past, a vast number of bacterial species have been reported as potential lipases producers. In this study, the lipases-producing bacterial species were isolated from an oil spillage area in the conventional night market. Isolated species were identified as Bacillus species by biochemical tests which indicate their predominant establishment, and further screened on the agar solid surfaces using lipid and gelatin as the substrates. Out of the ten strains tested, four potential strains were subjected to comparison analysis of the lipolytic versus proteolytic activities. Strain 10 exhibited the highest lipolytic and proteolytic activity. In all the strains, the proteolytic activity is higher than the lipolytic activity except for strain 8, suggesting the possibility for substrate-based extracellular gene induction. The simultaneous secretion of both the lipase and protease is a mean of survival. The isolated bacterial species which harbour both lipase and protease enzymes could render potential industrial-based applications and solve environmental issues.

Polyethyleneimine-modified superparamagnetic Fe3O4 nanoparticles for lipase immobilization: Characterization and application

International Nuclear Information System (INIS)

Khoobi, Mehdi; Motevalizadeh, Seyed Farshad; Asadgol, Zahra; Forootanfar, Hamid; Shafiee, Abbas; Faramarzi, Mohammad Ali

2015-01-01

Magnetically separable nanospheres consisting of polyethyleneimine (PEI) and succinated PEI grafted on silica coated magnetite (Fe 3 O 4 ) were prepared and characterized using Fourier transform infrared spectroscopy, thermogravimetric analysis, X-ray diffraction, vibrating sample magnetometer, scanning electron microscopy and transmission electron microscopy. The prepared magnetic nanoparticles were then applied for physical adsorption or covalent attachment of Thermomyces lanuginosa lipase (TLL) via glutaraldehyde or hexamethylene diisocyanate. The reusability, storage, pH and thermal stabilities of the immobilized enzymes compared to that of free lipase were examined. The obtained results showed that the immobilized lipase on MNPs@PEI-GLU was the best biocatalyst which retained 80% of its initial activity after 12 cycles of application. The immobilized lipase on the selected support (MNPs@PEI-GLU) was also applied for the synthesis of ethyl valerate. Following 24 h incubation of the immobilized lipase on the selected support in n-hexane and solvent free media, the esterification percentages were 72.9% and 28.9%, respectively. - Graphical abstract: A schematic of the preparation of PEI- and succinated PEI-grafted Fe 3 O 4 MNPs (MNPs@PEI) and the immobilization of lipase by covalent bonding and adsorption. - Highlights: • Functionalized polyethylenimine-grafted magnetic nanoparticles were synthesized. • The prepared supports were fully characterized by various analysis methods. • Lipase was immobilized on the nanostructures by adsorption and covalent attachment. • Immobilized lipase produced ethyl valerate in solvent free medium

Aplicação de lipase e monoglicerídeo em pão de forma enriquecido com fibras Application of lipase and monoglyceride in fiber enriched pan bread

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Kelly Moreira Gandra

2008-03-01

Full Text Available Neste trabalho, estudou-se a aplicação da enzima lipase e do emulsificante monoglicerídeo em pão de forma enriquecido com fibras, com o objetivo de verificar a possibilidade de substituição do emulsificante pela enzima. Inicialmente, foi realizada a caracterização das matérias-primas principais (farinha e farelo de trigo. Os pães de forma foram elaborados pelo método de massa direta. Foi utilizado um planejamento experimental do tipo composto central rotacional com duas variáveis independentes: i dosagem de lipase; e ii dosagem de monoglicerídeo e, paralelamente, realizou-se um teste controle (sem adição de lipase e monoglicerídeo para comparação. As variáveis dependentes foram as características de qualidade dos pães: i volume específico; ii aceitação sensorial (aparência, textura, aroma e sabor; e iii vida de prateleira avaliada pela umidade do miolo e firmeza dos pães após 1, 4 e 7 dias do forneamento. Dentro das faixas estudadas, foi possível verificar que somente a umidade dos pães no quarto e sétimo dia após o processamento foi influenciada pela variação das dosagens de lipase e monoglicerídeo. Na avaliação sensorial, verificou-se que as notas médias atribuídas aos pães do teste controle foram inferiores à menor nota média dos ensaios do planejamento experimental, com exceção do sabor e aroma. Como não foi possível obter modelos matemáticos para todas as respostas, foram selecionados os ensaios 5 (1% monoglicerídeo, 7 (25 ppm lipase e 9 (25 ppm lipase e 1% monoglicerídeo do planejamento experimental, e o teste controle, para a avaliação dos resultados por análise de variância. Nas condições em que os ensaios foram conduzidos e para as faixas de lipase (0 a 50 ppm e monoglicerídeo (0 a 2% estudadas, verificou-se a possibilidade de substituir o monoglicerídeo por lipase em formulação de pão de forma enriquecido com fibras.In this work, the application of lipase and monoglyceride in

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.

Enzymatic Production of FAME Biodiesel with Soluble Lipases

DEFF Research Database (Denmark)

T. Gundersen, Maria; Heltborg, Carsten Kirstejn; Yang, V

Biodiesel is a viable alternative to fossil fuels, and biocatalysis is gaining interest as a greener process. We focus on converting oils to Fatty Acid Methyl Ester (FAME) using soluble lipases, which offer an advantage compared to immobilized enzymes by cost efficiency and ease of implementation...... the defined operating space concerning: temperature, water content, initial methanol concentration and enzyme content. The identified optimum range was experimentally evaluated, and model findings were confirmed. Another barrier in lipase use in biodiesel production is the higher melting point (m...

New ether-functionalized ionic liquids for lipase-catalyzed synthesis of biodiesel.

Science.gov (United States)

Zhao, Hua; Song, Zhiyan; Olubajo, Olarongbe; Cowins, Janet V

2010-09-01

Ionic liquids (ILs) are being explored as solvents for the enzymatic methanolysis of triglycerides. However, most available ILs (especially hydrophobic ones) have poor capability in dissolving lipids, while hydrophilic ILs tend to cause enzyme inactivation. Recently, we synthesized a new type of ether-functionalized ionic liquids (ILs) carrying anions of acetate or formate; they are capable of dissolving a variety of substrates and are also lipase-compatible (Green Chem., 2008, 10, 696-705). In the present study, we carried out the lipase-catalyzed transesterifications of Miglyol oil 812 and soybean oil in these novel ILs. These ILs are capable of dissolving oils at the reaction temperature (50 degrees C); meanwhile, lipases maintained high catalytic activities in these media even in high concentrations of methanol (up to 50% v/v). High conversions of Miglyol oil were observed in mixtures of IL and methanol (70/30, v/v) when the reaction was catalyzed by a variety of lipases and different enzyme preparations (free and immobilized), especially with the use of two alkylammonium ILs 2 and 3. The preliminary study on the transesterification of soybean oil in IL/methanol mixtures further confirms the potential of using oil-dissolving and lipase-stabilizing ILs in the efficient production of biodiesels.

A quantitative assay measuring the function of lipase maturation factor 1

Science.gov (United States)

Yin, Fen; Doolittle, Mark H.; Péterfy, Miklós

2009-01-01

Newly synthesized lipoprotein lipase (LPL) and related members of the lipase gene family require an endoplasmic reticulum maturation factor for attainment of enzyme activity. This factor has been identified as lipase maturation factor 1 (Lmf1), and mutations affecting its function and/or expression result in combined lipase deficiency (cld) and hypertriglyceridemia. To assess the functional impact of Lmf1 sequence variations, both naturally occurring and induced, we report the development of a cell-based assay using LPL activity as a quantitative reporter of Lmf1 function. The assay uses a cell line homozygous for the cld mutation, which renders endogenous Lmf1 nonfunctional. LPL transfected into the mutant cld cell line fails to attain activity; however, cotransfection of LPL with wild-type Lmf1 restores its ability to support normal lipase maturation. In this report, we describe optimized conditions that ensure the detection of a complete range of Lmf1 function (full, partial, or complete loss of function) using LPL activity as the quantitative reporter. To illustrate the dynamic range of the assay, we tested several novel mutations in mouse Lmf1. Our results demonstrate the ability of the assay to detect and analyze Lmf1 mutations having a wide range of effects on Lmf1 function and protein expression. PMID:19471043

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

Inhibition of pancreatic lipase and amylase by extracts of different spices and plants.

Science.gov (United States)

Sellami, Mohamed; Louati, Hanen; Kamoun, Jannet; Kchaou, Ali; Damak, Mohamed; Gargouri, Youssef

2017-05-01

The aim of this study is to search new anti-obesity and anti-diabetic agents from plant and spices crude extracts as alternative to synthetic drugs. The inhibitory effect of 72 extracts was evaluated, in vitro, on lipase and amylase activities. Aqueous extracts of cinnamon and black tea exhibited an appreciable inhibitory effect on pancreatic amylase with IC 50 values of 18 and 87 μg, respectively. Aqueous extracts of cinnamon and mint showed strong inhibitory effects against pancreatic lipase with IC 50 of 45 and 62 μg, respectively. The presence of bile salts and colipase or an excess of interface failed to restore the lipase activity. Therefore, the inhibition of pancreatic lipase, by extracts of spices and plants, belongs to an irreversible inhibition. Crude extract of cinnamon showed the strongest anti-lipase and anti-amylase activities which offer a prospective therapeutic approach for the management of diabetes and obesity.

Analysis of Comparative Sequence and Genomic Data to Verify Phylogenetic Relationship and Explore a New Subfamily of Bacterial Lipases.

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Malihe Masomian

Full Text Available Thermostable and organic solvent-tolerant enzymes have significant potential in a wide range of synthetic reactions in industry due to their inherent stability at high temperatures and their ability to endure harsh organic solvents. In this study, a novel gene encoding a true lipase was isolated by construction of a genomic DNA library of thermophilic Aneurinibacillus thermoaerophilus strain HZ into Escherichia coli plasmid vector. Sequence analysis revealed that HZ lipase had 62% identity to putative lipase from Bacillus pseudomycoides. The closely characterized lipases to the HZ lipase gene are from thermostable Bacillus and Geobacillus lipases belonging to the subfamily I.5 with ≤ 57% identity. The amino acid sequence analysis of HZ lipase determined a conserved pentapeptide containing the active serine, GHSMG and a Ca(2+-binding motif, GCYGSD in the enzyme. Protein structure modeling showed that HZ lipase consisted of an α/β hydrolase fold and a lid domain. Protein sequence alignment, conserved regions analysis, clustal distance matrix and amino acid composition illustrated differences between HZ lipase and other thermostable lipases. Phylogenetic analysis revealed that this lipase represented a new subfamily of family I of bacterial true lipases, classified as family I.9. The HZ lipase was expressed under promoter Plac using IPTG and was characterized. The recombinant enzyme showed optimal activity at 65 °C and retained ≥ 97% activity after incubation at 50 °C for 1h. The HZ lipase was stable in various polar and non-polar organic solvents.

Fat digestion in the stomach: stability of lingual lipase in the gastric environment.

Science.gov (United States)

Fink, C S; Hamosh, P; Hamosh, M

1984-03-01

Digestion of dietary fat starts in the stomach, where lingual lipase hydrolyzes triglycerides to free fatty acids and partial glycerides at pH 3.0-6.0. Lingual lipase is secreted continuously from lingual serous glands and accumulates in the stomach between meals, when gastric pH is less than 3.0. We have, therefore, examined the resistance of lingual lipase to low pH and its possible protection by dietary components present in the stomach contents. Partially purified rat lingual lipase (7-15 micrograms enzyme protein) was preincubated at 37 degrees C for 10-60 min at pH 1.0-6.0 before incubation for assay of lipolytic activity, hydrolysis of tri-[3H]olein at pH 5.4. The data show that partially purified rat lingual lipase preparations are stable at 37 degrees C in the pH range of 2.5-6.0. Enzyme activity, however, is rapidly and irreversibly lost during preincubation at pH 1.0-2.4 for 10-30 min. Protein (gelatin 1% or albumin 1% or 2.5%) cannot prevent the inactivation of lingual lipase at low pH. The large molecular species (molecular weight greater than 500,000) of lingual lipase (thought to be an aggregate of enzyme with lipids) is slightly more resistant to inactivation than the 46,000 dalton preparation, suggesting that lipids might protect the enzyme from inactivation. Indeed, about 60% of the initial lipase activity is preserved during incubation at pH 2.0 in the presence of 50 mM lecithin or 10 mM triolein. The data indicate that triglycerides which are hydrolyzed by this enzyme as well as phospholipids that are not hydrolyzed can prevent the inactivation of the enzyme.(ABSTRACT TRUNCATED AT 250 WORDS)

LIPASES PRODUZIDAS POR LEVEDURAS: CATALISADORES PODEROSOS PARA APLICAÇÕES INDUSTRIAIS

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Luiza Lux Lock

2007-12-01

Full Text Available O termo “enzimas lipolíticas” refere-se a lípases e hidrolases éster carboxílico. A produção de lipase é ampla entre as leveduras, mas poucas são capazes de produzir lipases com características interessantes e em quantidades suficientes para serem industrialmente úteis. A literatura relativa a lipases produzidas por Candida rugosa, Yarrowia (Candida lipolytica, Candida antarctica e outras leveduras produtoras de lípases é revisada. O uso de lípases recombinantes é discutido, com ênfase na utilização de sistemas de expressão heteróloga e desenho de quimeras. Finalmente, as três abordagens que visam à melhora da produção de lipase ou a modificação da seletividade do substrato da enzima (engenharia do meio, do biocatalisador e da proteína são discutidos.

m-DOPA addition in MAPLE immobilization of lipase for biosensor applications

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Valeria Califano

2015-12-01

Full Text Available Matrix Assisted Pulsed Laser Evaporation (MAPLE is a thin film deposition technique which uses a pulsed laser beam impinging, inside a high vacuum chamber, on a frozen target containing the guest molecules in a volatile matrix to induce fast “evaporation” of the matrix, and ejection of the guest molecules. Lipase, an enzyme acting as a catalyst in hydrolysis of lipids, is widely used in biosensors for detection of triglycerides in blood serum. A key action to this purpose is lipase immobilization on a substrate. In a recent paper, we have shown that MAPLE technique is able to deposit lipase on a substrate in an active form. Here we show that addition to the guest/matrix target of a small amount of m-DOPA (3-(3,4-dihydroxyphenyl-2-methyl-l-alanine in order to improve adhesion and protect lipase secondary structure, also allows the lowering the laser pulse energy required for matrix evaporation and therefore the risk of damaging the enzyme.

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.

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.

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.

Lipase-catalyzed ring-opening polymerization of lactones to polyesters and its mechanistic aspects.

Science.gov (United States)

Namekawa, S; Suda, S; Uyama, H; Kobayashi, S

1999-01-01

Lipase catalysis induced a ring-opening polymerization of lactones with different ring-sizes. Small-size (four-membered) and medium-size lactones (six- and seven-membered) as well as macrolides (12-, 13-, 16-, and 17-membered) were subjected to lipase-catalyzed polymerization. The polymerization behaviors depended primarily on the lipase origin and the monomer structure. The macrolides showing much lower anionic polymerizability were enzymatically polymerized faster than epsilon-caprolactone. The granular immobilized lipase derived from Candida antartica showed extremely efficient catalysis in the polymerization of epsilon-caprolactone. Single-step terminal functionalization of the polyester was achieved by initiator and terminator methods. The enzymatic polymerizability of lactones was quantitatively evaluated by Michaelis-Menten kinetics.

Characterization of neutral lipase BT-1 isolated from the labial gland of Bombus terrestris males.

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Jana Brabcová

Full Text Available BACKGROUND: In addition to their general role in the hydrolysis of storage lipids, bumblebee lipases can participate in the biosynthesis of fatty acids that serve as precursors of pheromones used for sexual communication. RESULTS: We studied the temporal dynamics of lipolytic activity in crude extracts from the cephalic part of Bombus terrestris labial glands. Extracts from 3-day-old males displayed the highest lipolytic activity. The highest lipase gene expression level was observed in freshly emerged bumblebees, and both gene expression and lipase activity were lower in bumblebees older than 3 days. Lipase was purified from labial glands, further characterized and named as BT-1. The B. terrestris orthologue shares 88% sequence identity with B. impatiens lipase HA. The molecular weight of B. terrestris lipase BT-1 was approximately 30 kDa, the pH optimum was 8.3, and the temperature optimum was 50°C. Lipase BT-1 showed a notable preference for C8-C10 p-nitrophenyl esters, with the highest activity toward p-nitrophenyl caprylate (C8. The Michaelis constant (Km and maximum reaction rate (Vmax for p-nitrophenyl laurate hydrolysis were Km = 0.0011 mM and Vmax = 0.15 U/mg. CONCLUSION: This is the first report describing neutral lipase from the labial gland of B. terrestris. Our findings help increase understanding of its possible function in the labial gland.

Elevação da lipase e amilase no doente crítico: estudo retrospectivo Increased lipase and amilase levels in critically ill patients: retrospective study

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Margarida Ferreira

2008-12-01

Full Text Available OBJETIVOS: A elevação da lipase e amilase séricas são frequentemente encontradas em doentes internados em unidade de terapia intensiva sem que exista doença pancreática prévia, constituindo um desafio diagnóstico e terapêutico. Baseados nesta evidência os autores propuseram-se a determinar a incidência de hiperlipasemia assintomática nos doentes críticos, fatores desencadeantes e evolução clínica destes doentes. MÉTODOS: Estudo retrospectivo dos doentes internados na unidade de terapia intensiva de 1 de janeiro a 31 de dezembro de 2006, excluídas internações por pancreatite aguda, história de patologia pancreática, insuficiência renal ou falta de dados. Pacientes foram distribuídos em dois grupos (com e sem hiperlipasemia e feita comparação considerando diversas variáveis clínicas, laboratoriais, imagiológicas. Análise estatística: SPSS 13; testes t de Student e qui² (IC 95%, com significância estatística se p38 ºC (pOBJECTIVES: Elevated lipase and amylase are commonly found in patients in intensive care unit without a previously recognized pancreatic illness, constituting a diagnostic and therapeutic challenge. The authors therefore proposed to determine the frequency of asymptomatic high serum lipase in critically ill patients, involved risk factors and outcome. METHODS: Retrospective study of patients admitted in an intensive care unit from January 1 to December 31, 2006, excluding admissions for acute pancreatitis, history of pancreatic disease, renal insufficiency or lacking of data. Patients were divided in two groups (with and without high serum lipase that were compared for clinical, laboratory and radiological variables. Statistical analysis: SPSS 13; Student's t test and Chi-square test (CI 95% with statistical significance if p< 0.05. RESULTS: 102 patients were included with high serum lipase was present in 39.2% of patients, mean lipase of 797U/L. Patients with high serum lipase had longer hospital

Purification and characterization of a new cold active lipase, EnL A ...

African Journals Online (AJOL)

SONU

2015-06-03

Jun 3, 2015 ... palm oil mill effluent dump sites, Pedavegi, West Godavari Dist, A.P. India and was ... carried out with the lipase production medium optimized using ..... Non edible Castor Oil by Immobilized Lipase from Bacillus aerius.

Enzymatic activity of a novel halotolerant lipase from Haloarcula hispanica 2TK2

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Ozgen Melis

2016-06-01

Full Text Available A strain of Haloarcula hispanica isolated from Tuzkoy salt mine, Turkey exhibited extracellular lipolytic activity. Important parameters such as carbon sources and salt concentration for lipase production were investigated. Optimal conditions for the enzyme production from Haloarcula hispanica 2TK2 were determined. It was observed that the lipolytic activity of Haloarcula hispanica was stimulated by some of the carbon sources. The high lipase acitivity values were obtained in the presence of 2% (v/v walnut oil (6.16 U/ml, 1% (v/v fish oil (5.07 U/ml, 1% (v/v olive oil (4.52 U/ml and 1% (w/v stearic acid (4.88 U/ml at 4M NaCl concentration. Lipase was partially purified by ammonium sulfate precipitation and ultrafiltration. Optimal temperature and pH values were determined as 45°C and 8.0, respectively. Lipase activity decreased with the increasing salt concentration, but 85% activity of the enzyme was maintained at 5M NaCl concentration. The enzyme preserved 41% of its relative activity at 90°C. The partially purified lipase maintained its activity in the presence of surfactants such as Triton X-100 and SDS. Therefore, the lipase which is an extremozyme may have potential applications especially in detergent industry.

Pancreatic lipase inhibitory constituents from Morus alba leaves and optimization for extraction conditions.

Science.gov (United States)

Jeong, Ji Yeon; Jo, Yang Hee; Kim, Seon Beom; Liu, Qing; Lee, Jin Woo; Mo, Eun Jin; Lee, Ki Yong; Hwang, Bang Yeon; Lee, Mi Kyeong

2015-06-01

The leaves of Morus alba (Moraceae) have been traditionally used for the treatment of metabolic diseases including diabetes and hyperlipidemia. Thus, inhibitory effect of M. alba leaves on pancreatic lipase and their active constituents were investigated in this study. Twenty phenolic compounds including ten flavonoids, eight benzofurans, one stilbene and one chalcones were isolated from the leaves of M. alba. Among the isolated compounds, morachalcone A (20) exerted strong pancreatic lipase inhibition with IC50 value of 6.2 μM. Other phenolic compounds containing a prenyl group showed moderate pancreatic lipase inhibition with IC50 value of <50 μM. Next, extraction conditions with maximum pancreatic lipase inhibition and phenolic content were optimized using response surface methodology with three-level-three-factor Box-Behnken design. Our results suggested the optimized extraction condition for maximum pancreatic lipase inhibition and phenolic content as ethanol concentration of 74.9%; temperature 57.4 °C and sample/solvent ratio, 1/10. The pancreatic lipase inhibition and total phenolic content under optimized condition were found to be 58.5% and 26.2 μg GAE (gallic acid equivalent)/mg extract, respectively, which were well matched with the predicted value. Copyright © 2015 Elsevier Ltd. All rights reserved.

The effect of interaction between Lipoprotein Lipase and ApoVLDL-II ...

African Journals Online (AJOL)

Body weight, abdominal fat weight and serum biochemical levels were determined from lean and fat chicken breeds at 12 weeks of age. Single nucleotide polymorphism (SNP) in apoVLDL-II and lipoprotein lipase genes was screened by PCR-SSCP and detected by direct sequencing. Lipoprotein lipase gene frequency ...

Significantly Elevated Serum Lipase in Pregnancy with Nausea and Vomiting: Acute Pancreatitis or Hyperemesis Gravidarum?

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Amanda Johnson

2015-01-01

Full Text Available Hyperemesis gravidarum is a severe manifestation of nausea and vomiting of pregnancy and it is associated with weight loss and metabolic abnormalities. It is known that abnormal laboratory values, including mildly elevated serum lipase level, could be associated with hyperemesis gravidarum. However, in this case report details of two women with hyperemesis gravidarum but with significantly elevated serum lipase levels were discussed. These patients presented with severe nausea and vomiting but without abdominal pain. They were found to have severely elevated lipase levels over 1,000 units/liter. In the absence of other findings of pancreatitis, they were treated with conservative measures for hyperemesis gravidarum, with eventual resolution to normal lipase levels. Although significantly elevated lipase level in pregnant patients with nausea and vomiting is a concern for acute pancreatitis, these two cases of significantly elevated serum lipase without other clinical findings of pancreatitis led to this report that serum lipase could be quite elevated in hyperemesis gravidarum and that it might not be an accurate biochemical marker for acute pancreatitis. Imaging studies are thus necessary to establish the diagnosis of acute pancreatitis.

Controlled lid-opening in Thermomyces lanuginosus lipase- An engineered switch for studying lipase function

DEFF Research Database (Denmark)

Skjold-Jørgensen, Jakob; Vind, Jesper; Moroz, Olga V.

2017-01-01

Here, we present a lipase mutant containing a biochemical switch allowing a controlled opening and closing of the lid independent of the environment. The closed form of the TlL mutant shows low binding to hydrophobic surfaces compared to the binding observed after activating the controlled switch...... inducing lid-opening. We directly show that lipid binding of this mutant is connected to an open lid conformation demonstrating the impact of the exposed amino acid residues and their participation in binding at the water-lipid interface. The switch was created by introducing two cysteine residues......-D measurements revealed a seven-fold increase in binding rate for the unlocked lipase. The TlL_locked mutant shows structural changes across the protein important for understanding the mechanism of lid-opening and closing. Our experimental results reveal sites of interest for future mutagenesis studies aimed...

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.

Relevant pH and lipase for in vitro models of gastric digestion.

Science.gov (United States)

Sams, Laura; Paume, Julie; Giallo, Jacqueline; Carrière, Frédéric

2016-01-01

The development of in vitro digestion models relies on the availability of in vivo data such as digestive enzyme levels and pH values recorded in the course of meal digestion. The variations of these parameters along the GI tract are important for designing dynamic digestion models but also static models for which the choice of representative conditions of the gastric and intestinal conditions is critical. Simulating gastric digestion with a static model and a single set of parameters is particularly challenging because the variations in pH and enzyme concentration occurring in the stomach are much broader than those occurring in the small intestine. A review of the literature on this topic reveals that most models of gastric digestion use very low pH values that are not representative of the fed conditions. This is illustrated here by showing the variations in gastric pH as a function of meal gastric emptying instead of time. This representation highlights those pH values that are the most relevant for testing meal digestion in the stomach. Gastric lipolysis is still largely ignored or is performed with microbial lipases. In vivo data on gastric lipase and lipolysis have however been collected in humans and dogs during test meals. The biochemical characterization of gastric lipase has shown that this enzyme is rather unique among lipases: (i) stability and activity in the pH range 2 to 7 with an optimum at pH 4-5.4; (ii) high tensioactivity that allows resistance to bile salts and penetration into phospholipid layers covering TAG droplets; (iii) sn-3 stereospecificity for TAG hydrolysis; and (iv) resistance to pepsin. Most of these properties have been known for more than two decades and should provide a rational basis for the replacement of gastric lipase by other lipases when gastric lipase is not available.

The genotypic diversity and lipase production of some thermophilic bacilli from different genera

OpenAIRE

Koc, Melih; Cokmus, Cumhur; Cihan, Arzu Coleri

2015-01-01

Abstract Thermophilic 32 isolates and 20 reference bacilli were subjected to Rep-PCR and ITS-PCR fingerprinting for determination of their genotypic diversity, before screening lipase activities. By these methods, all the isolates and references could easily be differentiated up to subspecies level from each other. In screening assay, 11 isolates and 7 references were found to be lipase producing. Their extracellular lipase activities were measured quantitatively by incubating in both tributy...

Effects of methanol on lipases: molecular, kinetic and process issues in the production of biodiesel.

Science.gov (United States)

Lotti, Marina; Pleiss, Jürgen; Valero, Francisco; Ferrer, Pau

2015-01-01

The biotechnological production of biodiesel is based on transesterification/esterification reactions between a source of fatty acids and a short-chain alcohol, usually methanol, catalysed by enzymes belonging to the class known as lipases. Several lipases used in industrial processes, although stable in the presence of other organic solvents, are inactivated by methanol at or below the concentration optimal for biodiesel production, making it necessary to use stepwise methanol feeding or pre-treatment of the enzyme. In this review article we focus on what is currently know about methanol inactivation of lipases, a phenomenon which is not common to all lipase enzymes, with the goal of improving the biocatalytic process. We suggest that different mechanisms can lead to inactivation of different lipases, in particular substrate inhibition and protein unfolding. Attempts to improve the performances of methanol sensitive lipases by mutagenesis as well as process engineering approaches are also summarized. Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Production of extracellular lipase by the phytopathogenic fungus Fusarium solani FS1

OpenAIRE

Maia, Maria de Mascena Diniz; Morais, Marcia Maria Camargo de; Morais Jr., Marcos Antonio de; Melo, Eduardo Henrique Magalhães; Lima Filho, José Luiz de

1999-01-01

A Brazilian strain of Fusarium solani was tested for extracellular lipase production in peptone-olive oil medium. The fungus produced 10,500 U.l-1 of lipase after 72 hours of cultivation at 25oC in shake-flask at 120 rpm in a medium containing 3% (w/v) peptone plus 0.5% (v/v) olive oil. Glucose (1% w/v) was found to inhibit the inductive effect of olive oil. Peptone concentrations below 3% (w/v) resulted in a reduced lipase production while increased olive oil concentration (above 0.5%) did n...

Modeling of lipase catalyzed ring-opening polymerization of epsilon-caprolactone.

Science.gov (United States)

Sivalingam, G; Madras, Giridhar

2004-01-01

Enzymatic ring-opening polymerization of epsilon-caprolactone by various lipases was investigated in toluene at various temperatures. The determination of molecular weight and structural identification was carried out with gel permeation chromatography and proton NMR, respectively. Among the various lipases employed, an immobilized lipase from Candida antartica B (Novozym 435) showed the highest catalytic activity. The polymerization of epsilon-caprolactone by Novozym 435 showed an optimal temperature of 65 degrees C and an optimum toluene content of 50/50 v/v of toluene and epsilon-caprolactone. As lipases can degrade polyesters, a maximum in the molecular weight with time was obtained due to the competition of ring opening polymerization and degradation by specific chain end scission. The optimum temperature, toluene content, and the variation of molecular weight with time are consistent with earlier observations. A comprehensive model based on continuous distribution kinetics was developed to model these phenomena. The model accounts for simultaneous polymerization, degradation and enzyme deactivation and provides a technique to determine the rate coefficients for these processes. The dependence of these rate coefficients with temperature and monomer concentration is also discussed.

Diagnostic value of post-heparin lipase testing in detecting common genetic variants in the LPL and LIPC genes

NARCIS (Netherlands)

van Hoek, Mandy; Dallinga-Thie, Geesje M.; Steyerberg, Ewout W.; Sijbrands, Eric J. G.

2009-01-01

Post-heparin lipoprotein lipase and hepatic lipase activities are used to identify primary disorders of triglyceride and HDL-cholesterol metabolism. Their ability to identify common variants in the lipoprotein lipase (LPL) and hepatic lipase (LIPC) genes is unclear. To investigate the ability of

Different Covalent Immobilizations Modulate Lipase Activities of Hypocrea pseudokoningii

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Marita G. Pereira

2017-09-01

Full Text Available Enzyme immobilization can promote several advantages for their industrial application. In this work, a lipase from Hypocrea pseudokoningii was efficiently linked to four chemical supports: agarose activated with cyanogen bromide (CNBr, glyoxyl-agarose (GX, MANAE-agarose activated with glutaraldehyde (GA and GA-crosslinked with glutaraldehyde. Results showed a more stable lipase with both the GA-crosslinked and GA derivatives, compared to the control (CNBr, at 50 °C, 60 °C and 70 °C. Moreover, all derivatives were stabilized when incubated with organic solvents at 50%, such as ethanol, methanol, n-propanol and cyclohexane. Furthermore, lipase was highly activated (4-fold in the presence of cyclohexane. GA-crosslinked and GA derivatives were more stable than the CNBr one in the presence of organic solvents. All derivatives were able to hydrolyze sardine, açaí (Euterpe oleracea, cotton seed and grape seed oils. However, during the hydrolysis of sardine oil, GX derivative showed to be 2.3-fold more selectivity (eicosapentaenoic acid (EPA/docosahexaenoic acid (DHA ratio than the control. Additionally, the types of immobilization interfered with the lipase enantiomeric preference. Unlike the control, the other three derivatives preferably hydrolyzed the R-isomer of 2-hydroxy-4-phenylbutanoic acid ethyl ester and the S-isomer of 1-phenylethanol acetate racemic mixtures. On the other hand, GX and CNBr derivatives preferably hydrolyzed the S-isomer of butyryl-2-phenylacetic acid racemic mixture while the GA and GA-crosslink derivatives preferably hydrolyzed the R-isomer. However, all derivatives, including the control, preferably hydrolyzed the methyl mandelate S-isomer. Moreover, the derivatives could be used for eight consecutive cycles retaining more than 50% of their residual activity. This work shows the importance of immobilization as a tool to increase the lipase stability to temperature and organic solvents, thus enabling the possibility of

GDSL lipases modulate immunity through lipid homeostasis in rice.

Science.gov (United States)

Gao, Mingjun; Yin, Xin; Yang, Weibing; Lam, Sin Man; Tong, Xiaohong; Liu, Jiyun; Wang, Xin; Li, Qun; Shui, Guanghou; He, Zuhua

2017-11-01

Lipids and lipid metabolites play important roles in plant-microbe interactions. Despite the extensive studies of lipases in lipid homeostasis and seed oil biosynthesis, the involvement of lipases in plant immunity remains largely unknown. In particular, GDSL esterases/lipases, characterized by the conserved GDSL motif, are a subfamily of lipolytic enzymes with broad substrate specificity. Here, we functionally identified two GDSL lipases, OsGLIP1 and OsGLIP2, in rice immune responses. Expression of OsGLIP1 and OsGLIP2 was suppressed by pathogen infection and salicylic acid (SA) treatment. OsGLIP1 was mainly expressed in leaf and leaf sheath, while OsGLIP2 showed high expression in elongating internodes. Biochemical assay demonstrated that OsGLIP1 and OsGLIP2 are functional lipases that could hydrolyze lipid substrates. Simultaneous down-regulation of OsGLIP1 and OsGLIP2 increased plant resistance to both bacterial and fungal pathogens, whereas disease resistance in OsGLIP1 and OsGLIP2 overexpression plants was significantly compromised, suggesting that both genes act as negative regulators of disease resistance. OsGLIP1 and OsGLIP2 proteins mainly localize to lipid droplets and the endoplasmic reticulum (ER) membrane. The proper cellular localization of OsGLIP proteins is indispensable for their functions in immunity. Comprehensive lipid profiling analysis indicated that the alteration of OsGLIP gene expression was associated with substantial changes of the levels of lipid species including monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG). We show that MGDG and DGDG feeding could attenuate disease resistance. Taken together, our study indicates that OsGLIP1 and OsGLIP2 negatively regulate rice defense by modulating lipid metabolism, thus providing new insights into the function of lipids in plant immunity.

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.

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.

Liquid lipases for enzymatic concentration of n-3 polyunsaturated fatty acids in monoacylglycerols via ethanolysis: Catalytic specificity and parameterization.

Science.gov (United States)

He, Yongjin; Li, Jingbo; Kodali, Sitharam; Balle, Thomas; Chen, Bilian; Guo, Zheng

2017-01-01

This work examined catalytic specificity and fatty acid selectivity of five liquid lipases C. antarctica lipase A and B (CAL-A/B), and lipase TL (T. lanuginosus), Eversa Transfrom and NS in ethanolysis of fish oil with the aim to concentrate n-3 PUFAs into monoacylglycerols (MAGs) products. Lipase TL, Eversa Transform & NS entail a much faster reaction and produce higher MAGs yield (>30%); whereas CAL-A obtains the highest concentration of n-3 PUFAs/DHA/EPA into MAGs products (88.30%); followed by lipase NS (81.02%). 13 C NMR analysis indicates that CAL-B and lipase TL are sn-1,3 specific; but CAL-A and lipase Eversa Transform are non-regiospecific or weak sn-2 specific; which plausibly explains high enrichment effect of the latter two lipases. All liquid lipases are observed reusable for a certain times (lipase Eversa Transform up to 12 times), demonstrating their competitive advantage over immobilized form for industrial application because of their higher activity and cheaper operation cost. Copyright © 2016 Elsevier Ltd. All rights reserved.

Microbial lipase mediated by health beneficial modification of cholesterol and flavors in food products: A review.

Science.gov (United States)

Sharma, Ranjana; Sharma, Nivedita

2017-06-14

The tremendous need of lipase in varied applications in biotechnological increases its economical value in food and allied industries. Lipase has an impressive number of applications viz. enhancements of flavor in food products (Cheese, butter, alcoholic beverages, milk chocolate and diet control food stuffs), detergent industry in removing oil, grease stain, organic chemical processing, textile industry, oleochemical industry, cosmetic industry and also as therapeutic agents in pharmaceutical industries. This communication extends the frontier of lipase catalyzed benefits to human body by lowering serum cholesterol and enhancement of flavor in different food products. Among all, multiple innovations going on in the field of lipase applications are widening its scope in food industries consistently. Therefore in the present work an effort has been made to explore the utilization of lipase in the field of food product enhancement. Supplementation of food products with lipase results in modification of its physical, chemical and biochemical properties by enhancing its therapeutic activity. Lipases are the most important enzymes used in food industries. They are utilized as industrial catalysts for lipid hydrolysis. Because of lipases hydrolysis nature it is widely exploited to catalyze lipids or fats in different food products and enhancement of food flavors. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

PPARγ regulates exocrine pancreas lipase.

Science.gov (United States)

Danino, Hila; Naor, Ronny Peri-; Fogel, Chen; Ben-Harosh, Yael; Kadir, Rotem; Salem, Hagit; Birk, Ruth

2016-12-01

Pancreatic lipase (triacylglycerol lipase EC 3.1.1.3) is an essential enzyme in hydrolysis of dietary fat. Dietary fat, especially polyunsaturated fatty acids (PUFA), regulate pancreatic lipase (PNLIP); however, the molecular mechanism underlying this regulation is mostly unknown. As PUFA are known to regulate expression of proliferator-activated receptor gamma (PPARγ), and as we identified in-silico putative PPARγ binding sites within the putative PNLIP promoter sequence, we hypothesized that PUFA regulation of PNLIP might be mediated by PPARγ. We used in silico bioinformatics tools, reporter luciferase assay, PPARγ agonists and antagonists, PPARγ overexpression in exocrine pancreas AR42J and primary cells to study PPARγ regulation of PNLIP. Using in silico bioinformatics tools we mapped PPARγ binding sites (PPRE) to the putative promoter region of PNLIP. Reporter luciferase assay in AR42J rat exocrine pancreas acinar cells transfected with various constructs of the putative PNLIP promoter showed that PNLIP transcription is significantly enhanced by PPARγ dose-dependently, reaching maximal levels with multi PPRE sites. This effect was significantly augmented in the presence of PPARγ agonists and reduced by PPARγ antagonists or mutagenesis abrogating PPRE sites. Over-expression of PPARγ significantly elevated PNLIP transcript and protein levels in AR42J cells and in primary pancreas cells. Moreover, PNLIP expression was up-regulated by PPARγ agonists (pioglitazone and 15dPGJ2) and significantly down-regulated by PPARγ antagonists in non-transfected rat exocrine pancreas AR42J cell line cells. PPARγ transcriptionally regulates PNLIP gene expression. This transcript regulation resolves part of the missing link between dietary PUFA direct regulation of PNLIP. Copyright © 2016 Elsevier B.V. All rights reserved.

Novel Lipases: Expression and Improvement for Applied Biocatalysis = Nuevas lipasas: expresión y mejoras para biocatálisis aplicadatalysi Novel Lipases: Expression and Improvement for Applied Biocatalysis = Nuevas lipasas: expresión y mejoras para biocatálisis aplicada

OpenAIRE

Infanzón Ramos, Belén

2017-01-01

[eng] This thesis is focused in the identification and improvement of lipases for biotechnological application. The importance of lipases is increasing in several industries. However, the commercial use of lipases is still a drawback in the economics of the lipase-based industrial applications. There are many tools for improving and adapting the enzyme properties to the desired requirements of a process that could lead lipase catalysis through a cost-effective process. In this context, the m...

Influence of dietary recombinant microbial lipase on performance and quality characteristics of rainbow trout, Oncorhynchus mykiss

DEFF Research Database (Denmark)

Samuelsen, Troels; Isaksen, Mai; McLean, Ewen

2001-01-01

In order to assess whether supplementary lipase affected growth and body composition of trout, four diets were produced, consisting of (A) feed containing high (2083 mg kg(-1)), (B) low (208.3 mg kg(-1)) concentrations of lipase, (C) heat-treated (inactivated) lipase (2083 mg kg(-1)), and (D......) a basal control diet. Rainbow trout (n = 40/tank; initial wt. 23.22 +/- 4.81 g; length 124.7 +/- 6.35 mm) were fed, according to commercial feed tables, 6 days/week for 202 days. Retained activity of supplemental lipase was verified by monitoring free fatty acid appearance (FAA), which was significantly...... higher(P Lipase addition had no effect(P > 0.05) on growth, fillet proximate composition, hepatosomatic, cardiac, or gut indices, and carcass percentage. However, lipase supplementation influenced the mono-unsaturated fatty acid profiles of the fillet (P

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.

Effect of immobilized lipase supplementation of diets on the performance of the Japanese quails

International Nuclear Information System (INIS)

Abu-Taleb, A.M.; Ezzat, I.E.; Saleh, M.

2004-01-01

In the present study, lipase was immobilized onto two different supports, agarose and gelatin. Some physico-chemical properties of the free and immobilized lipase such as optimum temperature, optimum ph and storage stability were studied. Storage of the enzymes for 2 months showed that the free enzyme lost its activity, while the immobilized on the gelatin showed better resistance towards ph and temperature variations than that immobilized onto agarose. Four experiments were conducted to test the effect of the immobilized lipase supplementation on the productive performance of the Japanese quails. During the first 3 weeks, the addition of lipase to poultry diets caused an increase in the body weight gain of birds than the enzyme-free diet. An obvious improvement in quail day egg production during the laying period was observed with the groups fed on a diet supplemented with 3000 and 2000 I U of immobilized lipase per kilogram feed. Blood cholesterol was not affected with lipase addition, while total lipids were significantly increased. Significant reduction was also observed in thyroid hormones (T 3 and T 4 ) as compared with the control group

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.

Quantitative approach to track lipase producing Pseudomonas sp. S1 in nonsterilized solid state fermentation.

Science.gov (United States)

Sahoo, R K; Subudhi, E; Kumar, M

2014-06-01

Proliferation of the inoculated Pseudomonas sp. S1 is quantitatively evaluated using ERIC-PCR during the production of lipase in nonsterile solid state fermentation an approach to reduce the cost of enzyme production. Under nonsterile solid state fermentation with olive oil cake, Pseudomonas sp. S1 produced 57·9 IU g(-1) of lipase. DNA fingerprints of unknown bacterial isolates obtained on Bushnell Haas agar (BHA) + tributyrin exactly matched with that of Pseudomonas sp. S1. Using PCR-based enumeration, population of Pseudomonas sp. S1 was proliferated from 7·6 × 10(4) CFU g(-1) after 24 h to 4·6 × 10(8) CFU g(-1) after 96 h, which tallied with the maximum lipase activity as compared to control. Under submerged fermentation (SmF), Pseudomonas sp. S1 produced maximum lipase (49 IU ml(-1) ) using olive oil as substrate, while lipase production was 9·754 IU ml(-1) when Pseudomonas sp. S1 was grown on tributyrin. Optimum pH and temperature of the crude lipase was 7·0 and 50°C. Crude enzyme activity was 71·2% stable at 50°C for 360 min. Pseudomonas sp. S1 lipase was also stable in methanol showing 91·6% activity in the presence of 15% methanol, whereas 75·5 and 51·1% of activity were retained in the presence of 20 and 30% methanol, respectively. Thus, lipase produced by Pseudomonas sp. S1 is suitable for the production of biodiesel as well as treatment of oily waste water. This study presents the first report on the production of thermophilic organic solvent tolerant lipase using agro-industry waste in nonsterile solid state fermentation. Positive correlation between survival of Pseudomonas sp. S1 and lipase production under nonsterile solid state fermentation was established, which may emphasize the need to combine molecular tools and solid state fermentation in future studies. Our study brings new insights into the lipase production in cost-effective manner, which is an industrially relevant approach. © 2014 The Society for Applied Microbiology.

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.

Sequential Detection of Thermophilic Lipase and Protease by Zymography.

Science.gov (United States)

Kurz, Liliana; Hernández, Zully; Contreras, Lellys M; Wilkesman, Jeff

2017-01-01

Lipase and protease present in cell-free fractions of thermophilic Bacillus sp. cultures were analyzed by polyacrylamide gel (PAG) electrophoresis. After run, the gel is electrotransferred to another PAG copolymerized with glycerol tributyrate, olive oil, and gelatin. This multi-substrate gel was incubated first for lipase detection, until bands appeared, and then stained with Coomassie for protease detection. Advantages of this sequential procedure are the detection of two different enzyme activities on a single PAG, beside time and resource saving.