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Sample records for pichia guilliermondii indigenous

  1. Spoilage yeasts in Patagonian winemaking: molecular and physiological features of Pichia guilliermondii indigenous isolates Levaduras contaminantes en vinos patagónicos: características moleculares y fisiológicas de los aislamientos indígenas de Picchia guilliermondii

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    C. A. Lopes

    2009-09-01

    Full Text Available Yeasts belonging to the genus Dekkera/Brettanomyces, especially the species Dekkera bruxellensis, have long been associated with the production of volatile phenols responsible for off-flavour in wines. According to recent reports, the species Pichia guilliermondii could also produce these compounds at the initial stages of fermentation. Based on the abundance of P. guilliermondii in Patagonian winemaking, we decided to study the relevance of indigenous isolates belonging to this species as wine spoilage yeast. Twenty-three indigenous isolates obtained from grape surfaces and red wine musts were analyzed in their capacity to produce volatile phenols on grape must. The relationship between molecular Random Amplified Polymorphic DNA (RAPD and physiological (killer biotype patterns detected in indigenous populations of P. guilliermondii and volatile phenol production was also evaluated. Different production levels of 4-ethylphenol, 4-vinylguaiacol and 4-ethylguaiacol were detected among the isolates; however, the values were always lower than those produced by the D. bruxellensis reference strain in the same conditions. High levels of 4-vinylphenol were detected among P. guilliermondii indigenous isolates. The combined use of RAPD and killer biotype allowed us to identify the isolates producing the highest volatile phenol levels.Las levaduras del género Dekkera/Brettanomyces, sobre todo la especie Dekkera bruxellensis, siempre han sido asociadas con la producción de fenoles volátiles responsables de aromas desagradables en los vinos. Recientemente, se ha demostrado que la especie Pichia guilliermondii también es capaz de producir estos compuestos, particularmente durante las etapas iniciales de la fermentación. Dada la abundancia de P. guilliermondii en las bodegas de la Patagonia, se decidió evaluar la importancia de algunos aislamientos indígenas de esta especie como levaduras alterantes de vinos regionales. Se evaluó la capacidad de

  2. Purification of Candida guilliermondii and Pichia ohmeri killer toxin as an active agent against Penicillium expansum.

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    Coelho, Alexandre Rodrigo; Tachi, Masahico; Pagnocca, Fernando Carlos; Nobrega, Gisele Maria Andrade; Hoffmann, Fernando Leite; Harada, Ken-Ichi; Hirooka, Elisa Yoko

    2009-01-01

    An antifungal assay with cell-free culture supernatant of Pichia ohmeri 158 and Candida guilliermondii P3 was tested against Penicillium expansum strain #2 at 25 degrees C by measuring hyphal length and percentage conidia germination. C. guilliermondii was more effective against P. expansum conidia germination (58.15% inhibition), while P. ohmeri showed higher inhibition of mycelial growth (66.17%), indicating a probable mechanism associated with killer activity. This killer toxin (molecular mass expansum:% inhibition rose from 42.16 to 90.93% (C. guilliermondii) and 39.32 to 91.12% (P. ohmeri) (p Penicillium activity.

  3. Mitotically unstable polyploids in the yeast Pichia guilliermondii.

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    Klinner, U; Böttcher, F

    1992-01-01

    Attempts to obtain triploids or tetraploids of P. guilliermondii by sexual hybridization led to mitotically stable hybrids. However, their DNA content per cell was not higher than in diploids. The results of random spore analysis demonstrate that these hybrids were in fact aneuploids which obviously suffered drastic chromosome losses immediately after mating. This phenomenon could have been caused either by aneuploidy already present in the parental strains or it might have been due to a general inability of P. guilliermondii to maintain a polyploid genome.

  4. A PUBLISHED KINETIC MODEL EXPLAINS THE VARIATION IN NITROGEN CONTENT OF Pichia guilliermondii DURING ITS BATCH CULTIVATION ON DIESEL OIL

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    BORZANI W.

    1999-01-01

    Full Text Available Variation in nitrogen content of Pichia guilliermondii during its batch cultivation on media containing diesel oil as the main carbon source may be explained by means of a kinetic model proposed earlier to interpret the kinetics of nitrogen consumption during the process.

  5. Enhanced volatile phenols in wine fermented with Saccharomyces cerevisiae and spoiled with Pichia guilliermondii and Dekkera bruxellensis.

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    Sáez, J S; Lopes, C A; Kirs, V C; Sangorrín, M P

    2010-08-01

    To investigate whether the presence of Pichia guilliermondii impacts on the production of volatile phenols from mixed wine fermentations with Dekkera bruxellensis and Saccharomyces cerevisiae. Four inoculation strategies were performed in small-scale fermentations involving P. guilliermondii, D. bruxellensis and S. cerevisiae using Syrah grape juice supplemented with 100 mg l(-1) of p-coumaric acid. High pressure liquid chromatography was used for the quantification or volatile phenols. Significant high levels of 4-ethylphenol and 4-ethylguaicol (720 and 545 microg l(-1), respectively), as well as the highest levels of 4-vinylphenol (>4500 microg l(-1)), were observed when P. guilliermondii species was inoculated from the beginning of the fermentation. The metabolic interaction occurring between the high vinylphenol producer species P. guilliermondii and D. bruxellensis exhibiting a high vinylphenol reductase activity resulted in an increased production of volatile phenols in wine. Pichia guilliermondii must be considered a very important spoilage yeast in the wine industry capable of producing large amounts of volatile phenols.

  6. Identification and characterization of Dekkera bruxellensis, Candida pararugosa, and Pichia guilliermondii isolated from commercial red wines

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    Jensen, Susanne Langgård; Umiker, Nicole L.; Arneborg, Nils

    2009-01-01

    Yeast isolates from commercial red wines were characterized with regards to tolerances to molecular SO2, ethanol, and temperature as well as synthesis of 4-ethyl-phenol/4-ethyl-guaiacol in grape juice or wine. Based on rDNA sequencing, nine of the 11 isolates belonged to Dekkera bruxellensis (B1a......, B1b, B2a, E1, F1a, F3, I1a, N2, and P2) while the other two were Candida pararugosa (Q2) and Pichia guilliermondii (Q3). Strains B1b, Q2, and Q3 were much more resistant to molecular SO2 in comparison to the other strains of Dekkera. These strains were inoculated (103-104 cfu/ml) along with lower...

  7. Physical Interactions between Yeast Pichia guilliermondii and Post-Harvest Fruit Pathogen Penicillium expansum

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    SRI WIDYASTUTI

    2008-03-01

    Full Text Available Attachment of yeast cells or bacteria on fungal hyphae have been observed in various antagonisms between microorganisms. Physical interactions between yeast Pichia guilliermondii and postharvest fruit pathogen Penicillium expansum in culture were studied in detail using light and transmission electron microscope to give better understanding on their mode of antagonism. Both organisms were co-cultured for 24-hr on potato dextrose agar. Light microscopy observations on the co-culture showed that the yeast cells attached firmly on the fungal hyphae. This attachment was inhibited by several substances such as enzymes degrading protein (protease or trypsin, a respiration inhibitor (sodium azide, an acid (hydrochloric acid or an alkali (sodium hydroxide. Although autoclaved hyphae did not affect the attachment, but boiled enzymes and autoclaved yeast cells totally abolished the attachment. These evidences suggested that the attachment might be an active process mediated by certain protein from live yeast cells. Transmission electron micrographs on the ultrastructure of the co-culture revealed that the hyphae showed abnormalities in their structure and organelles, and a degree of obvious damage. Physical interactions observed in this study could be contributed to the mechanism of antagonism between P. guilliermondii and P. expansum.

  8. Molecular typing of the yeast species Dekkera bruxellensis and Pichia guilliermondii recovered from wine related sources.

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    Martorell, Patricia; Barata, André; Malfeito-Ferreira, Manuel; Fernández-Espinar, M Teresa; Loureiro, Virgilio; Querol, Amparo

    2006-01-15

    A total of 63 strains of Dekkera bruxellensis and 32 strains of Pichia guilliermondii isolated from wine related environments were identified by restriction analysis of the 5.8S-ITS region of the rDNA. These strains were subjected to intraspecific discrimination using mtDNA restriction and RAPD-PCR analysis. The isolates identified as D. bruxellensis yielded 3 different molecular patterns of mtDNA restriction using the endonuclease HinfI. The pattern A was the most frequent (58 strains) among strains from different sources, regions and countries. Pattern B (4 strains) and C (one strain) were determined in isolates from Portuguese wines. The discrimination among the pattern A strains was achieved by a RAPD-PCR assay with 3 primers (OPA-2, OPA-3 and OPA-9). A total of 12 haplotypes were obtained with the combination of the patterns provided by the 3 OPAs. The pattern 2 was the most frequent and extensively distributed being found in strains from different countries and from different sources like wine, barrique wood and insects. The strains of P. guilliermondii were characterized with restriction of mtDNA using the endonuclease HinfI yielding 7 different restriction patterns. These patterns were associated with different efficiencies of 4-ethylphenol production. Patterns A to D corresponded to 19 strains producing low levels of 4-ethylphenol (50 mg/l), when grown in synthetic media supplemented with 100 mg/l of p-coumaric acid. The high degree of polymorphism observed shows that intraspecific typing is essential for accurate yeast dissemination studies in wine related environments.

  9. Identification and characterization of Dekkera bruxellensis, Candida pararugosa, and Pichia guilliermondii isolated from commercial red wines.

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    Jensen, Susanne L; Umiker, Nicole L; Arneborg, Nils; Edwards, Charles G

    2009-12-01

    Yeast isolates from commercial red wines were characterized with regards to tolerances to molecular SO(2), ethanol, and temperature as well as synthesis of 4-ethyl-phenol/4-ethyl-guaiacol in grape juice or wine. Based on rDNA sequencing, nine of the 11 isolates belonged to Dekkera bruxellensis (B1a, B1b, B2a, E1, F1a, F3, I1a, N2, and P2) while the other two were Candida pararugosa (Q2) and Pichia guilliermondii (Q3). Strains B1b, Q2, and Q3 were much more resistant to molecular SO(2) in comparison to the other strains of Dekkera. These strains were inoculated (10(3)-10(4)cfu/ml) along with lower populations of Saccharomyces (10(5)cfu/ml. In wine, Q3 never entered logarithmic growth and quickly died in contrast to Q2 which survived >40 days after inoculation. B1b grew well in wine incubated at 21 degrees C while slower growth was observed at 15 degrees C. Neither Q2 nor Q3 produced 4-ethyl-phenol or 4-ethyl-guaiacol, unlike B1b. However, lower concentrations of volatile phenols were present in wine incubated at 15 degrees C compared to 21 degrees C.

  10. Mutations and environmental factors affecting regulation of riboflavin synthesis and iron assimilation also cause oxidative stress in the yeast Pichia guilliermondii.

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    Boretsky, Yuriy R; Protchenko, Olga V; Prokopiv, Tetiana M; Mukalov, Igor O; Fedorovych, Daria V; Sibirny, Andriy A

    2007-10-01

    Iron deficiency causes oversynthesis of riboflavin in several yeast species, known as flavinogenic yeasts. However, the mechanisms of such regulation are not known. We found that mutations causing riboflavin overproduction and iron hyperaccumulation (rib80, rib81 and hit1), as well as cobalt excess or iron deficiency all provoke oxidative stress in the Pichia guilliermondii yeast. Iron content in the cells, production both of riboflavin and malondialdehyde by P. guilliermondii wild type and hit1 mutant strains depend on a type of carbon source used in cultivation media. The data suggest that the regulation of riboflavin biosynthesis and iron assimilation in P. guilliermondii are linked with cellular oxidative state.

  11. Isolation and characterisation of 8-hydroxy-3Z,5Z-tetradecadienoic acid, a putative intermediate in Pichia guilliermondii gamma-decalactone biosynthesis from ricinoleic acid.

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    Iacazio, G; Martini, D; Faure, B; N'Guyen, M H

    2002-03-19

    During a screening procedure for the discovery of a strong gamma-decalactone producer from ricinoleic acid, we observed that the yeast Pichia guilliermondii accumulated transiently 8-hydroxy-3Z,5Z-tetradecadienoic acid 1 during gamma-decalactone biosynthesis in the stationary phase of growth. The structural elucidation of 1 was based on nuclear magnetic resonance, infrared, ultraviolet and gas chromatography-mass spectrometry experiments. The occurrence of 1 is discussed in relation with previously proposed gamma-decalactone biosynthetic pathways.

  12. The ability of the antagonist yeast Pichia guilliermondii strain Z1 to suppress green mould infection in citrus fruit

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    Rachid Lahlali

    2014-12-01

    Full Text Available In previous studies it was shown that Pichia guilliermondii strain Z1, isolated from healthy Moroccan citrus Valencia-Late oranges, was effective against Penicillium italicum. Here the effectiveness of strain Z1 was assessed against Penicillium digitatum, the causal agent of green mould, under different temperature (5-25°C and relative humidity (RH (45-100% regimes for its reliable and largescale application in packinghouse. All main effects and interactions were significant (P80%. Its applications as a formulated product significantly reduced the incidence of infected fruit (55% and the percentage of infected wounds (47% compared to the only pathogen control treatment. However, disease control with formulated product was significantly less than that obtained with thiabendazole (30% or strain Z1 culturable cells (35%. These results highlight that strain Z1 is an effective biological control agent for control of green mould under varying environmental conditions, and control may be optimized by combining its use with other environmentally-safe post-harvest treatments or improved formulation.

  13. Deficiency in frataxin homologue YFH1 in the yeast Pichia guilliermondii leads to missregulation of iron acquisition and riboflavin biosynthesis and affects sulfate assimilation.

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    Pynyaha, Yuriy V; Boretsky, Yuriy R; Fedorovych, Daria V; Fayura, Lubov R; Levkiv, Andriy I; Ubiyvovk, Vira M; Protchenko, Olha V; Philpott, Caroline C; Sibirny, Andriy A

    2009-12-01

    Pichia guilliermondii is a representative of yeast species that overproduce riboflavin (vitamin B2) in response to iron deprivation. P. guilliermondii YFH1 gene coding for frataxin homologue, eukaryotic mitochondrial protein involved in iron trafficking and storage, was identified and deleted. Constructed P. guilliermondii Δyfh1 mutant grew very poorly in a sucrose-containing synthetic medium supplemented with sulfate or sulfite as a sole sulfur source. Addition of sodium sulfide, glutathione, cysteine, methionine, N-acetyl-L-cysteine partially restored growth rate of the mutant suggesting that it is impaired in sulfate assimilation. Cellular iron content in Δyfh1 mutant was ~3-3.5 times higher as compared to the parental strain. It produced 50-70 times more riboflavin in iron sufficient synthetic media relative to the parental wildtype strain. Biomass yield of the mutant in the synthetic glutathione containing medium supplemented with glycerol as a sole carbon source was 1.4- and 2.6-fold increased as compared to sucrose and succinate containing media, respectively. Oxygen uptake of the Δyfh1 mutant on sucrose, glycerol or succinate, when compared to the parental strain, was decreased 5.5-, 1.7- and 1.5-fold, respectively. Substitution of sucrose or glycerol in the synthetic iron sufficient medium with succinate completely abolished riboflavin overproduction by the mutants. Deletion of the YFH1 gene caused hypersensitivity to hydrogen peroxide and exogenously added riboflavin and led to alterations in superoxide dismutase activities. Thus, deletion of the gene coding for yeast frataxin homologue has pleiotropic effect on metabolism in P. guilliermondii.

  14. Co-culturing of Pichia guilliermondii enhanced volatile flavor compound formation by Zygosaccharomyces rouxii in the model system of Thai soy sauce fermentation.

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    Wah, Thin Thin; Walaisri, Supawan; Assavanig, Apinya; Niamsiri, Nuttawee; Lertsiri, Sittiwat

    2013-01-01

    The roles of salt-tolerant yeasts such as Zygosaccharomyces rouxii, Candida versatilis, and Candida etchellsii in the production of volatile flavor compounds (VFCs) in soy sauce fermentation have been well documented. However, the knowledge of VFC production by other salt-tolerant yeasts is still limited. In this work, the roles of Z. rouxii and Pichia guilliermondii strains in VFC production were investigated in moromi medium as a model system for soy sauce fermentation. Inoculation of a single culture of either Z. rouxii or P. guilliermondii as well as co-cultures of these two yeasts into moromi medium showed increased numbers of viable yeast at around 0.7 to 1.9 log CFU/mL after 7days of cultivation at 30°C. During cultivation, both single and co-cultures displayed survival over a 7-day time period, compared with the controls (no culture added). Overall, yeast inoculation enhanced the production of VFCs in the moromi media with higher amounts of ethanol, alcohols, furanones, esters, aldehyde, acid, pyrone and phenols, known as important characteristic flavor compounds in soy sauce. Moreover, the co-culture produced more alcohols, furanones, esters, maltol and benzoic acid than the single culture of Z. rouxii.

  15. Bioremediation of chromium by the yeast Pichia guilliermondii: toxicity and accumulation of Cr (III) and Cr (VI) and the influence of riboflavin on Cr tolerance.

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    Ksheminska, Helena; Jaglarz, Anita; Fedorovych, Daria; Babyak, Lyubov; Yanovych, Dmytro; Kaszycki, Pawel; Koloczek, Henryk

    2003-01-01

    A comparative study has been made on the sensitivity of the yeast Pichia guilliermondii to Cr (III) and Cr (VI) as well as on the Cr uptake potential at growth-inhibitory concentrations of chromium. The strains used in the study were either isolated from natural sources or obtained from a laboratory strain collection. The results show that most of the natural strains were more tolerant to chromium and were able to grow in the presence of 5 mM Cr (III) or 0.5 mM Cr (VI), that is at concentrations which substantially inhibited the growth of laboratory strains. The cellular Cr content after treatment was similar for both strain types and ranged from 1.2-4.0 mg/g d.w. and 0.4-0.9 mg/g d.w., for Cr (III) and Cr (VI) forms, respectively, however, in one case of a natural strain it reached the value of 10 mg Cr (III)/g dry mass. Natural-source strains were grouped into four groups based on the yeasts' differential response to Cr (III) and Cr (VI). Hexavalent Cr-resistant mutants of a P. giuilliermondii laboratory strain, which revealed markedly changed capabilities of chromium accumulation, were obtained by means of UV-induced mutagenesis. Cr (VI) treatment triggered oversynthesis of riboflavin and the addition of exogenous riboflavin increased P. guilliermondii resistance to both Cr (III) and Cr (VI). Electrophoretic protein profiles revealed the induction and/or suppression of several proteins in response to toxic Cr (VI) levels.

  16. [Properties of 2,5-diamino-4-oxy-6-ribosylaminopyrimidine-5'- phosphate reductase, a enzyme of the second stage of flavinogenesis in Pichia guilliermondii yeasts].

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    Logvinenko, E M; Shavlovskiĭ, G M; Zakal'skiĭ, A E; Kontorovskaia, N Iu

    1989-01-01

    2,5-Diamino-4-oxy-6-ribosylaminopyrimidine-5'-phosphate reductase has been isolated from cells of Pichia guilliermondii and subjected to 20-fold purification by treating extracts with streptomycin sulphate, frationating proteins (NH4)2SO4 at 45-75% of saturation and chromatography on blue sepharose CL-6B. The use of gel filtration through Sephadex G-150 and chromatography on DEAE-cellulose proved to be less effective for the enzyme purification. It has been established that it is 2,5-diamino-4-oxy-6-ribosylaminopyrimidine-5-phosphate but not its dephosphorylated form that is the substrate of the given reductase; Km is equal to 7.10(-5) M. The reaction proceeds in the presence of NADPH or NADH. The enzyme affinity to NADPH (Km = 4.7.10(-5) M) is approximately one order higher than that to NADPH (Km = 5.5.10(-4) M). The enzyme manifests the optimum of action at pH 7.2 and the temperature of 37 degrees C; the molecular weight is 140 kD. EDTA as well as flavins in the concentration of 1.10(-3) M exert no effect on the reductase activity. The enzyme is labile at 4 degrees C and is inactivated in the frozen state at -15 degrees C. The 2.5-diamino-4-oxy-6-ribosylaminopyrimidine-5'-phosphate reductase has been also revealed in Torulopsis candida, Debaryomyces klöckeri, Schwanniomyces occidentalis, Eremothecium ashbyii (flavinogenic species) and Candida utilis. Aspergillus nidulans, Neurospora crassa (nonflavinogenic species). The synthesis of this enzyme contrary to other enzymes of the riboflavin biosynthesis is not regulated in flavinogenic yeast by iron ions.

  17. Novel transporters from Kluyveromyces marxianus and Pichia guilliermondii expressed in Saccharomyces cerevisiae enable growth on L-arabinose and D-xylose.

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    Knoshaug, Eric P; Vidgren, Virve; Magalhães, Frederico; Jarvis, Eric E; Franden, Mary Ann; Zhang, Min; Singh, Arjun

    2015-10-01

    Genes encoding L-arabinose transporters in Kluyveromyces marxianus and Pichia guilliermondii were identified by functional complementation of Saccharomyces cerevisiae whose growth on L-arabinose was dependent on a functioning L-arabinose transporter, or by screening a differential display library, respectively. These transporters also transport D-xylose and were designated KmAXT1 (arabinose-xylose transporter) and PgAXT1, respectively. Transport assays using L-arabinose showed that KmAxt1p has K(m) 263 mM and V(max) 57 nM/mg/min, and PgAxt1p has K(m) 0.13 mM and V(max) 18 nM/mg/min. Glucose, galactose and xylose significantly inhibit L-arabinose transport by both transporters. Transport assays using D-xylose showed that KmAxt1p has K(m) 27 mM and V(max) 3.8 nM/mg/min, and PgAxt1p has K(m) 65 mM and V(max) 8.7 nM/mg/min. Neither transporter is capable of recovering growth on glucose or galactose in a S. cerevisiae strain deleted for hexose and galactose transporters. Transport kinetics of S. cerevisiae Gal2p showed K(m) 371 mM and V(max) 341 nM/mg/min for L-arabinose, and K(m) 25 mM and V(max) 76 nM/mg/min for galactose. Due to the ability of Gal2p and these two newly characterized transporters to transport both L-arabinose and D-xylose, one scenario for the complete usage of biomass-derived pentose sugars would require only the low-affinity, high-throughput transporter Gal2p and one additional high-affinity general pentose transporter, rather than dedicated D-xylose or L-arabinose transporters. Additionally, alignment of these transporters with other characterized pentose transporters provides potential targets for substrate recognition engineering.

  18. Condition Optimization of Lipase Production from Pichia guilliermondii ypy17%季也蒙毕赤酵母ypy17产脂肪酶条件的优化

    Institute of Scientific and Technical Information of China (English)

    李晓宇; 骆焱平; 柳志强; 邱波

    2011-01-01

    A lipase producing yeast Pichia guilliermonda ypyl7 was isolated from the coastal water of Yangpu in Hainan. Plackett-Burman design and response surface methodology (RSM) were used to optimize the production conditions of lipase. The results showed that olive oil, initial pH and K2HPO4 were the most significant factors influencing lipase production. The optimized composition of medium was olive oil 31.57 g/L, (NH4)2SO4 10.00 g/L, NaCl 30.00 g/L, K2HPO4 1.16 g/L, initial pH 7.30. After optimization, the lipase activity was 18.20 U/mL.%从海南洋浦近海岸分离到1株产脂肪酶的酵母菌株,经鉴定为季也蒙毕赤酵母(Pichia guilliermondii),利用Plackett-Burman设计对发酵条件进行了筛选.结果表明,橄榄油、初始pH值和K2HPO4浓度对脂肪酶生产具有显著的影响;利用响应面法对3个因子进行了优化,获得了最佳发酵条件为橄榄油31.57 g/L,硫酸铵10.00g/L,NaCl 30.00 g/L,K2HPO4 1.16 g/L,初始pH值7.30,28℃培养72 h,脂肪酶活力达到18.20 U/mL.

  19. Occurrence and Diversity of Pichia spp. in Marine Environments

    Institute of Scientific and Technical Information of China (English)

    LI Jing; CHI Zhenming; WANG Xianghong; WANG Lin; SHENG Jun; GONG Fang

    2008-01-01

    A total of 328 yeast strains from seawater, sediments, mud of saltems, the guts of marine fish and marine algae were obtained. The results of routine identification and molecular methods show that five yeast strains obtained in this study belonged to Pichia spp., including Pichia guilliermondii luv-small, Pichia ohmeri YF04d, Pichiafermentans YF12b, Pichia burtonii YF11A and Pichia anomala YF07b. Further studies revealed that Pichia anomala YF07b could produce killer toxin against pathogenic yeasts in crabs while Pichia guilliermondii luv-small could produce high activity of extracellular inulinase. It is advisable to test if Pichia ohmeri YF04d obtained in this study is related to central-venous-catheter-associated infection.

  20. 21 CFR 173.160 - Candida guilliermondii.

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    2010-04-01

    ... CONSUMPTION Enzyme Preparations and Microorganisms § 173.160 Candida guilliermondii. The food additive Candida... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Candida guilliermondii. 173.160 Section 173.160 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED)...

  1. Candida guilliermondii as the aetiology of candidosis Candida guilliermondii como agente de candidose

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    Alessandro Comarú Pasqualotto

    2006-06-01

    Full Text Available Candida guilliermondii is one of the components of human microbiota. This yeast has been infrequently associated with human infections, which may be related to its low pathogenicity. The aim of this study was to provide clinical and epidemiological data for patients infected with C. guilliermondii at Santa Casa Complexo Hospitalar, Brazil. From October 1997 to October 2003, C. guilliermondii was isolated from clinical samples from 11 patients. Three patients were excluded because the isolation of the yeast represented colonisation. Specimens from the eight patients included in the study corresponded to blood (n = 5, ascitis fluid (n = 2, and oesophagus biopsy (n = 1. Three patients (37.5% had major immunosuppressed conditions, including solid organ transplantation, AIDS, and leukaemia. Previous use of antibiotics occurred in 87.5%. Main invasive medical procedures were central venous catheter (50.0%, abdominal surgery (25.0%, and peritoneal dialysis (50.0%. No susceptibility data was obtained. Although risk factors for candidaemia were similar amongst patients infected by with C. guilliermondii or other Candida species, mortality associated with C. guilliermondii was significantly lower.Candida guilliermondii é um dos componentes da microbiota humana e infecções associadas com esta levedura têm sido incomuns, o que pode ser atribuído a sua baixa patogenicidade. O objetivo deste trabalho foi documentar aspectos clínico-epidemiológicos em pacientes que tiveram C. guilliermondii isolada a partir de amostras biológicas. O estudo foi conduzido na Santa Casa Complexo Hospitalar, Brasil. Durante outubro de 1997 e outubro de 2003, C. guilliermondii foi isolada de 11 pacientes, três dos quais foram excluídos por se apresentarem apenas colonizados. Espécimes clínicos corresponderam a sangue (n = 5, líquido de ascite (n = 2 e biópsia de esôfago (n = 1. Três pacientes eram imunodeprimidos, incluindo transplante de órgãos sólidos, SIDA e

  2. Effects of indigenous yeasts on physicochemical and microbial properties of Korean soy sauce prepared by low-salt fermentation.

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    Song, Young-Ran; Jeong, Do-Youn; Baik, Sang-Ho

    2015-10-01

    This study deals with understanding the effects of salt reduction on both the physicochemical and microbiological properties of soy sauce fermentation and also the application of indigenous yeast starters to compensate for undesirable changes occurring in salt-reduced processes. Fermentation was tested in situ at a Korean commercial soy sauce processing unit. Salt reduction resulted in higher acidity as well as lower pH and contents of residual sugar and ethanol. Moreover, undesired flavor characteristics, due to a lack of distinctive compounds, was observed. In addition, putrefactive Staphylococcus and Enterococcus spp. were present only during salt-reduced fermentation. To control these adverse effects, a single or mixed culture of two indigenous yeasts, Torulaspora delbrueckii and Pichia guilliermondii, producing high ethanol and 3-methyl-1-butanol, respectively, were tested. Overall, all types of yeast applications inhibited undesirable bacterial growth despite salt reduction. Of the starter cultures tested, the mixed culture resulted in a balance of more complex and richer flavors with an identical flavor profile pattern to that obtained from high salt soy sauce. Hence, this strategy using functional yeast cultures offers a technological option to manufacture salt-reduced soy sauce while preserving its typical sensory characteristics without affecting safety.

  3. Growth of Candida guilliermondii FTI 20037 on mixed substrate

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    Patrick V. Gurgel

    1998-01-01

    Full Text Available Candida guilliermondii FTI 20037 was grown on a mixed substrate comprising glucose and xylose. Inocula were grown using xylose or glucose as carbon source. Results showed that xylose utilization was delayed until glucose was utilized. Inoculum prepared on glucose showed a lag phase in xylose consumption. Cell mass production was higher when glucose was utilized during fermentation.

  4. [Comparative study of the protein makeup in diploid and haploid forms of Saccharomyces and Pichia].

    Science.gov (United States)

    Temina, A V; Tolstorukov, I I; Korogodin, V I; Gololobov, A D

    1979-01-01

    The rates of growth, biomass accumulation, and electrophoretic spectra of mobile cytoplasmic proteins were studied with nonisogenous haploid and diploid cultures of Saccharomyces cerevisiae and Pichia guilliermondii as well as with isogenous haploid-diploid pairs of Saccharomyces cerevisiae and Pichia pinus. On a mineral medium with glucose, differences in these parameters in various yeast strains were found to be due to the genotype of a strain rather than to ploidy: nonisogenous haploid and diploid cultures displayed considerable and random variability of these properties while no differences were found in isogenous haploid-diploid pairs. Studies on solubility of protein fractions in various solvents made it possible to reveal differences connected with ploidy, namely: both in nonisogenous and isogenous haploid-diploid systems, the content of the water-soluble fraction decreased in diploid cultures.

  5. Microbiological and physicochemical characterisation of caxiri, an alcoholic beverage produced by the indigenous Juruna people of Brazil.

    Science.gov (United States)

    Santos, Claudia Cristina Auler do Amaral; Almeida, Euziclei Gonzaga de; Melo, Gilberto Vinícius Pereira de; Schwan, Rosane Freitas

    2012-05-15

    Caxiri is a traditional fermented alcoholic beverage produced from cassava and sweet potatoes by the indigenous Juruna or Yudjá people in Brazil. Our results showed that caxiri fermentation is invariably associated with the following: (i) an increase in the total microbial population, with yeast being the largest group detected; (ii) a decrease in reducing sugars, malic, tartaric, succinic, oxalic and propionic acid; and (iii) a final product characterised by a high content of ethanol and a high concentration of lactic acid. The microbial community dynamics were investigated by culture-based and culture-independent approaches. Fermentation was assisted by a complex microbial community that changed in structure and composition during the fermentative process. The bacterial population ranged from 3.05 to 5.33 log/mL, and the yeast population varied from 3.27 log CFU/mL to 7.34 log CFU/mL, showing that yeasts dominated the fermentation process after 48 h. A total of 343 colonies of bacteria and 205 colonies of yeasts were isolated and initially grouped by Amplified Ribosomal DNA Restriction Analysis (ARDRA) and by biochemical features. Phylogenetic analysis of the 16S rRNA gene sequences of representative isolates showed that the bacteria were mainly represented by endospore-forming low-G+C content Gram-positive bacilli (Bacillus spp.; 61.5% of the isolates), with Bacillus pumilus, Bacillus spp. (Bacillus cereus group), and Bacillus subtilis being the main species identified. The species Sphingomonas sp. and Pediococcus acidilactici were also found. The dominant yeast identified was Saccharomyces cerevisiae. Rhodotorula mucilaginosa, Pichia membranifaciens, Pichia guilliermondii and Cryptococcus luteolus were also found. According to the Polymerase Chain Reaction and Denaturing Gradient Gel Electrophoresis (PCR-DGGE) analysis, the microbial communities present during fermentation were probably from the raw materials, ambient or present on the utensils used during

  6. Production and characterization of an extracellular lipase from Candida guilliermondii

    Directory of Open Access Journals (Sweden)

    Anne Caroline Defranceschi Oliveira

    2014-12-01

    Full Text Available Extracellular lipases from the endophytic yeast Candida guilliermondii isolated from castor leaves (Ricinus communis L. were produced using low-cost raw materials such as agro-industrial residues and applying them in the esterification of oleic acid for evaluating their potential use in biodiesel production. After partial purification using ammonium sulfate, the enzyme was characterized and presented higher activity (26.8 ± 1.5 U mL-1 in the presence of 5 mmol L-1 NaCl at 30 ºC and pH 6.5. The production through submerged fermentation was formerly performed in 150 mL erlenmeyer flasks and, once the enzyme production was verified, assays in a 14 L bioreactor were conducted, obtaining 18 ± 1.4 U mL-1. The produced enzyme was applied in the oleic acid esterification under different solvents: hexane, cyclohexane or cyclohexanone and different acid:alcohol molar ratios. Higher ester conversion rate (81% was obtained using hexane and the molar ratio of 1:9 was the best conditions using methanol. The results suggest the potential for development of endophytic yeast in the production of biocatalyst through submerged fermentation using agroindustrial residues as culture medium.

  7. Enzymatic degradation of nitriles by a Candida guilliermondii UFMG-Y65.

    Science.gov (United States)

    Dias, J C; Rezende, R P; Rosa, C A; Lachance, M A; Linardi, V R

    2000-06-01

    Candida guilliermondii UFMG-Y65, isolated from a gold mine, was able to utilize different nitriles and the corresponding amides as sole source of nitrogen, at concentrations up to 2 M. Resting cells cultivated on YCB-acetonitrile medium showed nitrile hydrolyzing enzyme activities against acrylonitrile and benzonitrile. These enzymes were inducible and intracellular; the optimum pH was 7.0-8.0, and the optimum temperature 25 degrees C-30 degrees C. Liquid chromatographic analysis indicated that C. guilliermondii UFMG-Y65 metabolized 12 mM benzonitrile to 11 mM benzoic acid and 10 mM acrylonitrile to 7.9 mM acrylic acid. The results suggest that C. guilliermondii UFMG-Y65 may be useful for the bioproduction of amides and acids, and for the bioremediation of environments contaminated with nitriles.

  8. Disruption of Protein Mannosylation Affects Candida guilliermondii Cell Wall, Immune Sensing, and Virulence

    Science.gov (United States)

    Navarro-Arias, María J.; Defosse, Tatiana A.; Dementhon, Karine; Csonka, Katalin; Mellado-Mojica, Erika; Dias Valério, Aline; González-Hernández, Roberto J.; Courdavault, Vincent; Clastre, Marc; Hernández, Nahúm V.; Pérez-García, Luis A.; Singh, Dhirendra K.; Vizler, Csaba; Gácser, Attila; Almeida, Ricardo S.; Noël, Thierry; López, Mercedes G.; Papon, Nicolas; Mora-Montes, Héctor M.

    2016-01-01

    The fungal cell wall contains glycoproteins that interact with the host immune system. In the prominent pathogenic yeast Candida albicans, Pmr1 acts as a Golgi-resident ion pump that provides cofactors to mannosyltransferases, regulating the synthesis of mannans attached to glycoproteins. To gain insight into a putative conservation of such a crucial process within opportunistic yeasts, we were particularly interested in studying the role of the PMR1 homolog in a low-virulent species that rarely causes candidiasis, Candida guilliermondii. We disrupted C. guilliermondii PMR1 and found that loss of Pmr1 affected cell growth and morphology, biofilm formation, susceptibility to cell wall perturbing agents, mannan levels, and the wall composition and organization. Despite the significant increment in the amount of β1,3-glucan exposed at the wall surface, this positively influenced only the ability of the mutant to stimulate IL-10 production by human monocytes, suggesting that recognition of both mannan and β1,3-glucan, is required to stimulate strong levels of pro-inflammatory cytokines. Accordingly, our results indicate C. guilliermondii sensing by monocytes was critically dependent on the recognition of N-linked mannans and β1,3-glucan, as reported in other Candida species. In addition, chemical remotion of cell wall O-linked mannans was found to positively influence the recognition of C. guilliermondii by human monocytes, suggesting that O-linked mannans mask other cell wall components from immune cells. This observation contrasts with that reported in C. albicans. Finally, mice infected with C. guilliermondii pmr1Δ null mutant cells had significantly lower fungal burdens compared to animals challenged with the parental strain. Accordingly, the null mutant showed inability to kill larvae in the Galleria mellonella infection model. This study thus demonstrates that mannans are relevant for the C. guilliermondii-host interaction, with an atypical role for O

  9. Disruption of Protein Mannosylation Affects Candida guilliermondii Cell Wall, Immune Sensing, and Virulence

    Directory of Open Access Journals (Sweden)

    María J. Navarro-Arias

    2016-12-01

    Full Text Available The fungal cell wall contains glycoproteins that interact with the host immune system. In the prominent pathogenic yeast Candida albicans, Pmr1 acts as a Golgi-resident ion pump that provides cofactors to mannosyltransferases, regulating the synthesis of mannans attached to glycoproteins. To gain insight into a putative conservation of such a crucial process within opportunistic yeasts, we were particularly interested in studying the role of the PMR1 homolog in a low-virulent species that rarely causes candidiasis, Candida guilliermondii. We disrupted C. guilliermondii PMR1 and found that loss of Pmr1 affected cell growth and morphology, biofilm formation, susceptibility to cell wall perturbing agents, mannan levels, and the wall composition and organization. Despite there was a significant increment in the amount of β1,3-glucan exposed at the wall surface, this positively influenced only the ability of the mutant to stimulate IL-10 production by human monocytes, suggesting that recognition of both mannan and β1,3-glucan, is required to stimulate strong levels of pro-inflammatory cytokines. Accordingly, our results indicate C. guilliermondii sensing by monocytes was critically dependent on the recognition of N-linked mannans and β1,3-glucan, as reported in other Candida species. In addition, chemical remotion of cell wall O-linked mannans was found to positively influence the recognition of C. guilliermondii by human monocytes, suggesting that O-linked mannans mask other cell wall components from immune cells. This observation contrasts with that reported in C. albicans. Finally, mice infected with C. guilliermondii pmr1 null mutant cells had significantly lower fungal burdens compared to animals challenged with the parental strain. Accordingly, the null mutant showed inability to kill larvae in the Galleria mellonella infection model. This study thus demonstrates that mannans are relevant for the C. guilliermondii-host interaction, with

  10. Indigenous homelessness

    DEFF Research Database (Denmark)

    attempts at assimilation that disrupted Indigenous practices, languages, and cultures—including patterns of housing and land use—can be seen today in the disproportionate number of Indigenous people affected by homelessness in both rural and urban settings. Essays in this collection explore the meaning......Being homeless in one’s homeland is a colonial legacy for many Indigenous people in settler societies. The construction of Commonwealth nation-states from colonial settler societies depended on the dispossession of Indigenous peoples from their lands. The legacy of that dispossession and related...... and scope of Indigenous homelessness in the Canada, Australia, and New Zealand. They argue that effective policy and support programs aimed at relieving Indigenous homelessness must be rooted in Indigenous conceptions of home, land, and kinship, and cannot ignore the context of systemic inequality...

  11. Biodegradation Mechanisms of Patulin in Candida guilliermondii: An iTRAQ-Based Proteomic Analysis

    Directory of Open Access Journals (Sweden)

    Yong Chen

    2017-02-01

    Full Text Available Patulin, a potent mycotoxin, contaminates fruits and derived products worldwide, and is a serious health concern. Several yeast strains have shown the ability to effectively degrade patulin. However, the mechanisms of its biodegradation still remain unclear at this time. In the present study, biodegradation and involved mechanisms of patulin by an antagonistic yeast Candida guilliermondii were investigated. The results indicated that C. guilliermondii was capable of not only multiplying to a high population in medium containing patulin, but also effectively reducing patulin content in culture medium. Degradation of patulin by C. guilliermondii was dependent on the yeast cell viability, and mainly occurred inside cells. E-ascladiol was the main degradation product of patulin. An iTRAQ-based proteomic analysis revealed that the responses of C. guilliermondii to patulin were complex. A total of 30 differential proteins involved in 10 biological processes were identified, and more than two-thirds of the differential proteins were down-accumulated. Notably, a short-chain dehydrogenase (gi|190348612 was markedly induced by patulin at both the protein and mRNA levels. Our findings will provide a foundation to help enable the commercial development of an enzyme formulation for the detoxification of patulin in fruit-derived products.

  12. Biodegradation Mechanisms of Patulin in Candida guilliermondii: An iTRAQ-Based Proteomic Analysis

    Science.gov (United States)

    Chen, Yong; Peng, Huai-Min; Wang, Xiao; Li, Bo-Qiang; Long, Man-Yuan; Tian, Shi-Ping

    2017-01-01

    Patulin, a potent mycotoxin, contaminates fruits and derived products worldwide, and is a serious health concern. Several yeast strains have shown the ability to effectively degrade patulin. However, the mechanisms of its biodegradation still remain unclear at this time. In the present study, biodegradation and involved mechanisms of patulin by an antagonistic yeast Candida guilliermondii were investigated. The results indicated that C. guilliermondii was capable of not only multiplying to a high population in medium containing patulin, but also effectively reducing patulin content in culture medium. Degradation of patulin by C. guilliermondii was dependent on the yeast cell viability, and mainly occurred inside cells. E-ascladiol was the main degradation product of patulin. An iTRAQ-based proteomic analysis revealed that the responses of C. guilliermondii to patulin were complex. A total of 30 differential proteins involved in 10 biological processes were identified, and more than two-thirds of the differential proteins were down-accumulated. Notably, a short-chain dehydrogenase (gi|190348612) was markedly induced by patulin at both the protein and mRNA levels. Our findings will provide a foundation to help enable the commercial development of an enzyme formulation for the detoxification of patulin in fruit-derived products. PMID:28208714

  13. Development of an affordable typing method for Meyerozyma guilliermondii using microsatellite markers.

    Science.gov (United States)

    Wrent, Petra; Rivas, Eva-María; Peinado, José M; de Silóniz, María-Isabel

    2016-01-18

    Despite previously published methods, there is still a lack of rapid and affordable methods for genotyping the Meyerozyma guilliermondii yeast species. The development of microsatellite markers is a useful genotyping method in several yeast species. Using the Tandem Repeat Finder Software, a total of 19 microsatellite motifs (di-, tri-, and tetra- repetition) were found in silico in seven of the nine scaffolds published so far. Primer pairs were designed for all of them, although only four were used in this work. All microsatellite amplifications showed size polymorphism, and the results were identical when repeated. The combination of three microsatellite markers (sc15F/R, sc32 F/R and sc72 F/R) produced a different pattern for each of the Type Culture Collection strains of M. guilliermondii used to optimize the method. The three primer pairs can be used in the same PCR reaction, which reduces costs, in tandem with the fluorescent labeling of only the forward primer in each primer pair. Microsatellite typing was applied on 40 more M. guilliermondii strains. The results showed that no pattern is repeated between the different environmental niches. Four M. guilliermondii strains were only amplified with primer pair sc32 F/R, and subsequently identified as Meyerozyma caribbica by Taq I-RFLP of the 5.8S ITS rDNA. Most out-group species gave negative results even for physiologically similarly species such as Debaryomyces hansenii. The microsatellite markers used in this work were stable over time, which enables their use as a traceability tool.

  14. Phytase-producing capacity of yeasts isolated from traditional African fermented food products and PHYPk gene expression of Pichia kudriavzevii strains.

    Science.gov (United States)

    Greppi, Anna; Krych, Łukasz; Costantini, Antonella; Rantsiou, Kalliopi; Hounhouigan, D Joseph; Arneborg, Nils; Cocolin, Luca; Jespersen, Lene

    2015-07-16

    Phytate is known as a strong chelate of minerals causing their reduced uptake by the human intestine. Ninety-three yeast isolates from traditional African fermented food products, belonging to nine species (Pichia kudriavzevii, Saccharomyces cerevisiae, Clavispora lusitaniae, Kluyveromyces marxianus, Millerozyma farinosa, Candida glabrata, Wickerhamomyces anomalus, Hanseniaspora guilliermondii and Debaryomyces nepalensis) were screened for phytase production on solid and liquid media. 95% were able to grow in the presence of phytate as sole phosphate source, P. kudriavzevii being the best growing species. A phytase coding gene of P. kudriavzevii (PHYPk) was identified and its expression was studied during growth by RT-qPCR. The expression level of PHYPk was significantly higher in phytate-medium, compared to phosphate-medium. In phytate-medium expression was seen in the lag phase. Significant differences in gene expression were detected among the strains as well as between the media. A correlation was found between the PHYPk expression and phytase extracellular activity.

  15. Candida guilliermondii Complex Is Characterized by High Antifungal Resistance but Low Mortality in 22 Cases of Candidemia.

    Science.gov (United States)

    Marcos-Zambrano, Laura Judith; Puig-Asensio, Mireia; Pérez-García, Felipe; Escribano, Pilar; Sánchez-Carrillo, Carlos; Zaragoza, Oscar; Padilla, Belén; Cuenca-Estrella, Manuel; Almirante, Benito; Martín-Gómez, M Teresa; Muñoz, Patricia; Bouza, Emilio; Guinea, Jesús

    2017-07-01

    The objectives of our study were to describe the characteristics of patients with Candida guilliermondii candidemia and to perform an in-depth microbiological characterization of isolates and compare them with those of patients with C. albicans candidemia. We described the risk factors and outcomes of 22 patients with candidemia caused by the C. guilliermondii complex. Incident isolates were identified using molecular techniques, and susceptibility to fluconazole, anidulafungin, and micafungin was studied. Biofilm formation was measured using the crystal violet assay (biomass production) and the XTT reduction assay (metabolic activity), and virulence was studied using the Galleria mellonella model. Biofilm formation was compared with that observed for C. albicans The main conditions predisposing to infection were malignancy (68%), immunosuppressive therapy (59%), and neutropenia (18%). Clinical presentation of candidemia was less severe in patients infected by the C. guilliermondii complex than in patients infected by C. albicans, and 30-day mortality was lower in C. guilliermondii patients (13.6% versus 33.9%, respectively; P = 0.049). Isolates were identified as C. guilliermondiisensu stricto (n = 17) and Candida fermentati (n = 5). The isolates produced biofilms with low metabolic activity and moderate biomass. The G. mellonella model showed that C. guilliermondii was less virulent than C. albicans (mean of 6 days versus 1 day of survival, respectively; P candidemia caused by the C. guilliermondii complex had severe and debilitating underlying conditions. Overall, the isolates showed diminished susceptibility to fluconazole and echinocandins, although poor biofilm formation and the low virulence were associated with a favorable outcome. Copyright © 2017 American Society for Microbiology.

  16. Indigenous religions

    DEFF Research Database (Denmark)

    Geertz, Armin W.

    2009-01-01

    Dette essay diskuterer en publikation af James L. Cox med titlen From Primitive to Indigenous (2007). Bogen analyserer forskellige forfatteres holdninger til studiet af indfødte kulturers religioner. Cox's analyser tages op i dette essay og de problematiseres i forhold til mit eget arbejde....

  17. Primary Candida guilliermondii Infection of the Knee in a Patient without Predisposing Factors

    Directory of Open Access Journals (Sweden)

    Gun Woo Lee

    2012-01-01

    Full Text Available Isolated primary candidal infection of joint is extremely rare, with only a few reported cases. It occurs as a result of accidental implantations of fungus during traumatic procedures, such as surgery, and is usually reported in patients with predisposing factors such as immunosuppression, malignancy, and drug abuse. If left untreated, irreversible deformity and pain with severe osteoarticular destruction occur. Thus, early diagnosis and treatment are important. This paper presents a case of 72-year-old man with primary C. guilliermondii infection of knee joint without predisposing factors and previous traumatic procedures, who was misdiagnosed with advanced degenerative osteoarthritis. Our case is the second case of primary C. guilliermondii arthritis of knee to be reported in the English-language literature and the first to be successfully treated with total knee arthroplasty following IV amphotericin B and oral fluconazole. Primary candidal infection of joint is generally asymptomatic or involves only mild pain and swelling in the affected knee. Thus, although the majority of knee joint infections are of a pyogenic or tuberculous origin, if a patient complains of mild pain and swelling in the knee and has mild signs of infection, the possibility of fungal infection should be considered.

  18. AKTIVITAS INULINASE OLEH Pichia manshurica DAN FUSAN F4 PADA FERMENTASI BATCH DENGAN UMBI DAHLIA (Dahlia sp SEBAGAI SUBSTRAT

    Directory of Open Access Journals (Sweden)

    Wijanarka Wijanarka

    2013-11-01

    Full Text Available ACTIVITY OF INULINASE OF Pichia Manshuria AND FUSAN F4 ON BATCH FERMENTATION UDING DAHLIA TUBER (Dahlia sp AS A SUBSTRATE. A dahlia tuber is one of the common inulin rich crops. Inulin is formed by units of fructans, which are polymers of D-fructose. Inulinases (EC 3.2.1.7 catalyze the hydrolysis of inulin, producing fructooligosaccharides (FOS, inulooligosaccharides (IOS, pulullan, acetone, butanol and sorbitol, therefore dahlia tubers are used as growth media. The inulin hydrolyzing activity has been reported from various microbial strains Pichia manshurica and Fusan F4 which is the result of fusion protoplast. The objective of this study was to determine the activity of inulinase Pichia manshurica and Fusan F4 on the substrate dahlia tubers. Fusan F4 to increase inulinase activity compared with Pichia manshurica and to investigate the kinetics of specific growth rate (μ and time double (g from of Pichia manshurica and Fusan F4. The results showed that the exponential phase occurs at 0-12 hour without a lag phase. P. manshurica has a specific growth rate (μ of 0.18/hour with time double (g 3.90 hours and the inulinase enzyme activity of 0.56 IU, while for Fusan F4 consecutive has a value μ of 0.20/hour, g of 3.49 hours and the activity of 0.69 IU. The conclusion of this research is to improve Fusan F4 inulinase activity and the ability has to be better than the Pichia manshurica.Umbi dahlia merupakan salah satu umbi yang mengandung inulin. Inulin merupakan polimer fruktan yang dapat dipecah oleh enzim inulinase (E.C. 3.2.1.7 menjadi fruktosa. Fruktosa merupakan bahan baku dasar untuk pembuatan FOS, IOS, pulullan, aseton dan sorbitol, oleh karena itu umbi dahlia digunakan sebagai media pertumbuhan. Enzim inulinase ini secara indigenous dimiliki oleh Pichia manshurica dan Fusan F4 yang merupakan hasil fusi protoplas.Tujuan  penelitian ini adalah  untuk mengetahui aktivitas inulinase Pichia manshurica dan Fusan F4 pada substrat umbi dahlia

  19. Pichia stipitis genomics, transcriptomics, and gene clusters

    Science.gov (United States)

    Thomas W. Jeffries; Jennifer R. Headman Van Vleet

    2009-01-01

    Genome sequencing and subsequent global gene expression studies have advanced our understanding of the lignocellulose-fermenting yeast Pichia stipitis. These studies have provided an insight into its central carbon metabolism, and analysis of its genome has revealed numerous functional gene clusters and tandem repeats. Specialized physiological traits are often the...

  20. Improvement of biotechnological xylitol production by glucose during cultive of Candida guilliermondii in sugarcane bagasse hydrolysate

    Directory of Open Access Journals (Sweden)

    Débora Danielle Virgínio da Silva

    2007-03-01

    Full Text Available The effect of glucose on xylose-to-xylitol bioconversion by Candida guilliermondii was examined by adding it to sugarcane bagasse hydrolysate medium to obtain different glucose:xylose ratios (1:25, 1:12, 1:5 and 1:2.5. Under experimental conditions, increasing glucose:xylose ratio improved the assimilation of the xylose present in the hydrolysate by yeast, resulting in biomass increase, and in the formation of xylitol and glycerol/ethanol by-products. Maximum values of xylitol yield (0.59 g g-1 and volumetric productivity (0.53 g l-1.h-1 were obtained with glucose:xylose ratio of 1:5, resulting in the higher conversion efficiency (64.3%.O efeito da glicose na bioconversão de xilose em xilitol por Candida guilliermondii foi avaliado em hidrolisado hemicelulósico de bagaço de cana com diferentes relações glicose:xilose (1:25, 1:12, 1:5 and 1:2,5. Sob as condições experimentais, o aumento da relação glicose:xilose favoreceu a assimilação da xilose presente no hidrolisado, resultando em aumento da biomassa celular e aumento da formação de xilitol e dos sub-produtos glicerol e etanol. Os valores máximos do fator de conversão de xilose em xilitol (0,59 g g-1 e da produtividade volumétrica de xilitol (0,53 g l-1.h-1 foram obtidos com a relação glicose:xilose 1:5, resultando na maior eficiência de conversão (64,3%.

  1. PRODUCCIÓN DE XILITOL A PARTIR DE CASCARILLA DE ARROZ UTILIZANDO Candida guilliermondii XYLITOL PRODUCTION FROM RICE HUSK USING Candida guilliermondii

    Directory of Open Access Journals (Sweden)

    Marcela Villalba Cadavid

    2009-06-01

    Full Text Available En este estudio se empleó cascarilla de arroz como materia prima para la obtención de xilitol, previa hidrólisis durante 60 minutos con ácido sulfúrico al 4% p/v; a 121 ºC y 15 psig,; La xilosa producida se transformó en xilitol mediante su fermentación con Candida guilliermondii. Se estudió el efecto de las variables concentración inicial de xilosa, concentración de inóculo y relación volumen del medio/volumen del matraz, así como sus efectos combinados, sobre la producción de xilitol. Se encontró que las concentraciones iniciales de xilosa e inóculo más adecuadas, entre los valores ensayados, fueron 80 y 5,0 g/l, respectivamente. En estas condiciones, la concentración final de xilitol obtenida fue de 45,2 g/l, con una productividad volumétrica de 0,23 g/l•h y un rendimiento de 0,57 g/g.In this study was used rice husk, previosly hydrolyzed with diluted sulfuric acid at 121 ºC and 15 psig, with a residence time 60 min. The initial concentration of substrate, inoculum, and relationship between media volume/flask volume and their combined effects were studied on the production of xylitol. The initial concentrations of 80 g/l xylose and 5 g/l inoculums led the best xylitol production (45.2 g/l, productivity (0.23 g/l•h and yield (0.57 g/g.

  2. Reclassification of Candida guilliermondii FTI 20037 as Candida tropicalis based on molecular phylogenetic analysis Reclassificação de Candida guilliermondii FTI 20037 como Candida tropicalis baseada na análise filogenética molecular

    Directory of Open Access Journals (Sweden)

    Luanne Helena Augusto Lima

    2003-11-01

    Full Text Available Yeasts of the genus Candida are of clinical importance and also have many industrial applications, mainly in the food industry. The yeast Candida guilliermondii FTI 20037 has been extensively studied in order to establish a biotechnological process for the production of xylitol. The goal of this study was to verify the taxonomic classification of this strain based on the analysis of rDNA sequences and the xyl1 gene. DNA fragments from these sequences were amplified by PCR and BLAST analysis revealed strong identity with the corresponding sequences from Candida tropicalis. Based on these results, we propose that C. guilliermondii FTI 20037 must be reclassified as C. tropicalis.As leveduras do gênero Candida possuem tanto importância clínica como diversas aplicações industriais, principalmente na indústria de alimentos. A levedura Candida guilliermondii FTI 20037 tem sido exaustivamente estudada pois pretende-se utilizá-la no estabelecimento de um processo biotecnológico para a produção de xilitol. O objetivo deste trabalho foi verificar a classificação taxonômica desta levedura por análise de sequências do rDNA e do gene xyl1. Fragmentos correspondentes a estas regiões foram amplificados por PCR e a análise destas sequências por BLAST revelou alta identidade com sequências correspondentes de Candida tropicalis. Estes resultados nos levam a propor que C. guilliermondii FTI 20037 deva ser reclassificada como C. tropicalis.

  3. Evaluation of nutrient supplementation to charcoal-treated and untreated rice straw hydrolysate for xylitol production by Candida guilliermondii

    Directory of Open Access Journals (Sweden)

    Solange Inês Mussatto

    2005-05-01

    Full Text Available Xylitol was produced by Candida guilliermondii from charcoal-treated and untreated rice straw hemicellulosic hydrolysate with or without nutrients (ammonium sulphate, calcium chloride, rice bran extract. Both, xylitol yield and volumetric productivity decreased significantly when the nutrients were added to treated and untreated hydrolysates. In the treated hydrolysate, the efficiency of xylose conversion to xylitol was 79% when the nutrients were omitted. The results demonstrated that rice straw hemicellulosic hydrolysate treated with activated charcoal was a cheap source of xylose and other nutrients for xylitol production by C. guilliermondii. The non-necessity of adding nutrients to the hydrolysate media would be very advantageous since the process becomes less costly.Este trabalho avaliou a produção de xilitol pela levedura Candida guilliermondii, a partir de hidrolisado hemicelulósico de palha de arroz não tratado e tratado com carvão ativo, ambos suplementados ou não com nutrientes (sulfato de amônio, cloreto de cálcio e extrato de farelo de arroz. Os resultados mostraram que tanto o rendimento como a produtividade volumétrica em xilitol diminuíram quando os nutrientes foram adicionados em ambos hidrolisados, tratado e não tratado. Em hidrolisado tratado, a eficiência de conversão de xilose em xilitol foi de 79% quando em ausência de nutrientes. Estes resultados mostram que o hidrolisado hemicelulósico de palha de arroz tratado com carvão ativo é uma fonte barata de xilose e outros nutrientes, para a produção de xilitol por Candida guilliermondii. A não necessidade de adicionar nutrientes ao meio a base de hidrolisado é muito vantajosa, uma vez que o processo se torna mais econômico.

  4. Effect of Furfural, Vanillin and Syringaldehyde on Candida guilliermondii Growth and Xylitol Biosynthesis

    Science.gov (United States)

    Kelly, Christine; Jones, Opal; Barnhart, Christopher; Lajoie, Curtis

    Xylitol is a five-carbon sugar alcohol with established commercial use as an alternative sweetener and can be produced from hemicellulose hydrolysate. However, there are difficulties with microbiological growth and xylitol biosynthesis on hydrolysate because of the inhibitors formed from hydrolysis of hemicellulose. This research focused on the effect of furfural, vanillin, and syringaldehyde on growth of Candida guilliermondii and xylitol accumulation from xylose in a semi-synthetic medium in microwell plate and bioreactor cultivations. All three compounds reduced specific growth rate, increased lag time, and reduced xylitol production rate. In general, increasing concentration of inhibitor increased the severity of inhibition, except in the case of 0.5 g vanillin per liter, which resulted in a faster late batch phase growth rate and increased biomass yield. At concentrations of 1 g/1 or higher, furfural was the least inhibitory to growth, followed by syringaldehyde. Vanillin most severely reduced specific growth rate. All three inhibitors reduced xylitol production rate approximately to the same degree.

  5. 21 CFR 573.750 - Pichia pastoris dried yeast.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Pichia pastoris dried yeast. 573.750 Section 573.750 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS FOOD ADDITIVES PERMITTED IN FEED AND DRINKING WATER OF ANIMALS Food Additive Listing § 573.750 Pichia...

  6. Indigenous Education in Mexico: Indigenous Students' Voices

    Science.gov (United States)

    Despagne, Colette

    2013-01-01

    The purpose of this article is to investigate whether, despite a shift in political and educational discourses over the last decades that suggests that Indigenous cultures and languages are recognized, any real change has occurred in terms of Indigenous education in Mexico. It is possible that official bilingual intercultural education is still…

  7. Indigenous Education in Mexico: Indigenous Students' Voices

    Science.gov (United States)

    Despagne, Colette

    2013-01-01

    The purpose of this article is to investigate whether, despite a shift in political and educational discourses over the last decades that suggests that Indigenous cultures and languages are recognized, any real change has occurred in terms of Indigenous education in Mexico. It is possible that official bilingual intercultural education is still…

  8. Why Indigenous Nations Studies?

    Science.gov (United States)

    Porter, Robert; Yellow Bird, Michael

    2000-01-01

    The development of a new Indigenous Nations Studies program at the University of Kansas is described. Success depended on a critical mass of Indigenous and non-Indigenous faculty and students that had a sense of political and social justice and understood the need for institutional change. The biggest challenge was countering the entrenched…

  9. Indigenous Storytelling in Namibia

    DEFF Research Database (Denmark)

    Rodil, Kasper; Winschiers-Theophilus, Heike

    2016-01-01

    fairytales to outsiders with little relevance to the physical world, they are very functional and foundational for communities where storytelling is enacted. This paper debates concepts related to indigenous storytelling and its relevance to knowledge and learning for indigenous youths. In an attempt...... to understand indigenous youths’ own conception of storytelling the paper presents empirical data from a study with indigenous Khoisan children in Namibia. This is followed by a discussion of an effort of digitizing indigenous intangible cultural heritage in relation to technologies’ embodied bias...

  10. Sugarcane straw as a feedstock for xylitol production by Candida guilliermondii FTI 20037

    Directory of Open Access Journals (Sweden)

    Andrés Felipe Hernández-Pérez

    2016-06-01

    Full Text Available Abstract Sugarcane straw has become an available lignocellulosic biomass since the progressive introduction of the non-burning harvest in Brazil. Besides keeping this biomass in the field, it can be used as a feedstock in thermochemical or biochemical conversion processes. This makes feasible its incorporation in a biorefinery, whose economic profitability could be supported by integrated production of low-value biofuels and high-value chemicals, e.g., xylitol, which has important industrial and clinical applications. Herein, biotechnological production of xylitol is presented as a possible route for the valorization of sugarcane straw and its incorporation in a biorefinery. Nutritional supplementation of the sugarcane straw hemicellulosic hydrolyzate as a function of initial oxygen availability was studied in batch fermentation of Candida guilliermondii FTI 20037. The nutritional supplementation conditions evaluated were: no supplementation; supplementation with (NH42SO4, and full supplementation with (NH42SO4, rice bran extract and CaCl2·2H2O. Experiments were performed at pH 5.5, 30 °C, 200 rpm, for 48 h in 125 mL Erlenmeyer flasks containing either 25 or 50 mL of medium in order to vary initial oxygen availability. Without supplementation, complete consumption of glucose and partial consumption of xylose were observed. In this condition the maximum xylitol yield (0.67 g g-1 was obtained under reduced initial oxygen availability. Nutritional supplementation increased xylose consumption and xylitol production by up to 200% and 240%, respectively. The maximum xylitol volumetric productivity (0.34 g L-1 h-1 was reached at full supplementation and increased initial oxygen availability. The results demonstrated a combined effect of nutritional supplementation and initial oxygen availability on xylitol production from sugarcane straw hemicellulosic hydrolyzate.

  11. A Pan-Indigenous Vision of Indigenous Studies.

    Science.gov (United States)

    Masaquiza, Martina; B'alam, Pakal

    2000-01-01

    Identifies four groups with conflicting interests in indigenous studies programs and the nature of the conflicts. Proposes the formation of a pan-indigenous intellectual network. Describes the ideal indigenous studies program devoted to building a pan-indigenous infrastructure that indigenous nations would direct and use as a tool to strengthen…

  12. Utilización de Candida guilliermondii aislada del corozo chiquito (Bactris guineensis en la producción de xilitol

    Directory of Open Access Journals (Sweden)

    Irina Cristina Herazo Camaño

    2011-08-01

    Full Text Available Utilization of Candida guilliermondii isolated of corozo chiquito (Bactris guineensis IN the production of xylitol Resumen.  La levadura Candida guilliermondii es objeto de estudio debido a su capacidad de producir xilitol aprovechando compuestos hemicelulósicos ricos en xilosa, dado esto, la cepa Candida guilliermondii aislada del fruto del corozo chiquito (Bactris guineensis fue usada en este estudio con el fin de evaluar su capacidad para producir xilitol sobre un sustrato hidrolizado de cascarilla de arroz. El objetivo de este trabajo fue determinar los parámetros fermentativos como producción de xilitol, productividad volumétrica (Qp y rendimiento de sustrato en producto (Yp/s durante la fermentación con la cepa nativa Candida guilliermondii. Se emplearon 200 ml de medio de cultivo hidrolizado de cascarilla de arroz, el cual contenía una concentración de xilosa de 27,5 g/L. La fermentación se llevó a cabo bajo las siguientes condiciones: temperatura 30 ºC, pH del medio 5,8, agitación 120 rpm e inóculo adaptado de 3 g/L. Los resultados mostraron que después de 120 horas de fermentación se obtuvieron 2,6 g/L de xilitol con productividad volumétrica (Qp de 0,02 g/L-h y rendimiento de sustrato en producto (Yp/s de 0,13 g/g. De esta manera, la cepa nativa Candida guilliermondii, aislada del fruto de Corozo chiquito (Bactris guineensis, produjo xilitol bajo condiciones específicas de fermentación. Palabras clave: xilosa, Candida guilliermondii, fermentación, levadura.  Abstract. The yeast Candida guilliermondii has been studied due to its ability to produce xylitol in xylose-rich hemicellulosic compounds, Candida guilliermondii strain isolated from the fruit of Corozo chiquito (Bactris guineensis was used in this study to assess their ability to xylitol production on these substrates. The aim of this study was to determine the fermentation parameters such as xylitol production, volumetric productivity (Qp and yield of

  13. Untersuchungen zur Sphingolipid-Biosynthese in der Hefe Pichia ciferrii

    OpenAIRE

    Börgel, Daniel

    2008-01-01

    Im Rahmen der vorliegenden Dissertation sollte der Sphingolipid-Biosyntheseweg der Hefe Pichia ciferrii näher charakterisiert werden, um die Entwicklung einer fermentativen Route zur Sphingosin-Produktion zu ermöglichen. Darüber hinaus galt es patentierbare Selektionssysteme für diese Hefe zu etablieren. Durch Sequenzvergleiche mit nahe verwandten Hefen und das Ableiten degenerierter Primer wurden elf für die Sphingolipid-Biosynthese von Pichia ciferrii relevante Gene isoliert und sequenziert...

  14. Hansenula polymorpha (Pichia angusta): Biology and Applications

    Science.gov (United States)

    Kunze, Gotthard; Kang, Hyun Ah; Gellissen, Gerd

    Hansenula polymorpha (Pichia angusta) belongs to a limited number of methylotrophic yeast species. It is able to assimilate nitrate and can grow on a range of carbon sources. Furthermore, H. polymorpha is a thermo-tolerant microorganism with some strains growing at temperatures up to 50° C and more. These unusual characteristics render H. polymorpha attractive as a model organism to study the development and functions of peroxisomes and the biochemistry of nitrate assimilation. H. polymorpha provides an established platform for heterologous gene expression and is distinguished by an impressive track record as producer of recom-binant proteins that include commercially available pharmaceuticals like hepatitis B vaccine, insulin and the IFN α-2a

  15. Dynamics in bioprocess development for Pichia pastoris.

    Science.gov (United States)

    Spadiut, Oliver; Herwig, Christoph

    2014-01-01

    Pichia pastoris is a widely used host organism for the recombinant production of proteins. It is attracting increasing interest for the production of biopharmaceuticals, due to its capability of performing posttranslational modifications. Traditionally, production processes with P. pastoris describe fed-batch processes based on feed forward regimes with a constant specific growth rate. However, this strategy does not consider physiological changes of the organism, bearing the risk of overfeeding and thus harming the cells. Recently, we introduced the specific substrate uptake rate as a novel physiological parameter to design fed-batch strategies for P. pastoris. We showed that by doing dynamic batch experiments strain specific parameters, which are needed to set up respective feeding profiles, can be easily determined. Furthermore we proved that dynamics during feeding directly affects productivity and product purity. Here, we sum up our findings regarding dynamics in bioprocess development for P. pastoris.

  16. In vivo unnatural amino acid expression in the methylotrophic yeast Pichia pastoris

    Energy Technology Data Exchange (ETDEWEB)

    Young, Travis; Schultz, Peter G.

    2017-08-15

    The invention provides orthogonal translation systems for the production of polypeptides comprising unnatural amino acids in methylotrophic yeast such as Pichia pastoris. Methods for producing polypeptides comprising unnatural amino acids in methylotrophic yeast such as Pichia pastoris are also provided.

  17. In vivo unnatural amino acid expression in the methylotrophic yeast Pichia pastoris

    Energy Technology Data Exchange (ETDEWEB)

    Young, Travis [San Diego, CA; Schultz, Peter G [La Jolla, CA

    2014-02-11

    The invention provides orthogonal translation systems for the production of polypeptides comprising unnatural amino acids in methyltrophic yeast such as Pichia pastoris. Methods for producing polypeptides comprising unnatural amino acids in methyltrophic yeast such as Pichia pastoris are also provided.

  18. Indigenous Existentialism and the Body

    Directory of Open Access Journals (Sweden)

    Brendan Hokowhitu

    2011-04-01

    Full Text Available This article begins a discussion on indigenous existentialism. The theme developed as a result of engagement at the intersection between Indigenous Studies and Cultural Studies, and the realisation that cultural concepts often canonised within Indigenous Studies departments, such as tradition and authenticity (when exclusive, detract from the conception of indigenous culture as part of the immediate material reality of indigenous lives. In turn, when indigenous culture is too often defined only in relation to an imagined authentic past, indigenous existentialism is inhibited because indigenous people lack a conscious awareness of cultural immediacy. There is nothing more immediate than the body and, thus, I began to theorise indigenous existentialism through an analyses of the indigenous body, its genealogy, and its immediacy. To help me process this theorisation I engage with current Cultural Studies debates surrounding the analyses of the body. I conclude that an indigenous existentialism will recognise that the power of the body is still unknown.

  19. Antimicrobial peptides (AMPs) produced by Saccharomyces cerevisiae induce alterations in the intracellular pH, membrane permeability and culturability of Hanseniaspora guilliermondii cells.

    Science.gov (United States)

    Branco, Patrícia; Viana, Tiago; Albergaria, Helena; Arneborg, Nils

    2015-07-16

    Saccharomyces cerevisiae produces antimicrobial peptides (AMPs) during alcoholic fermentation that are active against several wine-related yeasts (e.g. Hanseniaspora guilliermondii) and bacteria (e.g. Oenococcus oeni). In the present study, the physiological changes induced by those AMPs on sensitive H. guilliermondii cells were evaluated in terms of intracellular pH (pHi), membrane permeability and culturability. Membrane permeability was evaluated by staining cells with propidium iodide (PI), pHi was determined by a fluorescence ratio imaging microscopy (FRIM) technique and culturability by a classical plating method. Results showed that the average pHi of H. guilliermondii cells dropped from 6.5 (healthy cells) to 5.4 (damaged cells) after 20 min of exposure to inhibitory concentrations of AMPs, and after 24 h 77.0% of the cells completely lost their pH gradient (∆pH=pHi-pHext). After 24h of exposure to AMPs, PI-stained (dead) cells increased from 0% to 77.7% and the number of viable cells fell from 1×10(5) to 10 CFU/ml. This means that virtually all cells (99.99%) became unculturable but that a sub-population of 22.3% of the cells remained viable (as determined by PI staining). Besides, pHi results showed that after 24h, 23% of the AMP-treated cells were sub-lethally injured (with 0pH<3). Taken together, these results indicated that this subpopulation was under a viable but non-culturable (VBNC) state, which was further confirmed by recuperation assays. In summary, our study reveals that these AMPs compromise the plasma membrane integrity (and possibly also the vacuole membrane) of H. guilliermondii cells, disturbing the pHi homeostasis and inducing a loss of culturability. Copyright © 2015 Elsevier B.V. All rights reserved.

  20. Optimization of the Expression of Equistatin in Pichia pastoris

    NARCIS (Netherlands)

    Outchkourov, N.S.; Stiekema, W.J.; Jongsma, M.A.

    2002-01-01

    To improve the expression of equistatin, a proteinase inhibitor from the sea anemone Actinia equina, in the yeast Pichia pastoris, we prepared gene variants with yeast-preferred codon usage and lower repetitive AT and GC content. The full gene optimization approximately doubled the level of steady-s

  1. Genetic engineering of Pichia stipitis for fermentation of xylose

    Science.gov (United States)

    Thomas W. Jeffries; N. Q. Shi; J. Y. Cho; P. Lu; K. Dahn; J. Hendrick; H. K. Sreenath

    1998-01-01

    A useful genetic system has been developed for the transformation of Pichia stipitis. This includes two selectable markers (URA3 and LEU2), integrating and autonomous replication vectors, a pop-out cassette that enables multiple targeted disruptions, and a genomic X-library for rapid cloning. Using this system we have cloned two genes for alcohol dehydrogenase (PsADH1...

  2. Enhancing ethanol production from cellulosic sugars using Scheffersomyces (Pichia) stipitis

    Science.gov (United States)

    Studies were performed on the effect of CaCO3 and CaCl2 supplementation to fermentation medium for ethanol production from xylose, glucose, or their mixtures using Scheffersomyces (Pichia) stipitis. Both of these chemicals were found to improve maximum ethanol concentration and ethanol productivity....

  3. Indigenous Partner Violence, Indigenous Sentencing Courts, and Pathways to Desistance.

    Science.gov (United States)

    Marchetti, Elena; Daly, Kathleen

    2016-09-13

    Mainstream sentencing courts do little to change the behavior of partner violence offenders, let alone members of more socially marginal groups. Indigenous offenders face a court system that has little relevance to the complexity of their relations and lived experiences. Assisted by respected Elders and Community Representatives, Australian Indigenous sentencing courts seek to create a more meaningful sentencing process that has a deeper impact on Indigenous offenders' attitudes and, ultimately, their behavior. Drawing from interviews with 30 Indigenous offenders, we explore the ways in which the courts can motivate Indigenous partner violence offenders on pathways to desistence.

  4. Designing Indigenous Language Revitalization

    Science.gov (United States)

    Hermes, Mary; Bang, Megan; Marin, Ananda

    2012-01-01

    Endangered Indigenous languages have received little attention within the American educational research community. However, within Native American communities, language revitalization is pushing education beyond former iterations of culturally relevant curriculum and has the potential to radically alter how we understand culture and language in…

  5. Designing Indigenous Language Revitalization

    Science.gov (United States)

    Hermes, Mary; Bang, Megan; Marin, Ananda

    2012-01-01

    Endangered Indigenous languages have received little attention within the American educational research community. However, within Native American communities, language revitalization is pushing education beyond former iterations of culturally relevant curriculum and has the potential to radically alter how we understand culture and language in…

  6. Aproveitamento de subproduto industrial de óleos vegetais para produção de riboflavina por Candida guilliermondii DM 644 Utilization industrial waste from vegetal oils riboflavine production by candida guilliermondii DM 644

    Directory of Open Access Journals (Sweden)

    Maria de Lourdes Andrade Pessoa

    2003-12-01

    Full Text Available A produção e consumo de alimentos industrializados têm aumentado a preocupação com suplementação e enriquecimento de alimentos com vitaminas e sais minerais, visando repor as possíveis perdas durante os processos de fabricação, principalmente das vitaminas hidrossolúveis, mais especificamente da vitamina B2 ou riboflavina. Assim sendo, a proposta deste trabalho foi utilizar como componente principal do meio, para produção da riboflavina, um subproduto do refino de óleos vegetais e o microrganismo Candida guillermondii DM 644. A produção da vitamina B2 foi realizada por fermentação em batelada utilizando Erlenmeyer. As condições empregadas foram agitação orbital, ausência de luz, 30°C, e 24h de incubação. A otimização da produção de riboflavina foi realizada através de Delineamento Fatorial Fracionário, para avaliar os efeitos da concentração de matéria graxa, fonte de nitrogênio, pH, velocidade de agitação, fonte de fósforo e extrato de levedura e as possíveis interações. A concentração máxima de riboflavina foi 19,12mg/mL. Os fatores mais importantes para produção de riboflavina foram a concentração de matéria graxa e a fonte de nitrogênio, enquanto que a fonte de fósforo e o extrato de leveduras não estimularam sua biossíntese. A máxima produção foi obtida com matéria graxa a 10g/L, uréia a 2,5g/L e pH 5,0. A velocidade de agitação (200 e 400rpm não interferiu no processo biotecnológico.The ever growing production and consumption of industrialized foods have increased the concern about supplementation and enrichment of food with vitamins and minerals, attempting the replenishment of the possible losses during their production processes, mainly the hydrosoluble vitamins, more specifically B2 vitamin or riboflavin. A subproduct of vegetal oil refining and the microorganism Candida guilliermondii DM 644 were used as the main substrate. The vitamin B2 was produced by the batch

  7. Open access to sequence: Browsing the Pichia pastoris genome

    Directory of Open Access Journals (Sweden)

    Graf Alexandra

    2009-10-01

    Full Text Available Abstract The first genome sequences of the important yeast protein production host Pichia pastoris have been released into the public domain this spring. In order to provide the scientific community easy and versatile access to the sequence, two web-sites have been installed as a resource for genomic sequence, gene and protein information for P. pastoris: A GBrowse based genome browser was set up at http://www.pichiagenome.org and a genome portal with gene annotation and browsing functionality at http://bioinformatics.psb.ugent.be/webtools/bogas. Both websites are offering information on gene annotation and function, regulation and structure. In addition, a WiKi based platform allows all users to create additional information on genes, proteins, physiology and other items of P. pastoris research, so that the Pichia community can benefit from exchange of knowledge, data and materials.

  8. Applications of recombinant Pichia pastoris in the healthcare industry

    Directory of Open Access Journals (Sweden)

    Daniel Weinacker

    2013-12-01

    Full Text Available Since the 1970s, the establishment and development of the biotech industry has improved exponentially, allowing the commercial production of biopharmaceutical proteins. Nowadays, new recombinant protein production is considered a multibillion-dollar market, in which about 25% of commercial pharmaceuticals are biopharmaceuticals. But to achieve a competitive production process is not an easy task. Any production process has to be highly productive, efficient and economic. Despite that the perfect host is still not discovered, several research groups have chosen Pichia pastoris as expression system for the production of their protein because of its many features. The attempt of this review is to embrace several research lines that have adopted Pichia pastoris as their expression system to produce a protein on an industrial scale in the health care industry.

  9. Characterization of wheat germin (oxalate oxidase) expressed by Pichia pastoris

    Science.gov (United States)

    Pan, Heng-Yen; Whittaker, Mei M.; Bouveret, Romaric; Berna, Anne; Bernier, François; Whittaker, James W.

    2007-01-01

    High-level secretory expression of wheat (Triticum aestivum) germin/oxalate oxidase was achieved in Pichia pastoris fermentation cultures as an α-mating factor signal peptide fusion, based on the native wheat cDNA coding sequence. The oxalate oxidase activity of the recombinant enzyme is substantially increased (7-fold) by treatment with sodium periodate, followed by ascorbate reduction. Using these methods, approximately 1 g (4×104 U) of purified, activated enzyme was obtained following eight days of induction of a high density Pichia fermentation culture, demonstrating suitability for large-scale production of oxalate oxidase for biotechnological applications. Characterization of the recombinant protein shows that it is glycosylated, with N-linked glycan attached at Asn47. For potential biomedical applications, a nonglycosylated (S49A) variant was also prepared which retains essentially full enzyme activity, but exhibits altered protein-protein interactions. PMID:17399681

  10. Indigenous microbiota and Leishmaniasis.

    Science.gov (United States)

    Lopes, M E M; Carneiro, M B H; Dos Santos, L M; Vieira, L Q

    2016-01-01

    Animals are colonized by their indigenous microbiota from the early days of life. The estimated number of associated bacterial cells in humans is around of 10(14) per individual, most of them in the gut. Several studies have investigated the microbiota-host relationship, and the use of germfree animals has been an important tool in these studies. These animals, when infected with a pathogen, have shown to be sometimes more resistant and other times more susceptible than conventional animals. Leishmaniasis is a worldwide public health problem and presents a spectrum of clinical manifestations. However, very few studies have addressed the role of the indigenous microbiota on the outcome of this disease. In this review, we will highlight and discuss the data available on the ways by which the microbiota can influence the outcome of the disease in murine experimental models of cutaneous infection with Leishmania.

  11. INDIGENISM AND EXCLUSION

    OpenAIRE

    José Angel Vera Noriega

    2006-01-01

    SUMMARYThe objective of the work is to carry out an analysis of the Western vision of the indigenous towns from the four types of exclusion of which Focault speaks (1978) when he talks about madness. The Social exclusion or work exclusion, family exclusion or emotional affective exclusion, the symbolic or linguistic exclusion and the playful or images exclusion. It is a reflection and transformation of ideas that allows coexisting worlds playing the power game in where the most important thin...

  12. INDIGENISM AND EXCLUSION

    OpenAIRE

    José Angel Vera Noriega

    2006-01-01

    SUMMARYThe objective of the work is to carry out an analysis of the Western vision of the indigenous towns from the four types of exclusion of which Focault speaks (1978) when he talks about madness. The Social exclusion or work exclusion, family exclusion or emotional affective exclusion, the symbolic or linguistic exclusion and the playful or images exclusion. It is a reflection and transformation of ideas that allows coexisting worlds playing the power game in where the most important thin...

  13. Indigenous Australian Education and Globalisation

    Science.gov (United States)

    Brady, Wendy

    1997-09-01

    This article focuses on the impact of colonisation and its associated impact on Indigenous teaching and learning. Western European institutions have dominated Indigenous ways of knowing and in Australia this has led to barriers which restrict the participation of Aboriginal people in education systems. Globally Indigenous people are attempting to bring into the introduced educational systems culturally appropriate teaching and learning practices so that a more holistic approach to education can become the norm rather than the exception. The relationship between Indigenous knowledge and western European concepts of knowledge and knowing need to placed in a framework of mutual interaction so that not only do Indigenous people benefit, but so do non-Indigenous educators and students.

  14. Anger in Australian Indigenous and Non-Indigenous Students

    Science.gov (United States)

    Boman, Peter; Mergler, Amanda; Furlong, Michael; Caltabiano, Nerina

    2014-01-01

    This descriptive pilot study examined the cultural differences in the dimensions of self-reported anger in Indigenous and non-Indigenous (Caucasian) students aged 10-13 years in Far North Queensland, Australia. The Multidimensional School Anger Inventory-Revised (MSAI-R) (Boman, Curtis, Furlong, & Smith, 2006) was used to measure affective,…

  15. Indigenous Language Immersion Schools for Strong Indigenous Identities

    Science.gov (United States)

    Reyhner, Jon

    2010-01-01

    Drawing on evidence from indigenous language immersion programs in the United States, this article makes the case that these immersion programs are vital to healing the negative effects of colonialism and assimilationist schooling that have disrupted many indigenous homes and communities. It describes how these programs are furthering efforts to…

  16. The Double Binds of Indigeneity and Indigenous Resistance

    Directory of Open Access Journals (Sweden)

    Francis Ludlow

    2016-07-01

    Full Text Available During the twentieth century, indigenous peoples have often embraced the category of indigenous while also having to face the ambiguities and limitations of this concept. Indigeneity, whether represented by indigenous people themselves or others, tends to face a “double bind”, as defined by Gregory Bateson, in which “no matter what a person does, he can’t win.” One exit strategy suggested by Bateson is meta-communication—communication about communication—in which new solutions emerge from a questioning of system-internal assumptions. We offer case studies from Ecuador, Peru and Alaska that chart some recent indigenous experiences and strategies for such scenarios.

  17. Antimicrobial peptides (AMPs) produced by Saccharomyces cerevisiae induce alterations in the intracellular pH, membrane permeability and culturability of Hanseniaspora guilliermondii cells

    DEFF Research Database (Denmark)

    Branco, Patrícia; Monteiro Lomba Viana, Tiago; Albergaria, Helena

    2015-01-01

    Hi results showed that after 24h, 23% of the AMP-treated cells were sub-lethally injured (with 0pHindicated that this subpopulation was under a viable but non-culturable (VBNC) state, which was further confirmed by recuperation assays. In summary, our study reveals...... on sensitive H. guilliermondii cells were evaluated in terms of intracellular pH (pHi), membrane permeability and culturability. Membrane permeability was evaluated by staining cells with propidium iodide (PI), pHi was determined by a fluorescence ratio imaging microscopy (FRIM) technique and culturability...... by a classical plating method. Results showed that the average pHi of H. guilliermondii cells dropped from 6.5 (healthy cells) to 5.4 (damaged cells) after 20min of exposure to inhibitory concentrations of AMPs, and after 24h 77.0% of the cells completely lost their pH gradient (∆pH=pHi-pHext). After 24h...

  18. Assessment of the Factors Contributing to the Growth or Spoilage of Meyerozyma guilliermondii in Organic Yogurt: Comparison of Methods for Strain Differentiation

    Directory of Open Access Journals (Sweden)

    Petra Wrent

    2015-08-01

    Full Text Available In this work we analyze the spoiling potential of Meyerozyma guilliermondii in yogurt. The analysis was based on contaminated samples sent to us by an industrial laboratory over two years. All the plain and fruit yogurt packages were heavily contaminated by yeasts, but only the last ones, containing fermentable sugars besides lactose, were spoiled by gas swelling. These strains were unable to grow and ferment lactose (as the type strain; they did grow on lactate plus galactose, fermented glucose and sucrose, and galactose (weakly, but did not compete with lactic acid bacteria for lactose. This enables them to grow in any yogurt, although only those with added jam were spoiled due to the fermentation of the fruit sugars. Fermentation, but not growth, was strongly inhibited at 8 °C. In consequence, in plain yogurt as well as in any yogurt maintained at low temperature, yeast contamination would not be detected by the consumer. The risk could be enhanced because the species has been proposed for biological control of fungal infections in organic agriculture. The combination of the IGS PCR-RFLP (amplification of the intergenic spacer region of rDNA followed by restriction fragment length polymorphism analysis method and mitochondrial DNA-RFLP makes a good tool to trace and control the contamination by M. guilliermondii.

  19. A Novel Production Method for High-Fructose Glucose Syrup from Sucrose-Containing Biomass by a Newly Isolated Strain of Osmotolerant Meyerozyma guilliermondii.

    Science.gov (United States)

    Khattab, Sadat Mohammad Rezq; Kodaki, Tsutomu

    2016-04-28

    One osmotolerant strain from among 44 yeast isolates was selected based on its growth abilities in media containing high concentrations of sucrose. This selected strain, named SKENNY, was identified as Meyerozyma guilliermondii by sequencing the internal transcribed spacer regions and partial D1/D2 large-subunit domains of the 26S ribosomal RNA. SK-ENNY was utilized to produce high-fructose glucose syrup (HFGS) from sucrose-containing biomass. Conversion rates to HFGS from 310-610 g/l of pure sucrose and from 75-310 g/l of sugar beet molasses were 73.5-94.1% and 76.2-91.1%, respectively. In the syrups produced, fructose yields were 89.4-100% and 96.5-100% and glucose yields were 57.6-82.5% and 55.3-79.5% of the theoretical values for pure sucrose and molasses sugars, respectively. This is the first report of employing M. guilliermondii for production of HFGS from sucrose-containing biomass.

  20. Indigenous innovation in China

    DEFF Research Database (Denmark)

    Jin, Jun; Slepniov, Dmitrij

    2012-01-01

    In the past two decades, China has earned the reputation of ‘manufacturing power house’ of the world. Chinese companies in their vast numbers have been very successful in exploiting their access to low-cost labour and have established themselves as unbeatable high volume low-cost manufacturing...... on their own in-house resources. The development and implementation of indigenous innovation solutions for these companies is an imperative which has not been adequately addressed in the literature. Therefore, by employing an explorative case of a Chinese company behind an innovative logistics concept...

  1. Indigenous Affairs = Asuntos Indigenas, 1997.

    Science.gov (United States)

    Indigenous Affairs, 1997

    1997-01-01

    This document contains the three 1997 English-language issues of Indigenous Affairs and the three corresponding issues in Spanish. (The last two quarterly issues were combined.) These periodicals provide a resource on the history, current conditions, and struggles for self-determination and human rights of indigenous peoples around the world.…

  2. Indigenous Affairs = Asuntos Indigenas, 1998.

    Science.gov (United States)

    Indigenous Affairs, 1998

    1998-01-01

    This document contains the four 1998 English-language issues of Indigenous Affairs and the four corresponding issues in Spanish. These periodicals provide a resource on the history, current conditions, and struggles for self-determination and human rights of indigenous peoples around the world. The first issue is a theme issue on the indigenous…

  3. Indigenous Affairs = Asuntos Indigenas, 1996.

    Science.gov (United States)

    Indigenous Affairs, 1996

    1996-01-01

    This document contains the four 1996 English-language issues of Indigenous Affairs and the four corresponding issues in Spanish. These newsletters provide a resource on the history, current conditions, and struggles for self-determination and human rights of indigenous peoples around the world. Articles on the United States and Canada (1) discuss…

  4. Indigenous Affairs = Asuntos Indigenas, 1998.

    Science.gov (United States)

    Indigenous Affairs, 1998

    1998-01-01

    This document contains the four 1998 English-language issues of Indigenous Affairs and the four corresponding issues in Spanish. These periodicals provide a resource on the history, current conditions, and struggles for self-determination and human rights of indigenous peoples around the world. The first issue is a theme issue on the indigenous…

  5. Indigenous Affairs = Asuntos Indigenas, 1996.

    Science.gov (United States)

    Indigenous Affairs, 1996

    1996-01-01

    This document contains the four 1996 English-language issues of Indigenous Affairs and the four corresponding issues in Spanish. These newsletters provide a resource on the history, current conditions, and struggles for self-determination and human rights of indigenous peoples around the world. Articles on the United States and Canada (1) discuss…

  6. Indigenous Affairs = Asuntos Indigenas, 1997.

    Science.gov (United States)

    Indigenous Affairs, 1997

    1997-01-01

    This document contains the three 1997 English-language issues of Indigenous Affairs and the three corresponding issues in Spanish. (The last two quarterly issues were combined.) These periodicals provide a resource on the history, current conditions, and struggles for self-determination and human rights of indigenous peoples around the world.…

  7. Indigenous rights, performativity and protest

    NARCIS (Netherlands)

    Hanna, Philippe; Langdon, Esther Jean; Vanclay, Frank

    2016-01-01

    Protests to claim rights are a common practice among Indigenous peoples of the world, especially when their interests conflict with those of nation states and/or multinational corporations regarding the use of their lands and resources. Drawing on a case study of the National Indigenous Mobilization

  8. Indigenous education and heritage revitalization

    NARCIS (Netherlands)

    Ke, Wen-Li

    2011-01-01

    The thesis (working title: 'Indigenous Education and Heritage Revitalization') focuses on the (possible) roles of tangible and intangible cultural heritage in the education of indigenous peoples in Taiwan, against the background of worldwide discussions and studies of the possibilities to create and

  9. Indigenization of urban mobility

    Science.gov (United States)

    Yang, Zimo; Lian, Defu; Yuan, Nicholas Jing; Xie, Xing; Rui, Yong; Zhou, Tao

    2017-03-01

    The identification of urban mobility patterns is very important for predicting and controlling spatial events. In this study, we analyzed millions of geographical check-ins crawled from a leading Chinese location-based social networking service (Jiepang.com), which contains demographic information that facilitates group-specific studies. We determined the distinct mobility patterns of natives and non-natives in all five large cities that we considered. We used a mixed method to assign different algorithms to natives and non-natives, which greatly improved the accuracy of location prediction compared with the basic algorithms. We also propose so-called indigenization coefficients to quantify the extent to which an individual behaves like a native, which depends only on their check-in behavior, rather than requiring demographic information. Surprisingly, the hybrid algorithm weighted using the indigenization coefficients outperformed a mixed algorithm that used additional demographic information, suggesting the advantage of behavioral data in characterizing individual mobility compared with the demographic information. The present location prediction algorithms can find applications in urban planning, traffic forecasting, mobile recommendation, and so on.

  10. Protecting indigenous rights. Guatemala.

    Science.gov (United States)

    1996-01-01

    Guatemala's recent ratification of the International Labor Organization (ILO) Convention regarding indigenous and tribal peoples (1989, No. 169) represents a commitment to guarantee the rights of the country's majority Mayan population. Ratifying governments are obligated to respect the traditional values and land rights of tribal and indigenous peoples and to consult with them on any decisions affecting their economic or social development. Ratification of this Convention was a key element in an eight-part UN-sponsored negotiation aimed at ending the civil war in Guatemala. Efforts are underway to promote dialogue between organized civil society and government. Negotiations in May 1996, conducted with ILO assistance, resulted in a socioeconomic agreement under which Guatemala will increase social investment in education, undertake agrarian reform, and institute tripartite consultation on all major social and economic issues. However, two key issues in the peace negotiations--the role of the army in civil society and constitutional reform--remain unresolved. The final global peace accord is expected to be signed in September 1996. UN organizations are already working to mobilize international support for transforming these agreements into political and social realities for the Guatemalan people.

  11. Indigenous Contributions to Sustainability

    Science.gov (United States)

    Barnhardt, R.

    2010-12-01

    Throughout the course of the Fourth International Polar Year(s), indigenous peoples have assumed a prominent role as significant partners in the pursuit of a broader and deeper understanding of the multifaceted dimensions of the human role in the Arctic region. Most salient in this partnership has been the substantial underlying differences in perspective, some political, some ideological, but most fundamental and intractable are the differences in world views, between those of the relative newcomers to the area (i.e. the miners, loggers, oil field workers, commercial fishermen, tourists, and even the occasional scientist), and the Native people with roots in the land that go back millennia. But no longer can these differences be cast in simplistic either/or terms, implying some kind of inherent dichotomy between those who live off the land vs. those tied to the cash economy, or traditional vs. modern technologies, or anecdotal vs. scientific evidence. These lines have been blurred with the realities that indigenous cultures are not static, and western structures are no longer dominant. Instead, we now have a much more fluid and dynamic situation in which once competing views of the world are striving toward reconciliation through new structures and frameworks that foster co-existence rather than domination and exploitation.

  12. Expression of Aspergillus niger 9891 Endoinulinase in Pichia pastoris

    Institute of Scientific and Technical Information of China (English)

    Wang Jianhua(王建华); Teng Da; Yao Yi; Yang Yalin; Zhang Fan

    2004-01-01

    An endoinulinase gene from Aspergillus niger 9891 (CGMCC0991) has been expressed in Pichia pastoris GS115 using pPIC9 vector. The recombinant endoinulinase was highly expressed and the optimization of the expression in a 7 liter of fermentor has been investigated. In fermented broth, the concentration of protein secreted is 2.15 mg/ml. The activity of endoinulinase is 1501 U/ml with sucrose as substrate and 291 U/ml with inulin as substrate, 105 and 273 times higher than that from the original strain respectively.

  13. Indigenous Educational Attainment in Canada

    Directory of Open Access Journals (Sweden)

    Catherine E. Gordon

    2014-06-01

    Full Text Available In this article, the educational attainment of Indigenous peoples of working age (25 to 64 years in Canada is examined. This diverse population has typically had lower educational levels than the general population in Canada. Results indicate that, while on the positive side there are a greater number of highly educated Indigenous peoples, there is also a continuing gap between Indigenous and non-Indigenous peoples. Data also indicate that the proportion with less than high school education declined, which corresponds with a rise of those with a PSE; the reverse was true in 1996. Despite these gains, however, the large and increasing absolute numbers of those without a high school education is alarming. There are intra-Indigenous differences: First Nations with Indian Status and the Inuit are not doing as well as non-Status and Métis peoples. Comparisons between the Indigenous and non-Indigenous populations reveal that the documented gap in post-secondary educational attainment is at best stagnant. Out of the data analysis, and based on the history of educational policy, we comment on the current reform proposed by the Government of Canada, announced in February of 2014, and propose several policy recommendations to move educational attainment forward.

  14. Indigenous Child Health in Brazil

    Science.gov (United States)

    del Pino Marchito, Sandra; Vitoy, Bernardino

    2016-01-01

    Abstract Improving the health status of indigenous children is a long-standing challenge. Several United Nations committees have identified the health of indigenous peoples as a human rights concern. Addressing the health of indigenous children cannot be separated from their social, cultural, and historic contexts, and any related health program must offer culturally appropriate services and a community perspective broad enough to address the needs of children and the local worlds in which they live. Evaluations of programs must, therefore, address process as well as impacts. This paper assesses interventions addressing indigenous children’s health in Brazil, ranging from those explicitly targeting indigenous children’s health, such as the targeted immunization program for indigenous peoples, as well as more generalized programs, including a focus upon indigenous children, such as the Integrated Management of Childhood Illness. The paper discusses the tensions and complexities of ethnically targeted health interventions as well as the conceptual and methodological challenge of measuring the processes employed and their impact. The lessons learned, especially the need for countries to more systematically collect data and evaluate impacts using ethnicity as an analytical category, are drawn out with respect to ensuring human rights for all within health sector responses. PMID:27781012

  15. Indigenous Nations' Responses to Climate Change

    Science.gov (United States)

    Grossman, Zoltan

    2008-01-01

    On August 1st, 2007, Indigenous nations from within the United States, Canada, Australia, and Aotearoa (New Zealand) signed a treaty to found the United League of Indigenous Nations. The Treaty of Indigenous Nations offers a historic opportunity for sovereign Indigenous governments to build intertribal cooperation outside the framework of the…

  16. Indigenous innovation in China

    DEFF Research Database (Denmark)

    Jin, Jun; Slepniov, Dmitrij

    2012-01-01

    on their own in-house resources. The development and implementation of indigenous innovation solutions for these companies is an imperative which has not been adequately addressed in the literature. Therefore, by employing an explorative case of a Chinese company behind an innovative logistics concept......In the past two decades, China has earned the reputation of ‘manufacturing power house’ of the world. Chinese companies in their vast numbers have been very successful in exploiting their access to low-cost labour and have established themselves as unbeatable high volume low-cost manufacturing...... champions. However, recently growing number of Chinese companies are seeking to create a foundation for growth and development based on innovation. As a result of this, many of them spread their operations to the countries of the traditional industrial ‘triad’ of North America, Europe and Japan to capture...

  17. INDIGENISM AND EXCLUSION

    Directory of Open Access Journals (Sweden)

    José Angel Vera Noriega

    2006-09-01

    Full Text Available SUMMARYThe objective of the work is to carry out an analysis of the Western vision of the indigenous towns from the four types of exclusion of which Focault speaks (1978 when he talks about madness. The Social exclusion or work exclusion, family exclusion or emotional affective exclusion, the symbolic or linguistic exclusion and the playful or images exclusion. It is a reflection and transformation of ideas that allows coexisting worlds playing the power game in where the most important thing is not the one who wins or that loses, but the imagination and the creativity to modify the order of the rules and norms for that power game. It is discussed and it is exemplified about the territory concept, medicine and religion. It is discussed in terms of fairness policies and equality in a pertinent and relevant dialog for the construction of possible alliances.

  18. Isolation of Pichia manshurica protoplast from Dahlia sp plant

    Directory of Open Access Journals (Sweden)

    Wijanarka Wijanarka

    2014-02-01

    Full Text Available Isolation of protoplasts is an important step in the fusion process. Protoplasts are cells that have eliminated the cell wall, but the cell membranes and organs can still function properly. Pichia manshurica is one of indogenous yeast that derived from Dahlia €™s plants. The success rate protoplast isolation was determined by various factors, include the age of the culture and the used of lytic enzymes. The purpose of this research is to get the perfect age of yeast culture that is ready to be harvested and also to get the appropriate concentration of Glucanex lytic enzymes which used for protoplast isolation. The yeast of Pichia manshurica grown on YPD broth medium and growth observed in turbidimetry. Observation of the growth of yeasts performed every 6 hours for 42 hours. Glucanex lytic enzyme concentration used for the isolation of protoplasts is 0 mg / mL (L0 = control, 2 mg / mL (L2 and 4 mg / mL (L4. The results showed that the age of the culture is right and ready for harvest at the age of 24 hours and Glucanex lytic enzyme concentration of 4 mg / mL (L4 is able to produce the best of protoplasts at 7.2 x 1010.

  19. Expression of human acidic fibroblast growth factor in Pichia pastoris

    Institute of Scientific and Technical Information of China (English)

    YU Ying; CAI Shaoxi; Harald G. WERIRICH; XIA Yuxian

    2003-01-01

    Pichia pastoris expression system is similar to that of the mammal cell in modification of expressed protein, including refolding and glycosylation. A human aFGF gene was cloned into the intracellular expression vector pPIC9K. The Pichia pastoriS KM71 strain was transformed with the recombined expression plasmid. Transgenic expression was observed after screening the transformants with G418. The expression and secretion of recombinant human aFGF (rhaFGF) into the culture medium were testified by ELISA assay. The yield peaked after two days of induction and was approximately 10 mg.L-1 in shake-flask fermentation medium. The recombinant proteins were purified by the combination of heparin-Sepharose affinity chromatography and gel filtration chromatography. Two proteins with relative molecular masses (Mr) of 17 000 and 35 000 were purified as a single band in SDS-PAGE, whose biological activities were determined by MTT assay. It is found that the protein with Mr of 1 7 000 is nonglycosylated haFGF, and that with Mr of 35 000 is glycosylated haFGF; and the latter has a lower biological activity than the former.

  20. Cyber-Indigeneity: Urban Indigenous Identity on Facebook

    Science.gov (United States)

    Lumby, Bronwyn

    2010-01-01

    This paper addresses understandings and theorising of identity in cyberspace. In particular, it focuses on the construction, maintenance and performance of urban Indigenous identities on the contemporary internet social space, Facebook.

  1. Cyber-Indigeneity: Urban Indigenous Identity on Facebook

    Science.gov (United States)

    Lumby, Bronwyn

    2010-01-01

    This paper addresses understandings and theorising of identity in cyberspace. In particular, it focuses on the construction, maintenance and performance of urban Indigenous identities on the contemporary internet social space, Facebook.

  2. INDIGENOUS LANGUAGE IMPLEMENTATION AND NATION ...

    African Journals Online (AJOL)

    NGOZI

    language teaching and use in the early child education stages can enhance nation building. .... Experience. 48 the second language learners (L2) and a multilingual nation. Is it ... banish the indigenous cultures and languages, but to erect a ...

  3. A Mitochondrial Autonomously Replicating Sequence from Pichia pastoris for Uniform High Level Recombinant Protein Production

    National Research Council Canada - National Science Library

    Karl Friehs; Jan-Philipp Schwarzhans; Tobias Luttermann; Daniel Wibberg; Anika Winkler; Wolfgang Hübner; Thomas Huser; Jörn Kalinowski

    2017-01-01

    Pichia pastoris is a non-conventional methylotrophic yeast that is widely used for recombinant protein production, typically by stably integrating the target gene into the genome as part of an expression cassette...

  4. Expression and analysis of the glycosylation properties of recombinant human erythropoietin expressed in Pichia pastoris

    OpenAIRE

    Ser Huy Teh; Mun Yik Fong; Zulqarnain Mohamed

    2011-01-01

    The Pichia pastoris expression system was used to produce recombinant human erythropoietin, a protein synthesized by the adult kidney and responsible for the regulation of red blood cell production. The entire recombinant human erythropoietin (rhEPO) gene was constructed using the Splicing by Overlap Extension by PCR (SOE-PCR) technique, cloned and expressed through the secretory pathway of the Pichia expression system. Recombinant erythropoietin was successfully expressed in P. pastoris. The...

  5. Indigenous health and socioeconomic status in India.

    Directory of Open Access Journals (Sweden)

    S V Subramanian

    2006-10-01

    Full Text Available BACKGROUND: Systematic evidence on the patterns of health deprivation among indigenous peoples remains scant in developing countries. We investigate the inequalities in mortality and substance use between indigenous and non-indigenous, and within indigenous, groups in India, with an aim to establishing the relative contribution of socioeconomic status in generating health inequalities. METHODS AND FINDINGS: Cross-sectional population-based data were obtained from the 1998-1999 Indian National Family Health Survey. Mortality, smoking, chewing tobacco use, and alcohol use were four separate binary outcomes in our analysis. Indigenous status in the context of India was operationalized through the Indian government category of scheduled tribes, or Adivasis, which refers to people living in tribal communities characterized by distinctive social, cultural, historical, and geographical circumstances.Indigenous groups experience excess mortality compared to non-indigenous groups, even after adjusting for economic standard of living (odds ratio 1.22; 95% confidence interval 1.13-1.30. They are also more likely to smoke and (especially drink alcohol, but the prevalence of chewing tobacco is not substantially different between indigenous and non-indigenous groups. There are substantial health variations within indigenous groups, such that indigenous peoples in the bottom quintile of the indigenous-peoples-specific standard of living index have an odds ratio for mortality of 1.61 (95% confidence interval 1.33-1.95 compared to indigenous peoples in the top fifth of the wealth distribution. Smoking, drinking alcohol, and chewing tobacco also show graded associations with socioeconomic status within indigenous groups. CONCLUSIONS: Socioeconomic status differentials substantially account for the health inequalities between indigenous and non-indigenous groups in India. However, a strong socioeconomic gradient in health is also evident within indigenous

  6. Functional analysis of alcohol dehydrogenase (ADH) genes in Pichia pastoris.

    Science.gov (United States)

    Karaoglan, Mert; Karaoglan, Fidan Erden; Inan, Mehmet

    2016-03-01

    To characterize the genes responsible for ethanol utilization in Pichia pastoris. ADH3 (XM_002491337) and ADH (FN392323) genes were disrupted in P. pastoris. The ADH3 mutant strain, MK115 (Δadh3), lost its ability to grow on minimal ethanol media but produced ethanol in minimal glucose medium. ADH3p was responsible for 92 % of total Adh enzyme activity in glucose media. The double knockout strain MK117 (Δadh3Δadh) also produced ethanol. The Adh activities of X33 and MK116 (Δadh) strains were not different. Thus, the ADH gene does not play a role in ethanol metabolism. The PpADH3 is the only gene responsible for consumption of ethanol in P. pastoris.

  7. [Overexpression of Penicillium expansum lipase gene in Pichia pastoris].

    Science.gov (United States)

    Yuan, Cai; Lin, Lin; Shi, Qiao-Qin; Wu, Song-Gang

    2003-03-01

    The alkaline lipase gene of Penicillium expansum (PEL) was coloned into the yeast integrative plasmid pPIC3.5K, which was then transformed into His4 mutant yeast GS115. Recombinant Pichia strains were obtained by minimal olive oil-methanol plates screening and confirmed by PCR. The expression producus of PEL gene was analysis by SDS-PAGE and olive oil plate, the result indicated that PEL gene was functionally overexpressed in Pichia pastoris and up to 95% of the secreted protein. Recombinant lipase had a molecular mass of 28kD, showing a range similar to that of PEL, could hydrolyze olive oil and formed clear halos in the olive oil plates. Four different strategies (different media, pH, glycerol and methanol concentration) were applied to optimize the cultivation conditions, the activity of lipase was up to 260 u/mL under the optimal cultivation conditions. It is pointed out that the absence of the expensive biotin and yeast nitrogen base in the medium increased the lipase production. The possible reason of this result is absence of yeast nitrogen base increased the medium pH during cultivation, and PEL shows a higher stability at this condition. The lipase activity of the supernatant from the culture grown at pH 7 was higher than the one from the culture in the same medium at pH 6.0 is due to the pH stability of PEL too. The results also showed that the methanol and glycerol concentration had a marked effect on the production of lipase.

  8. Enhancement of thermoalkaliphilic xylanase production by Pichia pastoris through novel fed-batch strategy in high cell-density fermentation

    National Research Council Canada - National Science Library

    Shang, Tingting; Si, Dayong; Zhang, Dongyan; Liu, Xuhui; Zhao, Longmei; Hu, Cong; Fu, Yu; Zhang, Rijun

    2017-01-01

    .... This study aims to enhance thermoalkaliphilic xylanase production in Pichia pastoris through fermentation parameters optimization and novel efficient fed-batch strategy in high cell-density fermentation...

  9. Secretory expression of human protein in the Yeast Pichia pastoris by controlled fermentor culture.

    Science.gov (United States)

    Murasugi, Akira

    2010-06-01

    The heterologous protein expression system of Pichia pastoris is now widely used for expression of many human proteins, because the efficiently expressed proteins will be correctly folded in Pichia pastoris cells and also efficiently secreted from the cells. Recombinant human serum albumin (rHSA) is efficiently secreted from Pichia pastoris. Nowadays, the expression of rHSA exceeds 10g in 1 L fermentor culture broth, and the protein is completely purified. Recombinant HSA expressed in Pichia pastoris was approved as a medicine by the authorities in 2007, and launched in 2008 in Japan. One of the insulin precursors (IP) was also successfully expressed in Pichia pastoris, and secreted up to 3.6g in 1 L medium using a multi-copy transformant. The insulin precursor could be efficiently converted to insulin, the final product, in vitro. Human growth hormone was also expressed in Pichia pastoris, and secreted up to 49 mg in 1 L medium. These proteins are also important for clinical applications. Midkine and pleiotrophin may be two of the candidates for clinical applications. Secretion signals, the copy number of an expression cassette in transformants, and culture conditions for fermentation were examined for efficient expression of these proteins in Pichia pastoris. The best signal was selected, and other factors were optimized. The amounts of native midkine and native pleiotrophin expressed were approximately 0.36g and 0.26g in 1 L medium, respectively. Expression of bile-salt stimulated lipase (BSSL) had been extremely low in the beginning of a fermentor culture experiment. However, approximately 1 g rBSSL in 1 L medium was finally expressed in a fermentor by unlimited feeding of glycerol for cell growth and optimization of other factors. BSSL from human milk and rBSSL from Pichia cells are glycosylated. The structure differences between these glycans are obvious. When humanization of Pichia glycans is established by genetic engineering, the Pichia pastoris expression

  10. Genome-scale metabolic reconstructions of Pichia stipitis and Pichia pastoris and in silico evaluation of their potentials

    Directory of Open Access Journals (Sweden)

    Caspeta Luis

    2012-04-01

    Full Text Available Abstract Background Pichia stipitis and Pichia pastoris have long been investigated due to their native abilities to metabolize every sugar from lignocellulose and to modulate methanol consumption, respectively. The latter has been driving the production of several recombinant proteins. As a result, significant advances in their biochemical knowledge, as well as in genetic engineering and fermentation methods have been generated. The release of their genome sequences has allowed systems level research. Results In this work, genome-scale metabolic models (GEMs of P. stipitis (iSS884 and P. pastoris (iLC915 were reconstructed. iSS884 includes 1332 reactions, 922 metabolites, and 4 compartments. iLC915 contains 1423 reactions, 899 metabolites, and 7 compartments. Compared with the previous GEMs of P. pastoris, PpaMBEL1254 and iPP668, iLC915 contains more genes and metabolic functions, as well as improved predictive capabilities. Simulations of physiological responses for the growth of both yeasts on selected carbon sources using iSS884 and iLC915 closely reproduced the experimental data. Additionally, the iSS884 model was used to predict ethanol production from xylose at different oxygen uptake rates. Simulations with iLC915 closely reproduced the effect of oxygen uptake rate on physiological states of P. pastoris expressing a recombinant protein. The potential of P. stipitis for the conversion of xylose and glucose into ethanol using reactors in series, and of P. pastoris to produce recombinant proteins using mixtures of methanol and glycerol or sorbitol are also discussed. Conclusions In conclusion the first GEM of P. stipitis (iSS884 was reconstructed and validated. The expanded version of the P. pastoris GEM, iLC915, is more complete and has improved capabilities over the existing models. Both GEMs are useful frameworks to explore the versatility of these yeasts and to capitalize on their biotechnological potentials.

  11. Adult Learning, Transformative Education, and Indigenous Epistemology

    Science.gov (United States)

    McEachern, Diane

    2016-01-01

    This chapter describes an innovative program that weaves together adult learning, transformative education, and indigenous epistemology in order to prepare Alaskan rural indigenous social service providers to better serve their communities.

  12. Reflecting on Research Practices and Indigenous Community ...

    African Journals Online (AJOL)

    Reflecting on Research Practices and Indigenous Community Benefits for Poverty ... of various research projects within indigenous communities that brought forth ... Best research practices as well as practices that do not yield much success ...

  13. Adult Learning, Transformative Education, and Indigenous Epistemology

    Science.gov (United States)

    McEachern, Diane

    2016-01-01

    This chapter describes an innovative program that weaves together adult learning, transformative education, and indigenous epistemology in order to prepare Alaskan rural indigenous social service providers to better serve their communities.

  14. Ethical dimension of indigenous knowledge systems | Mutula ...

    African Journals Online (AJOL)

    Ethical dimension of indigenous knowledge systems. ... by way of social exclusion, linguistic marginalization, erosion of cultural identity, preference for ... The World Summit on Information Society (WSIS) provided an opportunity for indigenous ...

  15. The World Indigenous Research Alliance (WIRA): Mediating and Mobilizing Indigenous Peoples' Educational Knowledge and Aspirations

    Science.gov (United States)

    Whitinui, Paul; McIvor, Onowa; Robertson, Boni; Morcom, Lindsay; Cashman, Kimo; Arbon, Veronica

    2015-01-01

    There is an Indigenous resurgence in education occurring globally. For more than a century Euro-western approaches have controlled the provision and quality of education to, and for Indigenous peoples. The World Indigenous Research Alliance (WIRA) established in 2012, is a grass-roots movement of Indigenous scholars passionate about making a…

  16. Reassembling the Indigenous Public Sphere

    Directory of Open Access Journals (Sweden)

    Jack Latimore

    2017-07-01

    Full Text Available This paper seeks to provide an initial theoretical grounding to assess a practical project: a new software application that attempts to be a beneficial resource in the field of Indigenous representation. As a starting point, we are concerned to provide a theoretical ground for considering the inherited and shifting spaces of Indigenous media representation. To this end, this paper reconsiders the strengths and weaknesses of debates surrounding the ‘Indigenous public sphere’. This is used as grounds for critically understanding the relations that constitute this field. Following this, we consider how a more materialist approach to publics might enable a productive reconceptualization, and in particular how digital media initiatives and shifting news markets may be contributing to change. Finally, drawing on this model, we outline both the ‘Wakul app’ project, and how this framework might inform an assessment of its impact.

  17. Toward an Integrative Framework of Indigenous Research

    DEFF Research Database (Denmark)

    Li, Peter Ping

    2012-01-01

    It has long been recognized that indigenous research should be helpful, if not essential, for an adequate understanding of local phenomena. The indigenous approach is consistent with, but extends beyond, the repeated calls for contextualizing management and organization research. However, the cha......It has long been recognized that indigenous research should be helpful, if not essential, for an adequate understanding of local phenomena. The indigenous approach is consistent with, but extends beyond, the repeated calls for contextualizing management and organization research. However...

  18. Indigenous Research on Chinese Management

    DEFF Research Database (Denmark)

    Li, Peter Ping; Leung, Kwok; Chen, Chao C.

    2012-01-01

    We attempt to provide a definition and a typology of indigenous research on Chinese management as well as outline the general methodological approaches for this type of research. We also present an integrative summary of the four articles included in this special issue and show how they illustrate...... our definition and typology of indigenous research on Chinese management, as well as the various methodological approaches we advocate. Further, we introduce a commentary on the four articles from the perspective of engaged scholarship, and also three additional articles included in this issue...

  19. More Like Ourselves: Indigenous Capitalism through Tourism

    Science.gov (United States)

    Bunten, Alexis Celeste

    2010-01-01

    Through a comparison of Indigenous-owned cultural tourism businesses in southeastern Alaska and New Zealand as well as secondary data examining Indigenous tourism across the Pacific, this article introduces the concept of "Indigenous capitalism" as a distinct strategy to achieve ethical, culturally appropriate, and successful Indigenous…

  20. More Like Ourselves: Indigenous Capitalism through Tourism

    Science.gov (United States)

    Bunten, Alexis Celeste

    2010-01-01

    Through a comparison of Indigenous-owned cultural tourism businesses in southeastern Alaska and New Zealand as well as secondary data examining Indigenous tourism across the Pacific, this article introduces the concept of "Indigenous capitalism" as a distinct strategy to achieve ethical, culturally appropriate, and successful Indigenous…

  1. Indigenous Studies and the Politics of Language

    Science.gov (United States)

    McGloin, Colleen; Carlson, Bronwyn L.

    2013-01-01

    Language use changes over time. In Indigenous contexts, language alters to suit the shifting nature of cultural expression as this might fit with Indigenous peoples' preference or as a consequence of changes to outdated and colonial modes of expression. For students studying in the discipline of Indigenous Studies, learning to use appropriate…

  2. Indigenous Affairs = Asuntos Indigenas, 1994-1995.

    Science.gov (United States)

    Indigenous Affairs, 1995

    1995-01-01

    This document consists of the eight issues of the IWGIA newsletter "Indigenous Affairs" published during 1994-95. Each issue is published in separate English and Spanish versions. The newsletter is published by the International Work Group for Indigenous Affairs (IWGIA), an organization that supports indigenous peoples in their efforts…

  3. Indigenous Affairs = Asuntos Indigenas, 1994-1995.

    Science.gov (United States)

    Indigenous Affairs, 1995

    1995-01-01

    This document consists of the eight issues of the IWGIA newsletter "Indigenous Affairs" published during 1994-95. Each issue is published in separate English and Spanish versions. The newsletter is published by the International Work Group for Indigenous Affairs (IWGIA), an organization that supports indigenous peoples in their efforts to gain…

  4. Indigenous Specializations: Dreams, Developments, Delivery and Vision

    Science.gov (United States)

    Richardson, Cathy; Thomas, Robina; Green, Jacquie; Ormiston, Todd

    2012-01-01

    This article documents the establishment of the Indigenous Specializations program in the School of Social Work at the University of Victoria. In the absence of funding for Indigenous programs, First Nations professors Robina Thomas and Jacquie Green developed the Indigenous Specializations program "off the side of their desk". This…

  5. Identification of indigenous yeast flora isolated from the five winegrape varieties harvested in Xiangning, China.

    Science.gov (United States)

    Sun, Yue; Guo, Jingjing; Liu, Fubing; Liu, Yanlin

    2014-03-01

    Inoculated fermentation by selected indigenous yeast strains from a specific location could provide the wine with unique regional sensory characteristics. The identification and differentiation of local yeasts are the first step to understand the function of yeasts and develop a better strain-selection program for winemaking. The indigenous yeasts in five grape varieties, Chardonnay, Cabernet Franc, Cabernet Sauvignon, Marselan, and Merlot cultivated in Xiangning, Shanxi, China were investigated. Eight species of seven genera including Aureobasidium pullulans, Candida zemplinina, Hanseniaspora uvarum, Hanseniaspora occidentalis, Issatchenkia terricola, Metschnikowia pulcherrima, Pichia kluyveri, and Saccharomyces cerevisiae were identified using Wallerstein Laboratory Nutrient medium with sequencing of the 26S rDNA D1/D2 domain. H. uvarum and S. cerevisiae were the predominant species, while most non-Saccharomyces species were present in the whole fermentation process at different levels among the grape varieties. The genotypes of S. cerevisiae from each microvinification were determined by using interdelta sequence analysis. The 102 isolates showed eight different genotypes, and genotype III was the predominant genotype found. The distribution of S. cerevisiae strains during the fermentation of Marselan was also studied. Six genotypes were observed among the 92 strains with different genotypes of competitiveness at different sampling stages. Genotype V demonstrated the potential for organizing starter strains and avoiding inefficient fermentation. In general, this study explored the yeast species in the grapes grown in Xiangning County and provided important information of relationship of local yeast diversity and its regional wine sensory characteristics.

  6. Culturally capable and culturally safe: Caseload care for Indigenous women by Indigenous midwifery students.

    Science.gov (United States)

    West, R; Gamble, J; Kelly, J; Milne, T; Duffy, E; Sidebotham, M

    2016-12-01

    Evidence is emerging of the benefits to students of providing continuity of midwifery care as a learning strategy in midwifery education, however little is known about the value of this strategy for midwifery students. To explore Indigenous students' perceptions of providing continuity of midwifery care to Indigenous women whilst undertaking a Bachelor of Midwifery. Indigenous Bachelor of Midwifery students' experiences of providing continuity of midwifery care to Indigenous childbearing women were explored within an Indigenous research approach using a narrative inquiry framework. Participants were three Indigenous midwifery students who provided continuity of care to Indigenous women. Three interconnected themes; facilitating connection, being connected, and journeying with the woman. These themes contribute to the overarching finding that the experience of providing continuity of care for Indigenous women creates a sense of personal affirmation, purpose and a validation of cultural identity in Indigenous students. Midwifery philosophy aligns strongly with the Indigenous health philosophy and this provides a learning platform for Indigenous student midwives. Privileging Indigenous culture within midwifery education programs assists students develop a sense of purpose and affirms them in their emerging professional role and within their community. The findings from this study illustrate the demand for, and pertinence of, continuity of care midwifery experiences with Indigenous women as fundamental to increasing the Indigenous midwifery workforce in Australia. Australian universities should provide this experience for Indigenous student midwives. Copyright © 2016 Australian College of Midwives. Published by Elsevier Ltd. All rights reserved.

  7. Biculturalism among Indigenous College Students

    Science.gov (United States)

    Miller, Colton D.

    2011-01-01

    "Indigenous" college students in both Canada and the United States have the lowest rates of obtaining postsecondary degrees, and their postsecondary dropout rates are higher than for any other minority (Freeman & Fox, 2005; Mendelson, 2004; Reddy, 1993). There has been very little research done to uncover possible reasons for such…

  8. Indigenous agroforestry in American Samoa

    Science.gov (United States)

    Malala (Mike) Misa; Agnes M. Vargo

    1993-01-01

    Agroforestry exists in American Samoa as a system where indigenous trees and natural vegetation used for food, fuelwood, crafts and medicine are incorporated with traditional staple crops and livestock on a set piece of land, usually a mountainous slope. Most agroforests are taro-based (Colocasia esculenta). While nutritional, cultural, social,...

  9. Indigenous Affairs = Asuntos Indigenas, 2000.

    Science.gov (United States)

    Indigenous Affairs, 2000

    2000-01-01

    This document contains the four English-language issues of Indigenous Affairs published in 2000 and four corresponding issues in Spanish. The Spanish issues contain all or some of the articles contained in the English issues plus additional articles on Latin America. These periodicals provide a resource on the history, current conditions, and…

  10. Indigenous Affairs = Asuntos Indigenas, 2000.

    Science.gov (United States)

    Indigenous Affairs, 2000

    2000-01-01

    This document contains the four English-language issues of Indigenous Affairs published in 2000 and four corresponding issues in Spanish. The Spanish issues contain all or some of the articles contained in the English issues plus additional articles on Latin America. These periodicals provide a resource on the history, current conditions, and…

  11. Rethinking resilience from indigenous perspectives.

    Science.gov (United States)

    Kirmayer, Laurence J; Dandeneau, Stéphane; Marshall, Elizabeth; Phillips, Morgan Kahentonni; Williamson, Karla Jessen

    2011-02-01

    The notions of resilience that have emerged in developmental psychology and psychiatry in recent years require systematic rethinking to address the distinctive cultures, geographic and social settings, and histories of adversity of indigenous peoples. In Canada, the overriding social realities of indigenous peoples include their historical rootedness to a specific place (with traditional lands, communities, and transactions with the environment) and the profound displacements caused by colonization and subsequent loss of autonomy, political oppression, and bureaucratic control. We report observations from an ongoing collaborative project on resilience in Inuit, Métis, Mi'kmaq, and Mohawk communities that suggests the value of incorporating indigenous constructs in resilience research. These constructs are expressed through specific stories and metaphors grounded in local culture and language; however, they can be framed more generally in terms of processes that include: regulating emotion and supporting adaptation through relational, ecocentric, and cosmocentric concepts of self and personhood; revisioning collective history in ways that valorize collective identity; revitalizing language and culture as resources for narrative self-fashioning, social positioning, and healing; and renewing individual and collective agency through political activism, empowerment, and reconciliation. Each of these sources of resilience can be understood in dynamic terms as emerging from interactions between individuals, their communities, and the larger regional, national, and global systems that locate and sustain indigenous agency and identity. This social-ecological view of resilience has important implications for mental health promotion, policy, and clinical practice.

  12. Expression and Purification of C-Peptide Containing Insulin Using Pichia pastoris Expression System

    Directory of Open Access Journals (Sweden)

    Mohammed N. Baeshen

    2016-01-01

    Full Text Available Increase in the incidence of Insulin Dependent Diabetes Mellitus (IDDM among people from developed and developing countries has created a large global market for insulin. Moreover, exploration of new methods for insulin delivery including oral or inhalation route which require very high doses would further increase the demand of cost-effective recombinant insulin. Various bacterial and yeast strains have been optimized to overproduce important biopharmaceuticals. One of the approaches we have taken is the production of recombinant human insulin along with C-peptide in yeast Pichia pastoris. We procured a cDNA clone of insulin from Origene Inc., USA. Insulin cDNA was PCR amplified and cloned into yeast vector pPICZ-α. Cloned insulin cDNA was confirmed by restriction analysis and DNA sequencing. pPICZ-α-insulin clone was transformed into Pichia pastoris SuperMan5 strain. Several Zeocin resistant clones were obtained and integration of insulin cDNA in Pichia genome was confirmed by PCR using insulin specific primers. Expression of insulin in Pichia clones was confirmed by ELISA, SDS-PAGE, and Western blot analysis. In vivo efficacy studies in streptozotocin induced diabetic mice confirmed the activity of recombinant insulin. In conclusion, a biologically active human proinsulin along with C-peptide was expressed at high level using Pichia pastoris expression system.

  13. Inactivation of a GAL4-Like Transcription Factor Improves Cell Fitness and Product Yield in Glycoengineered Pichia pastoris Strains

    Science.gov (United States)

    Argyros, Rebecca; Bukowski, John; Nelson, Stephanie; Sharkey, Nathan; Kim, Sehoon; Copeland, Victoria; Davidson, Robert C.; Chen, Ronghua; Zhuang, Jun; Sethuraman, Natarajan; Stadheim, Terrance A.

    2014-01-01

    With a completely reengineered and humanized glycosylation pathway, glycoengineered Pichia pastoris has emerged as a promising production host for the manufacture of therapeutic glycoproteins. However, the extensive genetic modifications have also negatively affected the overall fitness levels of the glycoengineered host cells. To make glycoengineered Pichia strains more compatible with a scalable industrial fermentation process, we sought to identify genetic solutions to broadly improve cell robustness during fermentation. In this study, we report that mutations within the Pichia pastoris ATT1 (PpATT1) gene (a homolog of the Saccharomyces cerevisiae GAL4 [ScGAL4] transcriptional activator) dramatically increased the cellular fitness levels of glycoengineered Pichia strains. We demonstrate that deletion of the PpATT1 gene enabled glycoengineered Pichia strains to improve their thermal tolerance levels, reduce their cell lysis defects, and greatly improve fermentation robustness. The extension of the duration of fermentation enabled the PpATT1-modified glycoengineered Pichia strains to increase their product yields significantly without any sacrifice in product quality. Because the ATT1 gene could be deleted from any Pichia strains, including empty hosts and protein-expressing production strains alike, we suggest that the findings described in this study are broadly applicable to any Pichia strains used for the production of therapeutic proteins, including monoclonal antibodies, Fc fusions, peptides, hormones, and growth factors. PMID:25344235

  14. Cloning and expression of Pectobacterium carotovorum endo-polygalacturonase gene in Pichia pastoris for production of oligogalacturonates

    Science.gov (United States)

    A bacterial endo-polygalacturonase (endo-PGase) gene from the plant pathogen Pectobacterium carotovorum was cloned into pGAPZaA vector and constitutively expressed in Pichia pastoris. The recombinant endo-PGase secreted by the Pichia clone showed a 1.7 fold increase when the culture medium included ...

  15. Moving toward reconciliation in indigenous child welfare.

    Science.gov (United States)

    Auger, Andrea

    2012-01-01

    The Touchstones of Hope reconciliation movement consists of principles (culture and language, self-determination, structural interventions, non discrimination, and holistic approach) that guide a reconciliation process of truth-telling, acknowledging, restoring and relating to reshape indigenous child welfare led by indigenous peoples and supported by their non-indigenous counterparts. This article describes a reconciliation movement in Canada grounded in Touchstones of Hope principles, involving a reconciliation process between indigenous and non-indigenous individuals, which has enabled culturally relevant concepts of child welfare and plans for child safety to emerge.

  16. Specific learning processes and indigenous teacher training

    Directory of Open Access Journals (Sweden)

    Adir Casaro Nascimento

    2012-11-01

    Full Text Available Indigenous teacher formation and the issue of specific learning processes, as a right of the indigenous peoples derived from the 1988 Constitution, aim at the re-signification of pedagogical practices in specific socio-cultural contexts and at the visibility of indigenous education. Taking indigenous children as a reference, or rather, the agents that produce knowledge within the context of their particularities and territorialities, the essay points to the necessity of constructing new theoretical bases and a pedagogy that gives visibility to other local epistemic logics produced by “power coloniality”. They are different from the dominant Western logic in the process of training indigenous educators.

  17. Overexpression of the riboflavin biosynthetic pathway in Pichia pastoris

    Directory of Open Access Journals (Sweden)

    Mattanovich Diethard

    2008-07-01

    Full Text Available Abstract Background High cell density cultures of Pichia pastoris grown on methanol tend to develop yellow colored supernatants, attributed to the release of free flavins. The potential of P. pastoris for flavin overproduction is therefore given, but not pronounced when the yeast is grown on glucose. The aim of this study is to characterize the relative regulatory impact of each riboflavin synthesis gene. Deeper insight into pathway control and the potential of deregulation is established by overexpression of the single genes as well as a combined deregulation of up to all six riboflavin synthesis genes. Results Overexpression of the first gene of the riboflavin biosynthetic pathway (RIB1 is already sufficient to obtain yellow colonies and the accumulation of riboflavin in the supernatant of shake flask cultures growing on glucose. Sequential deregulation of all the genes, by exchange of their native promoter with the strong and constitutive glyceraldehyde-3-phosphate dehydrogenase promoter (PGAP increases the riboflavin accumulation significantly. Conclusion The regulation of the pathway is distributed over more than one gene. High cell density cultivations of a P. pastoris strain overexpressing all six RIB genes allow the accumulation of 175 mg/L riboflavin in the supernatant. The basis for rational engineering of riboflavin production in P. pastoris has thus been established.

  18. Production of human intestinal trefoil factor in Pichia pastoris

    Institute of Scientific and Technical Information of China (English)

    SUN Yong; PENG Xi; Lü Shang-jun; ZHANG Yong; WANG Shi-liang

    2006-01-01

    Objective:To construct a Pichia pastoris (P. Pastoris) expression vector of human intestinal trefoil factor (hITF) and study its expression and purification procedures. Methods:hITF gene encoding mature peptide was modified with a polyhistidine tag sequence at the N-terminal, and then inserted into the P. Pastoris expression vector pGAPZαA at the ownstream of the α-mating factor signal. After gene sequencing, the recombinant pGAPZαA-hITF was transformed into the P. Pastoris strain X-33 with lithium chloride. rhITF was induced to constitutively express in shake flask, and then analyzed with Tricine SDS-PAGE and Western blotting. The obtained rhITF was isolated from the cultured supernatants y ammonium sulfate precipitation, Ni-NTA affinity chromatography, and ultrafiltration. Results:The correctness and integrity of rhITF were identified by restriction digestion and gene sequencing. rhITF was successfully expressed to 50 mg/L as a secretive protein. After purification, the purity was above 95%.Tricine SDS-PAGE and Western-blot analysis howed that rhITF presented as a single band with a molecular weight of 10 kDa, a little larger than 7 879 Da as assayed by mass spectrometry analysis. Conclusion:hITF P. Pastoris expression vector is successfully constructed and rhITF is expressed in P. Pastoris at commercially relevant level. This research lays foundation for the further functional tudying of hITF.

  19. Biotechnological advances towards an enhanced peroxidase production in Pichia pastoris.

    Science.gov (United States)

    Krainer, Florian W; Gerstmann, Michaela A; Darnhofer, Barbara; Birner-Gruenberger, Ruth; Glieder, Anton

    2016-09-10

    Horseradish peroxidase (HRP) is a high-demand enzyme for applications in diagnostics, bioremediation, biocatalysis and medicine. Current HRP preparations are isolated from horseradish roots as mixtures of biochemically diverse isoenzymes. Thus, there is a strong need for a recombinant production process enabling a steady supply with enzyme preparations of consistent high quality. However, most current recombinant production systems are limited at titers in the low mg/L range. In this study, we used the well-known yeast Pichia pastoris as host for recombinant HRP production. To enhance recombinant enzyme titers we systematically evaluated engineering approaches on the secretion process, coproduction of helper proteins, and compared expression from the strong methanol-inducible PAOX1 promoter, the strong constitutive PGAP promoter, and a novel bidirectional promoter PHTX1. Ultimately, coproduction of HRP and active Hac1 under PHTX1 control yielded a recombinant HRP titer of 132mg/L after 56h of cultivation in a methanol-independent and easy-to-do bioreactor cultivation process. With regard to the many versatile applications for HRP, the establishment of a microbial host system suitable for efficient recombinant HRP production was highly overdue. The novel HRP production platform in P. pastoris presented in this study sets a new benchmark for this medically relevant enzyme. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  20. Cloning and Identification of Methionine Synthase Gene from Pichia pastoris

    Institute of Scientific and Technical Information of China (English)

    Lan HUANG; Dong-Yang LI; Shao-Xiao WANG; Shi-Ming ZHANG; Jun-Hui CHEN; Xiang-Fu WU

    2005-01-01

    Methionine synthase (MS) is grouped into two classes. Class One MS (MetH) and Class Two MS (MetE) share no homology and differ in their catalytic model. Based on the conserved sequences of metE genes from different organisms, a segment of the metE gene was first cloned from Pichia pastoris genomic DNA by PCR, and its 5' and 3' regions were further cloned by 5'- and 3'-rapid amplification of cDNA ends (RACE), respectively. The assembled sequence reveals an open reading frame encoding a polypeptide of 768 residues, and the deduced product shares 76% identity with MetE of Saccharomyces cerevisiae. P. pastoris methionine synthase (PpMetE) consists of two domains common to MetEs. The active site is located in the C-terminal domain, in which the residues involved in the interaction of zinc with substrates are conserved. Homologous expression of PpMetE in P. pastoris was achieved, and the heterologous expression of PpMetE in the S. cerevisiae strain XJB3-1D that is MetE-defective restored the growth of the mutant on methionine-free minimal media. The gene sequence has been submitted to GenBank/EMBL/DDBJ under accession No. AY601648.

  1. Crystal Structure of Alcohol Oxidase from Pichia pastoris.

    Directory of Open Access Journals (Sweden)

    Christian Koch

    Full Text Available FAD-dependent alcohol oxidases (AOX are key enzymes of methylotrophic organisms that can utilize lower primary alcohols as sole source of carbon and energy. Here we report the crystal structure analysis of the methanol oxidase AOX1 from Pichia pastoris. The crystallographic phase problem was solved by means of Molecular Replacement in combination with initial structure rebuilding using Rosetta model completion and relaxation against an averaged electron density map. The subunit arrangement of the homo-octameric AOX1 differs from that of octameric vanillyl alcohol oxidase and other dimeric or tetrameric alcohol oxidases, due to the insertion of two large protruding loop regions and an additional C-terminal extension in AOX1. In comparison to other alcohol oxidases, the active site cavity of AOX1 is significantly reduced in size, which could explain the observed preference for methanol as substrate. All AOX1 subunits of the structure reported here harbor a modified flavin adenine dinucleotide, which contains an arabityl chain instead of a ribityl chain attached to the isoalloxazine ring.

  2. Characterization of Human Bone Alkaline Phosphatase in Pichia Pastoris

    Science.gov (United States)

    Malone, Christine C.; Ciszak, Eva; Karr, Laurel J.

    1999-01-01

    A soluble form of human bone alkaline phosphatase has been expressed in a recombinant strain of the methylotrophic yeast Pichia pastoris. We constructed a plasmid containing cDNA encoding for human bone alkaline phosphatase, with the hydrophobic carboxyl terminal portion deleted. Alkaline phosphatase was secreted into the medium to a level of 32mg/L when cultured in shake flasks, and enzyme activity was 12U/mg, as measured by a spectrophotometric assay. By conversion to a fermentation system, a yield of 880mg/L has been achieved with an enzyme activity of 968U/mg. By gel electrophoresis analysis, it appears that greater than 50% of the total protein in the fermentation media is alkaline phosphatase. Although purification procedures are not yet completely optimized, they are expected to include filtration, ion exchange and affinity chromatography. Our presentation will focus on the purification and crystallization results up to the time of the conference. Structural data should provide additional information on the role of alkaline phosphatase in normal bone mineralization and in certain bone mineralization anomalies.

  3. Biocontrol of postharvest Rhizopus decay of peaches with Pichia caribbica.

    Science.gov (United States)

    Xu, Baitian; Zhang, Hongyin; Chen, Keping; Xu, Qin; Yao, Yao; Gao, Hui

    2013-08-01

    A new yeast antagonist, Pichia caribbica, isolated in our laboratory from the soil collected from unsprayed orchards, was evaluated for its biocontrol capability against Rhizopus stolonifer on peaches and the possible mechanisms involved. The decay incidence and lesion diameter of Rhizopus decay of peaches treated by P. caribbica were significantly reduced compared with the control fruits, and the higher the concentration of P. caribbica, the better the efficacy of the biocontrol. Rapid colonization of the yeast in peach wounds stored at 25 °C was observed. In peaches, the activities of peroxidase (POD), catalase (CAT), and phenylalanine ammonia-lyase (PAL) were significantly induced by P. caribbica treatment compared to those of the control fruits. All these results indicated that P. caribbica has a great potential for the development of commercial formulations to control postharvest Rhizopus decay of peaches. Its modes of action were based on competition for space and nutrients with pathogens, inducement of activities of defense-related enzymes such as POD, CAT, and PAL of peaches.

  4. High-Level Expression of Recombinant Bovine Lactoferrin in Pichia pastoris with Antimicrobial Activity

    Science.gov (United States)

    Iglesias-Figueroa, Blanca; Valdiviezo-Godina, Norberto; Siqueiros-Cendón, Tania; Sinagawa-García, Sugey; Arévalo-Gallegos, Sigifredo; Rascón-Cruz, Quintín

    2016-01-01

    In this study, bovine lactoferrin (bLf), an iron-binding glycoprotein considered an important nutraceutical protein because of its several properties, was expressed in Pichia pastoris KM71-H under AOX1 promoter control, using pJ902 as the recombinant plasmid. Dot blotting analysis revealed the expression of recombinant bovine lactoferrin (rbLf) in Pichia pastoris. After Bach fermentation and purification by molecular exclusion, we obtained an expression yield of 3.5 g/L of rbLf. rbLf and predominantly pepsin-digested rbLf (rbLfcin) demonstrated antibacterial activity against Escherichia coli (E. coli) BL21DE3, Staphylococcus aureus (S. aureus) FRI137, and, in a smaller percentage, Pseudomonas aeruginosa (Ps. Aeruginosa) ATCC 27833. The successful expression and characterization of functional rbLf expressed in Pichia pastoris opens a prospect for the development of natural antimicrobial agents produced recombinantly. PMID:27294912

  5. High-Level Expression of Recombinant Bovine Lactoferrin in Pichia pastoris with Antimicrobial Activity

    Directory of Open Access Journals (Sweden)

    Blanca Iglesias-Figueroa

    2016-06-01

    Full Text Available In this study, bovine lactoferrin (bLf, an iron-binding glycoprotein considered an important nutraceutical protein because of its several properties, was expressed in Pichia pastoris KM71-H under AOX1 promoter control, using pJ902 as the recombinant plasmid. Dot blotting analysis revealed the expression of recombinant bovine lactoferrin (rbLf in Pichia pastoris. After Bach fermentation and purification by molecular exclusion, we obtained an expression yield of 3.5 g/L of rbLf. rbLf and predominantly pepsin-digested rbLf (rbLfcin demonstrated antibacterial activity against Escherichia coli (E. coli BL21DE3, Staphylococcus aureus (S. aureus FRI137, and, in a smaller percentage, Pseudomonas aeruginosa (Ps. Aeruginosa ATCC 27833. The successful expression and characterization of functional rbLf expressed in Pichia pastoris opens a prospect for the development of natural antimicrobial agents produced recombinantly.

  6. Characterization of a yam class IV chitinase produced by recombinant Pichia pastoris X-33.

    Science.gov (United States)

    Akond, Muhammad Ali; Matsuda, Yusuke; Ishimaru, Takayuki; Iwai, Ken; Saito, Akira; Kato, Akio; Tanaka, Shuhei; Kobayashi, Jun; Koga, Daizo

    2014-01-01

    A yam (Dioscorea opposita Thunb) class IV chitinase, whose genomic DNA was cloned by Mitsunaga et al. (2004), was produced by the recombinant Pichia pastoris X-33 in high yields such as 66 mg/L of culture medium. The chitinase was purified by column chromatography after Endoglycosidase H treatment and then characterized. It showed properties similar to the original chitinase E purified from the yam tuber reported by Arakane et al. (2000). This Pichia-produced chitinase also showed strong lytic activity against Fusarium oxysporum and Phytophthora nicotianae, wide pH and thermal stability, optimum activity at higher temperature such as 70 °C, and high substrate affinity, indicating that one can use this Pichia-produced yam chitinase as a bio-control agent.

  7. Molecular characterization and technological properties of wine yeasts isolated during spontaneous fermentation of Vitis vinifera L.cv. Narince grape must grown in ancient wine making area Tokat, Anatolia

    Directory of Open Access Journals (Sweden)

    Çelik Zeynep Dilan

    2017-01-01

    Full Text Available Narince is a native white grape variety of Vitis vinifera L grown in Tokat and produces rich and balanced wines often with a greenish yellow tint and delicate fruity flavour. Fermentation by indigenous yeasts may produce wines with complex oenological properties that are unique to specific region. In this study yeast population during alcoholic fermentation of Narince was investigated. Yeasts were identified by PCR-RFLP analysis of the 5.8 ITS rRNA region and sequence information for the D1/D2 domains of the 26S gene. Eight different species belonging to nine genera were identified as: Hanseniaspora uvarum, Hansenispora guilliermondii, Pichia kluyveri, Metschnikowiaspp., Pichia occidentalis, Torulaspora delbrueckii, Candida zemplinina, Lachancea thermotolerance and Saccharomyces cerevisiae. Hanseniaspora guilliermondii, Metschnikowiaspp., Pichia occidentalis and Pichia kluyveri were identified only in the early stage of fermentation. Selected yeasts tested for their physiological traits, ethanol, SO2, temperature, pH tolerance, H2S production, killer and enzymatic activity, fermentation rate, flocculation characteristic, foam, volatile acid and volatile compounds production. Among the yeasts, one,Lachancea thermotolerance and four Saccharomyces cerevisiae strain showed remarkable technological properties and results were compared with those obtained by using commercial starter culture.

  8. Pichia garciniae sp. nov., isolated from a rotten mangosteen fruit (Garcinia mangostana L., Clusiaceae).

    Science.gov (United States)

    Bhadra, Bhaskar; Begum, Zareena; Shivaji, Sisinthy

    2008-11-01

    Ascogenous yeasts were isolated from a decaying mangosteen fruit (Garcinia mangostana L., Clusiaceae). Based on colony morphology and RAPD analysis, the strains were grouped into four groups (G-I, G-II, G-III and G-IV). Phenotypic properties and analysis of the D1/D2 domain sequence of the 26S rRNA gene identified representative members of groups G-II, G-III and G-IV as Pichia manshurica (=Pichia galeiformis), Pichia fermentans and Pichia anomala, respectively. Representatives of group G-I, YS110(T) and YS111, showed similar phenotypic traits, 99.9 % similarity in the sequence of the D1/D2 domain of the 26S rRNA gene and ITS1-5.8S rRNA gene-ITS2 sequence and 92 % DNA-DNA relatedness. Hence, YS110(T) and YS111 are regarded as members of the same species. Based on D1/D2 domain and ITS region sequences, the nearest phylogenetic neighbours of YS110(T) and YS111 were identified as Pichia membranifaciens and P. manshurica. However, YS110(T) differs from these two nearest phylogenetic neighbours by >3 % in the D1/D2 domain sequence and by >18 % with respect to the ITS region sequence. In addition, YS110(T) and YS111 differ from P. membranifaciens NRRL Y-2026(T) and P. manshurica NRRL Y-27978(T) with respect to a number of phenotypic traits. The strains show 52-57 % DNA-DNA relatedness with P. membranifaciens NRRL Y-2026(T) and P. manshurica NRRL Y-27978(T). Strains YS110(T) and YS111 are proposed as two strains of a novel species, for which the name Pichia garciniae sp. nov. is proposed. The type strain is YS110(T) (=NRRL Y-48422(T) =CBS 10758(T)).

  9. Circle of Courage Infusion into the Alberta Indigenous Games 2011

    Science.gov (United States)

    Marchand, Dawn Marie

    2011-01-01

    Thousands of indigenous people from across North America came to the Enoch Cree Nation for the Alberta Indigenous Games, six days of sport, education, and cultural awakening. The vision of the Alberta Indigenous Games is to recognize the value and potential of Indigenous culture and the young people. Activities include sports, indigenous arts,…

  10. Indigenous actinorhizal plants of Australia

    Indian Academy of Sciences (India)

    Nishath K Ganguli; Ivan R Kennedy

    2013-11-01

    Indigenous species of actinorhizal plants of Casuarinaceae, Elaeagnaceae and Rhamnaceae are found in specific regions of Australia. Most of these plants belong to Casuarinaceae, the dominant actinorhizal family in Australia. Many of them have significant environmental and economical value. The other two families with their indigenous actinorhizal plants have only a minor presence in Australia. Most Australian actinorhizal plants have their native range only in Australia, whereas two of these plants are also found indigenously elsewhere. The nitrogen-fixing ability of these plants varies between species. This ability needs to be investigated in some of these plants. Casuarinas form a distinctive but declining part of the Australian landscape. Their potential has rarely been applied in forestry in Australia despite their well-known uses, which are being judiciously exploited elsewhere. To remedy this oversight, a programme has been proposed for increasing and improving casuarinas that would aid in greening more regions of Australia, increasing the soil fertility and the area of wild life habitat (including endangered species). Whether these improved clones would be productive with local strains of Frankia or they need an external inoculum of Frankia should be determined and the influence of mycorrhizal fungi on these clones also should be investigated.

  11. Toxicological evaluation of lactase derived from recombinant Pichia pastoris.

    Science.gov (United States)

    Zou, Shiying; He, Xiaoyun; Liu, Yifei; Chen, Delong; Luo, Yunbo; Huang, Kunlun; Zhang, Wei; Xu, Wentao

    2014-01-01

    A recombinant lactase was expressed in Pichia pastoris, resulting in enzymatic activity of 3600 U/mL in a 5 L fermenter. The lactase product was subjected to a series of toxicological tests to determine its safety for use as an enzyme preparation in the dairy industry. This recombinant lactase had the highest activity of all recombinant strains reported thus far. Acute oral toxicity, mutagenicity, genotoxic, and subchronic toxicity tests performed in rats and mice showed no death in any groups. The lethal dose 50% (LD50) based on the acute oral toxicity study is greater than 30 mL/kg body weight, which is in accordance with the 1500 L milk consumption of a 50 kg human daily. The lactase showed no mutagenic activity in the Ames test or a mouse sperm abnormality test at levels of up to 5 mg/plate and 1250 mg/kg body weight, respectively. It also showed no genetic toxicology in a bone marrow cell micronucleus test at levels of up to 1250 mg/kg body weight. A 90-day subchronic repeated toxicity study via the diet with lactase levels up to 1646 mg/kg (1000-fold greater than the mean human exposure) did not show any treatment-related significant toxicological effects on body weight, food consumption, organ weights, hematological and clinical chemistry, or histopathology compared to the control groups. This toxicological evaluation system is comprehensive and can be used in the safety evaluation of other enzyme preparations. The lactase showed no acute, mutagenic, genetic, or subchronic toxicity under our evaluation system.

  12. Toxicological evaluation of lactase derived from recombinant Pichia pastoris.

    Directory of Open Access Journals (Sweden)

    Shiying Zou

    Full Text Available A recombinant lactase was expressed in Pichia pastoris, resulting in enzymatic activity of 3600 U/mL in a 5 L fermenter. The lactase product was subjected to a series of toxicological tests to determine its safety for use as an enzyme preparation in the dairy industry. This recombinant lactase had the highest activity of all recombinant strains reported thus far. Acute oral toxicity, mutagenicity, genotoxic, and subchronic toxicity tests performed in rats and mice showed no death in any groups. The lethal dose 50% (LD50 based on the acute oral toxicity study is greater than 30 mL/kg body weight, which is in accordance with the 1500 L milk consumption of a 50 kg human daily. The lactase showed no mutagenic activity in the Ames test or a mouse sperm abnormality test at levels of up to 5 mg/plate and 1250 mg/kg body weight, respectively. It also showed no genetic toxicology in a bone marrow cell micronucleus test at levels of up to 1250 mg/kg body weight. A 90-day subchronic repeated toxicity study via the diet with lactase levels up to 1646 mg/kg (1000-fold greater than the mean human exposure did not show any treatment-related significant toxicological effects on body weight, food consumption, organ weights, hematological and clinical chemistry, or histopathology compared to the control groups. This toxicological evaluation system is comprehensive and can be used in the safety evaluation of other enzyme preparations. The lactase showed no acute, mutagenic, genetic, or subchronic toxicity under our evaluation system.

  13. Functional inclusion bodies produced in the yeast Pichia pastoris.

    Science.gov (United States)

    Rueda, Fabián; Gasser, Brigitte; Sánchez-Chardi, Alejandro; Roldán, Mònica; Villegas, Sandra; Puxbaum, Verena; Ferrer-Miralles, Neus; Unzueta, Ugutz; Vázquez, Esther; Garcia-Fruitós, Elena; Mattanovich, Diethard; Villaverde, Antonio

    2016-10-01

    Bacterial inclusion bodies (IBs) are non-toxic protein aggregates commonly produced in recombinant bacteria. They are formed by a mixture of highly stable amyloid-like fibrils and releasable protein species with a significant extent of secondary structure, and are often functional. As nano structured materials, they are gaining biomedical interest because of the combination of submicron size, mechanical stability and biological activity, together with their ability to interact with mammalian cell membranes for subsequent cell penetration in absence of toxicity. Since essentially any protein species can be obtained as IBs, these entities, as well as related protein clusters (e.g., aggresomes), are being explored in biocatalysis and in biomedicine as mechanically stable sources of functional protein. One of the major bottlenecks for uses of IBs in biological interfaces is their potential contamination with endotoxins from producing bacteria. To overcome this hurdle, we have explored here the controlled production of functional IBs in the yeast Pichia pastoris (Komagataella spp.), an endotoxin-free host system for recombinant protein production, and determined the main physicochemical and biological traits of these materials. Quantitative and qualitative approaches clearly indicate the formation of IBs inside yeast, similar in morphology, size and biological activity to those produced in E. coli, that once purified, interact with mammalian cell membranes and penetrate cultured mammalian cells in absence of toxicity. Structurally and functionally similar from those produced in E. coli, the controlled production of IBs in P. pastoris demonstrates that yeasts can be used as convenient platforms for the biological fabrication of self-organizing protein materials in absence of potential endotoxin contamination and with additional advantages regarding, among others, post-translational modifications often required for protein functionality.

  14. Responding to Indigenous Australian Sexual Assault

    Directory of Open Access Journals (Sweden)

    Janya McCalman

    2014-01-01

    Full Text Available Indigenous Australians experience a high prevalence of sexual assault, yet a regional sexual assault service found few Indigenous Australians accessed their services. This prompted exploration of how its services might be improved. A resultant systematic search of the literature is reported in this article. Seven electronic databases and seven websites were systematically searched for peer reviewed and gray literature documenting responses to the sexual assault of Indigenous Australians. These publications were then classified by response type and study type. Twenty-three publications met the inclusion criteria. They included studies of legal justice, media, and community-based and mainstream service responses for Indigenous survivors and perpetrators. We located program descriptions, measurement, and descriptive research, but no intervention studies. There is currently insufficient evidence to confidently prescribe what works to effectively respond to Indigenous Australian sexual assault. The study revealed an urgent need for researchers, Indigenous communities, and services to work together to develop the evidence base.

  15. Emerging Ideas for Innovation in Indigenous Education: A Research Synthesis of Indigenous Educative Roles in Mainstream and Flexi Schools

    Science.gov (United States)

    Shay, Marnee

    2017-01-01

    The Indigenous education agenda in Australia remains focused on mainstream schooling contexts. Although overlooked in Indigenous education discourse, flexi schools appear to be engaging with disproportionately high numbers of Indigenous students and staff. The educative roles of Indigenous peoples in broader Indigenous education discourse are…

  16. Taiwanese Indigenous Knowledge Categories and Their Distribution: A Survey of Indigenous Publications

    OpenAIRE

    Gu-Le-Le Lu; Mei-Mei Wu

    2015-01-01

    In recent years, indigenous knowledge has received significant attention in Taiwan. Yet, due to the lack of a clear definition and framework of indigenous knowledge, government ministries and social organizations at all levels face enormous challenges in legislation and policymaking concerning indigenous knowledge preservation, organization, and transmission. This research intends to analyze the scope of published indigenous knowledge contents in Taiwan. By taking a qualitative ap...

  17. Indigenous Law and Identity Politics in Mexico: Indigenous Men's and Women's Struggles for a Multicultural Nation

    OpenAIRE

    Hernández Castillo, Rosalva Aída

    2002-01-01

    The author approaches the debate on multiculturalism in Mexico emphasizing the tensions that exist among gendered perspectives on identity politics within the indigenous movement and also among Mexican intellectuals. It demonstrates how Indigenous women have played a very important role in confronting the uses of a discourse about indigenous women's rights as arguments against indigenous cultural rights and shows how they are not intrinsically opposed to each other. Their conception of multic...

  18. Indigenous knowledge of animal breeding and breeds

    OpenAIRE

    I. Kohler-Rollefson

    2004-01-01

    Indigenous knowledge of animal breeding (IK-AB) includes concepts and practices used to influence the genetic composition of herds. Indigenous selection is often based on preferences based on physical characteristics, vigor, social and economic insurance. This issue paper summarizes the value of indigenous knowledge and local breeds to achieve agricultural sustainability. Links to IK-AB information are also provided. Available in SANREM office, ES

  19. Plant Provocations: Botanical Indigeneity and (Decolonial Imaginations

    Directory of Open Access Journals (Sweden)

    Narendran Kumarakulasingam

    Full Text Available Abstract Abstract: This paper examines the possibilities and limitations of an emergent global discourse of indigeneity to offer an oppositional praxis in the face of the depredations of settler colonialism in post-apartheid South Africa. Self-conscious articulations of indigeneity, we argue, reveal the fraught relationship between increasingly hegemonic and narrow understandings of the indigenous and the carceral logic of apartheid. We examine this by focusing on the meanings and attachments forged through indigenous plants in two realms: the world of indigenous gardening practised by white suburban dwellers and that of subsistence farming undertaken by rural black women. This juxtaposition reveals that in contrast to the pervasive resurrection of colonial time that defines metropolitan indigenous gardening, the social relations of a subsistence cultivator challenge the confines of colonial temporality, revealing a creative mode of dissent structured around dreams, ancestral knowledge, and the commons. Our exploration of struggles around botanical indigeneity suggests that anticolonial modes of indigeneity do not necessarily inhere in recognisable forms and that studies of the indigenous need to proceed beyond those that bear familial resemblance to emergent global understandings.

  20. THE INDIGENOUS GROUPS AND THE BRAZILIAN SWEETS

    Directory of Open Access Journals (Sweden)

    Mártin César Tempass

    2008-12-01

    Full Text Available In the books of Gilberto Freyre and Câmara Cascudo, that influencied so much the literature about brazilian alimentation, the participation of indigenous groups in the national sweets formation process is negligencied. However, is possible to find in book´s “interlineations” of these two authors valuables informations about indigenous contributions to this process. Starting from these two authors and based in the culinary system notion, this paper quests to situate the role of indigenous groups in the brazilian sweets formation and numbers the possibles causes to invisibility of sweets by indigenous at the culinary formation process.

  1. Preparing to Be Allies: Narratives of Non-Indigenous Researchers Working in Indigenous Contexts

    Science.gov (United States)

    Brophey, Alison; Raptis, Helen

    2016-01-01

    Insensitive research approaches have resulted in damaged relationships between non-Indigenous researchers and Indigenous communities, prompting scholars and funding agencies to call for more culturally compatible research methods. This paper addresses the qualities, skills and knowledge developed by six non-Indigenous researchers as they…

  2. The Impact of Professional Development and Indigenous Education Officers on Australian Teachers' Indigenous Teaching and Learning

    Science.gov (United States)

    Craven, Rhonda G.; Yeung, Alexander Seeshing; Han, Feifei

    2014-01-01

    The study investigated the impact of professional development (PD) in Indigenous teaching on teachers' psychological and behavioural aspects, and Indigenous students' learning engagement. Adopting a multiple-indicator-multiple-indicator-cause model, frequency of PD was found to have positive paths to teachers' self-concept in Indigenous teaching…

  3. Motivation Matters: Profiling Indigenous and Non-Indigenous Students' Motivational Goals

    Science.gov (United States)

    Magson, Natasha R.; Craven, Rhonda G.; Nelson, Genevieve F.; Yeung, Alexander S.; Bodkin-Andrews, Gawaian H.; McInerney, Dennis M.

    2014-01-01

    This research explored gender and cross-cultural similarities and differences in the motivational profiles of Indigenous Papua New Guinean (PNG) and Indigenous and non-Indigenous Australians. Secondary students (N = 1,792) completed self-report motivational measures. Invariance testing demonstrated that the Inventory of School Motivation…

  4. Xylitol production from DEO hydrolysate of corn stover by Pichia stipitis YS-30

    Science.gov (United States)

    Rita C.L.B. Rodrigues; William R. Kenealy; Thomas W. Jeffries

    2011-01-01

    Corn stover that had been treated with vapor-phase diethyl oxalate released a mixture of mono-and oligosaccharides consisting mainly of xylose and glucose. Following overliming and neutralization, a D-xylulokinase mutant of Pichia stipitis, FPL-YS30 (xyl3 -Ä1), converted the stover hydrolysate into xylitol. This research examined the effects of phosphoric or gluconic...

  5. Quantitative metabolomics analysis of amino acid metabolism in recombinant Pichia pastoris under different oxygen availability conditions

    NARCIS (Netherlands)

    Carnicer, M.; Ten Pierick, A.; Van Dam, J.; Heijnen, J.J.; Albiol, J.; Van Gulik, W.; Ferrer, P.

    2012-01-01

    Background: Environmental and intrinsic stress factors can result in the global alteration of yeast physiology, as evidenced by several transcriptional studies. Hypoxia has been shown to have a beneficial effect on the expression of recombinant proteins in Pichia pastoris growing on glucose. Further

  6. Positive selection of novel peroxisome biogenesis-defective mutants of the yeast Pichia pastoris

    NARCIS (Netherlands)

    Johnson, Monique A.; Waterham, Hans R.; Ksheminska, Galyna P.; Fayura, Liubov R.; Cereghino, Joan Lin; Stasyk, Oleh V.; Veenhuis, Marten; Kulachkovsky, Aleksander R.; Sibirny, Andrei A.; Cregg, James M.

    1999-01-01

    We have developed two novel schemes for the direct selection of peroxisome-biogenesis-defective (pex) mutants of the methylotrophic yeast Pichia pastoris. Both schemes take advantage of our observation that methanol-induced pex mutants contain little or no alcohol oxidase (AOX) activity. AOX is a pe

  7. Quantitative metabolomics analysis of amino acid metabolism in recombinant Pichia pastoris under different oxygen availability conditions

    NARCIS (Netherlands)

    Carnicer, M.; Ten Pierick, A.; Van Dam, J.; Heijnen, J.J.; Albiol, J.; Van Gulik, W.; Ferrer, P.

    2012-01-01

    Background: Environmental and intrinsic stress factors can result in the global alteration of yeast physiology, as evidenced by several transcriptional studies. Hypoxia has been shown to have a beneficial effect on the expression of recombinant proteins in Pichia pastoris growing on glucose. Further

  8. Kodamaea (Pichia) ohmeri fungemia in a pediatric patient admitted in a public hospital

    NARCIS (Netherlands)

    De Barros, J.D.; Do Nascimento, S.M.; de Araujo, F.J.; Braz Rde, F.; Andrade, V.S.; Theelen, B.J.F.; Boekhout, T.; Illnait-Zaragozi, M.T.; Gouveia, M.N.; Fernandes, M.C.; Monteiro, M.G.; De Oliveira, M.T.

    2009-01-01

    Kodamaea (Pichia) ohmeri is a yeast species that has not been reported to be a frequent cause of human infections. The current report describes a case of fungemia caused by K. ohmeri in a 3-year-old female patient hospitalized in the public hospital Maria Alice Fernandes, Natal, RN, Brazil. The

  9. The Nature of Indigenized Englishes: Interference--Creativity--Universals.

    Science.gov (United States)

    Platt, John

    1989-01-01

    Examines the concept of indigenized Englishes and compares them with pidgins and creoles, focusing on attitudes about indigenized English, creative aspects of indigenized English, substratum influences, and universals. (Author/CB)

  10. Recognizing appropriate representation of indigenous knowledge in design practice

    National Research Council Canada - National Science Library

    Kelly, Meghan; Kennedy, Russell

    2016-01-01

    ... of indigenous culture. Incorporating indigenous symbols into visual communication design strategies impacts a wide range of stakeholders and therefore requires a sensitive approach with broad consultation in regard...

  11. Towards a critique of indigenous African religion

    Directory of Open Access Journals (Sweden)

    Johan Strijdom

    2011-06-01

    Full Text Available In this article, it is argued that a postcolonial critique of the colonial study of religion should not preclude a critique of indigenous African religion itself. The latter may be developed from a human rights perspective and a critique of exclusionary views of indigeneity. The argument is illustrated by means of specific case studies.

  12. Including People with Disabilities: An Indigenous Perspective

    Science.gov (United States)

    Bevan-Brown, Jill

    2013-01-01

    Being victims of racial prejudice, religious intolerance, poverty, disempowerment and language loss it could be expected that indigenous people would be supportive of the Inclusion Movement with its philosophy of valuing and acceptance of all people. This supposition is examined for Maori, the indigenous people of Aotearoa/New Zealand. In…

  13. Bilingual Discourse Markers in Indigenous Languages

    Science.gov (United States)

    Torres, Lourdes

    2006-01-01

    This review of research considers the occurrence and function of Spanish discourse markers and other particles in indigenous speech. I discuss important research that has examined these phenomena and refer to studies of bilingual discourse markers in other non-indigenous language contact situations to address unresolved issues concerning the form…

  14. Indigenous Rights and Schooling in Highland Chiapas.

    Science.gov (United States)

    Gonzalez, Margaret Freedson; Perez, Elias Perez

    1998-01-01

    Educational reforms in Mexico to preserve indigenous linguistic and cultural rights often originate in Mexico City and lack grassroots support. Although native language instruction improves literacy development and preserves culture, Native parents may reject it because Spanish is the language of status. However, some indigenous communities in…

  15. Positive Educational Responses to Indigenous Student Mobility

    Science.gov (United States)

    Hill, Angela; Lynch, Andrea; Dalley-Trim, Leanne

    2012-01-01

    Engaging positively with the mobility of Indigenous students has been the centre of a 5-year action research project in Queensland, Australia. Drawing on responses developed for other marginalised mobile populations, and with consideration for the extent of mobility amongst many Indigenous people in Australia, this paper focuses on the…

  16. Gambling: A Poison Chalice for Indigenous Peoples'

    Science.gov (United States)

    Dyall, Lorna

    2010-01-01

    Indigenous populations are now being encouraged to be involved in the business of gambling as an operator or if not given that status, are actively encouraged to participate in gambling activities. Research both published and unpublished show that different indigenous populations often have a higher prevalence of problem and pathological gambling…

  17. Indigenous Students in the Tertiary Education Sector

    Science.gov (United States)

    Bandias, Susan; Fuller, Don; Larkin, Steven

    2014-01-01

    Important recent objectives of indigenous education policy in Australia have been aimed at redressing indigenous economic and social disadvantage through increasing student retention, progression and completion rates in both compulsory and post-compulsory education. The two sectors of the tertiary education system, vocational education and…

  18. Performance in Basic Mathematics of Indigenous Students

    Science.gov (United States)

    Sicat, Lolita V.; David, Ma. Elena D.

    2016-01-01

    This analytical study analyzed the performance in Basic Mathematics of the indigenous students, the Aeta students (Grade 6) of Sta. Juliana Elementary School, Capas, Tarlac, and the APC students of Malaybalay City, Bukidnon. Results were compared with regular students in rural, urban, private, and public schools to analyze indigenous students'…

  19. Indigenous Documents Related to the Quincentenary.

    Science.gov (United States)

    Hale, Charles R., Comp.

    1993-01-01

    Three documents of the "500 Years of Resistance" movement, which held continental meetings of indigenous peoples in July 1990 and October 1991, reject the celebration of the Columbus Quincentenary and call for true democracy and human rights for the Americas' indigenous peoples and an end to neocolonialism and social inequality. (SV)

  20. Science, Metaphoric Meaning, and Indigenous Knowledge

    Science.gov (United States)

    Elliott, Frank

    2009-01-01

    Western cultural approaches to teaching science have excluded Indigenous knowledges and culturally favored many non-Aboriginal science students. By asking the question "What connections exist between Western science and Indigenous knowledge?" elements of epistemological (how do we determine what is real?) and ontological (what is real?)…

  1. Weak recognition: Indigenous rights in Chile

    Directory of Open Access Journals (Sweden)

    Claudio Fuentes

    2017-01-01

    Full Text Available After seventeen years of debate, the Chilean Congress approved the ilo 169 Convention on indigenous rights that compels the State to consult indigenous communities on issues that directly affect them. As the political and economic conditions were unfavourable, this political outcome is surprising. Indeed, the legal status of the indigenous people in Chile is weaker than in the rest of Latin America. This article explains this outcome through a detailed description of institutional changes as well as social pressures from the indigenous movement. These factors made right-wing sectors to adapt their discourses in order to accept Chile as a multicultural society. Moreover, a relevant part of the story is related to territorial differences among legislators. Discourse adaptation toward a soft recognition of indigenous rights is a likely outcome in a very conservative environmental setting.

  2. Curriculum and the production of indigenous subjects

    Directory of Open Access Journals (Sweden)

    Adir Casaro Nascimento

    2015-10-01

    Full Text Available The policy on school education has always been explicit in its intentions to produce identities for indigenous peoples. The Federal Constitution of 1988 broke with the assimilationist/integrationist/colonizing proposal of curricula imposed on indigenous people and recognizes the ethnic identities of indigenous people inserted within the context of their cultural relations and the right to their customs, values, traditions, languages and knowledge. The use of the mother tongue and of learning processes has been orienting categories in the curricula of their schools. With culture as the focal point of discussion, this text is supported on testimonies by indigenous teachers from Terena, Guarani and Kaiowá tribes, subjects living the ambiguities and conflicts as well as their identities and the identities of those looking for schooling in different communities. Despite the difficulties that the indigenous school still faces, indigenous movements question homogenous and colonizing schooling models. This school is part of their lives and plays its social role of working with knowledge without excluding cultures as producers of sense and meaning, their knowledge that guarantees the difference in curriculum, area of struggle, and in the production of indigenous subjects.

  3. Revolutionizing Environmental Education through Indigenous Hip Hop Culture

    Science.gov (United States)

    Gorlewski, Julie; Porfilio, Brad J.

    2012-01-01

    Based upon the life histories of six Indigenous hip hop artists of the Beat Nation artist collective, this essay captures how Indigenous hip hop has the potential to revolutionize environmental education. Hip hop provides Indigenous youth an emancipatory space to raise their opposition to neocolonial controls of Indigenous territories that…

  4. Indigenous Student Participation in Higher Education: Emergent Themes and Linkages

    Science.gov (United States)

    Aseron, Johnnie; Wilde, Simon; Miller, Adrian; Kelly, Stephen

    2013-01-01

    Educational processes directed at Indigenous peoples have long propagated a disparity between the educational successes of Indigenous and non-Indigenous students (May 1999), a contrast which can be acutely observed in Australia. It is not surprising, then, that the educational needs of Indigenous students have been poorly served, with the extant…

  5. Revolutionizing Environmental Education through Indigenous Hip Hop Culture

    Science.gov (United States)

    Gorlewski, Julie; Porfilio, Brad J.

    2012-01-01

    Based upon the life histories of six Indigenous hip hop artists of the Beat Nation artist collective, this essay captures how Indigenous hip hop has the potential to revolutionize environmental education. Hip hop provides Indigenous youth an emancipatory space to raise their opposition to neocolonial controls of Indigenous territories that…

  6. An Indigenously Developed Insecticidal Aerosol

    Directory of Open Access Journals (Sweden)

    R. N. Varma

    1969-10-01

    Full Text Available A total of 6 "Test" insecticidal aerosols (TA-I to VI indigenously produced were tested during the years 1966-67 as suitable replacements for imported aerosols.TA-I produced deep yellow staining and a yellowish spray mist. Its capacity was only 120 ml fluid. TA-III types II and III containing modified aerosol formulation with "Esso solvent 3245" and mineral turpentine oil (Burmah Shelland Freon 12 11 (all indigenouswere comparable to he "SRA" in insecticidial efficacy. The container was also manufactured in the country and it compared well with the "SRA" in construction, resistance against rough usage and mechanical function. They were both finally approved for introduction in the services as replacement for imported aerosols. TA-IV performed well in inscticidial assessment, but the aerosols formulation. TA-V and VI were similar to TA-III types II and III respectively.

  7. Doing Climate Science in Indigenous Communities

    Science.gov (United States)

    Pandya, R. E.; Bennett, B.

    2009-12-01

    Historically, the goal of broadening participation in the geosciences has been expressed and approached from the viewpoint of the majority-dominated geoscience community. The need for more students who are American Indian, Native Hawaiian, or Alaska Native is expressed in terms of the need to diversify the research community, and strategies to engage more students are often posed around the question “what can we do to get more indigenous students interested in coming to our institutions to do geosciences?” This approach can lead to neglecting indigenous ways of knowing, inadvertently prioritizes western values over traditional ones, and doesn’t necessarily honor tribal community’s desire to hold on to their talented youth. Further, while this approach has resulted in some modest success, the overall participation in geoscience by students from indigenous backgrounds remains low. Many successful programs, however, have tried an alternate approach; they begin by approaching the geosciences from the viewpoint of indigenous communities. The questions they ask center around how geosciences can advance the priorities of indigenous communities, and their approaches focus on building capacity for the geosciences within indigenous communities. Most importantly, perhaps, these efforts originate in Tribal communities themselves, and invite the geoscience research community to partner in projects that are rooted in indigenous culture and values. Finally, these programs recognize that scientific expertise is only one among many skills indigenous peoples employ in their relation with their homelands. Climate change, like all things related to the landscape, is intimately connected to the core of indigenous cultures. Thus, emerging concerns about climate change provide a venue for developing new, indigenous-centered, approaches to the persistent problem of broadening participation in the geoscience. This presentation will highlight three indigenous-led efforts in to

  8. Heterologous Expression of Mycobacterium tuberculosis Ag85B in Pichia pastoris

    Institute of Scientific and Technical Information of China (English)

    LIU Yan; PENG Jian-hong; MA Li-xin; JU Wei; QU Xue-ju; LIU Jun-yan

    2005-01-01

    The DNA fragment encoding mature Mycobacterium tuberculosis major secretory protein Ag85B was inserted into the Pichia pastoris secretory expression vector pHBM905A, under the control of the AOX1 promoter. The recombinant plasmid pHBM905A-85B linearized by Sal Ⅰ was introduced into Pichia pastoris strain GS115 by PEG1000 transformation method. After phenotype screening and PCR identification, the resulting GSll5-pHBM905A-85B strain was cultivated and induced with methanol. The recombinant Ag85B protein in secreted form was attained with molecular weight of 35×103 approximately detected by SDS-PAGE and Western blot. ELISA experiment proved that the protein had good antigen specificity. Secretory expression of recombinant M. tuberculosis Ag85B in P. pastoris will open a door to mass production of the protein in heterologous host and allow ready evaluation of its immunological function.

  9. Gene cloning, expression, purification and characterization of lipoprotein- associated phospholipase A2 in Pichia pastoris

    Institute of Scientific and Technical Information of China (English)

    Fu-junZHANG; Yi-pingWANG

    2005-01-01

    AIM To express and purify Lipoprotein -associated phospholipase A2 (Lp-PLA2), and to establish a screening model for Lp-PLA2 inhibitors through the recombinant Lp-PLA2. METHODS The full-length gene of Lp-PLA2 was cloned from the differentiated THP-1 cells by RT-PCR and PCR. The Lp-PLA2 gene was subcloned into the Pichia expression vector pPIC9 and introduced a sequence encoding a C-terminal stretch of six histidine residues at the same time. The recombinant plasmid was transformed into Pichia pastoris GS115 by spheroplasting and the gene was then integrated into the GS115 genome. Lp- PLA2 was expressed in the yeast strain GS115 by inducing with 0.5% methanol.

  10. Structural and functional characterization of S-adenosylmethionine (SAM) synthetase from Pichia ciferrii.

    Science.gov (United States)

    Yoon, Sangyoung; Lee, Wonkyu; Kim, Minsoo; Kim, T Doohun; Ryu, Yeonwoo

    2012-01-01

    S-adenosylmethionine synthetase (SAM-s) catalyzes the synthesis of S-adenosylmethionine (SAM), which is essential for methylation, transcription, proliferation, and production of secondary metabolites. Here SAM-s from Pichia ciferrii were selectively cloned using RNA CapFishing and rapid amplification of cDNA ends (RACE). The putative full-length cDNA of SAM-s encoded a 383 amino acid protein (42.6 kDa), which has highly conserved metal binding sites, a phosphate-binding site, and functionally important motifs. The corresponding enzyme was over-expressed in a heterologous host of Pichia pastoris, and then purified to a homogenous form. Enzyme kinetics, immunoblotting, circular dichroism (CD), high performance liquid chromatography (HPLC), and molecular modeling were conducted to characterize the SAM-s from P. ciferrii. Structural and functional studies of SAM-s will provide important insights for industrial applications.

  11. [Optimization on the production of analgesic peptide from Buthus martensii Karsch in Pichia pastoris].

    Science.gov (United States)

    Yang, Jin-ling; He, Hui-xia; Zhu, Hui-xin; Cheng, Ke-di; Zhu, Ping

    2009-01-01

    The technology of liquid fermentation for producing the recombinant analgesic peptide BmK AngM1 from Buthus martensii Karsch in Pichia pastoris was studied by single-factor and orthogonal test. The results showed that the optimal culture conditions were as follows: 1.2% methanol, 0.6% casamino acids, initial pH 6.0, and three times of basal inoculation volume. Under the above culture conditions, the expression level of recombinant BmK AngM1 in Pichia pastoris was above 500 mg x L(-1), which was more than three times of the control. The study has laid a foundation for the large-scale preparation of BmK AngM1 to meet the needs of theoretical research of BmK AngM1 and development of new medicines.

  12. Expression and analysis of the glycosylation properties of recombinant human erythropoietin expressed in Pichia pastoris

    Directory of Open Access Journals (Sweden)

    Ser Huy Teh

    2011-01-01

    Full Text Available The Pichia pastoris expression system was used to produce recombinant human erythropoietin, a protein synthesized by the adult kidney and responsible for the regulation of red blood cell production. The entire recombinant human erythropoietin (rhEPO gene was constructed using the Splicing by Overlap Extension by PCR (SOE-PCR technique, cloned and expressed through the secretory pathway of the Pichia expression system. Recombinant erythropoietin was successfully expressed in P. pastoris. The estimated molecular mass of the expressed protein ranged from 32 kDa to 75 kDa, with the variation in size being attributed to the presence of rhEPO glycosylation analogs. A crude functional analysis of the soluble proteins showed that all of the forms were active in vivo.

  13. Not all semantics: similarities and differences in reminiscing function and content between Indigenous and non-Indigenous Australians.

    Science.gov (United States)

    Nile, Emma; Van Bergen, Penny

    2015-01-01

    This study explored why and how Indigenous and non-Indigenous Australians remember the past. Indigenous Australians traditionally share a strong oral tradition in which customs, personal and cultural histories, and other narratives are passed across groups and between generations by word of mouth. Drawing on this tradition, in which inherent value is placed on sharing knowledge and maintaining connectedness with others, we hypothesised that Indigenous Australians would be more likely than non-Indigenous Australians to report reminiscing to fulfil social functions (but not self or directive functions). Furthermore, we hypothesised that Indigenous Australians would recall personal past experiences more elaborately than would non-Indigenous Australians. In Study 1, 33 Indigenous Australians and 76 non-Indigenous Australians completed Webster's Reminiscence Functions Scale. As predicted, Indigenous participants reported higher scores on subscales related to social functions than did non-Indigenous Australians: particularly "Teach/Inform" and "Intimacy Maintenance". They also scored higher on the "Identity" subscale. In Study 2, 15 Indigenous and 14 non-Indigenous Australians shared three memories from the distant and recent past. While Indigenous and non-Indigenous narratives did not differ in either emotion or elaboration, Indigenous Australians provided more memory context and detail by including a greater proportion of semantic memory content. Taken together, these findings suggest differences in both why and how Australians remember.

  14. Recombinants proteins for industrial uses: utilization of Pichia pastoris expression system

    OpenAIRE

    Claudia Rabert; Daniel Weinacker; Adalberto Pessoa Jr; Farías,Jorge G.

    2013-01-01

    The innovation in industrial process with impact in the efficient production is the major challenge for actual industry. A high numerous of enzymes are utilized in at different level of process; the search for new alternatives with better characteristic has become a field of study of great interest, the recombinant protein achievement in a different host system is an alternative widely assessed for production of this. The microorganism Pichia pastoris has been used like a successful expressio...

  15. Lower Temperature Cultures Enlarge the Effects of Vitreoscilla Hemoglobin Expression on Recombinant Pichia pastoris

    OpenAIRE

    Wei-Chang Fu; Shin-Yao Wang; Jyh-Ming Wu

    2012-01-01

    An heterologous expression of Vitreoscilla hemoglobin (VHb) for improving cell growth and recombinant protein production has been successfully demonstrated in various hosts, including Pichia pastoris. Lower temperature cultures can enhance target protein production in some studies of P. pastoris. In this study, the strategy of combining heterologous VHb expression and lower temperature cultures in P. pastoris showed that final cell density and viability of VHb+ strain at 23 °C were hi...

  16. Enhanced production of recombinant secretory proteins in Pichia pastoris by optimizing Kex2 P1' site

    OpenAIRE

    Song Yang; Ye Kuang; Hongbo Li; Yuehong Liu; Xiaoyan Hui; Peng Li; Zhiwu Jiang; Yulai Zhou; Yu Wang; Aimin Xu; Shiwu Li; Pentao Liu; Donghai Wu

    2013-01-01

    Pichia pastoris is one of the most widely used expression systems for the production of recombinant secretory proteins. Its universal application is, however, somewhat hampered by its unpredictable yields for different heterologous proteins, which is now believed to be caused in part by their varied efficiencies to traffic through the host secretion machinery. The yeast endoprotease Kex2 removes the signal peptides from pre-proteins and releases the mature form of secreted proteins, thus, pla...

  17. A multi-level study of recombinant Pichia pastoris in different oxygen conditions

    OpenAIRE

    Gasser Brigitte; Maaheimo Hannu; Jouhten Paula; Stadlmann Johannes; Graf Alexandra B; Dragosits Martin; Carnicer Marc; Baumann Kristin; Albiol Joan; Mattanovich Diethard; Ferrer Pau

    2010-01-01

    Abstract Background Yeasts are attractive expression platforms for many recombinant proteins, and there is evidence for an important interrelation between the protein secretion machinery and environmental stresses. While adaptive responses to such stresses are extensively studied in Saccharomyces cerevisiae, little is known about their impact on the physiology of Pichia pastoris. We have recently reported a beneficial effect of hypoxia on recombinant Fab secretion in P. pastoris chemostat cul...

  18. High-level expression of the native barley alpha-amylase/subtilisin inhibitor in Pichia pastoris

    DEFF Research Database (Denmark)

    Micheelsen, Pernille Ollendorff; Ostergaard, Peter Rahbek; Lange, Lene

    2008-01-01

    An expression system for high-level expression of the native Hordeum vulgare alpha-amylase/subtilisin inhibitor (BASI) has been developed in Pichia pastoris, using the methanol inducible alcohol oxidase 1 (AOX1) promoter. To optimize expression, two codon-optimized coding regions have been design...... and characterized by Edman degradation, liquid chromatography mass spectrometry and insoluble blue starch assay, and was shown to possess the same characteristics as wild-type protein purified from barley grains....

  19. Integrative Transformation System for the Metabolic Engineering of the Sphingoid Base-Producing Yeast Pichia ciferrii

    OpenAIRE

    Bae, Jung-Hoon; Sohn, Jung-Hoon; Park, Chang-Seo; Rhee, Joon-Shick; Choi, Eui-Sung

    2003-01-01

    We have developed an integrative transformation system for metabolic engineering of the tetraacetyl phytosphingosine (TAPS)-secreting yeast Pichia ciferrii. The system uses (i) a mutagenized ribosomal protein L41 gene of P. ciferrii as a dominant selection marker that confer resistance to the antibiotic cycloheximide and (ii) a ribosomal DNA (rDNA) fragment of P. ciferrii as a target for multicopy gene integration into the chromosome. A locus within the nontranscribed region located between 5...

  20. Toward systems metabolic engineering of Aspergillus and Pichia species for the production of chemicals and biofuels

    DEFF Research Database (Denmark)

    Caspeta, Luis; Nielsen, Jens

    2013-01-01

    trends in systems biology of Aspergillus and Pichia species, highlighting the relevance of these developments for systems metabolic engineering of these organisms for the production of hydrolytic enzymes, biofuels and chemicals from biomass. Metabolic engineering is moving from traditional methods...... for the production of hydrolytic enzymes, biofuels and chemicals from biomass. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim....

  1. A Rapid Method for Determining the Concentration of Recombinant Protein Secreted from Pichia pastoris

    Science.gov (United States)

    Sun, L. W.; Zhao, Y.; Niu, L. P.; Jiang, R.; Song, Y.; Feng, H.; feng, K.; Qi, C.

    2011-02-01

    Pichia secretive expression system is one of powerful eukaryotic expression systems in genetic engineering, which is especially suitable for industrial utilization. Because of the low concentration of the target protein in initial experiment, the methods and conditions for expression of the target protein should be optimized according to the protein yield repetitively. It is necessary to set up a rapid, simple and convenient analysis method for protein expression levels instead of the generally used method such as ultrafiltration, purification, dialysis, lyophilization and so on. In this paper, acetone precipitation method was chosen to concentrate the recombinant protein firstly after comparing with four different protein precipitation methods systematically, and then the protein was analyzed by SDS-Polyacrylamide Gel Electrophoresis. The recombinant protein was determined with the feature of protein band by the Automated Image Capture and 1-D Analysis Software directly. With this method, the optimized expression conditions of basic fibroblast growth factor secreted from pichia were obtained, which is as the same as using traditional methods. Hence, a convenient tool to determine the optimized conditions for the expression of recombinant proteins in Pichia was established.

  2. Biosynthesis of silver and zinc oxide nanoparticles using Pichia fermentans JA2 and their antimicrobial property

    Science.gov (United States)

    Chauhan, Ritika; Reddy, Arpita; Abraham, Jayanthi

    2015-01-01

    The development of eco-friendly alternative to chemical synthesis of metal nanoparticles is of great challenge among researchers. The present study aimed to investigate the biological synthesis, characterization, antimicrobial study and synergistic effect of silver and zinc oxide nanoparticles against clinical pathogens using Pichia fermentans JA2. The extracellular biosynthesis of silver and zinc oxide nanoparticles was investigated using Pichia fermentans JA2 isolated from spoiled fruit pulp bought in Vellore local market. The crystalline and stable metallic nanoparticles were characterized evolving several analytical techniques including UV-visible spectrophotometer, X-ray diffraction pattern analysis and FE-scanning electron microscope with EDX-analysis. The biosynthesized metallic nanoparticles were tested for their antimicrobial property against medically important Gram positive, Gram negative and fungal pathogenic microorganisms. Furthermore, the biosynthesized nanoparticles were also evaluated for their increased antimicrobial activities with various commercially available antibiotics against clinical pathogens. The biosynthesized silver nanoparticles inhibited most of the Gram negative clinical pathogens, whereas zinc oxide nanoparticles were able to inhibit only Pseudomonas aeruginosa. The combined effect of standard antibiotic disc and biosynthesized metallic nanoparticles enhanced the inhibitory effect against clinical pathogens. The biological synthesis of silver and zinc oxide nanoparticles is a novel and cost-effective approach over harmful chemical synthesis techniques. The metallic nanoparticles synthesized using Pichia fermentans JA2 possess potent inhibitory effect that offers valuable contribution to pharmaceutical associations.

  3. Pichia pastoris expressed EtMic2 protein as a potential vaccine against chicken coccidiosis.

    Science.gov (United States)

    Zhang, Jie; Chen, Peipei; Sun, Hui; Liu, Qing; Wang, Longjiang; Wang, Tiantian; Shi, Wenyan; Li, Hongmei; Xiao, Yihong; Wang, Pengfei; Wang, Fangkun; Zhao, Xiaomin

    2014-09-15

    Chicken coccidiosis caused by Eimeria species leads to tremendous economic losses to the avian industry worldwide. Identification of parasite life cycle specific antigens is a critical step in recombinant protein vaccine development against Eimeria infections. In the present study, we amplified and cloned the microneme-2 (EtMIC2) gene from Eimeria tenella wild type strain SD-01, and expressed the EtMic2 protein using Pichia pastoris and Escherichia coli expression systems, respectively. The EtMic2 proteins expressed by P. pastoris and E. coli were used as vaccines to immunize chickens and their protective efficacies were compared and evaluated. The results indicated that both P. pastoris and E. coli expressed EtMic2 proteins exhibited good immunogenicity in stimulating host immune responses and the Pichia expressed EtMic2 provided better protection than the E. coli expressed EtMic2 did by significantly increasing growth rate, inducing high specific antibody response, reducing the oocyst output and cecal lesions. Particularly, the Pichia expressed EtMic2 protein exhibited much better ability in inducing cell mediated immune response than the E. coli expressed EtMic2. Copyright © 2014 Elsevier B.V. All rights reserved.

  4. Genetic improvement in indigenous chicken of Ethiopia

    NARCIS (Netherlands)

    Woldegiorgiss, W.E.

    2015-01-01

    Abstract Wondmeneh Esatu Woldegiorgiss (2015). Genetic improvement in indigenous chicken of Ethiopia. PhD thesis, Wageningen University, the Netherlands This thesis considered various approaches to study the potential for improvement of village poultry production system using

  5. Phenotypic characterization of Ethiopian indigenous goat populations

    African Journals Online (AJOL)

    PROMOTING ACCESS TO AFRICAN RESEARCH ... A survey was conducted to study the morphological characteristics of indigenous goats in ... Both qualitative and quantitative traits were recorded on 158 goats randomly sampled from six ...

  6. Curriculum enrichment through indigenous Zulu games | Roux |

    African Journals Online (AJOL)

    Curriculum enrichment through indigenous Zulu games. ... Abstract. In the South African context, education has emerged from missionary, colonial and. Afrikaner ... qualitative data (focus group discussions, interviews, video and audio tape

  7. Soil sustainability and indigenous soil management practices ...

    African Journals Online (AJOL)

    Soil sustainability and indigenous soil management practices among food crop farmers in Ogun State, Nigeria. ... Journal of Environmental Extension ... describe and analyse the current soil management practices among food crop farmers in ...

  8. SEMIOTICS IN INDIGENOUS DANCE PERFORMANCES: EKELEKE ...

    African Journals Online (AJOL)

    Dean SPGS NAU

    Introduction. Indigenous .... Charles. Peirce in his contextual analysis of representation in its physical .... essential information for understanding a dance context and also ..... based the Sense Making Theory, the Ekeleke dancers motivate.

  9. INDIGENOUS KNOWLEDGE IN THE MANAGEMENT OF A ...

    African Journals Online (AJOL)

    Apusigah

    appreciation of the role of indigenous knowledge in the process. Specifically in ... subsequent generations. Also .... Passing on knowledge and skills to the next .... such ethics need not come from the god; rather, society can cultivate them. Fifth ...

  10. Traditional uses of indigenous tree species

    African Journals Online (AJOL)

    Mo

    extinction. It is thus important to promote sustainable management and use of Uganda's indigenous tree species ... Except for picking, the rest of the methods used during harvesting are destructive to the entire plant. ... Thus there is a danger of.

  11. Emancipatory Indigenous Knowledge Systems: implications for ...

    African Journals Online (AJOL)

    Erna Kinsey

    implications for environmental education in South Africa. M.W. Maila ... This failure can be attributed purely to the lower status accorded to this type of knowledge in society. .... further argue that “indigenous information systems are dynamic and.

  12. Inhibitory effect of acetic acid on bioconversion of xylose in xylitol by Candida guilliermondii in sugarcane bagasse hydrolysate Efeito inibitório do ácido acético na bioconversão de xilose em xilitol por Candida guilliermondii em hidrolisado de bagaço de cana

    Directory of Open Access Journals (Sweden)

    Débora D.V. Silva

    2004-09-01

    Full Text Available Sugarcane bagasse hydrolysate (initial acetic acid concentration = 3.5g/L, was used as a fermentation medium for conversion of xylose into xylitol by the yeast Candida guilliermondii FTI 20037. Acetic acid (2.0g/L was added to the medium at different times of fermentation, with the aim of evaluating its effects on the bioconversion process. The addition of acetic acid to the medium after 12h of fermentation resulted in the strongest inhibition of the yeast metabolism. In this case, the xylose consumption and cell growth were, respectively, 23.22 and 11.24% lower than when acid was added to the medium at the beginning of fermentation. As a consequence of the inhibitory effect, lower values of the xylitol yield (0.39g/g and productivity (0.22g/L.h were observed, corresponding to a reduction of 36 and 48%, respectively, in relation to the values obtained with the addition of acetic acid after other fermentation times. The results obtained allowed to conclude that, under the experimental conditions employed in this work, the inhibitory effect of acetic acid on the xylose-xylitol bioconversion depends on the fermentation time when this acid was added, and not only on its concentration in the medium.Hidrolisado de bagaço de cana-de-açúcar contendo uma concentração inicial de ácido acético de 3,5g/L foi utilizado como meio de fermentação para a bioconversão de xilose em xilitol pela levedura Candida guilliermondii FTI 20037. Ácido acético (2,0g/L foi adicionado ao meio em diferentes tempos de fermentação, com o objetivo de avaliar o efeito deste ácido neste bioprocesso. O maior efeito inibitório deste ácido na bioconversão de xilose em xilitol pela levedura ocorreu quando este foi adicionado ao meio após 12h de fermentação. Nesta condição observou-se uma redução de 23,22% e 11,24%, respectivamente, no consumo de xilose e no crescimento celular em relação à fermentação em que a adição deste ácido ocorreu no tempo inicial

  13. Exporting by Migrants and Indigenous Entrepreneurs

    DEFF Research Database (Denmark)

    Ashourizadeh, Shayegheh; Schøtt, Thomas; Pişkinsüt Şengüler, Ece

    2016-01-01

    Migrants may become entrepreneurs in their host countries. They may utilize their dual embeddedness in both the home country and the host country, and also use transnational links to gain a competitive advantage in exporting compared to indigenous entrepreneurs. Migrant entrepreneurs’ advantage may...... around the world was surveyed in the Global Entrepreneurship Monitor, which reports on migration and exporting. Hierarchical linear modeling shows that migrant entrepreneurs export more than indigenous entrepreneurs, especially in the first generation, and especially among educated and male migrants...

  14. EFFECTS OF IMMOBILIZATION IN Ba-ALGINATE ON NITRILE-DEPENDENT OXYGEN UPTAKE RATES OF CANDIDA GUILLIERMONDII Efeitos da imobilização em Ba-alginato na taxa de remoção de oxigênio nitrila-dependente pelas células de Candida guilliermondii

    Directory of Open Access Journals (Sweden)

    João Carlos Teixeira Dias

    2001-10-01

    Full Text Available Yeast cells immobilized by entrapment in Ba-alginate gel were investigated for growth pattern and respiratory activity. The oxygen uptake rates (OUR of cells entrapped in gels with 4% alginate were 5.2 and 23% lower than the OUR of 2% alginate and free cells, respectively. The mass-transfer resistance offered by the matrix and growth of the entrapped cells determine a gradient of nutrients throughout the gel which is responsible for both a lower specific growth rate of immobilized cells with respect to that of free ones, and a heterogeneous biomass distribution, with progressively increasing cellular density from the inside to the outside of the matrix. Gel-matrix polymer concentration affected the maximum oxygen uptake of immobilized growing yeast cells.Foram estudados os efeitos da imobilização no crescimento e na atividade respiratória das células de Candida guilliermondii UFMG-Y65 imobilizadas por encapsulamento, utilizando-se gel de Ba-alginato. As taxas máximas de utilização de oxigênio apresentadas pelas células imobilizadas em matriz contendo 4% de alginato foram 5,2 e 23% inferiores às taxas apresentadas pelas células imobilizadas em 2% de alginato e livres, respectivamente A resistência à transferência de massa oferecida pela matriz e o crescimento das células foram responsáveis pela baixa taxa de crescimento. Como conseqüência, observou-se uma distribuição heterogênea da biomassa, com aumento da densidade celular progressivamente do interior para regiões periféricas da matriz. A concentração do polímero afetou a taxa de utilização máxima de oxigênio pelas células da levedura imobilizada.

  15. Immunisation issues for Indigenous Australian children.

    Science.gov (United States)

    Menzies, Robert; Andrews, Ross

    2014-10-01

    Vaccination has provided major benefits to the health of indigenous children in the face of continuing poorer socioeconomic conditions but several issues have been identified for improvement. While indigenous children are vaccinated at high rates for the standard schedule vaccines, vaccination is more commonly delayed. Coverage for 'targeted' vaccines is substantially lower, and data on coverage for indigenous adolescents is non-existent. Improved identification of indigenous clients by immunisation providers and the expansion of the childhood register are required. The progressive removal of early-acting Haemophilus influenzae type b vaccines from schedules for indigenous children because of an international shortage raises the risk of disease re-emergence and highlights the need for vigilant surveillance including carriage. The expanded use of existing vaccines (influenza) and early adoption of new vaccines (higher valency pneumococcal conjugates) are needed to maximise benefits, in particular the potential to impact on non-invasive disease such as otitis media and non-bacteraemic pneumonia that are so prevalent in indigenous children.

  16. Codon optimization, promoter and expression system selection that achieved high-level production of Yarrowia lipolytica lipase in Pichia pastoris.

    Science.gov (United States)

    Zhou, Wen-Jing; Yang, Jiang-Ke; Mao, Lin; Miao, Li-Hong

    2015-04-01

    Lipase (EC 3.1.1.3) stands amongst the most important and promising biocatalysts for industrial applications. In this study, in order to realize a high-level expression of the Yarrowia lipolytica lipase gene in Pichia pastoris, we optimized the codon of LIP2 by de novo gene design and synthesis, which significantly improved the lipase expression when compared to the native lip2 gene. We also comparatively analyzed the effects of the promoter types (PAOX1 and PFLD1) and the Pichia expression systems, including the newly developed PichiaPink system, on lipase production and obtained the optimal recombinants. Bench-top scale fermentation studies indicated that the recombinant carrying the codon-optimized lipase gene syn-lip under the control of promoter PAOX1 has a significantly higher lipase production capacity in the fermenter than other types of recombinants. After undergoing methanol inducible expression for 96h, the wet cell weight of Pichia, the lipase activity and the protein content in the fermentation broth reached their highest values of 262g/L, 38,500U/mL and 2.82g/L, respectively. This study has not only greatly facilitated the bioapplication of lipase in industrial fields but the strategies utilized, such as de novo gene design and synthesis, the comparative analysis among promoters and different generations of Pichia expression systems will also be useful as references for future work in this field.

  17. Bioethanol Production from Indigenous Algae

    Directory of Open Access Journals (Sweden)

    Madhuka Roy

    2015-02-01

    Full Text Available Enhanced rate of fossil fuel extraction is likely to deplete limited natural resources over short period of time. So search for alternative fuel is only the way to overcome this problem of upcoming energy crisis. In this aspect biofuel is a sustainable option. Agricultural lands cannot be compromised for biofuel production due to the requirement of food for the increasing population. Certain species of algae can produce ethanol during anaerobic fermentation and thus serve as a direct source for bioethanol production. The high content of complex carbohydrates entrapped in the cell wall of the microalgae makes it essential to incorporate a pre-treatment stage to release and convert these complex carbohydrates into simple sugars prior to the fermentation process. There have been researches on production of bioethanol from a particular species of algae, but this work was an attempt to produce bioethanol from easily available indigenous algae. Acid hydrolysis was carried out as pre-treatment. Gas Chromatographic analysis showed that 5 days’ fermentation by baker’s yeast had yielded 93% pure bioethanol. The fuel characterization of the bioethanol with respect to gasoline showed comparable and quite satisfactory results for its use as an alternative fuel.DOI: http://dx.doi.org/10.3126/ije.v4i1.12182International Journal of Environment Volume-4, Issue-1, Dec-Feb 2014/15, page: 112-120  

  18. Gut indigenous microbiota and epigenetics

    Directory of Open Access Journals (Sweden)

    Boris Arkadievich Shenderov

    2012-03-01

    Full Text Available This review introduces and discusses data regarding fundamental and applied investigations in mammalian epigenomics and gut microbiota received over the last 10 years. Analysis of these data enabled the author first to come to the conclusion that the multiple low molecular weight substances of indigenous gut microbiota origin should be considered one of the main endogenous factors actively participating in epigenomic mechanisms that responsible for the mammalian genome reprogramming and post-translated modifications. Gut microecological imbalance coursed by various biogenic and abiogenic agents and factors can produce the different epigenetic abnormalities and the onset and progression of metabolic diseases associated. The author substantiates the necessity to create an international project ‘Human Gut Microbiota and Epigenomics’ that facilitates interdisciplinary collaborations among scientists and clinicians engaged in host microbial ecology, nutrition, metagenomics, epigenomics and metabolomics investigations as well as in diseases prevention and treatment. Some priority scientific and applied directions in the current omic technologies coupled with gnotobiological approaches are suggested that can open a new era in characterizing the role of the symbiotic microbiota small metabolic and signal molecules in the host epigenomics. Although discussed subject is only at an early stage its validation can open novel approaches in drug discovery studies.

  19. [Eating characteristics of Chilean indigenous and non-indigenous adolescent girls].

    Science.gov (United States)

    Araneda, Jacqueline; Amigo, Hugo; Bustos, Patricia

    2010-03-01

    During childhood and adolescence, eating habits become established which are instrumental in determining eating behavior later in life. Various authors have described the acculturation of the Mapuche people toward Western culture. The objective of this study was to analyze the eating characteristics of indigenous and non-indigenous adolescent girls in the Araucania Region of Chile. A cross-sectional design was used with a probabilistic sample of 281 adolescents comprised of 139 indigenous and 142 non-indigenous girls attending 168 elementary schools. A modified food frequency questionnaire was applied, designed to obtain information about eating habits and consumption of Mapuche foods. The eating schedules are similar in both ethnic groups, with dinner being the meal that is least consumed. Total snack consumption per week has a mean of 7 with an interquartile range (IQR) of 5 to 10 without any differences between ethnic groups; of these snacks, only 2 were healthy (IQR = 1 to 3). The indigenous girls had a higher probability of consumption of native foods including mote (boiled wheat) (OR = 2.00; IC = 0.93-4.29), muday (fermented cereal alcohol) (OR = 3.45; IC = 1.90-6.27), and yuyo (field mustard) (OR = 4.40; IC = 2.06-9.39). The study's conclusion is that the the eating habits and behavior of indigenous adolescents are similar to those of non-indigenous girls, though the former still consume more indigenous foods.

  20. Indigenous Economies, Theories of Subsistence, and Women: Exploring the Social Economy Model for Indigenous Governance

    Science.gov (United States)

    Kuokkanen, Rauna

    2011-01-01

    The significance of traditional economies in indigenous communities goes beyond the economic realm--they are more than just livelihoods providing subsistence and sustenance to individuals or communities. The centrality of traditional economies to indigenous identity and culture has been noted by numerous scholars. However, today one can detect a…

  1. Is disaster “normal” for indigenous people? Indigenous knowledge and coping practices

    NARCIS (Netherlands)

    Hilhorst, Dorothea; Baart, Judith; Haar, van der Gemma; Leeftink, Floor Maria

    2015-01-01

    Purpose – The purpose of this paper is to contribute to debates on the value of indigenous knowledge for disaster risk reduction. Recent international policy papers advocate the importance of indigenous knowledge and calls for its recognition. The paper aims to explore these issues in the everyda

  2. Is disaster “normal” for indigenous people? Indigenous knowledge and coping practices

    NARCIS (Netherlands)

    Hilhorst, Dorothea; Baart, Judith; Haar, van der Gemma; Leeftink, Floor Maria

    2015-01-01

    Purpose – The purpose of this paper is to contribute to debates on the value of indigenous knowledge for disaster risk reduction. Recent international policy papers advocate the importance of indigenous knowledge and calls for its recognition. The paper aims to explore these issues in the everyda

  3. The Invisible Hand of Pedagogy in Australian Indigenous Studies and Indigenous Education

    Science.gov (United States)

    Rhea, Zane Ma; Russell, Lynette

    2012-01-01

    The Australian Learning and Teaching Council (ALTC)-funded project "Exploring Problem-Based Learning Pedagogy as Transformative Education in Indigenous Australian Studies" raised a number of issues that resonated with concerns we have had as professionals engaged in teaching and researching Australian Indigenous studies and Indigenous…

  4. The politics of indigeneity: Indigenous strategies for inclusion in climate change negotiations

    Directory of Open Access Journals (Sweden)

    Doolittle Amity

    2010-01-01

    Full Text Available Indigenous environmental activists have clearly articulated their views on global climate change policy. The content of these views was explored during the 10-day 2008 World Conservation Congress (WCC in Barcelona. Data were primarily collected through interviews and participant observation. In addition, policy statements and declarations made by indigenous environmental activists from 2000 to 2009 were analysed to place the perspectives of indigenous leaders and environmental activists in the context of their decade-long struggle to gain negotiating power at the United Nations Framework Convention on Climate Change. This study examines the rhetorical strategies indigenous leaders from around the world use to gain political recognition and legitimacy in climate change negotiations. Two core principles, relating to a particular representation of indigenous environmental knowledge are identified as fundamental rhetorical tools. These are a belief that the earth is a living being with rights and the conviction that it is the responsibility of indigenous peoples to protect the earth from over-exploitation. However, reference to indigenous environmental knowledge is not the only rhetorical mechanism used by indigenous leaders in the climate debates. When faced with specific United Nations policies to combat climate change that could have a profound impact on their land rights, some indigenous leaders adopt a more confrontational response. Fearing that new polices would reinforce historical trends of marginalisation, indigenous leaders seeking recognition in climate change debates speak less about their ecological knowledge and responsibility to the earth and more about their shared histories of political and economic marginalisation and land dispossession, experienced first through colonialism and more recently through globalisation.

  5. Feeding ecology of indigenous and non-indigenous fish species within the family Sphyraenidae.

    Science.gov (United States)

    Kalogirou, S; Mittermayer, F; Pihl, L; Wennhage, H

    2012-06-01

    The feeding ecology of two common indigenous (Sphyraena viridensis and Sphyraena sphyraena) and one abundant non-indigenous sphyraenid species, Sphyraena chrysotaenia, of Indo-Pacific Ocean origin, was investigated in an area of the eastern Mediterranean Sea. The stomach contents of 738 individuals of varying size, collected during the period December 2008 to August 2009, were examined. The dietary analyses revealed that all three species were specialized piscivores with a diet consisting of >90% fish, both by number and mass. Concurrent sampling of the fish assemblage made it possible to calculate selectivity as well as diet breadth and overlap of these strict piscivores. Even though several prey species were found in the stomachs of the three predators examined, selectivity towards Atherina boyeri was highly significant. For all species examined, >70% of the diet by mass was made up by three indigenous species of commercial value: Spicara smaris, Boops boops and A. boyeri. Diet breadth and size of prey increased with increasing body size for all predators. With increased body size, the diet overlap between indigenous and non-indigenous species decreased. This could be attributed to increased diet breadth and the specific life-history characteristics of indigenous species developing into larger individuals. During winter, the condition factor of the non-indigenous species was significantly lower than that of the indigenous, indicating that winter conditions in the Mediterranean Sea may limit its further expansion north and westward. With this study, the gap in knowledge of the feeding preferences of the most abundant piscivorous species found in coastal areas of the study region is filled. Additionally, the results indicate that non-indigenous species familial affiliation to indigenous ones does not facilitate invasion success.

  6. Optimization of the functional expression of Coprinus cinereus peroxidase in Pichia pastoris by varying the host and promoter.

    Science.gov (United States)

    Kim, Su-Jin; Lee, Jeong-Ah; Kim, Yong-Hwan; Song, Bong-Keun

    2009-09-01

    Peroxidase from Coprinus cinereus (CiP) has attracted attention for its high specific activity and broad substrate spectrum compared with other peroxidases. In this study, the functional expression of this peroxidase was successfully achieved in the methylotrophic yeast Pichia pastoris. The expression level of CiP was increased by varying the microbial hosts and the expression promoters. Since a signal sequence, such as the alpha mating factor of Saccharomyces cerevisiae, was placed preceding the cDNA of the CiP coding gene, expressed recombinant CiP (rCiP) was secreted into the culture broth. The Mut+ Pichia pastoris host showed a 3-fold higher peroxidase activity, as well as 2-fold higher growth rate, compared with the Muts Pichia pastoris host. Furthermore, the AOX1 promoter facilitated a 5-fold higher expression of rCiP than did the GAP promoter.

  7. Promoting the occupational health of indigenous farmworkers.

    Science.gov (United States)

    Farquhar, Stephanie; Samples, Julie; Ventura, Santiago; Davis, Shelley; Abernathy, Michelle; McCauley, Linda; Cuilwik, Nancy; Shadbeh, Nargess

    2008-06-01

    In the United States, approximately 78% of agricultural farmworkers are immigrants. In Oregon, a growing number of these farmworkers are indigenous and speak an indigenous language as their primary language. This group of farmworkers suffers from linguistic, cultural and geographic isolation and faces a unique set of challenges yet little has been done to identify their health needs. Using data from focus groups, partners from this community-based participatory research project examined indigenous farmworkers' concerns regarding occupational injury and illness, experiences of discrimination and disrespect, and language and cultural barriers. The data revealed examples of disrespect and discrimination based on the languages and cultures of indigenous farmworkers, and a lack of basic occupational health and safety information and equipment. For example, participants mentioned that occupational safety information was inaccessible because it was rarely provided in indigenous languages, and participants felt there were no legal means to protect farmworkers from occupational hazards. Community-based strategies designed to address the occupational health status of farmworkers must consider the unique circumstances of those farmworkers who do not speak Spanish or English.

  8. The brazilian indigenous planetary-observatory

    Science.gov (United States)

    Afonso, G. B.

    2003-08-01

    We have performed observations of the sky alongside with the Indians of all Brazilian regions that made it possible localize many indigenous constellations. Some of these constellations are the same as the other South American Indians and Australian aborigines constellations. The scientific community does not have much of this information, which may be lost in one or two generations. In this work, we present a planetary-observatory that we have made in the Park of Science Newton Freire-Maia of Paraná State, in order to popularize the astronomical knowledge of the Brazilian Indians. The planetary consists, essentially, of a sphere of six meters in diameter and a projection cylinder of indigenous constellations. In this planetary we can identify a lot of constellations that we have gotten from the Brazilian Indians; for instance, the four seasonal constellations: the Tapir (spring), the Old Man (summer), the Deer (autumn) and the Rhea (winter). A two-meter height wooden staff that is posted vertically on the horizontal ground similar to a Gnomon and stones aligned with the cardinal points and the soltices directions constitutes the observatory. A stone circle of ten meters in diameter surrounds the staff and the aligned stones. During the day we observe the Sun apparent motions and at night the indigenous constellations. Due to the great community interest in our work, we are designing an itinerant indigenous planetary-observatory to be used in other cities mainly by indigenous and primary schools teachers.

  9. Effect of aeration on D-xylose fermentation by Pachysolen tannophilus, Pichia stipitis, Kluyveromyces marxianus and Candida shehatae

    Energy Technology Data Exchange (ETDEWEB)

    Delgenes, J.P.; Moletta, R.; Navarro, J.M.

    1986-12-01

    The fermentation of D-xylose by Pachysolen tannophilus Y2460, Pichia stipitis Y7124, Kluyveromyces marxianus Y2415 and Candida shehatae Y12878 was investigated in aerobic, anaerobic and microaerophilic batch cultures. The aeration rate greatly influenced the fermentations; growth, rate of ethanol production and oxidation of ethanol are affected. Of the strains tested, Pichia stipitis appears superior; under anaerobic conditions it converts D-xylose (20 g/l) to ethanol with a yield of 0.04 g/l and it exhibits the highest ethanol specific productivity (3.5 g of ethanol per g dry cell per day) under microaerophilic conditions. 10 references.

  10. Effects of Non-indigenous Australian Human Resource Managers' Gender and Attitudes and Applicants' Ethnicity and Gender on Resume Evaluations

    National Research Council Canada - National Science Library

    Hughes, Lauren; Davidson, Graham R

    2011-01-01

    In this web-based survey, 60 non-Indigenous Australian human resource (HR) professionals reviewed four fictitious job resum s for an Indigenous male, Indigenous female, non-Indigenous male and non- Indigenous female...

  11. Upscale of recombinant α-L-rhamnosidase production by Pichia pastoris MutS strain

    Directory of Open Access Journals (Sweden)

    Kristína eMarkošová

    2015-10-01

    Full Text Available Pichia pastoris is currently one of the most preferred microorganisms for recombinant enzyme production due to its efficient expression system. The advantages include the production of high amounts of recombinant proteins containing the appropriate posttranslational modifications and easy cultivation conditions. α-L-Rhamnosidase is a biotechnologically important enzyme in food and pharmaceutical industry, used for example in debittering of citrus fruit juices, rhamnose pruning from naringin or enhancement of wine aromas, creating a demand for the production of an active and stable enzyme. The production of recombinant α-L-rhamnosidase cloned in the MutS strain of Pichia pastoris KM71H was optimized. The encoding gene is located under the control of the AOX promoter, which is induced by methanol whose concentration is instrumental for these strain types. Fermentation was upscaled in bioreactors employing various media and several methanol-feeding strategies. It was found that fed batch with BSM media was more effective compared to BMMH media due to lower cost and improved biomass formation. In BSM medium, the dry cell weight reached approximately 60 g/L, while in BMMH it was only 8.3 g/L, without additional glycerol, which positively influenced the amount of enzyme produced. New methanol feeding strategy, based on the level of dissolved oxygen was developed in this study. This protocol that is entirely independent on methanol monitoring was up scaled to a 19.5-L fermenter with 10-L working volume with the productivity of 13.34 mgprot/L/h and specific activity of α-L-rhamnosidase of 82 U/mg. The simplified fermentation protocol was developed for easy and effective fermentation of Pichia pastoris MutS based on dissolved oxygen monitoring in the induction phase of an enzyme production.

  12. 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.

  13. Indigenous information as tool for consolidating and promoting ...

    African Journals Online (AJOL)

    Indigenous information as tool for consolidating and promoting natural resources ... Log in or Register to get access to full text downloads. ... with the view to identifying the impacts of indigenous information methods on resource conservation.

  14. Analysis of alcohol dependence in indigenous peoples in Northern Siberia.

    Directory of Open Access Journals (Sweden)

    Michail Savchenko

    2015-06-01

    More severe course of alcoholism among indigenous population of North of Siberia leads to the destruction of traditional lifestyles and reduction of the indigenous population in the northern territories of the Russian Federation.

  15. The Effect of Physics in the Development of Indigenous Technology ...

    African Journals Online (AJOL)

    FIRST LADY

    An International Multi-Disciplinary Journal , Ethiopia. Vol. ... development of indigenous technology (production of local talking drum and canoe) in Akwa ... made good (indigenous technologies) to conform to modern standards has therefore ...

  16. Conducting Indigenous Research in Western Knowledge Spaces: Aligning Theory and Methodology

    Science.gov (United States)

    Singh, Myra; Major, Jae

    2017-01-01

    Walking simultaneously in two worlds as an Indigenous researcher, navigating Indigenous and Western epistemologies/methodologies can have its challenges. Indigenous methodologies have become an important element of qualitative research and have been increasingly taken up by both Indigenous and non-Indigenous researchers. Indigenous methodologies…

  17. Conducting Indigenous Research in Western Knowledge Spaces: Aligning Theory and Methodology

    Science.gov (United States)

    Singh, Myra; Major, Jae

    2017-01-01

    Walking simultaneously in two worlds as an Indigenous researcher, navigating Indigenous and Western epistemologies/methodologies can have its challenges. Indigenous methodologies have become an important element of qualitative research and have been increasingly taken up by both Indigenous and non-Indigenous researchers. Indigenous methodologies…

  18. Indigenous teacher training within an intercultural perspective

    Directory of Open Access Journals (Sweden)

    Maria Aparecida Bergamaschi

    2012-12-01

    Full Text Available Current analysis discusses indigenous teacher training foregrounded on the activities that involve teachers, Kaingang chiefs and government officials responsible for this policy in the state of Rio Grande do Sul, Brazil. The Specialization Course in Professional Education integrated to Fundamental Education, within the modality Education for Young People and Adults – A differentiated proposal for Amerindians, is the main objective of current study. The course is run by the Federal University of Rio Grande do Sul and funded by the Secretary for Professional Education and Technology of the Brazilian Ministry of Education. Studies related to the research ‘Amerindian Education and Interculturality’ underlie the above-mentioned specialization course within a wider context of the formation of indigenous teachers and indigene school education.

  19. Re-vitalizing an indigenous language

    DEFF Research Database (Denmark)

    Hansen, Annette Skovsted

    2014-01-01

    languages to match standards defined in nation-building and, thereby, enabled latent possibilities for indigenous populations to re-vitalize their languages in connection with the United Nations Year for Indigenous Peoples in 1993, and the first United Nations Decade for Indigenous Peoples, 1995......–2004. This article focuses on dictionaries of the languages of the Ainu populations in the borderlands between the nation-states Japan and Russia. The main argument is that the Ainu Cultural Promotion Act promulgated in 1997 had a significant impact on the production and purpose of Ainu dictionaries....... The dictionaries prior to 1997 functioned, predominantly, as records, which contributed to the increased visibility of Ainu populations inside and outside Japan in the immediate national interests of Japan, whereas the dictionaries published after 1997 are intended to enable the active use of Ainu language today...

  20. New opportunities by synthetic biology for biopharmaceutical production in Pichia pastoris

    Science.gov (United States)

    Vogl, Thomas; Hartner, Franz S; Glieder, Anton

    2013-01-01

    Biopharmaceuticals are an integral part of modern medicine and pharmacy. Both, the development and the biotechnological production of biopharmaceuticals are highly cost-intensive and require suitable expression systems. In this review we discuss established and emerging tools for reengineering the methylotrophic yeast Pichia pastoris for biopharmaceutical production. Recent advancements of this industrial expression system through synthetic biology include synthetic promoters to avoid methanol induction and to fine-tune protein production. New platform strains and molecular cloning tools as well as in vivo glycoengineering to produce humanized glycoforms have made P. pastoris an important host for biopharmaceutical production. PMID:23522654

  1. Low-pressure airlift fermenter for single cell protein production: II. Continuous culture of Pichia yeast

    Energy Technology Data Exchange (ETDEWEB)

    Chen, N.Y.; Srinivasan, S.; Leavitt, R.I.; Coty, V.F.; Kondis, E.F.

    1987-03-01

    Experiments using Pichia yeast grown on n-paraffins have been conducted in laboratory 10-l airlift fermenters and in a 640-l module of commercial scale. Results confirmed the design concept with low-pressure air. However, in the absence of mass transport constraints, the build up of toxic factors in the fermenter appeared to a major variable limiting cell productivity. Foaming in the large fermenter also presented a serious problem, which must be solved before low-pressure airlift fermenters become practical. 14 references.

  2. Response Surface Optimization of Bioethanol Production from Sugarcane Molasses by Pichia veronae Strain HSC-22

    Directory of Open Access Journals (Sweden)

    Hamed I. Hamouda

    2015-01-01

    Full Text Available Pichia veronae strain HSC-22 (accession number KP012558 showed a good tolerance to relatively high temperature, ethanol and sugar concentrations. Response surface optimization based on central composite design of experiments predicted the optimal values of the influencing parameters that affect the production of bioethanol from sugarcane molasses to be as follows: initial pH 5, 25% (w : v initial molasses concentration, 35°C, 116 rpm, and 60 h. Under these optimum operating conditions the maximum bioethanol production on a batch fermenter scale was recorded as 32.32 g/L with 44% bioethanol yield.

  3. Expression and Characterization of HIV-1 Envelope Glycoprotein in Pichia Pastoris

    Institute of Scientific and Technical Information of China (English)

    ZHAO Li-hui; YU Xiang-hui; JIANG Chun-lai; WU Yong-ge; SHEN Jia-cong; KONG Wei

    2008-01-01

    To obtain a sufficient amount of glycoprotein for further studying the structure and function of HIV-1 envelope glycoprotein, amplified and modified HIV-1 envelope glycoprotein gene which recombined subtypes(850amino acids) from Guangxi in China was inserted into Pichiapastoris expression vector pPICZaB; then the recombinant plasmid was transported into the yeast cells to induce the expression of Env protein with methanol. The results of SDS-PAGE and Western blot indicate that the envelope glycoprotein could be expressed in Pichia pastoris with productions of a 120000 glycoprotein and a 41000 glycoprotein, which showed satisfactory immunogenicity by indirect ELISA.

  4. Recent advances in the production of recombinant subunit vaccines in Pichia pastoris.

    Science.gov (United States)

    Wang, Man; Jiang, Shuai; Wang, Yefu

    2016-04-01

    Recombinant protein subunit vaccines are formulated using defined protein antigens that can be produced in heterologous expression systems. The methylotrophic yeast Pichia pastoris has become an important host system for the production of recombinant subunit vaccines. Although many basic elements of P. pastoris expression system are now well developed, there is still room for further optimization of protein production. Codon bias, gene dosage, endoplasmic reticulum protein folding and culture condition are important considerations for improved production of recombinant vaccine antigens. Here we comment on current advances in the application of P. pastoris for the synthesis of recombinant subunit vaccines.

  5. Production of ethanol from hemicellulose fraction of cocksfoot grass using pichia stipitis

    DEFF Research Database (Denmark)

    Njoku, Stephen Ikechukwu; Iversen, Jens Asmus; Uellendahl, Hinrich;

    2013-01-01

    liquid hydrolysate to ethanol is essential for economically feasible cellulosic ethanol processes. Fermentation of the separated hemicellulose liquid hydrolysates obtained after the WEx pretreatment was done by Pichia stipitis CBS 6054 (Scheffersomyces stipitis). Results: The fermentation of the WEx...... liquid hydrolysate from the pretreatment at higher severity (180°C, 15 min, 87 psi oxygen and 190°C, 15 min, 0.2% sulfuric acid) was fully inhibited probable by the presence of higher concentrations of inhibitory compounds such as furfural, HMF and acetic acid. The ethanol yield among other WEx...

  6. Expression and purification of soluble porcine cystatin 11 in Pichia pastoris.

    Science.gov (United States)

    Fan, Kuohai; Jiang, Junbing; Wang, Zhirui; Fan, Ruicheng; Yin, Wei; Sun, Yaogui; Li, Hongquan

    2014-11-01

    Cystatin 11 (CST11) belongs to the cystatin type 2 family of cysteine protease inhibitors and exhibits antimicrobial activity in vitro. In this study, we describe the expression and purification of recombinant porcine CST11 in the Pichia pastoris system. We then assess its antimicrobial activity against Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, and Bacillus subtilis by liquid growth inhibition assay. Kinetic studies indicate that the recombinant porcine CST11 has high potency against E. coli and S. aureus. Scanning electronic microscope analysis showed that CST11 might be targeting the bacterial membrane and, thus, could potentially be developed as a therapeutic agent for inhibiting microbe infection without the risk of antibiotic resistance.

  7. High-expression of recombinant human concensus interferon-α by Pichia pastoris

    Institute of Scientific and Technical Information of China (English)

    HAO Yuyou; SHI Qiqi; HE Yun; ZHUANG Yingping; WANG Yonghong; ZHANG Siliang; CHU Ju; LIU Zhimin

    2007-01-01

    The present work focused on the high expression of recombinant human consensus interferon-α (cIFN) by Pichia pastoris.The cycle of glycerol feeding,the strategy of methanol feeding and the optimum pH for protein induction were studied.The optimized strategies were a 4-h glycerol-feeding period,induction pH being kept at 5.0 and methanol concentration being kept under 5 g/L.The maximum dry cell weight,cIFN production and bioactivity obtained were 168,1.24 g/L and 5.4 × 107 U/mL,respectively.

  8. Characterization of the Genetic System of the Xylose-Fermenting Yeast Pichia stipitis

    Science.gov (United States)

    Melake; Passoth; Klinner

    1996-10-01

    High mutant frequencies indicated that the wild-type strains of Pichia stipitis are haploid. Sporulation ability of these clones pointed to a homothallic life cycle. Mating was induced by cultivation under nutritionally poor conditions on malt extract medium. Conjugation was followed immediately by sporulation. However, hybrids could be rescued by transferring the nascent zygotes to complete medium before meiosis had started. Under rich nutritional conditions, hybrids were mitotically stable and did not sporulate. The segregation pattern of auxotrophic markers of diploid zygotes indicated regular meiosis, although asci contained preferentially spore dyads.

  9. Dismantling the divide between indigenous and scientific knowledge

    OpenAIRE

    Agrawal, A

    1995-01-01

    Metadata only record In the past few years scholarly discussions have characterized indigenous knowledge as a significant resource for development. This article interrogates the concept of indigenous knowledge and the strategies its advocates present to promote development. The article suggests that both the concept of indigenous knowledge and its role in development, are problematic issues as currently conceptualized. To productively engage indigenous knowledge in development, we must go ...

  10. The use of indigenous knowledge in development: problems and challenges

    OpenAIRE

    Briggs, J.

    2005-01-01

    The use of indigenous knowledge has been seen by many as an alternative way of promoting development in poor rural communities in many parts of the world. By reviewing much of the recent work on indigenous knowledge, the paper suggests that a number of problems and tensions has resulted in indigenous knowledge not being as useful as hoped for or supposed. These include problems emanating from a focus on the (arte)factual; binary tensions between western science and indigenous knowledge system...

  11. Indigenous Participation in Intercultural Education: Learning from Mexico and Tanzania

    OpenAIRE

    Santos H. Alvarado Dzul; Francisco J. Rosado-May; Susanne Kissmann; Gemma Burford; Marie K. Harder

    2012-01-01

    Intercultural education seeks to create a forum for integrating Western scientific knowledge and indigenous knowledge to address local and global challenges such as biocultural diversity conservation, natural resource management, and social justice for indigenous peoples. Intercultural education is based on learning together with, rather than learning about or from, indigenous communities. In the best examples, problem-based learning dissolves the dichotomy between indigenous and nonindigenou...

  12. Collection and characterization of several indigenous vegetables in Indonesia

    OpenAIRE

    Bambang, P.

    2007-01-01

    This presentation describes the various characteristics and potentials of indigenous vegetables. Indigenous vegetables have nutritional value, and are a source of pro-vitamin A, C, calcium and iron. They are also a source of fiber and antioxidants. Indonesian indigenous vegetables are abundant and can be planted in a number of locations. They are also usually consumed only by local people. The objectives of this study were to collect and characterize several indigenous vegetables in Indonesia...

  13. Representing Mayas: Indigenous Authorities and Citizenship Demands in Guatemala

    NARCIS (Netherlands)

    Rasch, E.D.

    2011-01-01

    In this article, I analyze how indigenous authorities in Guatemala negotiate citizenship at the local level within the larger context of indigenous claim making in Latin America. I argue that the construction of citizenship at the local level is not only framed by models imposed on indigenous commun

  14. Indigenous Knowledge and Library Work in Sierra Leone

    Science.gov (United States)

    Kargbo, John Abdul

    2006-01-01

    Indigenous knowledge is vital information that is sadly diminishing at an alarming rate in Sierra Leone. There is, therefore, an urgent need to collect it before much of it is completely lost. This article explores the concept of indigenous knowledge and indigenous knowledge systems with a particular focus on Sierra Leone. Definitions and…

  15. The indigenous space and marginalized peoples in the United Nations

    DEFF Research Database (Denmark)

    Dahl, Jens

    an historical overview and first-hand account of the indigenous involvement with the UN with an analysis of global indigenous identity as a relativist and constructed term rather than a positivist definitional concept, Dahl addresses how indigenous peoples have implemented the UN achievements at home....

  16. Indigenous Representation and Alternative Schooling: Prioritising an Epistemology of Relationality

    Science.gov (United States)

    Keddie, Amanda

    2014-01-01

    This paper draws on a case study of a small alternative Indigenous school in Queensland, Australia. From the perspective of several of the school's Indigenous Elders, the paper foregrounds the significance of group differentiation at the school on the basis of Indigenous representation. However, it also considers how such…

  17. Community-Based Indigenous Digital Storytelling with Elders and Youth

    Science.gov (United States)

    Iseke, Judy; Moore, Sylvia

    2011-01-01

    Indigenous digital storytelling and research are as much about the process of community relationships as they are about the development of digital products and research outcomes. Indigenous researchers, digital storytelling producers, and academics work in different communities with research collaborators who are indigenous community members,…

  18. Situating Indigenous Student Mobility within the Global Education Research Agenda

    Science.gov (United States)

    Prout, Sarah; Hill, Angela

    2012-01-01

    In Australia, as in other global contexts, Indigenous student education outcomes are well below those of their non-Indigenous counterparts. A more robust understanding of, and responsiveness to, Indigenous temporary mobilities is a critical step to redressing such educational inequalities. This paper draws together learnings from the papers in…

  19. Using indigenous knowledge to improve agricultural and natural resource management

    OpenAIRE

    DeWalt, B.

    1994-01-01

    Metadata only record More effective and creative interactions between indigenous and scientific knowledge systems is needed. This paper describes the strengths and weaknesses of both scientific and indigenous knowledge systems by drawing on examples. The author then draws on those examples to indicate in what situations we should look for guidance and ideas from indigenous knowledge systems.

  20. Reflecting Visions. New Perspectives on Adult Education for Indigenous Peoples.

    Science.gov (United States)

    King, Linda, Ed.

    This book contains 14 papers: "Indigenous Peoples and Adult Education: A Growing Challenge" (Rodolfo Stavenhagen); "Indigenous Peoples: Progress in the International Recognition of Human Rights and the Role of Education" (Julian Burger); "Adult Learning in the Context of Indigenous Societies" (Linda King); "Linguistic Rights and the Role of…

  1. Indigenous Digital Storytelling in Video: Witnessing with Alma Desjarlais

    Science.gov (United States)

    Iseke, Judy M.

    2011-01-01

    Indigenous digital storytelling in video is a way of witnessing the stories of Indigenous communities and Elders, including what has happened and is happening in the lives and work of Indigenous peoples. Witnessing includes acts of remembrance in which we look back to reinterpret and recreate our relationship to the past in order to understand the…

  2. The Work-Study Experience of Indigenous Undergraduates in Taiwan

    Science.gov (United States)

    Chen, Shan-Hua

    2014-01-01

    Due to the large number of universities in Taiwan and the increased availability of scholarships for disadvantaged students, the number of college students from indigenous families has been on the rise in recent years. However, many indigenous students still find it necessary to work part-time. In this study, indigenous students were interviewed…

  3. Indigenous Education 1991-2000: Documents, Outcomes and Governments

    Science.gov (United States)

    Gunstone, Andrew

    2012-01-01

    There is often a disparity in Indigenous Affairs between many documents, such as policies, reports and legislation, and outcomes. This article explores this difference through analysing the policy area of Indigenous education during the period of 1991 to 2000. I examine three key documents relating to Indigenous education. These are the…

  4. Non-indigenous marine and estuarine species in The Netherlands

    NARCIS (Netherlands)

    Wolff, W.J.

    2005-01-01

    An overview is presented of non-indigenous marine and estuarine plant and animal species recorded from The Netherlands. In this list both exotic species from outside NW Europe and non-indigenous species from elsewhere in NW Europe are enumerated. Species that have been suggested to be non-indigenous

  5. The Impact of Immigration on Bilingualism among Indigenous American Peoples

    Science.gov (United States)

    Ahler, Janet Goldenstein

    2007-01-01

    Early federal government policies for American indigenous people alternated between extermination and assimilation. Imposing the colonists' and immigrants' language on indigenous people was important for achieving the latter. In the 1970-90's, federally funded grants for bilingual education for indigenous schools were offered to accommodate Native…

  6. Absence of disparities in anthropometric measures among Chilean indigenous and non-indigenous newborns

    Science.gov (United States)

    2010-01-01

    Background Studies throughout North America and Europe have documented adverse perinatal outcomes for racial/ethnic minorities. Nonetheless, the contrast in newborn characteristics between indigenous and non-indigenous populations in Latin America has been poorly characterized. This is due to many challenges, including a lack of vital registration information on ethnicity. The objective of this study was to analyze trends in anthropometric measures at birth in Chilean indigenous (Mapuche) and non-indigenous children over a 5-year period. Methods We examined weight and length at birth using information available through a national data base of all birth records for the years 2000 through 2004 (n = 1,166.513). Newborns were classified ethnically according to the origins of the parents' last names. Result The average birthweight was stable over the 5 year period with variations of less than 20 g in each group, and with mean values trivially higher in indigenous newborns. The proportion weighing less than 2500 g at birth increased modestly from 5.2% to 5.6% in non-indigenous newborns whereas the indigenous births remained constant at 5.2%. In multiple regression analyses, adjusting flexibly for gestational age and maternal characteristics, the occurrence of an indigenous surname added only 14 g to an average infant's birthweight while holding other factors constant. Results for length at birth were similar, and adjusted time trend variation in both outcomes was trivially small after adjustment. Anthropometric indexes at birth in Chile are quite favorable by international standards. Conclusion There is only a trivial degree of ethnic disparity in these values, in contrast to conditions for ethnic minorities in other countries. Moreover, these values remained roughly constant over the 5 years of observation in this study. PMID:20598150

  7. Expression of a cold-adapted fish trypsin in Pichia pastoris.

    Science.gov (United States)

    Macouzet, Martin; Simpson, Benjamin K; Lee, Byong H

    2005-06-01

    Trypsin is a highly valuable protease that has many industrial and biomedical applications. The growing demand for non-animal sources of the enzyme and for trypsins with special properties has driven the interest to clone and express this protease in microorganisms. Reports about expression of recombinant trypsins show wide differences in the degree of success and are contained mainly in patent applications, which disregard the difficulties associated with the developments. Although the yeast Pichia pastoris appears to be the microbial host with the greatest potential for the production of trypsin, it has shown problems when expressing cold-adapted fish trypsins (CAFTs). CAFTs are considered of immense value for their comparative advantage over other trypsins in a number of food-processing and biotechnological applications. Thus, to investigate potential obstacles related to the production of CAFTs in P. pastoris, the cunner fish trypsin (CFT) was cloned in different Pichia expression vectors. The vectors were constructed targeting both internal and secreted expression and keeping the CFT native signal peptide. Western-blotting analysis confirmed the expression with evident differences for each construct, observing a major effect of the leader peptide sequence on the expression patterns. Immobilized nickel affinity chromatography yielded a partially purified recombinant CFT, which exhibited trypsin-specific activity after activation with bovine enterokinase.

  8. Biomarkers to evaluate the effects of temperature and methanol on recombinant Pichia pastoris.

    Science.gov (United States)

    Zepeda, Andrea B; Figueroa, Carolina A; Abdalla, Dulcineia S P; Maranhão, Andrea Q; Ulloa, Patricio H; Pessoa, Adalberto; Farías, Jorge G

    2014-01-01

    Pichia pastoris is methylotrophic yeast used as an efficient expression system for heterologous protein production. In order to evaluate the effects of temperature (10 and 30 °C) and methanol (1 and 3% (v/v)) on genetically-modified Pichia pastoris, different biomarkers were evaluated: Heat stress (HSF-1 and Hsp70), oxidative stress (OGG1 and TBARS) and antioxidant (GLR). Three yeast cultures were performed: 3X = 3% methanol-10 °C, 4X = 3% methanol-30 °C, and 5X = 1% methanol-10°C. The expression level of HIF-1α, HSF-1, HSP-70 and HSP-90 biomarkers were measured by Western blot and in situ detection was performed by immunocytochemistry. Ours results show that at 3% methanol -30 °C there is an increase of mitochondrial OGG1 (mtOGG1), Glutathione Reductase (GLR) and TBARS. In addition, there was a cytosolic expression of HSF-1 and HSP-70, which indicates a deprotection against nucleolar fragmentation (apoptosis). On the other hand, at 3% methanol -10 °C and 1% and at methanol -10 °C conditions there was nuclear expression of OGG1, lower levels of TBARS and lower expression of GLR, cytosolic expression of HSF-1 and nuclear expression HSP-70. In conclusion, our results suggest that 3% methanol-30 °C is a condition that induces a strong oxidative stress and risk factors of apoptosis in modified-genetically P. pastoris.

  9. Expression of lignin peroxidase H2 from Phanerochaete chrysosporium by multi-copy recombinant Pichia strain

    Institute of Scientific and Technical Information of China (English)

    WANG Wei; WEN Xianghua

    2009-01-01

    The lipH2 gene, encoding the expression of lignin peroxidase, was cloned from Phanerochaete chrysosporium BKM-F-1767 and expressed in Pichia pastoris X-33, a yeast.The cDNA of LiPH2 was generated from total RNA extracted from P.chrysosporium by PCR with primers that do not contain a P.chrysosporium lignin peroxidase secretion signal.The gene was then successfully inserted into the expression vector pPICZα, resulting in the recombinant vector pPICZα-lipH2.The transformation was conducted in two ways.One was using the wild Pichia pastoris as the recipients, which results in the recombinant P.pastoris with single or low lipH2 gene copy.The second was using P.pastoris and single or low lipH2 gene copy as the recipients, which results in the recombinant P.pastoris with multi-copies of lipH2 genes.This study first expressed the gene lipH2 in P.pastoris and achieved the successful expression of the LiPH2 depending upon the generation of a recombinant strain that contains multiple copies.The lignin peroxidase activity reached a maximum of 15 U/L after 12 h induction.

  10. Overexpression and biochemical characterization of a thermostable phytase from Bacillus subtilis US417 in Pichia pastoris.

    Science.gov (United States)

    Hmida-Sayari, Aïda; Elgharbi, Fatma; Farhat, Ameny; Rekik, Hatem; Blondeau, Karine; Bejar, Samir

    2014-09-01

    The overexpression of the native gene encoding the thermostable Bacillus subtilis US417 phytase using Pichia pastoris system is described. The phytase gene, in which the sequence encoding the signal peptide was replaced by that of the α-factor of Saccharomyces cerevisiae, was placed under the control of the methanol-inducible promoter of the alcohol oxidase 1 gene and expressed in Pichia pastoris. Small-scale expression experiments and activity assays were used to screen positive colonies. A recombinant strain was selected and produces 43 and 227 U/mL of phytase activity in shake flasks and in high-cell-density fermentation, respectively. The purified phytase was glycosylated protein and varied in size (50-65 kDa). It has a molecular mass of 43 kDa when it was deglycosylated. The purified r-PHY maintains 100% of its activity after 10 min incubation at 75 °C and pH 7.5. This thermostable phytase, which is also active over broad pH ranges, may be useful as feed additives, since it can resist the temperature used in the feed-pelleting process.

  11. [Effects of mixed carbon sources on glucose oxidase production by recombinant Pichia pastoris].

    Science.gov (United States)

    Shen, Yina; Gu, Lei; Zhang, Juan; Chen, Jian; Du, Guocheng

    2013-07-01

    Glucose oxidase (GOD) is an important industrial enzyme with many potential applications. In order to increase the production and productivity of GOD by recombinant Pichia pastoris GS115, we investigated the feeding strategies of mixed carbon sources during induction phase, based on results of the optimization of initial cell and methanol concentration on GOD production. The optimal initial cell and methanol concentration were 100 g/L and 18 g/L. During induction phase, the mixed-carbon-sources strategies showed that glycerol, sorbitol or mannitol co-feeding with methanol could enhance GOD production. With mannitol co-feeding (20:1(W/W)), the maximum GOD production and maximum GOD productivity reached 711.3 U/mL and 4.60 U/(mL x h) after an induction period of 156 h. Compared to the control, the enhancements of GOD production and productivity were 66.3% and 67.9%, respectively. Meanwhile, we found an appropriate mannitol co-feeding strategy that would not inhibit the expression of promote. The activity of alcohol oxidase was 8.8 U/g, which was enhanced by 69.2% compared to the control (5.2 U/g). We can use the same optimization process to improve the production of other proteins from recombinant Pichia pastoris by changing the fermentation parameters.

  12. A visual method for direct selection of high-producing Pichia pastoris clones

    Directory of Open Access Journals (Sweden)

    Liu Sheng

    2011-03-01

    Full Text Available Abstract Background The methylotrophic yeast, Pichia pastoris, offers the possibility to generate a high amount of recombinant proteins in a fast and easy way to use expression system. Being a single-celled microorganism, P. pastoris is easy to manipulate and grows rapidly on inexpensive media at high cell densities. A simple and direct method for the selection of high-producing clones can dramatically enhance the whole production process along with significant decrease in production costs. Results A visual method for rapid selection of high-producing clones based on mannanase reporter system was developed. The study explained that it was possible to use mannanase activity as a measure of the expression level of the protein of interest. High-producing target protein clones were directly selected based on the size of hydrolysis holes in the selected plate. As an example, the target gene (9elp-hal18 was expressed and purified in Pichia pastoris using this technology. Conclusions A novel methodology is proposed for obtaining the high-producing clones of proteins of interest, based on the mannanase reporter system. This system may be adapted to other microorganisms, such as Saccharomyces cerevisiae for the selection of clones.

  13. Performance evaluation of Pichia kluyveri, Kluyveromyces marxianus and Saccharomyces cerevisiae in industrial tequila fermentation.

    Science.gov (United States)

    Amaya-Delgado, L; Herrera-López, E J; Arrizon, Javier; Arellano-Plaza, M; Gschaedler, A

    2013-05-01

    Traditionally, industrial tequila production has used spontaneous fermentation or Saccharomyces cerevisiae yeast strains. Despite the potential of non-Saccharomyces strains for alcoholic fermentation, few studies have been performed at industrial level with these yeasts. Therefore, in this work, Agave tequilana juice was fermented at an industrial level using two non-Saccharomyces yeasts (Pichia kluyveri and Kluyveromyces marxianus) with fermentation efficiency higher than 85 %. Pichia kluyveri (GRO3) was more efficient for alcohol and ethyl lactate production than S. cerevisiae (AR5), while Kluyveromyces marxianus (GRO6) produced more isobutanol and ethyl-acetate than S. cerevisiae (AR5). The level of volatile compounds at the end of fermentation was compared with the tequila standard regulation. All volatile compounds were within the allowed range except for methanol, which was higher for S. cerevisiae (AR5) and K. marxianus (GRO6). The variations in methanol may have been caused by the Agave tequilana used for the tests, since this compound is not synthesized by these yeasts.

  14. Indigenous Australian art in intercultural contact zones

    Directory of Open Access Journals (Sweden)

    Eleonore Wildburger

    2009-06-01

    Full Text Available This article comments on Indigenous Australian art from an intercultural perspective. The painting Bush Tomato Dreaming (1998, by the Anmatyerre artist Lucy Ngwarai Kunoth serves as model case for my argument that art expresses existential social knowledge. In consequence, I will argue that social theory and art theory together provide tools for intercultural understanding and competence.

  15. Strangulation injury from indigenous rocking cradle

    Directory of Open Access Journals (Sweden)

    Saha Abhijeet

    2010-01-01

    Full Text Available Indigenously made rocking cradle is frequently used in rural India. We report strangulation from an indigenously made rocking cradle in an 11-month-old female child. The unique mode of injury and its mechanism have been discussed. Strangulation is an important cause of homicidal and suicidal injury in adults but in children it is usually accidental leading to death due to asphyxia as a result of partial hanging. In western countries, it is the third most common cause of accidental childhood deaths, 17% of them being due to ropes and cords. It ranks fourth amongst the causes of unintentional injury in children less than 1 year of age following roadside accidents, drowning and burns. However, in India, strangulation injury is under reported although indigenous rocking cradles are very commonly used in rural India, and they are even more dangerous than the cribs and adult beds as there are no safety mechanisms therein. We report a case of accidental strangulation following suspension from an indigenously made rocking cradle. The unique mode of injury has prompted us to report this case.

  16. Indigenous Learning Preferences and Interactive Technologies

    Science.gov (United States)

    Kitchenham, Andrew

    2017-01-01

    This three-year research study examined the influence of interactive technologies on the math achievement of Indigenous students in Years 4, 5, 6 and 7 technology-equipped classrooms in a rural elementary school in British Columbia, Canada. Using a mixed-methods approach, the researcher conducted semistructured interviews and collected math…

  17. Indigenous Participation in VET: Understanding the Research

    Science.gov (United States)

    Ackehurst, Maree; Polvere, Rose-Anne; Windley, Georgina

    2017-01-01

    Indigenous educational and employment disadvantage is a much researched and discussed subject. The latest Prime Minister's Closing the Gap report (DPM&C 2017) shows that, while the gap is slowly decreasing in regard to participation in tertiary education, reducing employment disparity, particularly in remote areas, lags behind. This is despite…

  18. Control of indigenous pathogenic bacteria in seafood

    DEFF Research Database (Denmark)

    Huss, Hans Henrik

    1997-01-01

    The pathogenic bacteria indigenous to the aquatic and general environment are listed. Their distribution in nature, prevalence in seafood and the possibilities for growth of these organisms in various types of products are outlined These data, combined with what is known regarding the epidemiology...

  19. Policy and Indigenous Languages in Australia

    Science.gov (United States)

    McKay, Graham

    2011-01-01

    The use of Indigenous languages has been declining over the period of non-Aboriginal settlement in Australia as a result of repressive policies, both explicit and implicit. The National Policy on Languages (Lo Bianco, 1987) was the high point of language policy in Australia, given its national scope and status and its attempt to encompass all…

  20. Indigenous Languages and Mathematics in Elementary Schools

    Science.gov (United States)

    Edmonds-Wathen, Cris; Owens, Kay; Sakopa, Priscilla; Bino, Vagi

    2014-01-01

    Indigenous languages are used for instruction in elementary schools in Papua New Guinea, but teachers have generally received their own education in English. The challenges of identifying terminology to use in mathematics include many-to-one correspondences between English and the vernacular languages, and different grammatical structures.…

  1. Indigenous Youth Migration and Language Contact

    Science.gov (United States)

    Wyman, Leisy T.

    2013-01-01

    Few studies ethnographically detail how Indigenous young people's mobility intersects with sociolinguistic transformation in an interconnected world. Drawing on a decade-long study of youth and language contact, I analyze Yup'ik young people's migration in relation to emerging language ideologies and patterns of language use in "Piniq,"…

  2. Indilinga: African Journal of Indigenous Knowledge Systems

    African Journals Online (AJOL)

    The name Indilinga: stands for the "circular orientation" of indigenous African communities which ... Development of a juice using a mixture of Amaranthus thunbergii for ... AmaXhosa beadwork (Iintsimbi) for identity affirmation and economic ... food, nutrition security and sustainable livelihoods: a case study of Mkhambathini ...

  3. Indigenous Fallow Management on Yap Island

    Science.gov (United States)

    M.V.C. Falanruw; Francis Ruegorong

    2002-01-01

    On Yap Island, indigenous management of the fallow in shifting agriculture has resulted in the development of site-stable taro patch and tree garden agroforestry systems. These systems are relatively sustainable and supportive of household economies , with some surplus for local market sales. however, a broad range of crops whose harvest is complementary to those...

  4. indigenous communication, religion and education as determinants ...

    African Journals Online (AJOL)

    Mugumbate

    The data obtained were analyzed using. Analysis of Variance (ANOVA) and t-test to determine the difference ... aDepartment of Social Work, Faculty of Education, University of Ibadan,. Ibadan, Nigeria. ... gave examples of folk media as puppet show, folk drama, ... been done on the use of indigenous communication as an.

  5. Indigenous environmental values as human values

    Directory of Open Access Journals (Sweden)

    Monica Gratani

    2016-12-01

    Full Text Available The claim that in natural resource management (NRM a change from anthropocentric values and ethics to eco-centric ones is necessary to achieve sustainability leads to the search for eco-centric models of relationship with the environment. Indigenous cultures can provide such models; hence, there is the need for multicultural societies to further include their values in NRM. In this article, we investigate the environmental values placed on a freshwater environment of the Wet Tropics by a community of indigenous Australians. We discuss their environmental values as human values, and so as beliefs that guide communities’ understanding of how the natural world should be viewed and treated by humans. This perspective represents a step forward in our understanding of indigenous environmental values, and a way to overcome the paradigm of indigenous values as valued biophysical attributes of the environment or processes happening in landscapes. Our results show that the participant community holds biospheric values. Restoring these values in the NRM of the Wet Tropics could contribute to sustainability and environmental justice in the area.

  6. Locally Situated Digital Representation of Indigenous Knowledge

    DEFF Research Database (Denmark)

    Winschiers-Theophilus, Heike; Jensen, Kasper Løvborg; Rodil, Kasper

    2012-01-01

    Digital re-presentation of indigenous knowledge remains an absurdity as long as we fail to deconstruct the prevalent design paradigm and techniques continuously re-framing technology within a western epistemology. This paper discusses key challenges in attempts of co-constructing a digital...

  7. Desiderata: Towards Indigenous Models of Vocational Psychology

    Science.gov (United States)

    Leong, Frederick T. L.; Pearce, Marina

    2011-01-01

    As a result of a relative lack of cross-cultural validity in most current (Western) psychological models, indigenous models of psychology have recently become a popular approach for understanding behaviour in specific cultures. Such models would be valuable to vocational psychology research with culturally diverse populations. Problems facing…

  8. Control of indigenous pathogenic bacteria in seafood

    DEFF Research Database (Denmark)

    Huss, Hans Henrik

    1997-01-01

    The pathogenic bacteria indigenous to the aquatic and general environment are listed. Their distribution in nature, prevalence in seafood and the possibilities for growth of these organisms in various types of products are outlined These data, combined with what is known regarding the epidemiology...

  9. Desiderata: Towards Indigenous Models of Vocational Psychology

    Science.gov (United States)

    Leong, Frederick T. L.; Pearce, Marina

    2011-01-01

    As a result of a relative lack of cross-cultural validity in most current (Western) psychological models, indigenous models of psychology have recently become a popular approach for understanding behaviour in specific cultures. Such models would be valuable to vocational psychology research with culturally diverse populations. Problems facing…

  10. Cultural tourism and identity : rethinking indigeneity

    NARCIS (Netherlands)

    Tomaselli, K.G.

    2012-01-01

    Studies of cultural tourism and indigenous identity are fraught with questions concerning exploitation, entitlement, ownership and authenticity. Unease with the idea of leveraging a group identity for commercial gain is ever-present. This anthology articulates some of these debates from a multitude

  11. Strangulation injury from indigenous rocking cradle.

    Science.gov (United States)

    Saha, Abhijeet; Batra, Prerna; Bansal, Anuradha

    2010-07-01

    Indigenously made rocking cradle is frequently used in rural India. We report strangulation from an indigenously made rocking cradle in an 11-month-old female child. The unique mode of injury and its mechanism have been discussed. Strangulation is an important cause of homicidal and suicidal injury in adults but in children it is usually accidental leading to death due to asphyxia as a result of partial hanging. In western countries, it is the third most common cause of accidental childhood deaths, 17% of them being due to ropes and cords. It ranks fourth amongst the causes of unintentional injury in children less than 1 year of age following roadside accidents, drowning and burns. However, in India, strangulation injury is under reported although indigenous rocking cradles are very commonly used in rural India, and they are even more dangerous than the cribs and adult beds as there are no safety mechanisms therein. We report a case of accidental strangulation following suspension from an indigenously made rocking cradle. The unique mode of injury has prompted us to report this case.

  12. Antimicrobial agents deriving from indigenous plants.

    Science.gov (United States)

    Avrelija, Cencic; Walter, Chingwaru

    2010-01-01

    Phytonutrients in many indigenous plants are receiving a lot of attention as they are important in antimicrobial and anticancer therapies. Tropical areas, especially India, South America and Africa, are the main sources of patentable plant products and have indigenous populations with well developed traditional medicinal knowledge. Phytochemicals, including carotenoids, phenolics, alkaloids, nitrogen-containing compounds, and organosulfur compounds, are receiving much attention as they impart important health benefits. This article gives an insight into some important phytochemicals, and analyses the ethical issues on property rights of plant products. Many patent applications have been lodged, and quite a number have been granted. Pharmaceutical industries are engaging in massive speculative bioprospecting on plant based phytochemicals and products, usually resulting in conflicts with indigenous populations. More focus is given here-in to Tylosema esculentum (marama) plant, found in drier parts of Southern Africa and known to contain high quantities of essential phytochemicals. Important phytochemicals in marama include fatty acid (mainly oleic acid, linoleic acid, linolenic acid, behenic acid), protein and phenolic acid components. The marama plant has high potential as a source of medical and cosmetic products. If conflicts surrounding property rights on plant based products are resolved, phytochemicals can be a good source of income for indigenous populations in areas where such plants are found.

  13. The Nagoya Protocol and Indigenous Peoples

    Directory of Open Access Journals (Sweden)

    Maria Yolanda Teran

    2016-07-01

    Full Text Available This article is about Indigenous peoples’ involvement in the Nagoya Protocol negotiations from 2006 to 2010, as well as in its implementation to stop biopiracy in order to protect Pachamama, Mother Earth, and to ensure our survival and the survival of coming generations. The Nagoya Protocol is an international instrument that was adopted in Nagoya, Japan in October 2010 by the Conference of Parties (COP 10 and ratified by 51 countries in Pyeongchang, South Korea in October 2014 at COP 12. This protocol governs access to genetic resources and the fair and equitable sharing of benefits arising from their utilization (access and benefit sharing [ABS]. It has several articles related to Indigenous peoples and traditional knowledge, as well as: The interrelation and inseparable nature between genetic resources and traditional knowledge; The diversity of circumstances surrounding traditional knowledge ownership, including by country; The identification of traditional knowledge owners; The declaration of Indigenous peoples' human rights; and The role of women in the biodiversity process. In addition, this protocol lays out obligations on access, specifically participation in equitable benefit sharing, the accomplishment of prior and informed consent, and the mutually agreed terms and elaboration of a national legal ABS framework with the participation of Indigenous peoples and local communities in order to have well-defined roles, responsibilities, and times of negotiations.

  14. LIFE AND DEATH AMONGST INDIGENOUS PEOPLES

    Directory of Open Access Journals (Sweden)

    Jane Felipe Beltrão

    2015-06-01

    Full Text Available Considering the frequent rights violations perpetrated against indigenous peoples, which affect people and territories, compromising their lives and even their right to mourn the dead, it is imperative to understand the care and concerns of the indigenous towards life and death. Thus, we propose to analyze ethnographic narratives about the Apinayé, Ka'apor, Tapirapé, Tembé, Tenetehara, Terena and Asurini, in order to discuss the caring of people, considering the context of funerary rituals. The texts analyzed are able to reveal: (1 the existence (or not of the practice; (2 the specific contexts in which the funeral rites are (or not practiced; and (3 the meanings that the practice gain in ethnically differentiated societies. The narratives of indigenous peoples are included in order to attempt to make the peoples that nowadays find themselves accused by both the media and (reportedly pro-life organizations “be heard”. Therefore, using the classical literature we study the heritage of ritual practices, which besides confering dignity to the dead, indicate that life is the greater good among indigenous peoples.

  15. Handbook of Indigenous Foods Involving Alkaline Fermentation

    NARCIS (Netherlands)

    Sarkar, P.K.; Nout, M.J.R.

    2014-01-01

    This book details the basic approaches of alkaline fermentation, provides a brief history, and offers an overview of the subject. The book discusses the diversity of indigenous fermented foods involving an alkaline reaction, as well as the taxonomy, ecology, physiology, and genetics of predominant m

  16. High-Level Expression of Endo-β-N-Acetylglucosaminidase H from Streptomyces plicatus in Pichia pastoris and Its Application for the Deglycosylation of Glycoproteins

    OpenAIRE

    Fei Wang; Xiaojuan Wang; Xiaolan Yu; Ling Fu; Yunyun Liu; Lixin Ma; Chao Zhai

    2015-01-01

    Endo-β-N-acetylglucosaminidase H (Endo H, EC3.2.1.96) is a glycohydrolase that is widely used in the study of glycoproteins. The present study aimed to assess the effect of high-level endo-β-N-acetylglucosaminidase H expression in Pichia pastoris. The DNA coding sequence of this enzyme was optimized based on the codon usage bias of Pichia pastoris and synthesized through overlapping PCR. This novel gene was cloned into a pHBM905A vector and introduced into Pichia pastoris GS115 for secretary ...

  17. Glucose-methanol co-utilization in Pichia pastoris studied by metabolomics and instationary 13C flux analysis

    NARCIS (Netherlands)

    Jorda, J.; Suarez Mendez, C.A; Carnicer, M.; Ten Pierick, A.; Heijnen, J.J.; Van Gulik, W.M.; Ferrer, P.; Albiol, J.; Wahl, S.A.

    2013-01-01

    Background Several studies have shown that the utilization of mixed carbon feeds instead of methanol as sole carbon source is beneficial for protein production with the methylotrophic yeast Pichia pastoris. In particular, growth under mixed feed conditions appears to alleviate the metabolic burden r

  18. Fermentation kinetics for xylitol production by a Pichia stipitis D-xylulokinase mutant previously grown in spent sulfite liquor

    Science.gov (United States)

    Rita C.L.B. Rodrigues; Chenfeng Lu; Bernice Liu; Thomas W. Jeffries

    2008-01-01

    Spent sulfite pulping liquor (SSL) contains lignin, which is present as lignosulfonate, and hemicelluloses that are present as hydrolyzed carbohydrates. To reduce the biological oxygen demand of SSL associated with dissolved sugars, we studied the capacity of Pichia stipitis FPL-YS30 (xyl3[delta]) to convert these sugars into useful products. FPL-YS30 produces a...

  19. Glucose-methanol co-utilization in Pichia pastoris studied by metabolomics and instationary 13C flux analysis

    NARCIS (Netherlands)

    Jorda, J.; Suarez Mendez, C.A; Carnicer, M.; Ten Pierick, A.; Heijnen, J.J.; Van Gulik, W.M.; Ferrer, P.; Albiol, J.; Wahl, S.A.

    2013-01-01

    Background Several studies have shown that the utilization of mixed carbon feeds instead of methanol as sole carbon source is beneficial for protein production with the methylotrophic yeast Pichia pastoris. In particular, growth under mixed feed conditions appears to alleviate the metabolic burden r

  20. Glucose-methanol co-utilization in Pichia pastoris studied by metabolomics and instationary 13C flux analysis

    NARCIS (Netherlands)

    Jorda, J.; Suarez Mendez, C.A; Carnicer, M.; Ten Pierick, A.; Heijnen, J.J.; Van Gulik, W.M.; Ferrer, P.; Albiol, J.; Wahl, S.A.

    2013-01-01

    Background Several studies have shown that the utilization of mixed carbon feeds instead of methanol as sole carbon source is beneficial for protein production with the methylotrophic yeast Pichia pastoris. In particular, growth under mixed feed conditions appears to alleviate the metabolic burden

  1. Geographic distribution of isolated indigenous societies in Amazonia and the efficacy of indigenous territories.

    Directory of Open Access Journals (Sweden)

    Dylan C Kesler

    Full Text Available The headwaters of the Amazon Basin harbor most of the world's last indigenous peoples who have limited contact with encroaching colonists. Knowledge of the geographic distribution of these isolated groups is essential to assist with the development of immediate protections for vulnerable indigenous settlements. We used remote sensing to document the locations of 28 isolated villages within the four Brazilian states of Acre, Amazonas, Roraima, and Rondônia. The sites were confirmed during previous over-flights and by image evidence of thatched-roof houses; they are estimated to host over 1,700 individuals. Locational data were used to train maximum entropy models that identified landscape and anthropogenic features associated with the occurrence of isolated indigenous villages, including elevation, proximity to streams of five different orders, proximity to roads and settlements, proximity to recent deforestation, and vegetation cover type. Isolated villages were identified at mid elevations, within 20 km of the tops of watersheds and at greater distances from existing roads and trails. We further used model results, combined with boundaries of the existing indigenous territory system that is designed to protect indigenous lands, to assess the efficacy of the existing protected area network for isolated peoples. Results indicate that existing indigenous territories encompass all of the villages we identified, and 50% of the areas with high predicted probabilities of isolated village occurrence. Our results are intended to help inform policies that can mitigate against future external threats to isolated peoples.

  2. Geographic distribution of isolated indigenous societies in Amazonia and the efficacy of indigenous territories.

    Science.gov (United States)

    Kesler, Dylan C; Walker, Robert S

    2015-01-01

    The headwaters of the Amazon Basin harbor most of the world's last indigenous peoples who have limited contact with encroaching colonists. Knowledge of the geographic distribution of these isolated groups is essential to assist with the development of immediate protections for vulnerable indigenous settlements. We used remote sensing to document the locations of 28 isolated villages within the four Brazilian states of Acre, Amazonas, Roraima, and Rondônia. The sites were confirmed during previous over-flights and by image evidence of thatched-roof houses; they are estimated to host over 1,700 individuals. Locational data were used to train maximum entropy models that identified landscape and anthropogenic features associated with the occurrence of isolated indigenous villages, including elevation, proximity to streams of five different orders, proximity to roads and settlements, proximity to recent deforestation, and vegetation cover type. Isolated villages were identified at mid elevations, within 20 km of the tops of watersheds and at greater distances from existing roads and trails. We further used model results, combined with boundaries of the existing indigenous territory system that is designed to protect indigenous lands, to assess the efficacy of the existing protected area network for isolated peoples. Results indicate that existing indigenous territories encompass all of the villages we identified, and 50% of the areas with high predicted probabilities of isolated village occurrence. Our results are intended to help inform policies that can mitigate against future external threats to isolated peoples.

  3. Reclaiming Indigenous identities: Culture as strength against suicide among Indigenous youth in Canada.

    Science.gov (United States)

    Barker, Brittany; Goodman, Ashley; DeBeck, Kora

    2017-06-16

    In Canada, Indigenous youth suicide represents one of several health disparities burdening Indigenous populations, and like many other of these disparities, can be understood as an expression of societal, historical, cultural and familial trauma. As the number of Indigenous youth who take their own lives every year in Canada continues to far exceed national averages, it appears that conventional suicide prevention efforts remain ineffective among this population. A growing body of research argues that conventional interventions, largely rooted in Western individual-level behavioural change frameworks, are culturally discordant with Indigenous paradigms. In response, some Indigenous communities are turning to cultural revitalization as a holistic community-driven response to suicide prevention and treatment. The following commentary explores the emerging evidence base for "culture as treatment" - a novel approach to suicide that emphasizes the significance of interconnectedness in healing, alongside the revitalization of traditional values to reclaim community wellness. In doing so, we seek to contribute to a changing discourse surrounding Indigenous youth suicide by acknowledging culture as strength against this national crisis.

  4. Chromosomal profile of indigenous pig (Sus scrofa

    Directory of Open Access Journals (Sweden)

    P. Guru Vishnu

    2015-02-01

    Full Text Available Aim: The objective of this study was to investigate the chromosomal profile of indigenous pigs by computing morphometric measurements. Materials and Methods: A cytogenetic study was carried out in 60 indigenous pigs to analyze the chromosomal profile by employing the short term peripheral blood lymphocyte culture technique. Results: The modal chromosome number (2n in indigenous pigs was found to be 38 and a fundamental number of 64 as in the exotic. First chromosome was the longest pair, and thirteenth pair was the second largest while Y-chromosome was the smallest in the karyotype of the pig. The mean relative length, arm ratio, centromeric indices and morphological indices of chromosomes varied from 1.99±0.01 to 11.23±0.09, 1.04±0.05 to 2.95±0.02, 0.51±0.14 to 0.75±0.09 and 2.08±0.07 to 8.08±0.15%, respectively in indigenous pigs. Sex had no significant effect (p>0.05 on all the morphometric measurements studied. Conclusion: The present study revealed that among autosomes first five pairs were sub metacentric, next two pairs were sub telocentric (6-7, subsequent five pairs were metacentric (8-12 and remaining six pairs were telocentric (13-18, while both allosomes were metacentric. The chromosomal number, morphology and various morphometric measurements of the chromosomes of the indigenous pigs were almost similar to those established breeds reported in the literature.

  5. Indigenous knowledge and science in a globalized age

    Science.gov (United States)

    Regmi, Jagadish; Fleming, Michelle

    2012-06-01

    This forum explores and expands on Ben-Zvi Assaraf, Eshach, Orion, and Alamour's article titled "Cultural Differences and Students' Spontaneous Models of the Water Cycle: A Case Study of Jewish and Bedouin Children in Israel" by examining how indigenous knowledge is appropriated in science classrooms; how students from indigenous students' experiences are more complex than many non-indigenous students; and how science and globalization complicates the preservation of indigenous knowledge. In this forum we suggest that research on indigenous knowledge be examined through the lens of the locally situated contexts and the extent to which globalization hinders this kind of knowledge in the name of value neutral scientific knowledge. We finally suggest that research in indigenous communities has to be more intentional and respectful, and teachers need to rethink how useful and meaningful science learning can be for indigenous students.

  6. Using Modern Technologies to Capture and Share Indigenous Astronomical Knowledge

    CERN Document Server

    Nakata, N M; Warren, J; Byrne, A; Pagnucco, M; Harley, R; Venugopal, S; Thorpe, K; Neville, R; Bolt, R

    2014-01-01

    Indigenous Knowledge is important for Indigenous communities across the globe and for the advancement of our general scientific knowledge. In particular, Indigenous astronomical knowledge integrates many aspects of Indigenous Knowledge, including seasonal calendars, navigation, food economics, law, ceremony, and social structure. We aim to develop innovative ways of capturing, managing, and disseminating Indigenous astronomical knowledge for Indigenous communities and the general public for the future. Capturing, managing, and disseminating this knowledge in the digital environment poses a number of challenges, which we aim to address using a collaborative project involving experts in the higher education, library, and industry sectors. Using Microsoft's WorldWide Telescope and Rich Interactive Narratives technologies, we propose to develop software, media design, and archival management solutions to allow Indigenous communities to share their astronomical knowledge with the world on their terms and in a cult...

  7. REM: A Collaborative Framework for Building Indigenous Cultural Competence.

    Science.gov (United States)

    Power, Tamara; Virdun, Claudia; Sherwood, Juanita; Parker, Nicola; Van Balen, Jane; Gray, Joanne; Jackson, Debra

    2016-09-01

    The well-documented health disparities between the Australian Indigenous and non-Indigenous population mandates a comprehensive response from health professionals. This article outlines the approach taken by one faculty of health in a large urban Australian university to enhance cultural competence in students from a variety of fields. Here we outline a collaborative and deeply respectful process of Indigenous and non-Indigenous university staff collectively developing a model that has framed the embedding of a common faculty Indigenous graduate attribute across the curriculum. Through collaborative committee processes, the development of the principles of "Respect; Engagement and sharing; Moving forward" (REM) has provided both a framework and way of "being and doing" our work. By drawing together the recurring principles and qualities that characterize Indigenous cultural competence the result will be students and staff learning and bringing into their lives and practice, important Indigenous cultural understanding.

  8. CONTEMPORARY INDIGENOUS LITERATURE: FORMS AND CONTENTS IN THE POETRY AND PROSE OF THE II LITERARY PARTY OF INDIGENOUS POETICS.

    Directory of Open Access Journals (Sweden)

    Deborah Goldemberg

    2010-06-01

    Full Text Available By analyzing the forms and contents of the presentations made by indigenous performers and writers at the I Literary Party of Indigenous Poetics, this article exposes the challenges faced by traditional genre theories in tackling indigenous narratives and analyses how this “crisis” contributes to widening hierarchical and Western biased conceptions. On a stage open to contemporary indigenous expression, as is the literary party, the concepts of performance and storytelling, with the social function of maintaining tradition, continuous learning and transformation, better define this indigenous expression.

  9. A Community Engaged Dental Curriculum: A Rural Indigenous Outplacement Programme.

    Science.gov (United States)

    Abuzar, Menaka A; Owen, Julie

    2016-04-26

    Indigenous people worldwide suffer from poor oral health as compared to non-Indigenous citizens. One of the approaches to bring about improvement in Indigenous oral health is to enhance the service provision by implementing oral health outplacement programmes. A case study of such a programme for dental students in Australia reports how an educational institution can successfully engage with an Indigenous oral health service to provide learning experiences to the students as well as deliver much needed services to the community. The assessment of this ongoing outplacement programme over the period of 2008-14, based on students' feedback, highlights some of the key beneficial outcomes. Students agreed that the Indigenous outplacement programme improved their understanding of Indigenous issues (mean ± SD: 4.10±0.8; 5 refers to strongly agree on 5-point scale) and increased the possibility that they will practise in Indigenous health (3.66±1.0). They were pleased with the assistance received by clinical supervisors and clinic staff at the Indigenous dental clinic (4.28±0.8). This programme has demonstrated that structured student outplacements are valuable in building relations across cultures especially with Indigenous communities. It has also shown that university engagement with the public health sector can be beneficial to both institutions. Significance for public healthAn oral health outreach programme is one of the suggested approaches to effectively address the endemic issues of poor oral health among Indigenous people around the world. An Indigenous dental clinical outplacement in Australia provides an example of beneficial outcomes of such an approach. It provides dental students with an opportunity to experience the health issues related to Australian Indigenous communities and prepare future graduates to work comfortably in the public health care system. Indigenous people also develop trust and feel comfortable in receiving oral health care services

  10. Phytase-producing capacity of yeasts isolated from traditional African fermented food products and PHYPk gene expression of Pichia kudriavzevii strains

    DEFF Research Database (Denmark)

    Greppi, Anna; Krych, Lukasz; Costantini, Antonella

    2015-01-01

    Phytate is known as a strong chelate of minerals causing their reduced uptake by the human intestine. Ninety-three yeast isolates from traditional African fermented food products, belonging to nine species (Pichia kudriavzevii, Saccharomyces cerevisiae, Clavispora lusitaniae, Kluyveromyces...

  11. Molecular Cloning of Phytase Gene from ASUIA279 and Its Expression in Pichia pastoris System

    Directory of Open Access Journals (Sweden)

    M. H. Mahamad Maifiah

    2011-12-01

    Full Text Available Phytases catalyze the hydrolysis of phytate (myo-inositol hexakisphosphate, one of the major storage form of phosphate in plants, with subsequent release of myo-inositol, phosphate and phytate-bound minerals. Non-ruminant animals such as chicken, swine and fish can't use the organic phosphorus and minerals from their diet because there is no phytase activity in their digestive tract. Phytate degrading enzyme is added to the animal feed diet to improve phosphorus availability from the dietary phytate and at the same time this lessen the phosphate pollution level in areas of intensive animal production as the phosphate would not be excreted out to the environment. ASUIA279, a bacterial strain isolated from Malaysian soil has potentially shown good phytase activity. In the present work, the gene encoding for phytase has been amplified from the plasmid DNA of recombinant ASUIA279(5 obtained from a previous study (unpublished data by a polymerase chain reaction (PCR methodology. The amplified phytase gene was extracted, purified then cloned into the pPICZA plasmid and transformed into Pichia pastorisX-33 strain for enzyme production.ABSTRAK: Fitase pemangkinan hidrolisis fitat (myo-inositol hexakisphosphate, merupakan salah satu cara penyimpanan utama fosfat dalam tumbuhan, dengan pelepasan berturut myo-inositol , fosfat dan galian terikat fitat. Haiwan bukan ruminan seperti ayam, khinzir dan ikan tidak dapat memanfaatkan fosforus organik serta galian yang diperolehi daripada makanan kerana tidak mempunyai aktiviti fitase di dalam saluran pencernaan mereka. Enzim pengecilan fitase dicampurkan ke dalam pemakanan haiwan untuk mempertingkatkan keperolehan fosforus dari fitat diet. Pada masa yang sama ia dapat mengurangkan tahap pencemaran fosfat di kawasan yang terdapat penternakan haiwan secara intensif agar fosfat tidak dikumuhkan ke persekitaran. ASUIA279, satu strain bakteria yang diasingkan daripada tanih di Malaysiamenunjukkan aktiviti fitase

  12. Western and indigenous knowledges in intercultural bilingual education in Ecuador

    Directory of Open Access Journals (Sweden)

    Carmen Martinez Novo

    2016-12-01

    Full Text Available This article analyzes the relationships between indigenous and Western forms of knowledge in intercultural bilingual education (EIB in Ecuador. The work shows that the tensions between both forms of knowledge reflect differences in the ways that indigenous leaders and communities understand the role of formal education. The article also argues that the allies of indigenous movements (religious groups, academics, NGOs, and others and indigenous communities have different educational aims. Whereas allies and indigenous leaders see EIB as a tool to preserve indigenous language and culture, indigenous parents and students seek access to western forms of knowledge such as Spanish literacy, English, and computers. Communities do not underestimate indigenous knowledges based on oral narratives and elder wisdom. However, the article argues that indigenous individuals do not think that the school system is the right place for the reproduction of this wisdom. These knowledges are typically shared outside of school premises in the domains of the family and community. The author gathered the data for this article with a group of indigenous researchers through a collaborative methodology. Thus, this study contributes with internal and critical points of view that complement those of other studies. 

  13. Indigenous, colonist, and government Impacts on Nicaragua's Bosawas Reserve.

    Science.gov (United States)

    Stocks, Anthony; McMahan, Benjamin; Taber, Peter

    2007-12-01

    We studied the impacts of colonists, two groups of indigenous residents (Miskitu and Mayangna), and management by the Nicaraguan Ministry of Environment and Natural Resources (MARENA) on the forest of the Bosawas International Biosphere Reserve. Indigenous people and colonists subsist on the natural resources of the reserve, and MARENA is responsible for protecting the area from colonization and illicit exploitation. Using geostatistical procedures and Landsat images at three different time periods, we compared per capita deforestation and boundary stabilization in areas with colonists and areas with indigenous peoples. We also examined whether the Mayangna deforested less than the Miskitu and whether the Nicaraguan government has effectively defended the Bosawas boundary against the advance of the agricultural frontier. In addition, we analyzed the current distribution of land uses within the reserve and its contiguous indigenous areas with a supervised classification of current land cover. Indigenous demarcations protected the forest successfully, whereas the Bosawas boundary itself did not inhibit colonization and consequent deforestation. Indigenous farmers deforested significantly less per capita than colonists, and the two indigenous groups in Bosawas did not differ significantly in their effects on the forest. Our results show that indigenous common-property institutions and indigenous defense of homeland have been powerful factors in protecting the forests of Bosawas and that the difficult evolution of a nested cross-scale governance system in Bosawas-under pressure from indigenous peoples-is probably the key to the forest's survival thus far.

  14. Citrobacter amalonaticus Phytase on the Cell Surface of Pichia pastoris Exhibits High pH Stability as a Promising Potential Feed Supplement

    OpenAIRE

    Cheng Li; Ying Lin; Yuanyuan Huang; Xiaoxiao Liu; Shuli Liang

    2014-01-01

    Phytase expressed and anchored on the cell surface of Pichia pastoris avoids the expensive and time-consuming steps of protein purification and separation. Furthermore, yeast cells with anchored phytase can be used as a whole-cell biocatalyst. In this study, the phytase gene of Citrobacter amalonaticus was fused with the Pichia pastoris glycosylphosphatidylinositol (GPI)-anchored glycoprotein homologue GCW61. Phytase exposed on the cell surface exhibits a high activity of 6413.5 U/g, with an ...

  15. Tuberculosis in indigenous children in the Brazilian Amazon

    Directory of Open Access Journals (Sweden)

    Caroline Gava

    2013-02-01

    Full Text Available OBJECTIVE: Assess the epidemiological aspects of tuberculosis in Brazilian indigenous children and actions to control it. METHODS: An epidemiological study was performed with 356 children from 0 to 14 years of age in Rondônia State, Amazon, Brazil, during the period 1997-2006. Cases of TB reported to the Notifiable Diseases Surveillance System were divided into indigenous and non-indigenous categories and analyzed according to sex, age group, place of residence, clinical form, diagnostic tests and treatment outcome. A descriptive analysis of cases and hypothesis test (χ² was carried out to verify if there were differences in the proportions of illness between the groups investigated. RESULTS: A total of 356 TB cases were identified (125 indigenous, 231 non-indigenous of which 51.4% of the cases were in males. In the indigenous group, 60.8% of the cases presented in children aged 0-4 years old. The incidence mean was much higher among indigenous; in 2001, 1,047.9 cases/100,000 inhabitants were reported in children aged < 5 years. Pulmonary TB was reported in more than 80% of the cases, and in both groups over 70% of the cases were cured. Cultures and histopathological exams were performed on only 10% of the patients. There were 3 cases of TB/HIV co-infection in the non-indigenous group and none in the indigenous group. The case detection rate was classified as insufficient or fair in more than 80% of the indigenous population notifications, revealing that most of the diagnoses were performed based on chest x-ray. CONCLUSIONS: The approach used in this study proved useful in demonstrating inequalities in health between indigenous and non-indigenous populations and was superior to the conventional analyses performed by the surveillance services, drawing attention to the need to improve childhood TB diagnosis among the indigenous population.

  16. Critical Indigenous Studies: From Difference to Density

    Directory of Open Access Journals (Sweden)

    Chris Andersen

    2011-04-01

    Full Text Available Proponents of the discipline of Native Studies (in its various guises have attempted to produce a methodologically and theoretically distinctive body of scholarship to justify its existence in the field of academia. Critiquing Duane Champagne’s recent article published in a flagship journal for North American Native Studies, I argue that while establishing Native Studies as a discipline has little or nothing to do with securing Native Studies departments on university campuses, a place nonetheless exists for these departments. Marrying Native Studies literature on the importance of producing tribally specific knowledge with Australian-based Whiteness Studies literature focusing on the utility of indigeneity for denaturalising white privilege, I argue that the discipline of Native Studies should justify itself departmentally by teaching about the complex forms of local indigeneity upon which white privilege is reproduced.

  17. INDIGENOUS HOUSEHOLDS, REMITTANCES AND LIFE QUALITY

    Directory of Open Access Journals (Sweden)

    Ignacio César Cruz Islas

    2014-07-01

    Full Text Available Mexican migration to other countries, primarily United States, is a phenomenon that has been studied from different approaches. It is an important flow of people who, for decades, has left Mexico in search of employment opportunities and higher income. This is due to the weakness of opportunities structure present in Mexico, predominantly in rural areas, as well as budget constraints that prevent households to improve their living conditions. Remittances from other countries, in turn, are an alternative for families to address the lack of employment opportunities and income in their homeland, as well as life-deficit conditions. To see how remittances impact on living conditions of indigenous population, in this paper we analyze living conditions of indigenous households.

  18. Nuclear rocket using indigenous Martian fuel NIMF

    Science.gov (United States)

    Zubrin, Robert

    1991-01-01

    In the 1960's, Nuclear Thermal Rocket (NTR) engines were developed and ground tested capable of yielding isp of up to 900 s at thrusts up to 250 klb. Numerous trade studies have shown that such traditional hydrogen fueled NTR engines can reduce the inertial mass low earth orbit (IMLEO) of lunar missions by 35 percent and Mars missions by 50 to 65 percent. The same personnel and facilities used to revive the hydrogen NTR can also be used to develop NTR engines capable of using indigenous Martian volatiles as propellant. By putting this capacity of the NTR to work in a Mars descent/acent vehicle, the Nuclear rocket using Indigenous Martian Fuel (NIMF) can greatly reduce the IMLEO of a manned Mars mission, while giving the mission unlimited planetwide mobility.

  19. Social networks among Indigenous peoples in Mexico.

    Science.gov (United States)

    Skoufias, Emmanuel; Lunde, Trine; Patrinos, Harry Anthony

    2010-01-01

    We examine the extent to which social networks among indigenous peoples in Mexico have a significant effect on a variety of human capital investment and economic activities, such as school attendance and work among teenage boys and girls, and migration, welfare participation, employment status, occupation, and sector of employment among adult males and females. Using data from the 10 percent population sample of the 2000 Population and Housing Census of Mexico and the empirical strategy that Bertrand, Luttmer, and Mullainathan (2000) propose, which allows us to take into account the role of municipality and language group fixed effects, we confirm empirically that social network effects play an important role in the economic decisions of indigenous people, especially in rural areas. Our analysis also provides evidence that better access to basic services such as water and electricity increases the size and strength of network effects in rural areas.

  20. Globalisation And Local Indigenous Education In Mexico

    Science.gov (United States)

    Reinke, Leanne

    2004-11-01

    Globalisation is often viewed as a threat to cultural and linguistic diversity and therefore is a central concern of educational practices and policy. The present study challenges this common view by demonstrating that local communities can use global means to support and enhance their specific practices and policies. An historical exploration of education policy in Mexico reveals that there has been a continuing struggle by indigenous peoples to maintain locally relevant modes of teaching. Indigenous peoples have increasingly used technology to maintain their languages and local cultural practices. Such accentuation of the local in a global context is exemplified by the people of Chiapas: They live in subsistence-type communities, yet their recent education movements and appeals to international solidarity (such as in the Zapatista rebellion) have employed computer-aided technologies.

  1. Are Supernovae Recorded in Indigenous Astronomical Traditions?

    CERN Document Server

    Hamacher, Duane W

    2014-01-01

    Novae and supernovae are rare astronomical events that would have had an influence on the sky-watching peoples who witnessed them. Although several bright novae/supernovae have been visible during recorded human history, there are many proposed but no confirmed accounts of supernovae in oral traditions or material culture. Criteria are established for confirming novae/supernovae in oral and material culture, and claims from around the world are discussed to determine if they meet these criteria. Australian Aboriginal traditions are explored for possible descriptions of novae/supernovae. Although representations of supernovae may exist in Indigenous traditions, and an account of a nova in Aboriginal traditions has been confirmed, there are currently no confirmed accounts of supernovae in Indigenous oral or material traditions.

  2. Education for indigenous childhood at the Indigenous Reservation Napalpí (Chaco, Argentina. 1911-1936

    Directory of Open Access Journals (Sweden)

    Teresa Laura Artieda

    2016-01-01

    Full Text Available On this article we approach the education for indigenous childhood at the Indigenous Reservation napalpí (Chaco, Argentina between 1911 and 1936, where the first plan of the national state for the confinement and discipline of the subjected natives, members of the Qom, moqoit, shinpi’ peoples, was implemented in a highly conflicting scenario of military campaigns of the national state for controlling the territorial and political indigenous domains of the territory, the expansion of capitalism and the progressive proletarian condition of those populations in the regional farms. We analyze the schooling project for the indigenous childhood in the Reservation, we present some notes on its development during the first three decades of the twentieth century and the conceptions on childhood and the educating forms attributed to the indigenous populations.this work is registered on the social history of education, it deepens previous inquiries of our authorship and it integrates anthropological and regional history researches. Our corpus of data is based in state’s legislations, civil servants reports and national organizations memoirs.

  3. INDIGENOUS LANGUAGES & PORTUGUESE LANGUAGE IN INDIGENOUS COMMUNITIES IN THE SOUTHERN OF MATO GROSSO DO SUL

    Directory of Open Access Journals (Sweden)

    Marilze Tavares

    2016-12-01

    Full Text Available Most part of indigenous adults in the Guarani communities of Mato Grosso do Sul is bilingual and has one of the indigenous languages, Guarani Kaiowá or Guarani Ñandeva, as their mother tongue and Portuguese as a second language; only a few elderly and young children still who do not go to school speak only the mother tongue. In this paper, we try to verify which impression the speakers have for each of these languages and the importance they attribute to each one of them. Data analysis showed that the mother tongue is closely related to the expression of their traditional culture; in general, the indigenous claim their languages are being transmitted to new generations, and therefore preserved in an appropriate manner in the two communities. The Portuguese is also considered very important by all informants and the main motivation for its teaching/learning is the need to contact with the non-indigenous population. These results may help us understand issues related to the future of these indigenous languages and Portuguese language in the investigated communities.

  4. Poverty among the Indigenous Peoples of Nicaragua

    OpenAIRE

    Mark Jamieson

    1999-01-01

    This report is part of a larger project undertaken by the Inter-American Development Bank (IDB) designed to obtain information which will assist the planning of poverty alleviation programs in a number of countries in Latin America. The specific aim of this report is to identify the conditions of poverty experienced by indigenous peoples in Nicaragua, to outline as far as possible the structural causes of this poverty, and to recommend measures which might be taken to alleviate it.

  5. Locally Situated Digital Representation of Indigenous Knowledge

    DEFF Research Database (Denmark)

    Winschiers-Theophilus, Heike; Jensen, Kasper Løvborg; Rodil, Kasper

    2012-01-01

    Digital re-presentation of indigenous knowledge remains an absurdity as long as we fail to deconstruct the prevalent design paradigm and techniques continuously re-framing technology within a western epistemology. This paper discusses key challenges in attempts of co-constructing a digital......’s views are brought to light within the design interactions. A new digital reality is created at the periphery of the situated knowledge through continuous negotiations and joint meaning making....

  6. Rheumatic fever in indigenous Australian children.

    Science.gov (United States)

    Parnaby, Matthew G; Carapetis, Jonathan R

    2010-09-01

    Rheumatic heart disease (RHD) caused by acute rheumatic fever (ARF) is a disease of poverty, poor hygiene and poor living standards. RHD remains one of the major causes of childhood cardiac disease in developing nations. Within developed nations, there has been a dramatic drop in the prevalence of RHD because of the improvement of living standards, access to health care and the widespread availability of penicillin-based drugs. Despite a dramatic reduction of RHD in Australia overall, it continues to be a major contributor to childhood and adult cardiac disease in Indigenous communities throughout northern and central Australia. Currently, Australia has among the highest recorded rates of ARF and RHD in the world. The most accurate epidemiological data in Australia come from the Northern Territory's RHD control programme. In the Northern Territory, 92% of people with RHD are Indigenous, of whom 85% live in remote communities and towns. The incidence of ARF is highest in 5-14-year-olds, ranging from 150 to 380 per 100,000. Prevalence rates of RHD since 2000 have steadily increased to almost 2% of the Indigenous population in the Northern Territory, 3.2% in those aged 35-44 years. Living in remote communities is a contributing factor to ARF/RHD as well as a major barrier for adequate follow-up and care. Impediments to ARF/RHD control include the paucity of specialist services, rapid turnover of health staff, lack of knowledge of ARF/RHD by health staff, patients and communities, and the high mobility of the Indigenous population. Fortunately, the recently announced National Rheumatic Fever Strategy, comprising recurrent funding to the Northern Territory, Queensland and Western Australia for control programmes, as well as the creation of a National Coordination Unit suggest that RHD control in Australia is now a tangible prospect. For the disease to be eradicated, Australia will have to address the underpinning determinants of poverty, social and living conditions.

  7. Innovative MIOR Process Utilizing Indigenous Reservoir Constituents

    Energy Technology Data Exchange (ETDEWEB)

    Hitzman, D.O.; Bailey, S.A.; Stepp, A.K.

    2003-02-11

    This research program was directed at improving the knowledge of reservoir ecology and developing practical microbial solutions for improving oil production. The goal was to identify indigenous microbial populations which can produce beneficial metabolic products and develop a methodology to stimulate those select microbes with inorganic nutrient amendments to increase oil recovery. This microbial technology has the capability of producing multiple oil releasing agents. The potential of the system will be illustrated and demonstrated by the example of biopolymer production on oil recovery.

  8. Innovative MIOR Process Utilizing Indigenous Reservoir Constituents

    Energy Technology Data Exchange (ETDEWEB)

    Hitzman, D.O.; Stepp, A.K.; Dennis, D.M.; Graumann, L.R.

    2003-02-11

    This research program was directed at improving the knowledge of reservoir ecology and developing practical microbial solutions for improving oil production. The goal was to identify indigenous microbial populations which can produce beneficial metabolic products and develop a methodology to stimulate those select microbes with inorganic nutrient amendments to increase oil recovery. This microbial technology has the capability of producing multiple oil-releasing agents.

  9. The emergence of obesity among indigenous Siberians.

    Science.gov (United States)

    Snodgrass, J Josh; Leonard, William R; Sorensen, Mark V; Tarskaia, Larissa A; Alekseev, Vasili P; Krivoshapkin, Vadim

    2006-01-01

    Once considered a disease of affluence and confined to industrialized nations, obesity is currently emerging as a major health concern in nearly every country in the world. Available data suggest that the prevalence rate of obesity has reached unprecedented levels in most developing countries, and is increasing at a rate that far outpaces that of developed nations. This increase in obesity has also been documented among North American circumpolar populations and is associated with lifestyle changes related to economic development. While obesity has not been well studied among indigenous Siberians, recent anthropological studies indicate that obesity and its associated comorbidities are important health problems.The present study examines recent adult body composition data from four indigenous Siberian populations (Evenki, Ket, Buriat, and Yakut) with two main objectives: 1) to determine the prevalence of overweight and obesity among these groups, and 2) to assess the influence of lifestyle and socioeconomic factors on the development of excess body fat. The results of this study indicate that obesity has emerged as an important health issue among indigenous Siberians, and especially for women, whose obesity rates are considerably higher than those of men (12% vs. 7%). The present study investigated the association between lifestyle and body composition among the Yakut, and documented substantial sex differences in lifestyle correlates of obesity. Yakut men with higher incomes and who owned more luxury consumer goods were more likely to have excess body fat while, among Yakut women, affluence was not strongly associated with overweight and obesity.

  10. Alcoholic fermentation of xylose by immobilized Pichia stipitis in a fixed-bed pulsed bioreactor

    Energy Technology Data Exchange (ETDEWEB)

    Sanroman, A.; Chamy, R.; Nunez, M.J.; Lema, J.M. (Santiago de Compostela Univ. (Spain). Dept. of Chemical Engineering)

    1994-01-01

    To enhance the productivity of the fermentation of xylose by Pichia stipitis, it is important to use bioreactors that (a) allow a high concentration of yeast (this can be achieved by using immobilised yeasts), (b) reduce the diffusional limitations resulting from dead volume (due mainly to the supplied and produced gas), and (c) diminish the effect of inhibition by ethanol. The process of immobilising P. stipitis in [kappa]-carrageenan was amended through the subsequent treatment of the bioparticles with a hardening agent (Al[sup 3+]). The variables examined were the cellular mass/gel mass ratio, the concentration of the hardening solution, and the contact time between the bioparticles and the hardening agent. (author)

  11. Modeling the alcoholic fermentation of xylose by Pichia stipitis using a qualitative reasoning approach

    Energy Technology Data Exchange (ETDEWEB)

    Guerrin, F. (INRA (Institut National de la Recherche Agronomique), Biometrics and Artificial Intelligence Station, 31 - Castanet-Tolosan (France)); Delgenes, J.P. (INRA, Biotechnological Lab. for Food Industry Environment, 11 - Narbonne (France)); Moletta, R. (INRA, Biotechnological Lab. for Food Industry Environment, 11 - Narbonne (France))

    1994-03-01

    Qualitative Reasoning is a set of Artificial Intelligence theories, methods, and techniques that provide an answer to modeling problems in domains in which one can have a clear notion of how a system is functioning without being able to express it as classical mathematical equations, and where is posed the problem of using jointly quantitative and qualitative data, as well as processing a big amount of complex knowledge. SIMAO ('a System to Interpret Measurements And Observations') is an attempt to deal with such problems. Although primarily devised for heterogeneous data interpretation in hydroecology, it was thought possible to use SIMAO in a wider context, like biotechnological processes. Starting from specific problems posed by a batch fermentation, the D-xylose conversion into ethanol by the yeast Pichia stipitis, this paper descibes how was built and used a SIMAO model aimed at predicting the fermentation issue from initial conditions, i.e. set-points values and substrate concentration. (orig.)

  12. Automated pipeline for rapid production and screening of HIV-specific monoclonal antibodies using pichia pastoris.

    Science.gov (United States)

    Shah, Kartik A; Clark, John J; Goods, Brittany A; Politano, Timothy J; Mozdzierz, Nicholas J; Zimnisky, Ross M; Leeson, Rachel L; Love, J Christopher; Love, Kerry R

    2015-12-01

    Monoclonal antibodies (mAbs) that bind and neutralize human pathogens have great therapeutic potential. Advances in automated screening and liquid handling have resulted in the ability to discover antigen-specific antibodies either directly from human blood or from various combinatorial libraries (phage, bacteria, or yeast). There remain, however, bottlenecks in the cloning, expression and evaluation of such lead antibodies identified in primary screens that hinder high-throughput screening. As such, "hit-to-lead identification" remains both expensive and time-consuming. By combining the advantages of overlap extension PCR (OE-PCR) and a genetically stable yet easily manipulatable microbial expression host Pichia pastoris, we have developed an automated pipeline for the rapid production and screening of full-length antigen-specific mAbs. Here, we demonstrate the speed, feasibility and cost-effectiveness of our approach by generating several broadly neutralizing antibodies against human immunodeficiency virus (HIV).

  13. Expression of soluble recombinant lipoxygenase from Pleurotus sapidus in Pichia pastoris.

    Science.gov (United States)

    Kelle, Sebastian; Zelena, Katerina; Krings, Ulrich; Linke, Diana; Berger, Ralf G

    2014-03-01

    The first heterologous expression of an iron-containing lipoxygenase from a basidiomycete in Pichia pastoris is reported. Five different expression constructs of the lipoxygenase gene LOX1 from Pleurotus sapidus were cloned and successfully transferred into P. pastoris SMD1168, but only one pPIC9K vector construct was functionally expressed. In this construct the vector-provided α-factor signal sequence was replaced by insertion of a second Kozak sequence between the signal sequence and the LOX1 gene. His(+) transformants were screened for their level of resistance to geneticin (G418). Lox1 was expressed under different culture conditions and purified using the N-terminal His-tag. Relative enzyme activity increased significantly 48h after methanol induction and was highest with 2mll(-1) inducer. The recombinant enzyme showed an optimal lipoxygenase activity at pH 7 and 30-35°C and a vmax like the wild-type enzyme.

  14. Expression of a Deschampsia antarctica Desv. Polypeptide with Lipase Activity in a Pichia pastoris Vector

    Directory of Open Access Journals (Sweden)

    Claudia Rabert

    2014-02-01

    Full Text Available The current study isolated and characterized the Lip3F9 polypeptide sequence of Deschampsia antarctica Desv. (GeneBank Accession Number JX846628, which was found to be comprised of 291 base pairs and was, moreover, expressed in Pichia pastoris X-33 cells. The enzyme was secreted after 24 h of P. pastoris culture incubation and through induction with methanol. The expressed protein showed maximum lipase activity (35 U/L with an optimal temperature of 37 °C. The lipase-expressed enzyme lost 50% of its specific activity at 25 °C, a behavior characteristic of a psychrotolerant enzyme. Recombinant enzyme activity was measured in the presence of ionic and non-ionic detergents, and a decrease in enzyme activity was detected for all concentrations of ionic and non-ionic detergents assessed.

  15. Ethanol fermentation of red oak acid prehydrolysate by the yeast Pichia stipitis CBS 5776

    Energy Technology Data Exchange (ETDEWEB)

    Tran, A.V.; Chambers, R.P.

    1986-07-01

    Xylose, the dominant sugar in red oak acid prehydrolysate, was fermented to ethanol. In batch cultures of xylose concentrations ranging from 10 to 100 g/l, Pichia stipitis CBS 5776 yielded 0.50-0.40 g ethanol per g xylose consumed. Model compounds of inhibitors generated in the acid prehydrolysis of red oak hemicelluloses, lignin and extractives, hindered the fermentation. Recycled yeasts and treatments with molecular sieve or mixed bed ion resins facilitated the ethanol fermentation of red oak acid prehydrolysate. A maximal ethanol concentration of 9.9 g/l was obtained from an acid prehydrolysate containing 21.7 g/l of xylose. Fermentation inhibitors derived from red oak lignin and extractives were identified. 20 references.

  16. The lipidome and proteome of microsomes from the methylotrophic yeast Pichia pastoris.

    Science.gov (United States)

    Klug, Lisa; Tarazona, Pablo; Gruber, Clemens; Grillitsch, Karlheinz; Gasser, Brigitte; Trötzmüller, Martin; Köfeler, Harald; Leitner, Erich; Feussner, Ivo; Mattanovich, Diethard; Altmann, Friedrich; Daum, Günther

    2014-02-01

    The methylotrophic yeast Pichia pastoris is a popular yeast expression system for the production of heterologous proteins in biotechnology. Interestingly, cell organelles which play an important role in this process have so far been insufficiently investigated. For this reason, we started a systematic approach to isolate and characterize organelles from P. pastoris. In this study, we present a procedure to isolate microsomal membranes at high purity. These samples represent endoplasmic reticulum (ER) fractions which were subjected to molecular analysis of lipids and proteins. Organelle lipidomics included a detailed analysis of glycerophospholipids, fatty acids, sterols and sphingolipids. The microsomal proteome analyzed by mass spectrometry identified typical proteins of the ER known from other cell types, especially Saccharomyces cerevisiae, but also a number of unassigned gene products. The lipidome and proteome analysis of P. pastoris microsomes are prerequisite for a better understanding of functions of this organelle and for modifying this compartment for biotechnological applications.

  17. Genome-scale metabolic model of Pichia pastoris with native and humanized glycosylation of recombinant proteins

    DEFF Research Database (Denmark)

    Irani, Zahra Azimzadeh; Kerkhoven, Eduard J.; Shojaosadati, Seyed Abbas;

    2016-01-01

    Pichia pastoris is used for commercial production of human therapeutic proteins, and genome-scale models of P. pastoris metabolism have been generated in the past to study the metabolism and associated protein production by this yeast. A major challenge with clinical usage of recombinant proteins...... produced by P. pastoris is the difference in N-glycosylation of proteins produced by humans and this yeast. However, through metabolic engineering, a P. pastoris strain capable of producing humanized N-glycosylated proteins was constructed. The current genome-scale models of P. pastoris do not address...... native nor humanized N-glycosylation, and we therefore developed ihGlycopastoris, an extension to the iLC915 model with both native and humanized N-glycosylation for recombinant protein production, but also an estimation of N-glycosylation of P. pastoris native proteins. This new model gives a better...

  18. Solution structure of recombinant somatomedin B domain from vitronectin produced in Pichia pastoris

    DEFF Research Database (Denmark)

    Kjærgaard, Magnus; Gårdsvoll, Henrik; Hirschberg, Daniel;

    2007-01-01

    The cysteine-rich somatomedin B domain (SMB) of the matrix protein vitronectin is involved in several important biological processes. First, it stabilizes the active conformation of the plasminogen activator inhibitor (PAI-1); second, it provides the recognition motif for cell adhesion via...... S2-cells, both yielding structurally and functionally homogeneous protein preparations. Importantly, the entire population of our purified, recombinant SMB has a solvent exposure, both as a free domain and in complex with PAI-1, which is indistinguishable from that of plasma-derived SMB as assessed...... released from plasma-derived vitronectin by CNBr cleavage. However, different disulfide patterns and three-dimensional structures for SMB were reported. In the present study, we have expressed recombinant human SMB by two different eukaryotic expression systems, Pichia pastoris and Drosophila melanogaster...

  19. Cloning and Expression of Recombinant Human Thymosin in Yeast Pichia pastoris

    Institute of Scientific and Technical Information of China (English)

    Cao Junxia(曹俊霞); Jin Liji; Duan Yanlong; An Lijia

    2003-01-01

    The gene of human thymosin alpha 1(hT(1)was synthesised according to favorite codons of Pichia pastoris by PCR. N-terminal 28 amino acid residues of 40S ribosomal protein (RP), S24E that is N-acetylserine were replaced by hT(1 for the constitution of hT(1-RP fusion gene in order to express acetyllated thymosin α1. And also,the Asn-Gly bond was designed to faciliate isolation of the target protein.The fusion gene was cloned into the expression vector, pPIC/9K. The constructs were transformed into HIS4 mutant strain GS115 by electroporation. Both SDS-PAGE analysis and Western blot analysis indicated that the fusion protein was expressed successfully.

  20. Characterization and high expression of recombinant Ustilago maydis xylanase in Pichia pastoris.

    Science.gov (United States)

    Han, Hongjuan; You, Shuang; Zhu, Bo; Fu, Xiaoyan; Sun, Baihui; Qiu, Jin; Yu, Chengye; Chen, Lei; Peng, Rihe; Yao, Quanhong

    2015-03-01

    A recombinant xylanase gene (rxynUMB) from Ustilago maydis 521 was expressed in Pichia pastoris, and the enzyme was purified and characterized. Phylogenetic analysis demonstrated that rxynUMB belongs to glycosyl hydrolase family 11. The Trp84, Trp95, Glu93, and Glu189 residues are proposed to be present at the active site. The apparent molecular mass of the recombinant xylananse was approximately 24 kDa, and the optimum pH and temperature were 4.3 and 50 °C, respectively. Xylanase activity was enhanced by 166 and 115% with Fe(2+) and Mn(2+), respectively. The biochemical properties of this recombinant xylanase suggest that it may be a useful candidate for a variety of commercial applications.

  1. Production in Pichia pastoris of protein-based polymers with small heterodimer-forming blocks.

    Science.gov (United States)

    Domeradzka, Natalia E; Werten, Marc W T; de Vries, Renko; de Wolf, Frits A

    2016-05-01

    Some combinations of leucine zipper peptides are capable of forming α-helical heterodimeric coiled coils with very high affinity. These can be used as physical cross-linkers in the design of protein-based polymers that form supramolecular structures, for example hydrogels, upon mixing solutions containing the complementary blocks. Such two-component physical networks are of interest for many applications in biomedicine, pharmaceutics, and diagnostics. This article describes the efficient secretory production of A and B type leucine zipper peptides fused to protein-based polymers in Pichia pastoris. By adjusting the fermentation conditions, we were able to significantly reduce undesirable proteolytic degradation. The formation of A-B heterodimers in mixtures of the purified products was confirmed by size exclusion chromatography. Our results demonstrate that protein-based polymers incorporating functional heterodimer-forming blocks can be produced with P. pastoris in sufficient quantities for use in future supramolecular self-assembly studies and in various applications.

  2. Xylitol production from DEO hydrolysate of corn stover by Pichia stipitis YS-30.

    Science.gov (United States)

    Rodrigues, Rita C L B; Kenealy, William R; Jeffries, Thomas W

    2011-10-01

    Corn stover that had been treated with vapor-phase diethyl oxalate released a mixture of mono- and oligosaccharides consisting mainly of xylose and glucose. Following overliming and neutralization, a D-xylulokinase mutant of Pichia stipitis, FPL-YS30 (xyl3-∆1), converted the stover hydrolysate into xylitol. This research examined the effects of phosphoric or gluconic acids used for neutralization and urea or ammonium sulfate used as nitrogen sources. Phosphoric acid improved color and removal of phenolic compounds. D-Gluconic acid enhanced cell growth. Ammonium sulfate increased cell yield and maximum specific cell growth rate independently of the acid used for neutralization. The highest xylitol yield (0.61 g(xylitol)/g(xylose)) and volumetric productivity (0.18 g(xylitol)/g(xylose )l) were obtained in hydrolysate neutralized with phosphoric acid. However, when urea was the nitrogen source the cell yield was less than half of that obtained with ammonium sulfate.

  3. Internal ribosome entry site mediates protein synthesis in yeast Pichia pastoris.

    Science.gov (United States)

    Liang, Shuli; Lin, Ying; Li, Cheng; Ye, Yanrui

    2012-05-01

    The imitation of translation, as mediated by internal ribosome entry sites, has not yet been reported in Pichia pastoris. An IRES element from Saccharomyces cerevisiae was demonstrated to direct the translation of a dicistronic mRNA in P. pastoris. The 5′-untranslated region of GPR1 mRNA, termed GPR, was cloned into a dual reporter construct containing an upstream Rhizomucor miehei lipase (RML) and a downstream β-galactosidase gene (lacZ) from Escherichia coli BL21. After being transformed into P. pastoris, the RML gene and lacZ were simultaneously expressed. The possibility of DNA rearrangement, spurious splicing, or cryptic promoter in the GPR sequence were eliminated, indicating that expression of a second ORF was IRES-dependent. These findings strongly suggested that the IRES-dependent translation initiation mechanism is conserved in P. pastoris and provides a useful means to express multiple genes simultaneously.

  4. Enhanced activity of Rhizomucor miehei lipase by deglycosylation of its propeptide in Pichia pastoris.

    Science.gov (United States)

    Liu, Yue; Xie, Wenping; Yu, Hongwei

    2014-02-01

    Many studies have demonstrated that the properties of enzymes expressed in eukaryotes can be affected by the position and extent of glycosylation on enzyme. In this study, two potential glycosylation sites (the 8th and the 58th asparagine) were identified and the effect of propeptide glycosylation on Rhizomucor miehei lipase (RML) expressed in Pichia pastoris was investigated. To better understand the effect of glycosylation on the activity of RML, three mutants (M1, generated by N8A; M2, generated by N58A; and M3, generated by N8A and N58A) were designed to generate deglycosylated enzymes. The results showed that deglycosylated RML exhibited a twofold higher activity compared to the wild type. However, it was also found that glycosylation on the propeptide was important for the removal of the propeptide by Kex2 protease and secretion of the enzyme. Thus, our study provided a further understanding into the role of glycosylation on enzyme function.

  5. Production of a Rhizopus oryzae lipase from Pichia pastoris using alternative operational strategies.

    Science.gov (United States)

    Surribas, Anna; Stahn, Rainer; Montesinos, José Luis; Enfors, Sven-Olof; Valero, Francisco; Jahic, Mehmedalija

    2007-06-30

    Different cultivation strategies have been compared for the production of Rhizopus oryzae lipase (ROL) from Pichia pastoris. Several drawbacks have been found using a methanol non-limited fed-batch. On the one hand, oxygen limitation appeared at early cell dry weights and, on the other hand, high cell death was observed. A temperature limited fed-batch has been proposed to solve both problems. However, in our case study a methanol non-limited fed-batch results in better productivities. Finally, a lower salt medium were used to overcome cell death problems and a temperature limited fed-batch was applied thereafter to solve oxygen transfer limitations. This combined strategy has resulted in lower productivities when compared to a methanol non-limited fed-batch. However the culture could be longer prolonged and a 1.3-fold purer final product was obtained mainly due to cell death reduction.

  6. Functional expression of Rhizopus oryzae lipase in Pichia pastoris: high-level production and some properties.

    Science.gov (United States)

    Minning, S; Schmidt-Dannert, C; Schmid, R D

    1998-12-11

    The mature lipase of the fungus Rhizopus oryzae (ROL) was functionally expressed and secreted in the methylotrophic yeast Pichia pastoris. In a batch cultivation, where methanol feeding was linked to the dissolved oxygen content in the cultivation solution, a lipase activity of 500,000 units per liter (60 mg active lipase per liter) of culture was achieved after initial glycerol feeding of the culture. Recombinant ROL lipase was purified to homogeneity by a simple two-step purification procedure and had a specific activity of 8571 U mg-1 (triolein, 30 degrees C, pH 8.1) which is comparable with the purified native enzyme. The properties of the recombinant lipase were similar to those reported both for the native lipase and for the enzyme expressed in Escherichia coli and refolded from inactive inclusion bodies.

  7. Genomic sequence of the xylose fermenting, insect-inhabitingyeast, Pichia stipitis

    Energy Technology Data Exchange (ETDEWEB)

    Jeffries, Thomas W.; Grigoriev, Igor; Grimwood, Jane; Laplaza,Jose M.; Aerts, Andrea; Salamov, Asaf; Schmutz, Jeremy; Lindquist, Erika; Dehal, Paramvir; Shapiro, Harris; Jin, Yong-Su; Passoth, Volkmar; Richardson, Paul M.

    2007-06-25

    Xylose is a major constituent of angiosperm lignocellulose,so its fermentation is important for bioconversion to fuels andchemicals. Pichia stipitis is the best-studied native xylose fermentingyeast. Genes from P. stipitis have been used to engineer xylosemetabolism in Saccharomycescerevisiae, and the regulation of the P.stipitis genome offers insights into the mechanisms of xylose metabolismin yeasts. We have sequenced, assembled and finished the genome ofP.stipitis. As such, it is one of only a handful of completely finishedeukaryotic organisms undergoing analysis and manual curation. Thesequence has revealed aspects of genome organization, numerous genes forbiocoversion, preliminary insights into regulation of central metabolicpathways, numerous examples of co-localized genes with related functions,and evidence of how P. stipitis manages to achieve redox balance whilegrowing on xylose under microaerobic conditions.

  8. Expression and Characterization of HGV E2 cDNA in Pichia pastoris

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    A 559 base pair fragment of cDNA locating at the putative E2 region of GBV-C/HGV was in-serted into Pichia pastoris expression vector pPICgK in the reading frame of α-factor secreting signal pep-tide. The recombinant expression plasmid pPIC9K-E2 was introduced into P. pastoris GSll5 with electro-poration and recombined with the host genome by homological recombination. The His+Mut+ recombinantyeasts were selected and cultivated in the BMMY medium. After 3 days induction with 0. 5% methanol,the target protein(E2) accumulated up to 30% of total proteins in the supernatant. The expressed E2 pro-tein was proved possessing antigenicity and high specificity with Western blot and ELISA probed with serafrom the immunized rabbits and the patients infected by GBV-C/HGV.

  9. Improving 3'-Hydroxygenistein Production in Recombinant Pichia pastoris Using Periodic Hydrogen Peroxide-Shocking Strategy.

    Science.gov (United States)

    Wang, Tzi-Yuan; Tsai, Yi-Hsuan; Yu, I-Zen; Chang, Te-Sheng

    2016-03-01

    3'-Hydroxygenistein can be obtained from the biotransformation of genistein by the engineered Pichia pastoris X-33 strain, which harbors a fusion gene composed of CYP57B3 from Aspergillus oryzae and a cytochrome P450 oxidoreductase gene (sCPR) from Saccharomyces cerevisiae. P. pastoris X-33 mutants with higher 3'-hydroxygenistein production were selected using a periodic hydrogen peroxide-shocking strategy. One mutant (P2-D14-5) produced 23.0 mg/l of 3'-hydroxygenistein, representing 1.87-fold more than that produced by the recombinant X-33. When using a 5 L fermenter, the P2-D14-5 mutant produced 20.3 mg/l of 3'- hydroxygenistein, indicating a high potential for industrial-scale 3'-hydroxygenistein production.

  10. [Expression Of DNA-Encoded Antidote to Organophosphorus Toxins in the Methylotrophic Yeast Pichia Pastoris].

    Science.gov (United States)

    Terekhov, S S; Bobik, T V; Mokrushina, Yu A; Stepanova, A V; Aleksandrova, N M; Smirnov, I V; Belogurov, A A; Ponomarenko, N A; Gabibov, A G

    2016-01-01

    A platform for the cloning and expression of active human butyrylcholinesterase (BuChE) in the yeast Pichia pastoris is first presented. Genetic constructs for BuChE gene expression, separately and in conjunction with a proline-rich peptide called proline-rich attachment domain (PRAD), are based on the vector pPICZαA. It is shown that the highest level of production is achieved in the expression of a BuChE gene without PRAD pPICZαA. It is found that one can obtain up to 125 mg of active enzyme from 1 L of culture medium at an optimal pH environment (pH 7.6), an optical seed culture density of 3 o.u., and an optimum methanol addition mode of (0.5% methanol in the first day and 0.2% thereafter from the second day).

  11. Expression and purification of a recombinant avidin with a lowered isoelectric point in Pichia pastoris.

    Science.gov (United States)

    Zocchi, Andrea; Jobé, Anna Marya; Neuhaus, Jean-Marc; Ward, Thomas R

    2003-12-01

    A recombinant glycosylated avidin (recGAvi) with an acidic isoelectric point was expressed and secreted by the methylotrophic yeast Pichia pastoris. The coding sequence for recGAvi was de novo synthesized based on the codon usage of P. pastoris. RecGAvi is secreted at approximately 330mg/L of culture supernatant. RecGAvi monomer displays a molecular weight of 16.5kDa, as assessed by ESI mass spectrometry. N-terminal amino acid sequencing indicates the presence of three additional amino acids (E-A-E), which contribute to further lowering the isoelectric point to 5.4. The data presented here demonstrate that the P. pastoris system is suitable for the production of recGAvi and that the recombinant avidin displays biotin-binding properties similar to those of the hen-egg white protein.

  12. Desenvolvimento de linhagem auxotrófica de Pichia pastoris para o metabolismo de leucina

    OpenAIRE

    Ocampo Betancur, Maritza

    2014-01-01

    A levedura Pichia pastoris tem sido amplamente utilizada na produção de proteínas recombinantes devido a características como fácil manipulação, rápido crescimento e capacidade de fazer modificações pós-traducionais mais similares às dos mamíferos. Apesar do vasto uso desta levedura como sistema de expressão, há poucas marcas de seleção disponíveis para sua manipulação genética. As marcas existentes podem ser auxotróficas (genes de vias biossintéticas como HIS4, ARG4, URA3, ADE1, dentre outro...

  13. Isolation, characterization and evaluation of the Pichia pastoris sorbitol dehydrogenase promoter for expression of heterologous proteins.

    Science.gov (United States)

    Periyasamy, Sankar; Govindappa, Nagaraj; Sreenivas, Suma; Sastry, Kedarnath

    2013-11-01

    Sorbitol is used as a non-repressive carbon source to develop fermentation process for Mut(s) recombinant clones obtained using the AOX1 promoter in Pichia pastoris. Sorbitol dehydrogenase is an enzyme in the carbohydrate metabolism that catalyzes reduction of D-fructose into D-sorbitol in the presence of NADH. The small stretch of 211bps upstream region of sorbitol dehydrogenase coding gene has all the promoter elements like CAAT box, GC box, etc. It is able to promote protein production under repressive as well as non-repressive carbon sources. In this study, the strength of the sorbitol dehydrogenase promoter was evaluated by expression of two heterologous proteins: human serum albumin and erythrina trypsin inhibitor. Sorbitol dehydrogenase promoter allowed constitutive expression of recombinant proteins in all carbon sources that were tested to grow P. pastoris and showed activity similar to GAP promoter. The sorbitol dehydrogenase promoter was active in all the growth phases of the P. pastoris.

  14. Methyl labeling and TROSY NMR spectroscopy of proteins expressed in the eukaryote Pichia pastoris

    Energy Technology Data Exchange (ETDEWEB)

    Clark, Lindsay; Zahm, Jacob A.; Ali, Rustam [University of Texas Southwestern Medical Center, Department of Biophysics (United States); Kukula, Maciej; Bian, Liangqiao [University of Texas at Arlington, Shimadzu Center for Advanced Analytical Chemistry (United States); Patrie, Steven M. [University of Texas Southwestern Medical Center, Department of Pathology (United States); Gardner, Kevin H. [CUNY Advanced Science Research Center, Structural Biology Initiative (United States); Rosen, Michael K.; Rosenbaum, Daniel M., E-mail: dan.rosenbaum@utsouthwestern.edu [University of Texas Southwestern Medical Center, Department of Biophysics (United States)

    2015-07-15

    {sup 13}C Methyl TROSY NMR spectroscopy has emerged as a powerful method for studying the dynamics of large systems such as macromolecular assemblies and membrane proteins. Specific {sup 13}C labeling of aliphatic methyl groups and perdeuteration has been limited primarily to proteins expressed in E. coli, preventing studies of many eukaryotic proteins of physiological and biomedical significance. We demonstrate the feasibility of efficient {sup 13}C isoleucine δ1-methyl labeling in a deuterated background in an established eukaryotic expression host, Pichia pastoris, and show that this method can be used to label the eukaryotic protein actin, which cannot be expressed in bacteria. This approach will enable NMR studies of previously intractable targets.

  15. Expression of enzymes for the usage in food and feed industry with Pichia pastoris.

    Science.gov (United States)

    Spohner, Sebastian C; Müller, Hagen; Quitmann, Hendrich; Czermak, Peter

    2015-05-20

    The methylotrophic yeast Pichia pastoris is an established protein expression host for the production of industrial enzymes. This yeast can be grown to very high cell densities and produces high titers of recombinant protein, which can be expressed intercellularly or be secreted to the fermentation medium. P. pastoris offers some advantages over other established expression systems especially in protein maturation. In food and feed production many enzymatically catalyzed processes are reported and the demand for new enzymes grows continuously. For instance the unique catalytic properties of enzymes are used to improve resource efficiency, maintain quality, functionalize food, and to prevent off-flavors. This review aims to provide an overview on recent developments in heterologous production of enzymes with P. pastoris and their application within the food sector.

  16. Heterologous expression and characterization of a laccase from Laccaria bicolor in Pichia pastoris

    Directory of Open Access Journals (Sweden)

    Bo Wang

    2016-01-01

    Full Text Available Synthetic dyes are known to be highly toxic to mammalian cells and mutagenic and carcinogenic to humans and, therefore, should be detoxified and removed from industrial effluents. Different approaches for removal and detoxication are extensively sought. Biochemical methods are considered the most economical and effective method of dye decolourization. In this research, the laccase gene from Laccaria bicolor was modified and expressed in Pichia pastoris. The properties of the recombinant laccase and its ability to degrade synthetic dyes were studied. The laccase activity was optimal at pH 2.2 and 50 °C. Its Km value was 0.187 mmol/L for ABTS [2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid]. The laccase obtained was shown to decolorize the synthetic dyes, malachite green, crystal violet and orange G, with ABTS as a mediator. These results indicated that the laccase obtained may be used to treat industrial effluents containing artificial dyes.

  17. The Effect of Initial Cell Concentration on Xylose Fermentation by Pichia stipitis

    Science.gov (United States)

    Agbogbo, Frank K.; Coward-Kelly, Guillermo; Torry-Smith, Mads; Wenger, Kevin; Jeffries, Thomas W.

    Xylose was fermented using Pichia stipitis CBS 6054 at different initial cell concentrations. A high initial cell concentration increased the rate of xylose utilization, ethanol formation, and the ethanol yield. The highest ethanol concentration of 41.0 g/L and a yield of 0.38 g/g was obtained using an initial cell concentration of 6.5 g/L. Even though more xylitol was produced when the initial cell concentrations were high, cell density had no effect on the final ethanol yield. A two-parameter mathematical model was used to predict the cell population dynamics at the different initial cell concentrations. The model parameters, a and b correlate with the initial cell concentrations used with an R 2 of 0.99.

  18. Expression, Characterization and Antimicrobial Ability of a Variant T4 Lysozyme in Pichia pastoris

    Institute of Scientific and Technical Information of China (English)

    Ning SUN; Sanfeng CHEN; Xiangming XIE; Yueju WANG; Gangqiang LI; Nan WANG; Dehu LIU

    2014-01-01

    T4 lysozyme was engineered with disulfide bonds and expressed in Pichia pastoris. The secreted proteins were purified and made into powder by lyophiliza-tion. Recombinant protein purity was more than 70% measured by HPLC. The lytic activity of variant T4-lysozyme was measured by the lysis of the cel wal of Xan-thomonas oryzae pv. oryzae, X. oryzae pv. oryzicola, Ralstonia solanacearum comb. nov, Clavibacter michiganensis subsp. michiganensis, X. campestris pv. mal-vacearum, Fusarium oxysporium sp. vasinfectum, Verticil ium dahliae kleb. Inhibition zone assay showed that variant T4 lysozyme significantly inhibited X. o. oryzicola and X. c. malvacearum. The antifungal activities of this protein against F. o. vasin-fectum and V. d. kleb were also analyzed.

  19. Synthesis and high expression of chitin deacetylase from Colletotrichum lindemuthianum in Pichia pastoris GS115.

    Science.gov (United States)

    Kang, Lixin; Chen, Xiaomei; Zhai, Chao; Ma, Lixin

    2012-09-01

    A gene, ClCDA, encoding chitin deacetylase from Colletotrichum lindemuthianum, was optimized according to the codon usage bias of Pichia pastoris and synthesized in vitro by overlap extension PCR. It was secretorily expressed in P. pastoris GS115 using the constitutive expression vector pHMB905A. The expression level reached the highest with 110 mg/l culture supernatant after 72 h of methanol induction, which comprised 77.27 U/mg chitin deacetylase activity. SDS-PAGE, mass spectrometry, and deglycosylation assays demonstrated that partial recombinant protein was glycosylated with an apparent molecular mass of 33 kDa. The amino acid sequences of recombinant proteins were confirmed by mass spectrometry.

  20. Bioproduction of benzaldehyde in a solid-liquid two-phase partitioning bioreactor using Pichia pastoris.

    Science.gov (United States)

    Jain, Ashu N; Khan, Tanya R; Daugulis, Andrew J

    2010-11-01

    The bioproduction of benzaldehyde from benzyl alcohol using Pichia pastoris was examined in a solid-liquid two-phase partitioning bioreactor (TPPB) to reduce substrate and product inhibition. Rational polymer selection identified Elvax 40W as an effective sequestering phase, possessing partition coefficients for benzyl alcohol and benzaldehyde of 3.5 and 35.4, respectively. The use of Elvax 40W increased the overall mass of benzaldehyde produced by approx. 300% in a 5 l bioreactor, relative to a single phase biotransformation. The two-phase system had a molar yield of 0.99, indicating that only minor losses occurred. These results provide a promising starting point for solid-liquid TPPBs to enhance benzaldehyde production, and suggest that multiple, targeted polymers may provide relief for transformations characterized by multiple inhibitory substrates/product/by-products.

  1. Effects of pretreatment methods for hazelnut shell hydrolysate fermentation with Pichia Stipitis to ethanol.

    Science.gov (United States)

    Arslan, Yeşim; Eken-Saraçoğlu, Nurdan

    2010-11-01

    In this study, we investigated the use of hazelnut shell as a renewable and low cost lignocellulosic material for bioethanol production for the first time. High lignin content of hazelnut shell is an important obstacle for such a biotransformation. Biomass hydrolysis with acids yields reducing sugar with several inhibitors which limit the fermentability of sugars. The various conditioning methods for biomass and hydrolysate were performed to overcome the toxicity and their effects on the subsequent fermentation of hazelnut shell hydrolysate by Pichia stipitis were evaluated with shaking flasks experiments. Hazelnut shells hydrolysis with 0.7M H(2)SO(4) yielded 49 gl(-1) total reducing sugars and fermentation inhibitors in untreated hydrolysate. First, it was shown that several hydrolysate detoxification methods were solely inefficient in achieving cell growth and ethanol production in the fermentation of hazelnut shell hydrolysates derived from non-delignified biomass. Next, different pretreatments of hazelnut shells were considered for delignification and employed before hydrolysis in conjunction with hydrolysate detoxification to improve alcohol fermentation. Among six delignification methods, the most effective pretreatment regarding to ethanol concentration includes the treatment of shells with 3% (w/v) NaOH at room temperature, which was integrated with sequential hydrolysate detoxification by overliming and then treatment with charcoal twice at 60 degrees C. This treatment brought about a total reduction of 97% in furans and 88.4% in phenolics. Almost all trialed treatments caused significant sugar loss. Under the best assayed conditions, ethanol concentration of 16.79gl(-1) was reached from a hazelnut shell hyrolysate containing initial 50g total reducing sugar l(-1) after partial synthetic xylose supplementation. This value is equal to 91.25% of ethanol concentration that was obtained from synthetic d-xylose under same conditions. The present study

  2. Solution structure of recombinant somatomedin B domain from vitronectin produced in Pichia pastoris.

    Science.gov (United States)

    Kjaergaard, Magnus; Gårdsvoll, Henrik; Hirschberg, Daniel; Nielbo, Steen; Mayasundari, Anand; Peterson, Cynthia B; Jansson, Anna; Jørgensen, Thomas J D; Poulsen, Flemming M; Ploug, Michael

    2007-09-01

    The cysteine-rich somatomedin B domain (SMB) of the matrix protein vitronectin is involved in several important biological processes. First, it stabilizes the active conformation of the plasminogen activator inhibitor (PAI-1); second, it provides the recognition motif for cell adhesion via the cognate integrins (alpha(v)beta(3), alpha(v)beta(5), and alpha(IIb)beta(3)); and third, it binds the complex between urokinase-type plasminogen activator (uPA) and its glycolipid-anchored receptor (uPAR). Previous structural studies on SMB have used recombinant protein expressed in Escherichia coli or SMB released from plasma-derived vitronectin by CNBr cleavage. However, different disulfide patterns and three-dimensional structures for SMB were reported. In the present study, we have expressed recombinant human SMB by two different eukaryotic expression systems, Pichia pastoris and Drosophila melanogaster S2-cells, both yielding structurally and functionally homogeneous protein preparations. Importantly, the entire population of our purified, recombinant SMB has a solvent exposure, both as a free domain and in complex with PAI-1, which is indistinguishable from that of plasma-derived SMB as assessed by amide hydrogen ((1)H/(2)H) exchange. This solvent exposure was only reproduced by one of three synthetic SMB products with predefined disulfide connectivities corresponding to those published previously. Furthermore, this connectivity was also the only one to yield a folded and functional domain. The NMR structure was determined for free SMB produced by Pichia and is largely consistent with that solved by X-ray crystallography for SMB in complex with PAI-1.

  3. Production of human pro-relaxin H2 in the yeast Pichia pastoris.

    Science.gov (United States)

    Cimini, D; Corte, K Della; Finamore, R; Andreozzi, L; Stellavato, A; Pirozzi, A V A; Ferrara, F; Formisano, R; De Rosa, M; Chino, M; Lista, L; Lombardi, A; Pavone, V; Schiraldi, C

    2017-01-14

    Initially known as the reproductive hormone, relaxin was shown to possess other therapeutically useful properties that include extracellular matrix remodeling, anti-inflammatory, anti-ischemic and angiogenic effects. All these findings make relaxin a potential drug for diverse medical applications. Its precursor, pro-relaxin, is an 18 kDa protein, that shows activity in in vitro assays. Since extraction of relaxin from animal tissues raises several issues, prokaryotes and eukaryotes were both used as expression systems for recombinant relaxin production. Most productive results were obtained when using Escherichia coli as a host for human relaxin expression. However, in such host, relaxin precipitated in the form of inclusion bodies and, therefore, required several expensive recovery steps as cell lysis, refolding and reduction. To overcome the issues related to prokaryotic expression here we report the production and purification of secreted human pro-relaxin H2 by using the methylotrophic yeast Pichia pastoris as expression host. The methanol inducible promoter AOX1 was used to drive expression of the native and histidine tagged forms of pro-relaxin H2 in dual phase fed-batch experiments on the 22 L scale. Both protein forms presented the correct structure, as determined by mass spectrometry and western blotting analyses, and demonstrated to be biologically active in immune enzymatic assays. The presence of the tag allowed to simplify pro-relaxin purification obtaining higher purity. This work presents a strategy for microbial production of recombinant human pro-relaxin H2 in Pichia pastoris that allowed the obtainment of biologically active pro-hormone, with a final concentration in the fermentation broth ranging between 10 and 14 mg/L of product, as determined by densitometric analyses.

  4. Heterologous expression of leader-less pga gene in Pichia pastoris: intracellular production of prokaryotic enzyme

    Directory of Open Access Journals (Sweden)

    Kyslík Pavel

    2010-02-01

    Full Text Available Abstract Background Penicillin G acylase of Escherichia coli (PGAEc is a commercially valuable enzyme for which efficient bacterial expression systems have been developed. The enzyme is used as a catalyst for the hydrolytic production of β-lactam nuclei or for the synthesis of semi-synthetic penicillins such as ampicillin, amoxicillin and cephalexin. To become a mature, periplasmic enzyme, the inactive prepropeptide of PGA has to undergo complex processing that begins in the cytoplasm (autocatalytic cleavage, continues at crossing the cytoplasmic membrane (signal sequence removing, and it is completed in the periplasm. Since there are reports on impressive cytosolic expression of bacterial proteins in Pichia, we have cloned the leader-less gene encoding PGAEc in this host and studied yeast production capacity and enzyme authenticity. Results Leader-less pga gene encoding PGAEcunder the control of AOX1 promoter was cloned in Pichia pastoris X-33. The intracellular overproduction of heterologous PGAEc(hPGAEc was evaluated in a stirred 10 litre bioreactor in high-cell density, fed batch cultures using different profiles of transient phases. Under optimal conditions, the average volumetric activity of 25900 U l-1 was reached. The hPGAEc was purified, characterized and compared with the wild-type PGAEc. The α-subunit of the hPGAEc formed in the cytosol was processed aberrantly resulting in two forms with C- terminuses extended to the spacer peptide. The enzyme exhibited modified traits: the activity of the purified enzyme was reduced to 49%, the ratios of hydrolytic activities with cephalexin, phenylacetamide or 6-nitro-3-phenylacetylamidobenzoic acid (NIPAB to penicillin G increased and the enzyme showed a better synthesis/hydrolysis ratio for the synthesis of cephalexin. Conclusions Presented results provide useful data regarding fermentation strategy, intracellular biosynthetic potential, and consequences of the heterologous expression of PGAEc

  5. Metabolic engineering of Pichia pastoris for the production of dammarenediol-II.

    Science.gov (United States)

    Liu, Xin-Bin; Liu, Min; Tao, Xin-Yi; Zhang, Zhong-Xi; Wang, Feng-Qing; Wei, Dong-Zhi

    2015-12-20

    Dammarenediol-II is the nucleus of dammarane-type ginsenosides, which are a group of active triterpenoids exhibiting various pharmacological activities. Based on the native triterpene synthetic pathway, a dammarenediol-II synthetic pathway was established in Pichia pastoris by introducing a dammarenediol-II synthase gene (PgDDS) from Panax ginseng, which is responsible for the cyclization of 2,3-oxidosqualene to dammarenediol-II in this study. To enhance productivity, a strategy of "increasing supply and reducing competitive consumption of 2,3-oxidosqualene" was used. To increase the supply of 2,3-oxidosqualene, we augmented expression of the ERG1 gene, which is responsible for 2,3-oxidosqualene synthesis. This significantly improved the yield of dammarenediol-II over 6.7-fold, from 0.030mg/g dry cell weight (DCW) to 0.203mg/g DCW. Subsequently, to reduce competition for 2,3-oxidosqualene from ergosterol biosynthesis without affecting the normal growth of P. pastoris, we targeted the ERG7gene, which is responsible for conversion of 2,3-oxidosqualene to lanosterol. This gene was downregulated by replacing its native promoter with a thiamine-repressible promoter, using a marker-recycling and gene-targeting Cre- lox71/66 system developed for P. pastoris herein. The yield of dammarenediol-II was further increased more than 3.6-fold, to 0.736mg/g DCW. Furthermore, the direct addition of 0.5g/L squalene into the culture medium further enhanced the yield of dammarenediol-II to 1.073mg/g DCW, which was 37.5-fold higher than the yield from the strain with the PgDDS gene introduction only. The P. pastoris strains engineered in this study constitute a good platform for further production of ginsenosides in Pichia species.

  6. Purification and characterization of the first recombinant bird pancreatic lipase expressed in Pichia pastoris: The turkey

    Directory of Open Access Journals (Sweden)

    Fendri Ahmed

    2011-01-01

    Full Text Available Abstract Background The turkey pancreatic lipase (TPL was purified from delipidated pancreases. Some biochemical properties and kinetic studies were determined using emulsified system and monomolecular film techniques. Those studies have shown that despite the accumulation of free fatty acids at the olive oil/water interface, TPL continues to hydrolyse efficiently the olive oil and the TC4 in the absence of colipase and bile salts, contrary to most classical digestive lipases which denaturate rapidly under the same conditions. The aim of the present study was to express TPL in the methylotrophic yeast Pichia pastoris in order to get a large amount of this enzyme exhibiting interesting biochemical properties, to purify and characterize the recombinant enzyme. Results The recombinant TPL was secreted into the culture medium and the expression level reached about 15 mg/l after 4 days of culture. Using Q-PCR, the number of expression cassette integrated on Pichia genomic DNA was estimated to 5. The purified rTPL, with molecular mass of 50 kDa, has a specific activity of 5300 U/mg on emulsified olive oil and 9500 U/mg on tributyrin. The optimal temperature and pH of rTPL were 37°C and pH 8.5. The stability, reaction kinetics and effects of calcium ions and bile salts were also determined. Conclusions Our results show that the expressed TPL have the same properties as the native TPL previously purified. This result allows us the use of the recombinant enzyme to investigate the TPL structure-function relationships.

  7. Response surface methodology based optimization of β-glucosidase production from Pichia pastoris.

    Science.gov (United States)

    Batra, Jyoti; Beri, Dhananjay; Mishra, Saroj

    2014-01-01

    The thermotolerant yeast Pichia etchellsii produces multiple cell bound β-glucosidases that can be used for synthesis of important alkyl- and aryl-glucosides. Present work focuses on enhancement of β-glucosidase I (BGLI) production in Pichia pastoris. In the first step, one-factor-at-a-time experimentation was used to investigate the effect of aeration, antifoam addition, casamino acid addition, medium pH, methanol concentration, and mixed feed components on BGLI production. Among these, initial medium pH, methanol concentration, and mixed feed in the induction phase were found to affect BGLI production. A 3.3-fold improvement in β-glucosidase expression was obtained at pH 7.5 as compared to pH 6.0 on induction with 1 % methanol. Addition of sorbitol, a non-repressing substrate, led to further enhancement in β-glucosidase production by 1.4-fold at pH 7.5. These factors were optimized with response surface methodology using Box-Behnken design. Empirical model obtained was used to define the optimum "operating space" for fermentation which was a pH of 7.5, methanol concentration of 1.29 %, and sorbitol concentration of 1.28 %. Interaction of pH and sorbitol had maximum effect leading to the production of 4,400 IU/L. The conditions were validated in a 3-L bioreactor with accumulation of 88 g/L biomass and 2,560 IU/L β-glucosidase activity.

  8. Optimal Conditions for Biomass and Recombinant Glycerol Kinase Production Using the Yeast Pichia pastoris

    Directory of Open Access Journals (Sweden)

    Sandro R. Valentini

    2011-01-01

    Full Text Available The extracellular glycerol kinase gene from Saccharomyces cerevisiae (GUT1 was cloned into the expression vector pPICZα A and integrated into the genome of the methylotrophic yeast Pichia pastoris X-33. The presence of the GUT1 insert was confirmed by PCR analysis. Four clones were selected and the functionality of the recombinant enzyme was assayed. Among the tested clones, one exhibited glycerol kinase activity of 0.32 U/mL, with specific activity of 0.025 U/mg of protein. A medium optimized for maximum biomass production by recombinant Pichia pastoris in shaker cultures was initially explored, using 2.31 % (by volume glycerol as the carbon source. Optimization was carried out by response surface methodology (RSM. In preliminary experiments, following a Plackett-Burman design, glycerol volume fraction (φ(Gly and growth time (t were selected as the most important factors in biomass production. Therefore, subsequent experiments, carried out to optimize biomass production, followed a central composite rotatable design as a function of φ(Gly and time. Glycerol volume fraction proved to have a significant positive linear effect on biomass production. Also, time was a significant factor (at linear positive and quadratic levels in biomass production. Experimental data were well fitted by a convex surface representing a second order polynomial model, in which biomass is a function of both factors (R²=0.946. Yield and specific activity of glycerol kinase were mainly affected by the additions of glycerol and methanol to the medium. The optimized medium composition for enzyme production was: 1 % yeast extract, 1 % peptone, 100 mM potassium phosphate buffer, pH=6.0, 1.34 % yeast nitrogen base (YNB, 4·10^–5 % biotin, 1 % methanol and 1 % glycerol, reaching 0.89 U/mL of glycerol kinase activity and 14.55 g/L of total protein in the medium after 48 h of growth.

  9. Cloning and expression of recombinant human platelet-derived growth factor-BB in Pichia Pink.

    Science.gov (United States)

    Babavalian, H; Latifi, A M; Shokrgozar, M A; Bonakdar, S; Tebyanian, H; Shakeri, F

    2016-07-31

    The PDGF-BB plays a key role in several pathogenesis diseases and it is believed to be an important mediator for wound healing. The recombinant human PDGF-BB is safe and effective to stimulate the healing of chronic, full thickness and lower extremity diabetic neurotrophic ulcers. In the present study, we attempted to produce a PDGF-BB growth factor and also, evaluate its functionality in cell proliferation in yeast host Pichia pink. Pichia pink yeast was used as a host for evaluation of the rhPDGF-BB expression. The coding sequence of PDGF-BB protein was synthesized after optimization and packed into the pGEM. Recombinant proteins were produced and purified. The construct of pPinkα-HC-pdgf was confirmed by sequence, the PDGF-BB protein was expressed and purified with using a nickel affinity chromatography column and then characterized by SDS-PAGE electrophoresis. The biological activity of PDGF-BB was estimated with using human fibroblast cell line. The measurement of protein concentration was determined by Bradford and human PDGF-BB ELISA kit. Purified rhPDGF-BB showed similar biological activity (as the standard PDGF-BB) and suggested that the recombinant protein has a successful protein expression (as well as considerable biological activity in P. pink host). The exact amount of recombinant PDGF-BB concentrations were measured by specific ELISA test which it was about 30 μg/ml. Our study suggested that efficiency of biological activity of PDGF-BB protein may be related to its conformational similarity with standard type and also, it practically may be important in wound healing and tissue regeneration.

  10. Early Vocabulary Development of Australian Indigenous Children: Identifying Strengths

    Directory of Open Access Journals (Sweden)

    Brad M. Farrant

    2014-01-01

    Full Text Available The current study sought to increase our understanding of the factors involved in the early vocabulary development of Australian Indigenous children. Data from the Longitudinal Study of Indigenous Children were available for 573 Indigenous children (291 boys who spoke English (M=37.0 months, SD=5.4 months, at wave 3. Data were also available for 86 children (51 boys who spoke an Indigenous language (M=37.1 months, SD=6.0 months, at wave 3. As hypothesised, higher levels of parent-child book reading and having more children’s books in the home were associated with better English vocabulary development. Oral storytelling in Indigenous language was a significant predictor of the size of children’s Indigenous vocabulary.

  11. Implementation of Indigenous Rights in Russia: Shortcomings and Recent Developments

    Directory of Open Access Journals (Sweden)

    Anna Koch

    2014-10-01

    Full Text Available After more than 20 years of active engagement in Indigenous issues, RAIPON, the umbrella organization of the Indigenous peoples of the North, Siberia, and the Far East, was ordered to suspend its activities by the Russian Ministry of Justice in November 2012. Eventually, this order was withdrawn provided that RAIPON changed its statute, which subsequently took place in early 2013. Why such sudden and definitive decisions? Apparently, the measures taken against RAIPON were due to its active engagement to defend Indigenous peoples' rights especially vis-à-vis the Russian extractive industry. A starting point for all possible explanations is thus the existing gap between the legal protection of Indigenous peoples' and its enforcement. The aims of this article are thus to gain a deeper understanding of the legal protection of Indigenous peoples’ rights in the Russian Federation, and to explore the interests and the politics lying behind the government attitude vis-à-vis Indigenous peoples.

  12. Mobility aspirations and indigenous belonging among Chakma students in Dhaka

    DEFF Research Database (Denmark)

    Visser, Jacco; Gerharz, Eva

    2016-01-01

    In recent decades, indigenous people from the Chittagong Hill Tracts (CHT) in South-east Bangladesh have experienced increased social and spatial mobility. This article investigates how indigenous students from the CHT region who have migrated to Dhaka redefine indigenous belonging. By highlighting...... the juxtaposition of different forms of mobility (physical and social) the paper responds to a recent trend which has only rarely been the subject of scholarly enquiry. In particular, it examines the experiences of mobility of individual students and explores the ways in which these students justify their quest...... forward in mainstream discourses as well as by transnational indigenous activist networks. These lead to feelings of alienation between indigenous students and their Bengali Bangladeshi peers, leaving students to increasingly draw on indigenous networks to achieve mobility...

  13. Crash and rebound of indigenous populations in lowland South America

    Science.gov (United States)

    Hamilton, Marcus J.; Walker, Robert S.; Kesler, Dylan C.

    2014-04-01

    Lowland South America has long been a battle-ground between European colonization and indigenous survival. Initial waves of European colonization brought disease epidemics, slavery, and violence that had catastrophic impacts on indigenous cultures. In this paper we focus on the demography of 238 surviving populations in Brazil. We use longitudinal censuses from all known indigenous Brazilian societies to quantify three demographic metrics: 1) effects of European contact on indigenous populations; 2) empirical estimates of minimum viable population sizes; and 3) estimates of post-contact population growth rates. We use this information to conduct population viability analysis (PVA). Our results show that all surviving populations suffered extensive mortality during, and shortly after, contact. However, most surviving populations exhibit positive growth rates within the first decade post-contact. Our findings paint a positive demographic outlook for these indigenous populations, though long-term survival remains subject to powerful externalities, including politics, economics, and the pervasive illegal exploitation of indigenous lands.

  14. A Comparison between Australian Football League (AFL Injuries in Australian Indigenous versus Non-indigenous Players

    Directory of Open Access Journals (Sweden)

    Jessica Orchard

    2013-09-01

    Full Text Available It has previously been shown that being of aboriginal descent is a risk factor for hamstring injuries in Australian football. The aim of this study was to review the Australian Football League (AFL injury database to determine whether there were any injuries where indigenous players had different relative risks to non-indigenous players. Analysis was conducted using data from the AFL injury database, which included data from 4,492 players over 21 years (1992–2012, covering 162,683 player-matches at AFL level, 91,098 matches at lower levels and 328,181 weeks (possible matches of exposure. Compared to non-indigenous players, indigenous players had a significantly higher risk of hamstring injuries (RR 1.52, 95% CI 1.32–1.73 and calf strains (RR 1.30, 95% CI 1.00–1.69. Conversely, indigenous players had a significantly lower risk of lumbar/thoracic spine injuries (RR 0.61, 95% CI 0.41–0.91, groin strains/osteitis pubis (RR 0.75, 95% CI 0.58–0.96 and Achilles tendon injuries (RR 0.32, 95% CI 0.12–0.86. The results for the above injuries were also significant in terms of games missed. There was no difference between overall risk of injury (RR 1.03, 95% CI 0.96–1.10 or missed games (RR 1.00, 95% CI 0.97–1.04. This suggests that indigenous AFL players have the same overall number of injuries and missed games, but a slightly different injury profile.

  15. Poor food and nutrient intake among Indigenous and non-Indigenous rural Australian children

    Directory of Open Access Journals (Sweden)

    Gwynn Josephine D

    2012-02-01

    Full Text Available Abstract Background The purpose of this study was to describe the food and nutrient intake of a population of rural Australian children particularly Indigenous children. Participants were aged 10 to 12 years, and living in areas of relative socio-economic disadvantage on the north coast of New South Wales. Methods In this descriptive cross-sectional study 215 children with a mean age of 11.30 (SD 0.04 years (including 82 Indigenous children and 93 boys completed three 24-hour food recalls (including 1 weekend day, over an average of two weeks in the Australian summer of late 2005. Results A high proportion of children consumed less than the Australian Nutrient Reference Values for fibre (74-84% less than Adequate Intake (AI, calcium (54-86% less than Estimated Average Requirement (EAR, folate and magnesium (36% and 28% respectively less than EAR among girls, and the majority of children exceeded the upper limit for sodium (68-76% greater than Upper Limit (UL. Energy-dense nutrient-poor (EDNP food consumption contributed between 45% and 49% to energy. Hot chips, sugary drinks, high-fat processed meats, salty snacks and white bread were the highest contributors to key nutrients and sugary drinks were the greatest per capita contributor to daily food intake for all. Per capita intake differences were apparent by Indigenous status. Consumption of fruit and vegetables was low for all children. Indigenous boys had a higher intake of energy, macronutrients and sodium than non-Indigenous boys. Conclusions The nutrient intake and excessive EDNP food consumption levels of Australian rural children from disadvantaged areas are cause for concern regarding their future health and wellbeing, particularly for Indigenous boys. Targeted intervention strategies should address the high consumption of these foods.

  16. AGROBUSINESS PERSPECTIVES IN THE INDIGENOUS DEVELOPMENT: CASE QUERETARO

    OpenAIRE

    Gerardo Gómez González, Elvia Xitlaly Gómez Calderón y Yuriena Gerenarda Gómez Calderón

    2008-01-01

    This document argueses about aspects of agribusiness in the indigenous development in the state of Queretaro, considers as an economic activity under taken in rural areas, principally related to the use of agricultural and forestry resources, with an efficient management of productive resources. The approaches outlined here are the result of a research, training and organization in which over half a year the representatives and indigenous leaders of the State Council of Indigenous People of t...

  17. The management of diabetes in indigenous Australians from primary care

    Directory of Open Access Journals (Sweden)

    Thomas Merlin C

    2007-10-01

    Full Text Available Abstract Background Indigenous Australians have high rates of diabetes and its complications. This study examines ethnic differences in the management of patients with type 2 diabetes in Australian primary care. Methods Diabetes management and outcomes in Indigenous patients enrolled in the NEFRON study (n = 144 was systematically compared with that in non-Indigenous patients presenting consecutively to the same practitioner (n = 449, and the NEFRON cohort as a whole (n = 3893. Results Indigenous Australians with diabetes had high rates of micro- and macrovascular disease. 60% of Indigenous patients had an abnormal albumin to creatinine ratio compared to 33% of non-Indigenous patients (p 1c ≥ 8.0%, observed in 55% of all Indigenous patients, despite the similar frequency use of oral antidiabetic agents and insulin. Smoking was also more common in Indigenous patients (38%vs 10%, p Conclusion Although seeing the same doctors and receiving the same medications, glycaemic and smoking cessation targets remain unfulfilled in Indigenous patients. This cross-sectional study confirms Aboriginal ethnicity as a powerful risk factor for microvascular and macrovascular disease, which practitioners should use to identify candidates for intensive multifactorial intervention.

  18. A community engaged dental curriculum: a rural Indigenous outplacement programme

    Directory of Open Access Journals (Sweden)

    Menaka A. Abuzar

    2016-04-01

    Full Text Available Background. Indigenous people worldwide suffer from poor oral health as compared to non-Indigenous citizens. One of the approaches to bring about improvement in Indigenous oral health is to enhance the service provision by implementing oral health outplacement programmes. A case study of such a programme for dental students in Australia reports how an educational institution can successfully engage with an Indigenous oral health service to provide learning experiences to the students as well as deliver much needed services to the community. Design and Methods. The assessment of this ongoing outplacement programme over the period of 2008-14, based on students’ feedback, highlights some of the key beneficial outcomes. Students agreed that the Indigenous outplacement programme improved their understanding of Indigenous issues (mean ± SD: 4.10±0.8; 5 refers to strongly agree on 5-point scale and increased the possibility that they will practise in Indigenous health (3.66±1.0. They were pleased with the assistance received by clinical supervisors and clinic staff at the Indigenous dental clinic (4.28±0.8. Conclusions. This programme has demonstrated that structured student outplacements are valuable in building relations across cultures especially with Indigenous communities. It has also shown that university engagement with the public health sector can be beneficial to both institutions.

  19. Optimization of the expression of surface antigen SAG1/2 of Toxoplasma gondii in the yeast Pichia pastoris.

    Science.gov (United States)

    Thiruvengadam, G; Init, I; Fong, M Y; Lau, Y L

    2011-12-01

    Surface antigens are the most abundant proteins found on the surface of the parasite Toxoplasma gondii. Surface antigen 1 (SAG1) and Surface antigen 2 (SAG2) remain the most important and extensively studied surface proteins. These antigens have been identified to play a role in host cell invasion, immune modulation, virulence attenuation. Recombinant SAG1/2 was cloned and expressed in yeast Pichia pastoris. We describe here optimization of critical parameters involved in high yield expression of the recombinant SAG1/2. Our results suggest that recombinant SAG1/2 were best expressed at 30ºC, pH 6 and 1% methanol as the carbon source by X33 Pichia cells. Additional optimizations included the downstream process such as ammonium sulphate precipitation and dialysis. The fusion protein was purified using Ni-NTA purification system with 80% recovery. The purified protein was 100% specific and sensitive in detection of toxoplasmosis.

  20. Heterologous expression of hydrophobins RodA and RodB from Aspergillus fumigatus in host Pichia pastoris

    DEFF Research Database (Denmark)

    Pedersen, Mona Højgaard; Borodina, Irina; Frisvad, Jens Christian

    and investigation using the expression host Pichia pastoris. Methods and materials: The genes encoding hydrophobins RodA and RodB were amplified by RT-PCR with gene-specific primers from the total RNA isolated from the spores of A. fumigatus (AF296 strain). The resulting cDNA was cloned into TOPO vectors using TOPO...... proteins with the signal sequence of alfa-mating factor from Saccharomyces cerevisia known to work well for protein secretion from P. pastoris and the pPICZB plasmids had proteins cloned with their native signal sequences. The plasmids were linearized, transformed into P. pastoris strain X33...... of the purification of hydrophobins and functional investigations are being carried out at the moment. Conclusion: Hydrophobins RodA and RodB from Aspergillus fumigatus were successfully expressed and secreted by yeast host Pichia pastoris....

  1. Indigenous Georgian Wine-Associated Yeasts and Grape Cultivars to Edit the Wine Quality in a Precision Oenology Perspective

    Science.gov (United States)

    Vigentini, Ileana; Maghradze, David; Petrozziello, Maurizio; Bonello, Federica; Mezzapelle, Vito; Valdetara, Federica; Failla, Osvaldo; Foschino, Roberto

    2016-01-01

    In Georgia, one of the most ancient vine-growing environment, the homemade production of wine is still very popular in every rural family and spontaneous fermentation of must, without addition of chemical preservatives, is the norm. The present work investigated the yeast biodiversity in five Georgian areas (Guria, Imereti, Kakheti, Kartli, Ratcha-Lechkhumi) sampling grapes and wines from 22 different native cultivars, in 26 vineyards and 19 family cellars. One hundred and eighty-two isolates were ascribed to 15 different species by PCR-ITS and RFLP, and partial sequencing of D1/D2 domain 26S rDNA gene. Metschnikowia pulcherrima (F’ = 0.56, I’ = 0.32), Hanseniaspora guilliermondii (F’ = 0.49, I’ = 0.27), and Cryptococcus flavescens (F’ = 0.31, I’ = 0.11) were the dominant yeasts found on grapes, whereas Saccharomyces cerevisiae showed the highest prevalence into wine samples. Seventy four isolates with fermentative potential were screened for oenological traits such as ethanol production, resistance to SO2, and acetic acid, glycerol and H2S production. Three yeast strains (Kluyveromyces marxianus UMY207, S. cerevisiae UMY255, Torulaspora delbrueckii UMY196) were selected and separately inoculated in vinifications experiments at a Georgian cellar. Musts were prepared from healthy grapes of local varieties, Goruli Mtsvane (white berry cultivar) and Saperavi (black berry cultivar). Physical (°Brix) and microbial analyses (plate counts) were performed to monitor the fermentative process. The isolation of indigenous S. cerevisiae yeasts beyond the inoculated strains indicated that a co-presence occurred during the vinification tests. Results from quantitative GC-FID analysis of volatile compounds revealed that the highest amount of fermentation flavors, such as 4-ethoxy-4-oxobutanoic acid (monoethyl succinate), 2-methylpropan-1-ol, ethyl 2-hydroxypropanoate, and 2-phenylethanol, were significantly more produced in fermentation conducted in Saperavi variety

  2. Large-scale production and purification of recombinant Galanthus nivalis agglutinin (GNA) expressed in the methylotrophic yeast Pichia pastoris.

    Science.gov (United States)

    Baumgartner, Philippe; Harper, Karen; Raemaekers, Romaan J M; Durieux, Alain; Gatehouse, Angharad M R; Davies, Howard V; Taylor, Mark A

    2003-08-01

    The gene coding for agglutinin from Galanthus nivalis (GNA) was expressed in, and secreted by, the methylotrophic yeast, Pichia pastoris. Transformants of P. pastoris were selected and a process to produce and purify gram quantities of recombinant GNA was developed. GNA was secreted at approximately 80 mg l(-1) at the 200 1 scale and was purified to 95% homogeneity using hydrophobic interaction chromatography. The recombinant protein was similar to the protein synthesised in plant with respect to structure and biological activity.

  3. Pichia pastoris Exhibits High Viability and a Low Maintenance Energy Requirement at Near-Zero Specific Growth Rates

    OpenAIRE

    Rebnegger, Corinna; Vos, Tim; Alexandra B. Graf; Valli, Minoska; Pronk, Jack T.; Daran-Lapujade, Pascale; Mattanovich, Diethard

    2016-01-01

    ABSTRACT The yeast Pichia pastoris is a widely used host for recombinant protein production. Understanding its physiology at extremely low growth rates is a first step in the direction of decoupling product formation from cellular growth and therefore of biotechnological relevance. Retentostat cultivation is an excellent tool for studying microbes at extremely low specific growth rates but has so far not been implemented for P. pastoris. Retentostat feeding regimes were based on the maintenan...

  4. Novel Strategy of Using Methyl Esters as Slow Release Methanol Source during Lipase Expression by mut+ Pichia pastoris X33

    OpenAIRE

    Arti Kumari; Rani Gupta

    2014-01-01

    One of the major issues with heterologous production of proteins in Pichia pastoris X33 under AOX1 promoter is repeated methanol induction. To obviate repeated methanol induction, methyl esters were used as a slow release source of methanol in lipase expressing mut+ recombinant. Experimental design was based on the strategy that in presence of lipase, methyl esters can be hydrolysed to release their products as methanol and fatty acid. Hence, upon break down of methyl esters by lipase, first ...

  5. Widening inequality in extreme macrosomia between Indigenous and non-Indigenous populations of Québec, Canada.

    Science.gov (United States)

    Auger, Nathalie; Park, Alison L; Zoungrana, Hamado; Fon Sing, Mélanie; Lo, Ernest; Luo, Zhong-Cheng

    2013-02-01

    To evaluate trends in macrosomia by severity in Indigenous vs. non-Indigenous populations of Québec, Canada. We used a retrospective cohort of 2,298,332 singleton live births in the province of Québec, 1981-2008. Indigenous births were identified by community of residence (First Nations, Inuit, non-Indigenous) and language spoken (First Nations, Inuit, French/English). High birth weight (HBW) and large-for-gestational-age (LGA) births were categorised by severity (moderate, very, extreme). Time trends in HBW and LGA, by severity, were estimated using odds ratios (OR) and rate differences for Indigenous vs. non-Indigenous births, adjusting for maternal characteristics. Relative to non-Indigenous, First Nations (but not Inuit) had higher rates of extreme HBW (1.3% vs. 0.1%) and extreme LGA birth (12.6% vs. 2.2%), and rates increased over time. First Nations had progressively elevated ORs with greater severity of macrosomia, and associations were strongest for extreme HBW >5,000 g (OR=12.4) and LGA >97th percentile (OR=7.2). Inequalities in extreme macrosomia between First Nations and non-Indigenous Quebecers are pronounced and widened between 1981 and 2008. Studies are needed to determine why macrosomia rates are increasing in Québec's First Nations, and how they compare with Indigenous sub-groups of demographically similar countries, including Australia and New Zealand. © 2013 The Authors. ANZJPH © 2013 Public Health Association of Australia.

  6. A simple Pichia pastoris fermentation and downstream processing strategy for making recombinant pandemic Swine Origin Influenza a virus Hemagglutinin protein.

    Science.gov (United States)

    Athmaram, T N; Singh, Anil Kumar; Saraswat, Shweta; Srivastava, Saurabh; Misra, Princi; Kameswara Rao, M; Gopalan, N; Rao, P V L

    2013-02-01

    The present Influenza vaccine manufacturing process has posed a clear impediment to initiation of rapid mass vaccination against spreading pandemic influenza. New vaccine strategies are therefore needed that can accelerate the vaccine production. Pichia offers several advantages for rapid and economical bulk production of recombinant proteins and, hence, can be attractive alternative for producing an effective influenza HA based subunit vaccine. The recombinant Pichia harboring the transgene was subjected to fed-batch fermentation at 10 L scale. A simple fermentation and downstream processing strategy is developed for high-yield secretory expression of the recombinant Hemagglutinin protein of pandemic Swine Origin Influenza A virus using Pichia pastoris via fed-batch fermentation. Expression and purification were optimized and the expressed recombinant Hemagglutinin protein was verified by sodium dodecyl sulfate polyacrylamide gel electrophoresis, Western blot and MALDI-TOF analysis. In this paper, we describe a fed-batch fermentation protocol for the secreted production of Swine Influenza A Hemagglutinin protein in the P. pastoris GS115 strain. We have shown that there is a clear relationship between product yield and specific growth rate. The fed-batch fermentation and downstream processing methods optimized in the present study have immense practical application for high-level production of the recombinant H1N1 HA protein in a cost effective way using P. pastoris.

  7. A dynamic method based on the specific substrate uptake rate to set up a feeding strategy for Pichia pastoris.

    Science.gov (United States)

    Dietzsch, Christian; Spadiut, Oliver; Herwig, Christoph

    2011-03-03

    Pichia pastoris is one of the most important host organisms for the recombinant production of proteins in industrial biotechnology. To date, strain specific parameters, which are needed to set up feeding profiles for fed batch cultivations, are determined by time-consuming continuous cultures or consecutive fed batch cultivations, operated at different parameter sets. Here, we developed a novel approach based on fast and easy to do batch cultivations with methanol pulses enabling a more rapid determination of the strain specific parameters specific substrate uptake rate qs, specific productivity qp and the adaption time (Δtimeadapt) of the culture to methanol. Based on qs, an innovative feeding strategy to increase the productivity of a recombinant Pichia pastoris strain was developed. Higher specific substrate uptake rates resulted in increased specific productivity, which also showed a time dependent trajectory. A dynamic feeding strategy, where the setpoints for qs were increased stepwise until a qs max of 2.0 mmol·g-1·h-1 resulted in the highest specific productivity of 11 U·g-1·h-1. Our strategy describes a novel and fast approach to determine strain specific parameters of a recombinant Pichia pastoris strain to set up feeding profiles solely based on the specific substrate uptake rate. This approach is generic and will allow application to other products and other hosts.

  8. A dynamic method based on the specific substrate uptake rate to set up a feeding strategy for Pichia pastoris

    Directory of Open Access Journals (Sweden)

    Herwig Christoph

    2011-03-01

    Full Text Available Abstract Background Pichia pastoris is one of the most important host organisms for the recombinant production of proteins in industrial biotechnology. To date, strain specific parameters, which are needed to set up feeding profiles for fed batch cultivations, are determined by time-consuming continuous cultures or consecutive fed batch cultivations, operated at different parameter sets. Results Here, we developed a novel approach based on fast and easy to do batch cultivations with methanol pulses enabling a more rapid determination of the strain specific parameters specific substrate uptake rate qs, specific productivity qp and the adaption time (Δtimeadapt of the culture to methanol. Based on qs, an innovative feeding strategy to increase the productivity of a recombinant Pichia pastoris strain was developed. Higher specific substrate uptake rates resulted in increased specific productivity, which also showed a time dependent trajectory. A dynamic feeding strategy, where the setpoints for qs were increased stepwise until a qs max of 2.0 mmol·g-1·h-1 resulted in the highest specific productivity of 11 U·g-1·h-1. Conclusions Our strategy describes a novel and fast approach to determine strain specific parameters of a recombinant Pichia pastoris strain to set up feeding profiles solely based on the specific substrate uptake rate. This approach is generic and will allow application to other products and other hosts.

  9. Indigenizing Student-Centred Learning: A Western Approach in an Indigenous Educational Institution

    Science.gov (United States)

    Kennedy, Chona Pineda

    2013-01-01

    This study investigated the alignment of the teaching and learning practices with a student-centred learning approach in an indigenous educational institution. The findings indicated that when a western concept is applied in the classroom, it is vital for it to be culturally relevant and appropriate to the cultural beliefs and values of the…

  10. The Need for Learning Arenas: Non-Indigenous Teachers Working in Indigenous School Contexts

    Science.gov (United States)

    Parding, Karolina

    2013-01-01

    Work contexts shape conditions for work. Teachers working in Indigenous school contexts face conditions different from teachers working in mainstream schools. Challenging working conditions for these teachers result in high teacher turnover, making it even more difficult for already disadvantaged students to progress. From a social justice…

  11. Indigenizing Student-Centred Learning: A Western Approach in an Indigenous Educational Institution

    Science.gov (United States)

    Kennedy, Chona Pineda

    2013-01-01

    This study investigated the alignment of the teaching and learning practices with a student-centred learning approach in an indigenous educational institution. The findings indicated that when a western concept is applied in the classroom, it is vital for it to be culturally relevant and appropriate to the cultural beliefs and values of the…

  12. Challenges in Applying Indigenous Evaluation Practices in Mainstream Grant Programs to Indigenous Communities

    Science.gov (United States)

    Grover, Jane Gray

    2008-01-01

    How can indigenous evaluators implement culturally competent models in First Nations communities while ensuring that government grant evaluation requirements are met? Through describing the challenges in one tribal community in the United States, this article will discuss how American Indian/Alaska Native substance abuse prevention programs are…

  13. Soil indigenous knowledge in North Central Namibia

    Science.gov (United States)

    Prudat, Brice; Bloemertz, Lena; Kuhn, Nikolaus J.

    2016-04-01

    Mapping and classifying soils is part of an important learning process to improve soil management practices, soil quality and increase productivity. In order to assess soil quality improvement related to an ongoing land reform in North-Central Namibia, the characteristics that determine soil quality in the local land use context were determined in this study. To do so, we collated the indigenous soil knowledge in North-Central Namibia where the Ovakwanyama cultivate pearl millet for centuries. Local soil groups are defined mostly based on their productivity potential, which varies depending on the rainfall pattern. The morphological criteria used by the farmers to differentiate the soil groups (colour, consistence) were supported by a conventional analysis of soil physical and chemical properties. Now, they can be used to develop a soil quality assessment toolbox adapted to the regional use. The characteristics of the tool box do not directly indicate soil quality, but refer to local soils groups. The quality of these groups is relatively homogenous at the local scale. Our results show that understanding of indigenous soil knowledge has great potential to improve soil quality assessment with regards to land use. The integration of this knowledge with the conventional soil analysis improves the local meaning of such a "scientific" assessment and thus facilitates dialog between farmers and agronomists, but also scientists working in different regions of the world, but in similar conditions. Overall, the integration of indigenous knowledge in international classification systems (e.g. WRB) as attempted in this study has thus a major potential to improve soil mapping in the local context.

  14. Create a new vision for indigenous development

    Energy Technology Data Exchange (ETDEWEB)

    Chavez Alba, Rafael; Sanchez Arancibia, Oscar Armando [TRANSIERRA S.A., Santa Cruz (Bolivia)

    2009-07-01

    Transierra is a Bolivian company created in the year 2000 with the goal of transporting natural gas from the fields of San Alberto and San Antonio, in Tarija, to the Rio Grande Gas Compression Plant in Santa Cruz, for export to Brazil. Transierra has implemented a Social Action Plan, which allowed it to execute more than 800 community projects for the benefit of over 40 thousand families living in it's area of influence, with the presence of 146 indigenous communities, generally lagging behind in economic and productive life in the region and country. The Support Program to Guarani Development Plans (PA-PDG) is part of the Social Plan and is part of a long-term agreement signed between Transierra and indigenous organizations. The program has implemented more than one hundred projects for productive development, health, education, cultural revaluation, and strengthening organizational infrastructure, generating huge benefits in improving the living conditions of thousands of families of the Guarani people. This year a unique initiative was created with 4 Indigenous Captains and with the support of the International Finance Corporation (World Bank Group), including Business Plans to promote sustainable economic growth, created productive economic cycles involving improvements to the production and productivity to enter the commercial distribution of local and national markets. These four initiatives have meant a shift in the implementation and is helping to generate new dynamics in production, in addition to capturing significant resources from public and private investment, laying the groundwork for the improvement of the incomes and quality of life of its beneficiaries. (author)

  15. Innovative MIOR Process Utilizing Indigenous Reservoir Constituents

    Energy Technology Data Exchange (ETDEWEB)

    Hitzman, D.O.; Stepp, A.K.

    2003-02-11

    This research program was directed at improving the knowledge of reservoir ecology and developing practical microbial solutions for improving oil production. The goal was to identify indigenous microbial populations which can produce beneficial metabolic products and develop a methodology to stimulate those select microbes with inorganic nutrient amendments to increase oil recovery. This microbial technology has the capability of producing multiple oil-releasing agents. The potential of the system will be illustrated and demonstrated by the example of biopolymer production on oil recovery.

  16. Adult Education and Indigenous Peoples in Latin America

    Science.gov (United States)

    Schmelkes, Sylvia

    2011-01-01

    This article describes the educational situation of indigenous peoples in Latin America, and in particular their scant participation in adult education activities. It analyses the historical, structural and institutional barriers to their greater involvement in adult education. The article proposes to look at indigenous demands on education as a…

  17. Maori University Graduates: Indigenous Participation in Higher Education

    Science.gov (United States)

    Theodore, Reremoana; Tustin, Karen; Kiro, Cynthia; Gollop, Megan; Taumoepeau, Mele; Taylor, Nicola; Chee, Kaa-Sandra; Hunter, Jackie; Poulton, Richie

    2016-01-01

    Maori, the indigenous population of New Zealand, are gaining university qualifications in greater numbers. This article describes the history of Maori university graduates, their current situation and the implications for indigenous futures. Section one provides a brief overview of historical policies and practices that, similar to those used on…

  18. The Limits of Cultural Competence: An Indigenous Studies Perspective

    Science.gov (United States)

    Carey, Michelle

    2015-01-01

    Taking the Universities Australia report, "National best practice framework for Indigenous cultural competency in Australian universities" (2011) as the starting point for its discussion, this paper examines the applicability of cultural competence in the design and delivery of Australian Indigenous Studies. It argues that both the…

  19. Factors Associated with Growth in Daily Smoking among Indigenous Adolescents

    Science.gov (United States)

    Whitlock, Les B.; Sittner Hartshorn, Kelley J.; McQuillan, Julia; Crawford, Devan M.

    2012-01-01

    North American Indigenous adolescents smoke earlier, smoke more, and are more likely to become regular smokers as adults than youth from any other ethnic group, yet we know very little about their early smoking trajectories. We use multilevel growth modeling across five waves of data from Indigenous adolescents (aged 10-13 years at Wave 1) to…

  20. Indigenous Youth and Bilingualism--Theory, Research, Praxis

    Science.gov (United States)

    McCarty, Teresa L.; Wyman, Leisy T.

    2009-01-01

    In this introduction, we situate the theme issue within a growing body of research on Indigenous youth language practices, communicative repertoires, and ideologies, articulating points of intersection in scholarship on Indigenous and immigrant youth bilingualism. Our geographic focus is North America. Ethnographic studies from the Far North to…

  1. Indigenous Language Revitalization, Promotion, and Education: Function of Digital Technology

    Science.gov (United States)

    Galla, Candace Kaleimamoowahinekapu

    2016-01-01

    Within the last two decades, there has been increased interest in how technology supports Indigenous language revitalization and reclamation efforts. This paper considers the effect technology has on Indigenous language learning and teaching, while conceptualizing how language educators, speakers, learners, and technology users holistically…

  2. [The indigenous population. Demographic expansion of ethnic groups].

    Science.gov (United States)

    Valdes De Montano, L M

    1989-01-01

    Analysis of the vital statistics of the 450 Mexican municipios in which 70% or more of the population speak indigenous languages permits a rough analysis of the demographic behavior of 23 different ethnic groups. Statistics of government departments dealing with indigenous affairs and the various indigenous organizations created since the 1970s provide other estimates of the size of the indigenous population. The census indicated a population of 5,181,038 speakers of indigenous languages aged 5 or older in 1980. The public administration estimated the 1980 indigenous population at 10 million, and the indigenous organizations estimated 12 to 18 million in 1980 and 15 million in 1989. Census and vital statistics data indicate that each indigenous group has had unique patterns of mortality and fertility in the past 30 years. Mortality declined significantly in some groups, but fertility overall has not declined to the same extent. In 1970 and 1980 respectively for the sample as a whole, the average crude birth rates were 42.4 and 37.3/1000 and the average crude death rates were 13.8 and 8.2/1000. The average annual growth rate was estimated at 2.9% for both years. 1980 growth rates ranged from 3.5% for the Maya and Mixtec to a low of 0.5% for the Tzotzil.

  3. Genetic and nutrition development of indigenous chicken in Africa

    DEFF Research Database (Denmark)

    Khobondo, J O; Muasya, T K; Miyumo, S

    2015-01-01

    This review gives insights into genetic and feeding regime development for indigenous chicken genetic resources. We highlight and combine confirming evidence of genetic diversity and variability using morphological and molecular techniques. We further discuss previous past and current genetic...... requirement for indigenous chicken and report nutritive contents of various local feedstuffs under various production systems. Various conservation strategies for sustainable utilization are hereby reviewed...

  4. Genetic and nutrition development of indigenous chicken in Africa

    DEFF Research Database (Denmark)

    Khobondo, J O; Muasya, T K; Miyumo, S

    2015-01-01

    This review gives insights into genetic and feeding regime development for indigenous chicken genetic resources. We highlight and combine confirming evidence of genetic diversity and variability using morphological and molecular techniques. We further discuss previous past and current genetic...... requirement for indigenous chicken and report nutritive contents of various local feedstuffs under various production systems. Various conservation strategies for sustainable utilization are hereby reviewed...

  5. Alternative Education Engaging Indigenous Young People: Flexi Schooling in Queensland

    Science.gov (United States)

    Shay, Marnee; Heck, Deborah

    2015-01-01

    This article will discuss some of the findings from a qualitative research project that explored the connections between alternative education and Indigenous learners. This study investigated how flexi school leaders reported they were supporting Indigenous young people to remain engaged in education. The results of the survey provide demographic…

  6. Indigenous housing and health in the Canadian North

    DEFF Research Database (Denmark)

    Christensen, Julia

    2016-01-01

    In this article, I explore the relationship between housing, home and health amongst Indigenous homeless people living in the Canadian North. In particular, I examine the ways in which Indigenous homemaking practices conflict with housing policy, and exacerbate individual pathways to homelessness...

  7. Extractive Industries and Indigenous Peoples: A Changing Dynamic?

    Science.gov (United States)

    O'Faircheallaigh, Ciaran

    2013-01-01

    Indigenous peoples and other rural or remote populations often bear the social and environmental cost of extractive industries while obtaining little of the wealth they generate. Recent developments including national and international recognition of Indigenous rights, and the growth of "corporate social responsibility" initiatives among…

  8. Indigenous Education and Grassroots Language Planning in the USA.

    Science.gov (United States)

    McCarty, Teresa L.; Watahomigie, Lucille J.

    1999-01-01

    Indigenous literacy affirms indigenous identity; connects native speakers to the culture and each other; and stimulates other, more diffuse forces for language maintenance. Collaborative, grassroots Native language programs in the United States, New Zealand, Hawaii, Canada, and Puerto Rico are described. Immersion and literacy programs include…

  9. Resetting relationships: archives and Indigenous human rights in Australia

    NARCIS (Netherlands)

    McKemmish, S.; Iacovino, L.; Ketelaar, E.; Castan, M.; Russell, L.

    2011-01-01

    This paper provides an analysis of the Indigenous human rights agenda and identifies its relevance to Australian archivists.1 Based on this analysis and exploration of how far existing archival programs address archives-related Indigenous human rights issues, it presents a road map and action agenda

  10. Walking in Both Worlds: Rethinking Indigenous Knowledge in the Academy

    Science.gov (United States)

    Arnold, Josie

    2017-01-01

    Six generations ago, my Celtic forebears came to Australia as convicts and invaders displacing Indigenous peoples. As a scholar today, I am interested in how Indigenous knowledge remains a challenge in Australian Universities even in this postmodern and postcolonial moment. This paper recognises the need to extend discussion about how Indigenous…

  11. Utilising PEARL to Teach Indigenous Art History: A Canadian Example

    Science.gov (United States)

    Robertson, Carmen

    2012-01-01

    This article explores the concepts advanced from the Australian Learning and Teaching Council (ALTC)-funded project, "Exploring Problem-Based Learning pedagogy as transformative education in Indigenous Australian Studies". As an Indigenous art historian teaching at a mainstream university in Canada, I am constantly reflecting on how to better…

  12. Cultural Resiliency and the Rise of Indigenous Media

    Directory of Open Access Journals (Sweden)

    Derek Moscato

    2016-04-01

    Full Text Available Valerie Alia’s book, The New Media Nation: Indigenous Peoples and Global Communication (New York: Berghahn Books, 2012, 270 pp., points the way to major communication breakthroughs for traditional communities around the world, in turn fostering a more democratic media discourse. From Canada to Japan, and Australia to Mexico, this ambitious and wide-reaching work examines a broad international movement that at once protects ancient languages and customs but also communicates to audiences across countries, oceans, and political boundaries. The publication is divided roughly into five sections: The emergence of a global vision for Indigenous communities scattered around the world; government policy obstacles and opportunities; lessons from Canada, where Indigenous media efforts have been particularly dynamic; the global surge in television, radio and other technological media advances; and finally the long-term prospects and aspirations for Indigenous media. By laying out such a comprehensive groundwork for the rise of global Indigenous media over a variety of formats, particularly over the past century, Alia shows how recent social media breakthroughs such as the highly successful #IdleNoMore movement—a sustained online protest by Canada’s First Nations peoples—have been in fact inevitable. The world’s Indigenous communities have leveraged media technologies to overcome geographic isolation, to foster new linkages with Indigenous populations globally, and ultimately to mitigate structural power imbalances exacerbated by non-Indigenous media and other institutions.

  13. Conversations on Indigenous Education, Progress, and Social Justice in Peru

    Science.gov (United States)

    Huaman, Elizabeth Alva Sumida

    2013-01-01

    This article attempts to contribute to our expanding definitions of Indigenous education within a globalized world. Additionally, the article critiques notions of progress modeled by powerful nation-states due to their histories based on the intended consequences of marginalizing Indigenous populations for the purposes of material gain. Last,…

  14. The indigenous space and marginalized peoples in the United Nations

    DEFF Research Database (Denmark)

    Dahl, Jens

    For more than 20 years, Jens Dahl has observed and now analyzed how a relatively independent space, the Indigenous Space, has been constructed within the confines of the United Nations. In the UN, indigenous peoples have achieved more than any other group of people, minorities included. The book...

  15. Illuminating the Lived Experiences of Research with Indigenous Communities

    Science.gov (United States)

    Burnette, Catherine E.; Sanders, Sara; Butcher, Howard K.; Salois, Emily Matt

    2011-01-01

    The historical exploitation experienced by indigenous people in the United States has left a number of negative legacies, including distrust toward research. This distrust poses a barrier to progress made through culturally sensitive research. Given the complex history of research with indigenous groups, the purpose of this descriptive…

  16. Stories from the Sky: Astronomy in Indigenous Knowledge

    CERN Document Server

    Hamacher, Duane W

    2014-01-01

    Indigenous Australian practices, developed and honed over thousands of years, weave science with storytelling. In this Indigenous science series, we'll look at different aspects of First Australians' traditional life and uncover the knowledge behind them - starting today with astronomy.

  17. Manna in winter: indigenous Americans, huckleberries, and blueberries

    Science.gov (United States)

    More than 35 species of blueberries (Vaccinium L.) and huckleberries (Vaccinium and Gaylussacia Kunth.) are indigenous to North America. The indigenous North American peoples, wise in the ways of survival, recognized the quality of these edible fruits and revered these plants. Beyond food needs, the...

  18. Indigenous Employment and Enterprise Agreements in Australian Universities

    Science.gov (United States)

    Brown, Cath

    2014-01-01

    Considering the benefits that enterprise agreements (EAs) can bring to Indigenous employees, this paper considers the question of whether respectful cultural policies that are aligned with reconciliation and included in EAs can be achieved to Close the Gap on reducing Indigenous disadvantage. A document analysis of EAs at eight Australian…

  19. Closing the Gap: Using Graduate Attributes to Improve Indigenous Education

    Science.gov (United States)

    Anderson, Peter J.; Atkinson, Bernadette

    2013-01-01

    Peter J. Anderson and Bernadette Atkinson teach Indigenous and Traditionally Education in a Global World as a fourth year unit in the Faculty of Education at Monash University, Clayton. This paper is a self reflective piece of work where they discuss the use of graduate attributes relating to Indigenous Education, put forward by the Australian…

  20. The Emerging Research on Indigenous Management in Asia

    DEFF Research Database (Denmark)

    Li, Peter Ping; Sekiguchi, Tomoki; Zhou, Kevin

    2016-01-01

    Introduction to a Special Issue of the Asia Pacific Journal Management with the focus on the theme of indigenous management research in Asia.......Introduction to a Special Issue of the Asia Pacific Journal Management with the focus on the theme of indigenous management research in Asia....

  1. Effective Practices in Teaching Indigenous Students with Conductive Hearing Loss

    Science.gov (United States)

    Partington, Gary; Galloway, Ann

    2005-01-01

    Hearing impairment due to conductive hearing loss can have a devastating effect on children's language development, and consequently educational outcomes, especially for Indigenous students, for whom there may be the additional issue of being educated in their second or third language. With appropriate interventions, however, Indigenous students…

  2. Cultural Resiliency and the Rise of Indigenous Media

    Directory of Open Access Journals (Sweden)

    Derek Moscato

    2016-04-01

    Full Text Available Valerie Alia’s book, The New Media Nation: Indigenous Peoples and Global Communication (New York: Berghahn Books, 2012, 270 pp., points the way to major communication breakthroughs for traditional communities around the world, in turn fostering a more democratic media discourse. From Canada to Japan, and Australia to Mexico, this ambitious and wide-reaching work examines a broad international movement that at once protects ancient languages and customs but also communicates to audiences across countries, oceans, and political boundaries. The publication is divided roughly into five sections: The emergence of a global vision for Indigenous communities scattered around the world; government policy obstacles and opportunities; lessons from Canada, where Indigenous media efforts have been particularly dynamic; the global surge in television, radio and other technological media advances; and finally the long-term prospects and aspirations for Indigenous media. By laying out such a comprehensive groundwork for the rise of global Indigenous media over a variety of formats, particularly over the past century, Alia shows how recent social media breakthroughs such as the highly successful #IdleNoMore movement—a sustained online protest by Canada’s First Nations peoples—have been in fact inevitable. The world’s Indigenous communities have leveraged media technologies to overcome geographic isolation, to foster new linkages with Indigenous populations globally, and ultimately to mitigate structural power imbalances exacerbated by non-Indigenous media and other institutions.

  3. Growth and Empowerment for Indigenous Australians in Substance Abuse Treatment

    Science.gov (United States)

    Berry, Stacey L.; Crowe, T. P.; Deane, F. P.; Billingham, M.; Bhagerutty, Y.

    2012-01-01

    This paper describes psychosocial outcomes of an Indigenous residential substance abuse rehabilitation centre in Australia, examines the sensitivity to change of the new Growth and Empowerment Measure (GEM), and explores the degree to which service users value cultural components of the treatment program. Participants were 57 Indigenous and 46…

  4. Indigenous Thought, Appropriation, and Non-Aboriginal People

    Science.gov (United States)

    Haig-Brown, Celia

    2010-01-01

    In this article, I explore the question, "What is the relationship between appropriation of Indigenous thought and what might be called "deep learning" based in years of education in Indigenous contexts." Beginning with an examination of meanings ascribed to cultural appropriation, I bring texts from Gee on secondary…

  5. Across the Colonial Divide: Conversations about Evaluation in Indigenous Contexts

    Science.gov (United States)

    Cavino, Hayley Marama

    2013-01-01

    This essay engages questions of evaluator role and indigenous peoples participation in evaluation within colonial and decolonization contexts. Specifically, I critique the Western emphasis on cultural competence and contrast the utility of "mainstream" evaluation approaches alongside three indigenous inquiry models (Te Kotahitanga,…

  6. "Ethnic" Assimilates "Indigenous": A Study in Intellectual Neocolonialism.

    Science.gov (United States)

    Stevenson, Winona

    1998-01-01

    Argues that Native American Studies does not belong under the rubric of Ethnic Studies, where unbalanced power relations result in the marginalization, silencing, and exploitation of issues unique to indigenous peoples. Examines the process by which "ethnicity" assimilates "indigenous" and how this process and its impact…

  7. Experiencing and Writing Indigeneity, Rurality and Gender: Australian Reflections

    Science.gov (United States)

    Ramzan, Bebe; Pini, Barbara; Bryant, Lia

    2009-01-01

    This paper has two interrelated aims. The first is to contribute to knowledge about rurality, gender and Indigeneity. This is undertaken by the first author, Bebe Ramzan, an Indigenous woman living in the Anangu Pitjantjatjara Yankunytjatjara (APY) Lands. Bebe shows similarities across rural and remote areas in Australia and details her knowledge…

  8. The Importance of Place in Indigenous Science Education

    Science.gov (United States)

    Sutherland, Dawn; Swayze, Natalie

    2012-01-01

    In this issue of Cultural Studies of Science Education, Mack and colleagues (Mack et al. "2011") seek to identify the necessary components of science education in Indigenous settings. Using a review of current research in informal science education in Indigenous settings, along with personal interviews with American educators engaged in…

  9. Mathematics Registers in Indigenous Languages: Experiences from South Africa

    Science.gov (United States)

    Schafer, Marc

    2010-01-01

    Through reporting on an initiative in South Africa that aimed to provide epistemological access to teachers and learners of mathematics (and science) through translating mathematical concepts into two indigenous languages, this paper argues for the urgent development of mathematical registers in indigenous languages for mathematics and …

  10. Methodological Metissage: An Interpretive Indigenous Approach to Environmental Education Research

    Science.gov (United States)

    Lowan-Trudeau, Greg

    2012-01-01

    This article discusses the development of a methodological metissage that combined Indigenous and interpretive traditions. This metissage was developed during a doctoral study conducted with Canadian environmental educators who incorporate Western and Indigenous knowledge and philosophy into their ecological identities and pedagogical praxis. It…

  11. The Importance of Place in Indigenous Science Education

    Science.gov (United States)

    Sutherland, Dawn; Swayze, Natalie

    2012-01-01

    In this issue of Cultural Studies of Science Education, Mack and colleagues (Mack et al. "2011") seek to identify the necessary components of science education in Indigenous settings. Using a review of current research in informal science education in Indigenous settings, along with personal interviews with American educators engaged in these…

  12. Mobile Technologies for Preservation of Indigenous Knowledge in Rural Communities

    DEFF Research Database (Denmark)

    Winschiers-Theophilus, Heike; Zaman, Tariq; Jensen, Kasper Løvborg

    2013-01-01

    In this paper we explore the opportunities of mobile technologies in three of our own development endeavors with rural communities, promoting the preservation of indigenous knowledge. We reflect upon and recognize the fact that the representation of indigenous knowledge will be transformed within...

  13. Cinders in Snow? Indigenous Teacher Identities in Formation

    Science.gov (United States)

    Reid, Jo-Anne; Santoro, Ninetta

    2006-01-01

    The identity work engaged in by Indigenous teachers in school settings is highlighted in a study of Australian Indigenous teachers. The construction of identity in home and community relationships intersects with and can counteract the take up of a preferred identity in the workplace. In this paper we analyse data from interviews with Indigenous…

  14. African Indigenous science in higher education in Uganda

    Science.gov (United States)

    Akena Adyanga, Francis

    This study examines African Indigenous Science (AIS) in higher education in Uganda. To achieve this, I use anticolonial theory and Indigenous knowledge discursive frameworks to situate the subjugation of Indigenous science from the education system within a colonial historical context. These theories allow for a critical examination of the intersection of power relations rooted in the politics of knowledge production, validation, and dissemination, and how this process has become a systemic and complex method of subjugating one knowledge system over the other. I also employ qualitative and autoethnographic research methodologies. Using a qualitative research method, I interviewed 10 students and 10 professors from two universities in Uganda. My research was guided by the following key questions: What is African Indigenous Science? What methodology would help us to indigenize science education in Uganda? How can we work with Indigenous knowledge and anticolonial theoretical discursive frameworks to understand and challenge the dominance of Eurocentric knowledge in mainstream education? My research findings revealed that AIS can be defined in multiple ways, in other words, there is no universal definition of AIS. However, there were some common elements that my participants talked about such as: (a) knowledge by Indigenous communities developed over a long period of time through a trial and error approach to respond to the social, economic and political challenges of their society. The science practices are generational and synergistic with other disciplines such as history, spirituality, sociology, anthropology, geography, and trade among others, (b) a cumulative practice of the use, interactions with and of biotic and abiotic organism in everyday life for the continued existence of a community in its' totality. The research findings also indicate that Indigenous science is largely lacking from Uganda's education curriculum because of the influence of colonial and

  15. Wholistic and Ethical: Social Inclusion with Indigenous Peoples

    Directory of Open Access Journals (Sweden)

    Kathleen E. Absolon

    2016-02-01

    Full Text Available This paper begins with a poem and is inclusive of my voice as Anishinaabekwe (Ojibway woman and is authored from my spirit, heart, mind and body. The idea of social inclusion and Indigenous peoples leave more to the imagination and vision than what is the reality and actuality in Canada. This article begins with my location followed with skepticism and hope. Skepticism deals with the exclusion of Indigenous peoples since colonial contact and the subsequent challenges and impacts. Hope begins to affirm the possibilities, strengths and Indigenous knowledge that guides wholistic cultural frameworks and ethics of social inclusion. A wholistic cultural framework is presented; guided by seven sacred teachings and from each element thoughts for consideration are guided by Indigenous values and principles. From each element this paper presents a wholistic and ethical perspective in approaching social inclusion and Indigenous peoples.

  16. Adapting Western research methods to indigenous ways of knowing.

    Science.gov (United States)

    Simonds, Vanessa W; Christopher, Suzanne

    2013-12-01

    Indigenous communities have long experienced exploitation by researchers and increasingly require participatory and decolonizing research processes. We present a case study of an intervention research project to exemplify a clash between Western research methodologies and Indigenous methodologies and how we attempted reconciliation. We then provide implications for future research based on lessons learned from Native American community partners who voiced concern over methods of Western deductive qualitative analysis. Decolonizing research requires constant reflective attention and action, and there is an absence of published guidance for this process. Continued exploration is needed for implementing Indigenous methods alone or in conjunction with appropriate Western methods when conducting research in Indigenous communities. Currently, examples of Indigenous methods and theories are not widely available in academic texts or published articles, and are often not perceived as valid.

  17. Using Modern Technologies to Capture and Share Indigenous Astronomical Knowledge

    Science.gov (United States)

    Nakata, Martin; Hamacher, Duane W.; Warren, John; Byrne, Alex; Pagnucco, Maurice; Harley, Ross; Venugopal, Srikumar; Thorpe, Kirsten; Neville, Richard; Bolt, Reuben

    2014-06-01

    Indigenous Knowledge is important for Indigenous communities across the globe and for the advancement of our general scientific knowledge. In particular, Indigenous astronomical knowledge integrates many aspects of Indigenous Knowledge, including seasonal calendars, navigation, food economics, law, ceremony, and social structure. Capturing, managing, and disseminating this knowledge in the digital environment poses a number of challenges, which we aim to address using a collaborative project emerging between experts in the higher education, library, archive and industry sectors. Using Microsoft's WorldWide Telescope and Rich Interactive Narratives technologies, we propose to develop software, media design, and archival management solutions to allow Indigenous communities to share their astronomical knowledge with the world on their terms and in a culturally sensitive manner.

  18. Gendering Aboriginalism: A Performative Gaze on Indigenous Australian Women

    Directory of Open Access Journals (Sweden)

    Katelyn Barney

    2010-03-01

    Full Text Available One of the most common Aboriginalist representations of Indigenous Australian people is, as Indigenous female performer Lou Bennett points out, ‘basically a man, out in the desert, black skin, flat nose with a lap-lap on, standing on one leg, resting against a spear’. Her comment raises many issues. In what ways are discourses of Aboriginalism gendered? How does Aboriginalism affect performance and specifically Aboriginal women performers? In exploring these questions, I examine Aboriginalist representations of Aboriginal women performers by white male scholars and the role of women anthropologists in the production of Aboriginalist discourse about Aboriginal women. Drawing on interviews with Indigenous women performers and musical examples of their songs, I explore the impact of Aboriginalism on non-Indigenous expectations of Indigenous Australian women performing in contemporary music contexts, the strategies performers use to work within and against these constructions and my own relationship to Aboriginalism.

  19. Gendering Aboriginalism : a performative gaze on indigenous Australian women

    Directory of Open Access Journals (Sweden)

    Barney, Katelyn

    2010-01-01

    Full Text Available One of the most common Aboriginalist representations of Indigenous Australian people is, as Indigenous female performer Lou Bennett points out, ‘basically a man, out in the desert, black skin, flat nose with a lap-lap on, standing on one leg, resting against a spear’. Her comment raises many issues. In what ways are discourses of Aboriginalism gendered? How does Aboriginalism affect performance and specifically Aboriginal women performers? In exploring these questions, I examine Aboriginalist representations of Aboriginal women performers by white male scholars and the role of women anthropologists in the production of Aboriginalist discourse about Aboriginal women. Drawing on interviews with Indigenous women performers and musical examples of their songs, I explore the impact of Aboriginalism on non-Indigenous expectations of Indigenous Australian women performing in contemporary music contexts, the strategies performers use to work within and against these constructions and my own relationship to Aboriginalism.

  20. Cloning, sequence analysis, expression of Cyathus bulleri laccase in Pichia pastoris and characterization of recombinant laccase

    Directory of Open Access Journals (Sweden)

    Garg Neha

    2012-10-01

    Full Text Available Abstract Background Laccases are blue multi-copper oxidases and catalyze the oxidation of phenolic and non-phenolic compounds. There is considerable interest in using these enzymes for dye degradation as well as for synthesis of aromatic compounds. Laccases are produced at relatively low levels and, sometimes, as isozymes in the native fungi. The investigation of properties of individual enzymes therefore becomes difficult. The goal of this study was to over-produce a previously reported laccase from Cyathus bulleri using the well-established expression system of Pichia pastoris and examine and compare the properties of the recombinant enzyme with that of the native laccase. Results In this study, complete cDNA encoding laccase (Lac from white rot fungus Cyathus bulleri was amplified by RACE-PCR, cloned and expressed in the culture supernatant of Pichia pastoris under the control of the alcohol oxidase (AOX1 promoter. The coding region consisted of 1,542 bp and encodes a protein of 513 amino acids with a signal peptide of 16 amino acids. The deduced amino acid sequence of the matured protein displayed high homology with laccases from Trametes versicolor and Coprinus cinereus. The sequence analysis indicated the presence of Glu 460 and Ser 113 and LEL tripeptide at the position known to influence redox potential of laccases placing this enzyme as a high redox enzyme. Addition of copper sulfate to the production medium enhanced the level of laccase by about 12-fold to a final activity of 7200 U L-1. The recombinant laccase (rLac was purified by ~4-fold to a specific activity of ~85 U mg-1 protein. A detailed study of thermostability, chloride and solvent tolerance of the rLac indicated improvement in the first two properties when compared to the native laccase (nLac. Altered glycosylation pattern, identified by peptide mass finger printing, was proposed to contribute to altered properties of the rLac. Conclusion Laccase of C. bulleri was

  1. Indigenous Emotional Intelligence Scale: Development and Validation

    Directory of Open Access Journals (Sweden)

    Olukayode Ayooluwa Afolabi

    2017-04-01

    Full Text Available In Nigeria, it seems there is sparse of indigenous measure of emotional intelligence. Therefore, this research fills a gap in literature by developing a valid and reliable indigenous scale. To achieve this, the author combined the Bar-On and Goleman’s models of emotional intelligence. In Study 1, 255 items were generated and were reduced to 198 based on expert advice. At the end of several analyses, 52 items were retained. These items were put in a questionnaire form and administered to 850 students in a university. The author determined the factorial validity of the scale using a sample of 834 participants who returned the scale. The scale (using varimax rotation method was then subjected to principal component analysis and 40 items were retained in a 7 well defined factor structure. The factors/dimensions include interpersonal skill (r = .77, empathetic response (r = .73, stress tolerance (r = .69, optimism (r = .75, assertiveness (r = .78, problem solving (r = .74 and flexibility (r = .80. The scale has convergent validity because of its positive relationship with empathy (r = .67 and social intelligence (r = .79 and negative relationships with aggression (r = -.41 and impulsiveness (r = -.32. A test re-test reliability of .79 was also established for the scale.

  2. Indigenous Students and the Learning of English

    Directory of Open Access Journals (Sweden)

    Shahrier Pawanchik

    2010-01-01

    Full Text Available Problem statement: The problem of students’ proficiency in English in the Malaysian primary schools is still debatable. Approach: Unless the problem of students’ proficiency is solved at the primary school level, it will fossilize and contribute toward students’ anxiety in the language at the secondary and tertiary levels. Results: This research study looked into English needs of the indigenous or ‘Orang Asli’ students in primary schools in the district of Rompin-Endau, Pahang. These indigenous students still lag in education and with the implementation of teaching of science and mathematics in English in primary schools, they will be burdened with language difficulties. The researchers identify that the students preferred learning skill is listening to the teachers’ explanation. And the task-based activity that can improve their proficiency is listening to songs and singing in English. Conclusion/Recommendations: Findings from this research could provide useful information for the curriculum developers at the Ministry of Education of Malaysia whether to revamp the present English curriculum or formulate a new curriculum to meet the English needs of the ‘Orang Asli’ students.

  3. Are Isolated Indigenous Populations Headed toward Extinction?

    Directory of Open Access Journals (Sweden)

    Robert S Walker

    Full Text Available At least 50 indigenous groups spread across lowland South America remain isolated and have only intermittent and mostly hostile interactions with the outside world. Except in emergency situations, the current policy of governments in Brazil, Colombia, and Peru towards isolated tribes is a "leave them alone" strategy, in which isolated groups are left uncontacted. However, these no-contact policies are based on the assumption that isolated populations are healthy and capable of persisting in the face of mounting external threats, and that they can maintain population viability in the long-term. Here, we test this assumption by tracking the sizes and movements of cleared horticultural areas made by 8 isolated groups over the last 10-14 years. We used deforestation data derived from remote sensing Landsat satellite sensors to identify clearings, and those were then validated and assessed with high-resolution imagery. We found only a single example of a relatively large and growing population (c. 50 cleared ha and 400 people, whereas all of the other 7 groups exhibited much smaller villages and gardens with no sizable growth through time. These results indicated that the smaller groups are critically endangered, and it prompts an urgent re-thinking of policies toward isolated populations, including plans for well-organized contacts that may help save lives and rescue isolated indigenous populations from imminent extinction.

  4. Indigenous people (in (and the Paraguayan Independence

    Directory of Open Access Journals (Sweden)

    Ana María RIBEIRO GUTIÉRREZ

    2011-04-01

    Full Text Available The independence of Paraguay, which started as a revolution against the power of Buenos Aires, the capital city of the viceroyalty of la Plata, did not produce any thoughts against native exploitation, and neither did it have relevant indigenous leaders or demands, although demographically speaking the indo-mestizo presence was higher than in the Banda Oriental. Paraguayan revolutionaries’ stance in relation to the indigenous population was conditioned in the first place by the strategic position of Jesuit Missions, and soon after by the policies applied by Dr. Gaspar de Francia, who after an early egalitarian impulse which favoured the gradual creation of a new unity, implemented integration and expulsion measures similar to those used during colonial times. The suppression in 1848 of the communal systems of the Guaraní people by Carlos Antonio López culminated a strategic integration within a «Paraguayan» identity. This decisive step in the shaping of the Paraguayan nation-state was completed by constructing Paraguay’s past as a Guaraní nation, thus establishing the starting point for all future Creole accounts of the nation.

  5. Indigenous Astronomies and Progress in Modern Astronomy

    CERN Document Server

    Ruggles, Clive

    2010-01-01

    From an anthropological point of view, the whole concept of a "path of progress" in astronomical discovery is anathema, since it implicitly downgrades other cultural perspectives, such as the many "indigenous cosmologies" that still exist in the modern world. By doing so, one risks provoking those who hold them and-as is most obvious in places such as Hawaii where the two "world-views" come into direct contact-reating avoidable resistance to that very progress. The problem is complicated by the existence of "fringe" and "new-age" views that are increasingly confused with, and even passed off as, indigenous perceptions. In a modern world where widespread public perceptions include many that are unscientific in the broadest sense of the term, I shall argue that there are actually a range of positive benefits for progress in scientific astronomy to be derived from the mutual awareness and comprehension of "genuine" cultural world-views whose goals-in common with those of modern science-are to make sense of the c...

  6. Indigenous people's detection of rapid ecological change.

    Science.gov (United States)

    Aswani, Shankar; Lauer, Matthew

    2014-06-01

    When sudden catastrophic events occur, it becomes critical for coastal communities to detect and respond to environmental transformations because failure to do so may undermine overall ecosystem resilience and threaten people's livelihoods. We therefore asked how capable of detecting rapid ecological change following massive environmental disruptions local, indigenous people are. We assessed the direction and periodicity of experimental learning of people in the Western Solomon Islands after a tsunami in 2007. We compared the results of marine science surveys with local ecological knowledge of the benthos across 3 affected villages and 3 periods before and after the tsunami. We sought to determine how people recognize biophysical changes in the environment before and after catastrophic events such as earthquakes and tsunamis and whether people have the ability to detect ecological changes over short time scales or need longer time scales to recognize changes. Indigenous people were able to detect changes in the benthos over time. Detection levels differed between marine science surveys and local ecological knowledge sources over time, but overall patterns of statistically significant detection of change were evident for various habitats. Our findings have implications for marine conservation, coastal management policies, and disaster-relief efforts because when people are able to detect ecological changes, this, in turn, affects how they exploit and manage their marine resources. © 2014 Society for Conservation Biology.

  7. Indigenous healing practice: ayahuasca. Opening a discussion.

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    Prue, Robert; Voss, Richard W

    2014-01-01

    This essay frames an invitation to pastoral counselors and pastoral theologians to examine connections and perhaps interactions between themselves and traditional shamanic healers who use ayahuasca in their healing ceremonies. Indigenous people in South America have used ayahuasca for centuries, and the ritual has become common among the mestizo populations in urban areas of the Amazon, particularly as a curing ritual for drug addiction (Dobkin de Rios, 1970; Moir, 1998). Like peyote in the United States (Calabrese, 1997) ayahuasca use amongst the indigenous people of the Amazon is a form of cultural psychiatry. A review of the literature reveals very little commentary or discussion of shamanic practice in Pastoral Counseling (Pastoral Theology). The scant literature identifies an antithetical relationship at best. The current authors wonder about the possibility of to including shamanic practices in the context of pastoral counseling? This essay seeks to provide some basic information about the ritual use of ayahuasca and to offer a rationale for pastoral counselors to engage in a dialogue about its utility.

  8. Heart failure among Indigenous Australians: a systematic review

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    Woods John A

    2012-11-01

    Full Text Available Abstract Background Cardiovascular diseases contribute substantially to the poor health and reduced life expectancy of Indigenous Australians. Heart failure is a common, disabling, progressive and costly complication of these disorders. The epidemiology of heart failure and the adequacy of relevant health service provision in Indigenous Australians are not well delineated. Methods A systematic search of the electronic databases PubMed, Embase, Web of Science, Cinahl Plus, Informit and Google Scholar was undertaken in April 2012 for peer-reviewed journal articles relevant to the topic of heart failure in Indigenous Australians. Additionally, a website search was done to identify other pertinent publications, particularly government reports. Results There was a paucity of relevant peer-reviewed research, and government reports dominated the results. Ten journal articles, 1 published conference abstract and 10 reports were eligible for inclusion. Indigenous Australians reportedly have higher morbidity and mortality from heart failure than their non-Indigenous counterparts (age-standardised prevalence ratio 1.7; age-standardised hospital separation ratio ≥3; crude per capita hospital expenditure ratio 1.58; age-adjusted mortality ratio >2. Despite the evident disproportionate burden of heart failure in Indigenous Australians, the accuracy of estimation from administrative data is limited by poor indigenous identification, inadequate case ascertainment and exclusion of younger subjects from mortality statistics. A recent journal article specifically documented a high prevalence of heart failure in Central Australian Aboriginal adults (5.3%, noting frequent undiagnosed disease. One study examined barriers to health service provision for Indigenous Australians in the context of heart failure. Conclusions Despite the shortcomings of available published data, it is clear that Indigenous Australians have an excess burden of heart failure. Emerging data

  9. "Until I Became a Professional, I Was Not, Consciously, Indigenous": One Intercultural Bilingual Educator's Trajectory in Indigenous Language Revitalization

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    Hornberger, Nancy H.

    2014-01-01

    Drawing from long-term ethnographic research in the Andes, this paper examines one Quechua-speaking Indigenous bilingual educator's trajectory as she traversed (and traverses) from rural highland communities of southern Peru through development as teacher, teacher educator, researcher, and advocate for Indigenous identity and language…

  10. Principals as Literacy Leaders with Indigenous Communities (PALLIC) Building Relationships: One School's Quest to Raise Indigenous Learners' Literacy

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    Riley, Tasha; Webster, Amanda

    2016-01-01

    In 2011 to 2012, 48 schools in the Northern Territory, South Australia and Queensland participated in the Principals as Literacy Leaders with Indigenous Communities (PALLIC) project. Central to this project was the establishment of positive working relationships between school principals and Indigenous community leaders in order to improve…

  11. Socioeconomic status as a factor in Indigenous and non-Indigenous children with hearing loss: analysis of national survey data.

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    Simpson, Andrea; Enticott, Joanne C; Douglas, Jacinta

    2016-09-23

    In this paper, the association between socioeconomic status and speech, language and communication outcomes for primary-school-going children with hearing loss using population survey data was analysed. The dataset used for analysis consisted of 289973 children in total, of which 3174 children had hearing loss. For all children, higher socioeconomic status was positively correlated with better speech, language and communication outcomes. A hearing loss was indicated for 1% of non-Indigenous children and 4.3% of Indigenous children. Non-Indigenous children with hearing loss were found to be fairly evenly distributed by socioeconomic status, whereas Indigenous children with hearing loss were found to be statistically significantly more likely to be living in the most disadvantaged socioeconomic areas. Socioeconomic status was found to affect developmental outcomes for all children, regardless of Indigenous and hearing loss status.

  12. Population structure of four Thai indigenous chicken breeds.

    Science.gov (United States)

    Mekchay, Supamit; Supakankul, Pantaporn; Assawamakin, Anunchai; Wilantho, Alisa; Chareanchim, Wanwisa; Tongsima, Sissades

    2014-03-27

    In recent years, Thai indigenous chickens have increasingly been bred as an alternative in Thailand poultry market. Due to their popularity, there is a clear need to improve the underlying quality and productivity of these chickens. Studying chicken genetic variation can improve the chicken meat quality as well as conserving rare chicken species. To begin with, a minimal set of molecular markers that can characterize the Thai indigenous chicken breeds is required. Using AFLP-PCR, 30 single nucleotide polymorphisms (SNPs) from Thai indigenous chickens were obtained by DNA sequencing. From these SNPs, we genotyped 465 chickens from 7 chicken breeds, comprising four Thai indigenous chicken breeds--Pradhuhangdum (PD), Luenghangkhao (LK), Dang (DA) and Chee (CH), one wild chicken--the red jungle fowls (RJF), and two commercial chicken breeds--the brown egg layer (BL) and commercial broiler (CB). The chicken genotypes reveal unique genetic structures of the four Thai indigenous chicken breeds. The average expected heterozygosities of PD=0.341, LK=0.357, DA=0.349 and CH=0.373, while the references RJF= 0.327, CB=0.324 and BL= 0.285. The F(ST) values among Thai indigenous chicken breeds vary from 0.051 to 0.096. The F(ST) values between the pairs of Thai indigenous chickens and RJF vary from 0.083 to 0.105 and the FST values between the Thai indigenous chickens and the two commercial chicken breeds vary from 0.116 to 0.221. A neighbour-joining tree of all individual chickens showed that the Thai indigenous chickens were clustered into four groups which were closely related to the wild RJF but far from the commercial breeds. Such commercial breeds were split into two closely groups. Using genetic admixture analysis, we observed that the Thai indigenous chicken breeds are likely to share common ancestors with the RJF, while both commercial chicken breeds share the same admixture pattern. These results indicated that the Thai indigenous chicken breeds may descend from the

  13. Human papillomavirus prevalence among indigenous and non-indigenous Australian women prior to a national HPV vaccination program

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    Condon John R

    2011-09-01

    Full Text Available Abstract Background Indigenous women in Australia have a disproportionate burden of cervical cancer despite a national cervical screening program. Prior to introduction of a national human papilloma virus (HPV vaccination program, we determined HPV genotype prevalence by Indigenous status and residence in remote areas. Methods We recruited women aged 17 to 40 years presenting to community-based primary health services for routine Pap screening across Australia. A liquid-based cytology (LBC cervical specimen was tested for HPV DNA using the AMPLICOR HPV-DNA test and a PGMY09/11-based HPV consensus PCR; positive specimens were typed by reverse hybridization. We calculated age-adjusted prevalence by weighting to relevant population data, and determined predictors of HPV-DNA positivity by age, Indigenous status and area of residence using logistic regression. Results Of 2152 women (655 Indigenous, prevalence of the high-risk HPV genotypes was similar for Indigenous and non-Indigenous women (HPV 16 was 9.4% and 10.5%, respectively; HPV 18 was 4.1% and 3.8%, respectively, and did not differ by age group. In younger age groups, the prevalence of other genotypes also did not differ, but in those aged 31 to 40 years, HPV prevalence was higher for Indigenous women (35% versus 22.5%; P Conclusion Although we found no difference in the prevalence of HPV16/18 among Australian women by Indigenous status or, for Indigenous women, residence in remote regions, differences were found in the prevalence of risk factors and some other HPV genotypes. This reinforces the importance of cervical screening as a complement to vaccination for all women, and the value of baseline data on HPV genotype prevalence by Indigenous status and residence for the monitoring of vaccine impact.

  14. Dadirri: Using a Philosophical Approach to Research to Build Trust between a Non-Indigenous Researcher and Indigenous Participants

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    Megan Marie Stronach

    2014-09-01

    Full Text Available Abstract: This article focuses on a philosophical approach employed in a PhD research project that set out to investigate sport career transition (SCT experiences of elite Indigenous Australian sportsmen. The research was necessary as little is known about the transition of this cohort to a life after sport, or their experiences of retirement. A key problem within the SCT paradigm is a presumption that an end to elite sport requires a process of adjustment that is common to all sportspeople—a rather narrow perspective that fails to acknowledge the situational complexity and socio-cultural diversity of elite athletes. With such a range of personal circumstances, it is reasonable to suppose that athletes from different cultural groups will have different individual SCT needs. The researcher is non-Indigenous and mature aged: she encountered a number of challenges in her efforts to understand Indigenous culture and its important sensitivities, and to build trust with the Indigenous male participants she interviewed. An Indigenous philosophy known as Dadirri, which emphasises deep and respectful listening, guided the development of the research design and methodology. Consistent with previous studies conducted by non-Indigenous researchers, an open-ended and conversational approach to interviewing Indigenous respondents was developed. The objective was for the voices of the athletes to be heard, allowing the collection of rich data based on the participants’ perspectives about SCT. An overview of the findings is presented, illustrating that Indigenous athletes experience SCT in complex and distinctive ways. The article provides a model for non-Indigenous researchers to conduct qualitative research with Indigenous people.

  15. Engineering Pichia pastoris for improved NADH regeneration: A novel chassis strain for whole-cell catalysis

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    Martina Geier

    2015-09-01

    Full Text Available Many synthetically useful reactions are catalyzed by cofactor-dependent enzymes. As cofactors represent a major cost factor, methods for efficient cofactor regeneration are required especially for large-scale synthetic applications. In order to generate a novel and efficient host chassis for bioreductions, we engineered the methanol utilization pathway of Pichia pastoris for improved NADH regeneration. By deleting the genes coding for dihydroxyacetone synthase isoform 1 and 2 (DAS1 and DAS2, NADH regeneration via methanol oxidation (dissimilation was increased significantly. The resulting Δdas1 Δdas2 strain performed better in butanediol dehydrogenase (BDH1 based whole-cell conversions. While the BDH1 catalyzed acetoin reduction stopped after 2 h reaching ~50% substrate conversion when performed in the wild type strain, full conversion after 6 h was obtained by employing the knock-out strain. These results suggest that the P. pastoris Δdas1 Δdas2 strain is capable of supplying the actual biocatalyst with the cofactor over a longer reaction period without the over-expression of an additional cofactor regeneration system. Thus, focusing the intrinsic carbon flux of this methylotrophic yeast on methanol oxidation to CO2 represents an efficient and easy-to-use strategy for NADH-dependent whole-cell conversions. At the same time methanol serves as co-solvent, inductor for catalyst and cofactor regeneration pathway expression and source of energy.

  16. Production of the sesquiterpenoid (+)-nootkatone by metabolic engineering of Pichia pastoris.

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    Wriessnegger, Tamara; Augustin, Peter; Engleder, Matthias; Leitner, Erich; Müller, Monika; Kaluzna, Iwona; Schürmann, Martin; Mink, Daniel; Zellnig, Günther; Schwab, Helmut; Pichler, Harald

    2014-07-01

    The sesquiterpenoid (+)-nootkatone is a highly demanded and highly valued aroma compound naturally found in grapefruit, pummelo or Nootka cypress tree. Extraction of (+)-nootkatone from plant material or its production by chemical synthesis suffers from low yields and the use of environmentally harmful methods, respectively. Lately, major attention has been paid to biotechnological approaches, using cell extracts or whole-cell systems for the production of (+)-nootkatone. In our study, the yeast Pichia pastoris initially was applied as whole-cell biocatalyst for the production of (+)-nootkatone from (+)-valencene, the abundant aroma compound of oranges. Therefore, we generated a strain co-expressing the premnaspirodiene oxygenase of Hyoscyamus muticus (HPO) and the Arabidopsis thaliana cytochrome P450 reductase (CPR) that hydroxylated extracellularly added (+)-valencene. Intracellular production of (+)-valencene by co-expression of valencene synthase from Callitropsis nootkatensis resolved the phase-transfer issues of (+)-valencene. Bi-phasic cultivations of P. pastoris resulted in the production of trans-nootkatol, which was oxidized to (+)-nootkatone by an intrinsic P. pastoris activity. Additional overexpression of a P. pastoris alcohol dehydrogenase and truncated hydroxy-methylglutaryl-CoA reductase (tHmg1p) significantly enhanced the (+)-nootkatone yield to 208mg L(-1) cell culture in bioreactor cultivations. Thus, metabolically engineered yeast P. pastoris represents a valuable, whole-cell system for high-level production of (+)-nootkatone from simple carbon sources.

  17. Heterologous expression of a Penicillium purpurogenum exo-arabinanase in Pichia pastoris and its biochemical characterization.

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    Mardones, Wladimir; Callegari, Eduardo; Eyzaguirre, Jaime

    2015-12-01

    Arabinan is a component of pectin, which is one of the polysaccharides present in lignocelluose. The enzymes degrading the main chain of arabinan are the endo- (EC 3.2.1.99) and exo-arabinanases (3.2.1.-). Only three exo-arabinanases have been biochemically characterized; they belong to glycosyl hydrolase family 93. In this work, the cDNA of an exo-arabinanase (Arap2) from Penicillium purpurogenum has been heterologously expressed in Pichia pastoris. The gene is 1310 bp long, has three introns and codes for a protein of 380 amino acid residues; the mature protein has a calculated molecular mass of 39 823 Da. The heterologously expressed Arap2 has a molecular mass in the range of 60-80 kDa due to heterogeneous glycosylation. The enzyme is active on debranched arabinan with optimum pH of 4-5.5 and optimal temperature of 40 °C, and has an exo-type action mode, releasing arabinobiose from its substrates. The expression profile of arap2 in corncob and sugar beet pulp follows a different pattern and is not related to the presence of arabinan. This is the first exo-arabinanase studied from P. purpurogenum and the first expressed in yeast. The availability of heterologous Arap2 may be useful for biotechnological applications requiring acidic conditions. Copyright © 2015 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

  18. Camelid-derived heavy-chain nanobody against Clostridium botulinum neurotoxin E in Pichia pastoris.

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    Baghban, Roghayyeh; Gargari, Seyed Latif Mousavi; Rajabibazl, Masoumeh; Nazarian, Shahram; Bakherad, Hamid

    2016-01-01

    Botulinum neurotoxins (BoNTs) result in severe and often fatal disease, botulism. Common remedial measures such as equine antitoxin and human botulism immunoglobulin in turn are problematic and time-consuming. Therefore, diagnosis and therapy of BoNTs are vital. The variable domain of heavy-chain antibodies (VHH) has unique features, such as the ability to identify and bind specifically to target epitopes and ease of production in bacteria and yeast. The Pichia pastoris is suitable for expression of recombinant antibody fragments. Disulfide bond formation and correct folds of protein with a high yield are some of the advantages of this eukaryotic host. In this study, we have expressed and purified the camelid VHH against BoNT/E in P. pastoris. The final yield of P. pastoris-expressed antibody was estimated to be 16 mg/l, which is higher than that expressed by Escherichia coli. The nanobody expressed in P. pastoris neutralized 4LD50 of the BoNT/E upon i.p. injection in 25% of mice. The nanobody expressed in E. coli extended the mice's survival to 1.5-fold compared to the control. This experiment indicated that the quality of expressed protein in the yeast is superior to that of the bacterial expression. Favorable protein folding by P. pastoris seems to play a role in its better toxin-binding property.

  19. Metabolic engineering of Pichia pastoris to produce ricinoleic acid, a hydroxy fatty acid of industrial importance.

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    Meesapyodsuk, Dauenpen; Chen, Yan; Ng, Siew Hon; Chen, Jianan; Qiu, Xiao

    2015-11-01

    Ricinoleic acid (12-hydroxyoctadec-cis-9-enoic acid) has many specialized uses in bioproduct industries, while castor bean is currently the only commercial source for the fatty acid. This report describes metabolic engineering of a microbial system (Pichia pastoris) to produce ricinoleic acid using a "push" (synthesis) and "pull" (assembly) strategy. CpFAH, a fatty acid hydroxylase from Claviceps purpurea, was used for synthesis of ricinoleic acid, and CpDGAT1, a diacylglycerol acyl transferase for the triacylglycerol synthesis from the same species, was used for assembly of the fatty acid. Coexpression of CpFAH and CpDGAT1 produced higher lipid contents and ricinoleic acid levels than expression of CpFAH alone. Coexpression in a mutant haploid strain defective in the Δ12 desaturase activity resulted in a higher level of ricinoleic acid than that in the diploid strain. Intriguingly, the ricinoleic acid produced was mainly distributed in the neutral lipid fractions, particularly the free fatty acid form, but with little in the polar lipids. This work demonstrates the effectiveness of the metabolic engineering strategy and excellent capacity of the microbial system for production of ricinoleic acid as an alternative to plant sources for industrial uses. Copyright © 2015 by the American Society for Biochemistry and Molecular Biology, Inc.

  20. Comparison of laccase production levels in Pichia pastoris and Cryptococcus sp. S-2.

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    Nishibori, Nahoko; Masaki, Kazuo; Tsuchioka, Hiroaki; Fujii, Tsutomu; Iefuji, Haruyuki

    2013-04-01

    The heterologous expression of the laccase gene from Trametes versicolor and Gaeumannomyces graminis was evaluated in the yeasts Pichia pastoris and Cryptococcus sp. S-2. The expression levels of both laccase genes in Cryptococcus sp. S-2 were considerably higher than those in P. pastoris. The codon usage of Cryptococcus sp. S-2 as well as the GC content were similar to those of T. versicolor and G. graminis. These results suggest that using a host with a similar codon usage for the expressed gene may improve protein expression. The use of Cryptococcus sp. S-2 as a host may be advantageous for the heterologous expression of genes with high GC content. Moreover, this yeast provides the same advantages as P. pastoris for the production of recombinant proteins, such as growth on minimal medium, capacity for high-density growth during fermentation, and capability for post-translational modifications. Therefore, we propose that Cryptococcus sp. S-2 be used as an expression host to improve enzyme production levels when other hosts have not yielded good results.

  1. Recombinant human N-acetylgalactosamine-6-sulfate sulfatase (GALNS) produced in the methylotrophic yeast Pichia pastoris

    Science.gov (United States)

    Rodríguez-López, Alexander; Alméciga-Díaz, Carlos J.; Sánchez, Jhonnathan; Moreno, Jefferson; Beltran, Laura; Díaz, Dennis; Pardo, Andrea; Ramírez, Aura María; Espejo-Mojica, Angela J.; Pimentel, Luisa; Barrera, Luis A.

    2016-01-01

    Mucopolysaccharidosis IV A (MPS IV A, Morquio A disease) is a lysosomal storage disease (LSD) produced by mutations on N-acetylgalactosamine-6-sulfate sulfatase (GALNS). Recently an enzyme replacement therapy (ERT) for this disease was approved using a recombinant enzyme produced in CHO cells. Previously, we reported the production of an active GALNS enzyme in Escherichia coli that showed similar stability properties to that of a recombinant mammalian enzyme though it was not taken-up by culture cells. In this study, we showed the production of the human recombinant GALNS in the methylotrophic yeast Pichia pastoris GS115 (prGALNS). We observed that removal of native signal peptide and co-expression with human formylglycine-generating enzyme (SUMF1) allowed an improvement of 4.5-fold in the specific GALNS activity. prGALNS enzyme showed a high stability at 4 °C, while the activity was markedly reduced at 37 and 45 °C. It was noteworthy that prGALNS was taken-up by HEK293 cells and human skin fibroblasts in a dose-dependent manner through a process potentially mediated by an endocytic pathway, without any additional protein or host modification. The results show the potential of P. pastoris in the production of a human recombinant GALNS for the development of an ERT for Morquio A. PMID:27378276

  2. Expression, solubilisation, and purification of a functional CMP-sialic acid transporter in Pichia pastoris.

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    Maggioni, Andrea; Hadley, Barbara; von Itzstein, Mark; Tiralongo, Joe

    2014-09-01

    Membrane proteins, including solute transporters play crucial roles in cellular function and have been implicated in a variety of important diseases, and as such are considered important targets for drug development. Currently the drug discovery process is heavily reliant on the structural and functional information discerned from high-resolution crystal structures. However, membrane protein structure determination is notoriously difficult, due in part to challenges faced in their expression, solubilisation and purification. The CMP-sialic acid transporter (CST) is considered to be an attractive target for drug discovery. CST inhibition reduces cancer cell sialylation and decreases the metastatic potential of cancer cells and to date, no crystal structure of the CST, or any other nucleotide sugar transporter exists. Here we describe the optimised conditions for expression in Pichia pastoris, solubilisation using n-nonyl β-d-maltopyranoside (NM) and single step purification of a functional CST. Importantly we show that despite being able to solubilise and purify the CST using a number of different detergents, only NM was able to maintain CST functionality.

  3. Optimization of media for bioethanol production by Pichia stipitis from sugarcane bagasse pretreated by dilute acid

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    Mohsen Ahi

    2014-04-01

    Full Text Available Introduction: Reduction of fossil fuels due to its increasing consumption caused the biofuels production as an important topic, today. Using resources that have not food application was regarded as the second generation biofuels and consisted of lignocelluloses. Since considerable amount of lignocellulosic material are pentoses, utilizing them is important for the production of biofuels. Materials and methods: Sugarcane bagasse was pretreated with dilute acid method. Pichia stipitis was used for the fermentation of released sugars. A L27 Taguchi orthogonal array was considered to optimize the fermentation process and increase the amount of ethanol. The eight factors with three levels considering nitrogen, phosphorus, zinc, sulfur, magnesium, and vitamins sources were considered in this study. Results: The analysis of the results shows that corn steep liquor, ammonium hydrogen phosphate, potassium di-hydrogen phosphate and magnesium sulfate have a significant effect on the production of ethanol, respectively. Confirmation of the optimal conditions shows that ethanol production was increased 97% relative to the mean of the observed results. The yield and productivity during 48 h of the fermentation were reached to 0.26 (g ethanol/g consumed sugar and 0.125g (L.h, respectively. Discussion and conclusion: At the optimum condition the production of ethanol from sugarcane bagasse hydrolysate had higher efficiency relative to previous studies. Results of medium optimization considering cheap resources showed showed an excellent potential toward an economical bioethanol production process.

  4. Production of Equine Infectious Anemia Virus (EIAV) antigen in Pichia pastoris.

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    de Arruda Coutinho, Luciana Cavalcanti; de Jesus, André Luiz Santos; de Paiva Fontes, Karin Florêncio Lins; Coimbra, Eliane Campos; Mariz, Filipe Colaço; de Freitas, Antonio Carlos; de Cássia Carvalho Maia, Rita; de Castro, Roberto Soares

    2013-08-01

    Equine Infectious Anemia (EIA) is a persistent lentivirus infection of horses which causes a chronic clinical condition with worldwide importance in veterinary medicine. The p26 protein is usually prepared for use as an antigen in serological tests for EIA diagnosis since it is a well-conserved gene sequence and very immunogenic. In view of the ability of yeast to make post-translational modifications of proteins, this study was carried out to allow Pichia pastoris to be used for the expression of a synthetic codon-optimized EIAV p26 gene. The gene was cloned into pPICZαA vector after appropriate enzymatic digestion. P. pastoris clones transformed with the pPICZαAp26 construction were induced to produce the recombinant p26 protein (rp26) under the regulation of alcohol oxidase 1 promoter by adding methanol to the culture medium. The p26 gene expression was detected by RT-PCR and the production of rp26 was confirmed by dot blotting, Western blotting, ELISA and AGID. The P. pastoris expression system was capable of producing a functional EIAV p26 protein that can be used directly in the functionality tests without requiring laborious purification or recovery steps. This is the first reported study of EIAV p26 protein production in yeast cells. Copyright © 2013 Elsevier B.V. All rights reserved.

  5. Cloning and Characterization of Two Pyruvate Decarboxylase Genes from Pichia stipitis CBS 6054

    Science.gov (United States)

    Lu, Ping; Davis, Brian P.; Jeffries, Thomas W.

    1998-01-01

    In Pichia stipitis, fermentative and pyruvate decarboxylase (PDC) activities increase with diminished oxygen rather than in response to fermentable sugars. To better characterize PDC expression and regulation, two genes for PDC (PsPDC1 and PsPDC2) were cloned and sequenced from P. stipitis CBS 6054. Aside from Saccharomyces cerevisiae, from which three PDC genes have been characterized, P. stipitis is the only organism from which multiple genes for PDC have been identified and characterized. PsPDC1 and PsPDC2 have diverged almost as far from one another as they have from the next most closely related known yeast gene. PsPDC1 contains an open reading frame of 1,791 nucleotides encoding 597 amino acids. PsPDC2 contains a reading frame of 1,710 nucleotides encoding 570 amino acids. An 81-nucleotide segment in the middle of the β domain of PsPDC1 codes for a unique segment of 27 amino acids, which may play a role in allosteric regulation. The 5′ regions of both P. stipitis genes include two putative TATA elements that make them similar to the PDC genes from S. cerevisiae, Kluyveromyces marxianus, and Hanseniaspora uvarum. PMID:9435065

  6. Heterologous expression of Pycnoporus cinnabarinus cellobiose dehydrogenase in Pichia pastoris and involvement in saccharification processes

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    Bey Mathieu

    2011-12-01

    Full Text Available Abstract Background Cellobiose dehydrogenase (CDH is an extracellular hemoflavoenzyme produced by lignocellulose-degrading fungi including Pycnoporus cinnabarinus. We investigated the cellulolytic system of P. cinnabarinus, focusing on the involvement of CDH in the deconstruction of lignocellulosic biomass. Results First, P. cinnabarinus growth conditions were optimized for CDH production. Following growth under cellulolytic conditions, the main components secreted were cellulases, xylanases and CDH. To investigate the contribution of P. cinnabarinus secretome in saccharification processes, the Trichoderma reesei enzymatic cocktail was supplemented with the P. cinnabarinus secretome. A significant enhancement of the degradation of wheat straw was observed with (i the production of a large amount of gluconic acid, (ii increased hemicellulose degradation, and (iii increased overall degradation of the lignocellulosic material. P. cinnabarinus CDH was heterologously expressed in Pichia pastoris to obtain large amounts of pure enzyme. In a bioreactor, the recombinant CDH (rCDH expression level reached 7800 U/L. rCDH exhibited values of biochemical parameters similar to those of the natural enzyme, and was able to bind cellulose despite the absence of a carbohydrate-binding module (CBM. Following supplementation of purified rCDH to T. reesei enzymatic cocktail, formation of gluconic acid and increased hemicellulose degradation were observed, thus confirming the previous results observed with P. cinnabarinus secretome. Conclusions We demonstrate that CDH offers an attractive tool for saccharification process enhancement due to gluconic acid production from raw lignocellulosic material.

  7. ¹³C-based metabolic flux analysis of recombinant Pichia pastoris.

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    Ferrer, Pau; Albiol, Joan

    2014-01-01

    Overexpression of a foreign protein may negatively affect several cell growth parameters, as well as cause cellular stress. Central (or core) metabolism plays a crucial role since it supplies energy, reduction equivalents, and precursor molecules for the recombinant product, cell's maintenance, and growth needs. However, the number of quantitative physiology studies of the impact of recombinant protein production on the central metabolic pathways of yeast cell factories has been traditionally rather limited, thereby hampering the application of rational strain engineering strategies targeting central metabolism.The development and application of quantitative physiology and modelling tools and methodologies is allowing for a systems-level understanding of the effect of bioprocess parameters such as growth rate, temperature, oxygen availability, and substrate(s) choice on metabolism, and its subsequent interactions with recombinant protein synthesis, folding, and secretion.Here, we review the recent developments and applications of (13)C-based metabolic flux analysis ((13)C-MFA) of Pichia pastoris and the gained understanding of the metabolic behavior of this yeast in recombinant protein production bioprocesses. We also discuss the potential of multilevel studies integrating (13)C-MFA with other omics analyses, as well as future perspectives on the metabolic modelling approaches to study and design metabolic engineering strategies for improved protein production.

  8. Heterologous Expression of Tulip Petal Plasma Membrane Aquaporins in Pichia pastoris for Water Channel Analysis▿

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    Azad, Abul Kalam; Sawa, Yoshihiro; Ishikawa, Takahiro; Shibata, Hitoshi

    2009-01-01

    Water channels formed by aquaporins (AQPs) play an important role in the control of water homeostasis in individual cells and in multicellular organisms. Plasma membrane intrinsic proteins (PIPs) constitute a subclass of plant AQPs. TgPIP2;1 and TgPIP2;2 from tulip petals are members of the PIP family. In this study, we overexpressed TgPIP2;1 and TgPIP2;2 in Pichia pastoris and monitored their water channel activity (WCA) either by an in vivo spheroplast-bursting assay performed after hypo-osmotic shock or by growth assay. Osmolarity, pH, and inhibitors of AQPs, protein kinases (PKs), and protein phosphatases (PPs) affect the WCA of heterologous AQPs in this expression system. The WCA of TgPIP2;2-expressing spheroplasts was affected by inhibitors of PKs and PPs, which indicates that the water channel of this homologue is regulated by phosphorylation in P. pastoris. From the results reported herein, we suggest that P. pastoris can be employed as a heterologous expression system to assay the WCA of PIPs and to monitor the AQP-mediated channel gating mechanism, and it can be developed to screen inhibitors/effectors of PIPs. PMID:19251885

  9. Heterologous expression of tulip petal plasma membrane aquaporins in Pichia pastoris for water channel analysis.

    Science.gov (United States)

    Azad, Abul Kalam; Sawa, Yoshihiro; Ishikawa, Takahiro; Shibata, Hitoshi

    2009-05-01

    Water channels formed by aquaporins (AQPs) play an important role in the control of water homeostasis in individual cells and in multicellular organisms. Plasma membrane intrinsic proteins (PIPs) constitute a subclass of plant AQPs. TgPIP2;1 and TgPIP2;2 from tulip petals are members of the PIP family. In this study, we overexpressed TgPIP2;1 and TgPIP2;2 in Pichia pastoris and monitored their water channel activity (WCA) either by an in vivo spheroplast-bursting assay performed after hypo-osmotic shock or by growth assay. Osmolarity, pH, and inhibitors of AQPs, protein kinases (PKs), and protein phosphatases (PPs) affect the WCA of heterologous AQPs in this expression system. The WCA of TgPIP2;2-expressing spheroplasts was affected by inhibitors of PKs and PPs, which indicates that the water channel of this homologue is regulated by phosphorylation in P. pastoris. From the results reported herein, we suggest that P. pastoris can be employed as a heterologous expression system to assay the WCA of PIPs and to monitor the AQP-mediated channel gating mechanism, and it can be developed to screen inhibitors/effectors of PIPs.

  10. Fructo-oligosaccharides production by the Gluconacetobacter diazotrophicus levansucrase expressed in the methylotrophic yeast Pichia pastoris.

    Science.gov (United States)

    Trujillo, L E.; Arrieta, J G.; Dafhnis, F; García, J; Valdés, J; Tambara, Y; Pérez, M; Hernández, L

    2001-02-01

    Levansucrase (LsdA) (EC 2.4.1.10) from Gluconacetobacter diazotrophicus (formerly Acetobacter diazotrophicus) yields high levels of fructo-oligosaccharides (FOS) from sucrose. A DNA fragment encoding the precursor LsdA lacking the first 57 amino acids was fused to the pho1 signal sequence under the control of the Pichia pastoris-alcohol oxidase 1 (AOX1) promoter. Methanol induction of a P. pastoris strain harboring a single copy of the lsdA expression cassette integrated in the genome resulted in the production of active levansucrase. After fermentation of the recombinant yeast, LsdA activity was detected in the periplasmic fraction (81%) and in the culture supernatant (18%) with an overall yield of 1% of total protein. The recombinant LsdA was glycosylated and displayed optimal pH and temperature for enzyme activity similar to those of the native enzyme, but thermal stability was increased. Neither fructosylpolymerase activity nor FOS production was affected. Incubation of recombinant LsdA in sucrose (500 g l(-1)) yielded 43% (w/w) of total sugar as 1-kestose, with a conversion efficiency about 70%. Intact recombinant yeast cells also converted sucrose to FOS although for a 30% efficiency.

  11. Generation of diploid Pichia pastoris strains by mating and their application for recombinant protein production

    Directory of Open Access Journals (Sweden)

    Chen Ming-Tang

    2012-07-01

    Full Text Available Abstract Background Yeast mating provides an efficient means for strain and library construction. However, biotechnological applications of mating in the methylotrophic yeast Pichia pastoris have been hampered because of concerns about strain stability of P. pastoris diploids. The aim of the study reported here is to investigate heterologous protein expression in diploid P. pastoris strains and to evaluate diploid strain stability using high cell density fermentation processes. Results By using a monoclonal antibody as a target protein, we demonstrate that recombinant protein production in both wild-type and glycoengineered P. pastoris diploids is stable and efficient during a nutrient rich shake flask cultivation. When diploid strains were cultivated under bioreactor conditions, sporulation was observed. Nevertheless, both wild-type and glycoengineered P. pastoris diploids showed robust productivity and secreted recombinant antibody of high quality. Specifically, the yeast culture maintained a diploid state for 240 h post-induction phase while protein titer and N-linked glycosylation profiles were comparable to that of a haploid strain expressing the same antibody. As an application of mating, we also constructed an antibody display library and used mating to generate novel full-length antibody sequences. Conclusions To the best of our knowledge, this study reports for the first time a comprehensive characterization of recombinant protein expression and fermentation using diploid P. pastoris strains. Data presented here support the use of mating for various applications including strain consolidation, variable-region glycosylation antibody display library, and process optimization.

  12. RP-HPLC determination of recombinant human interferon omega in the Pichia pastoris fermentation broth.

    Science.gov (United States)

    Liu, Hong; Pan, Hong-Chun; Peng, Li; Cai, Shao-Xi

    2005-07-15

    A rapid and valid reversed-phase high performance liquid chromatography (RP-HPLC) method for determination of recombinant human interferon omega (rhIFNomega) in the yeast Pichia pastoris fermentation broth was developed. The method is based on the hydrophobicity of rhIFNomega followed by RP-HPLC separation with UV detection. The chromatography analysis was performed on EC 250/4 NUCLEOSIL 300-5 C18 (250 mm x 4 mm i.d., 300 A, with a particle size of 5 microm) column. The compositions of the mobile phase A and B were 999:1 (v/v) water/TFA and 999:1 (v/v) acetonitrile/TFA at a flow rate of 1.0 ml min(-1). Detection was done by spectrophotometry at 280 nm and the column temperature was 30+/-1 degrees C. Calibration curve was linear (r=0.9986, n=7) in the range of 0.074-0.555 mg ml(-1) for rhIFNomega and the regression equation was y=2.02 x 10(6)x-1.27 x 10(5). Limit of detection for rhIFNomega was 0.053 mg ml(-1). The values of R.S.D. (%) of intra-day and inter-day precision were recovery rate of recovery experiment were <1.23 (n=3) and 97.97%.

  13. Integrative transformation system for the metabolic engineering of the sphingoid base-producing yeast Pichia ciferrii.

    Science.gov (United States)

    Bae, Jung-Hoon; Sohn, Jung-Hoon; Park, Chang-Seo; Rhee, Joon-Shick; Choi, Eui-Sung

    2003-02-01

    We have developed an integrative transformation system for metabolic engineering of the tetraacetyl phytosphingosine (TAPS)-secreting yeast Pichia ciferrii. The system uses (i) a mutagenized ribosomal protein L41 gene of P. ciferrii as a dominant selection marker that confer resistance to the antibiotic cycloheximide and (ii) a ribosomal DNA (rDNA) fragment of P. ciferrii as a target for multicopy gene integration into the chromosome. A locus within the nontranscribed region located between 5S and 26S rDNAs was selected as the integration site. A maximum frequency of integrative transformation of approximately 1,350 transformants/ microg of DNA was observed. To improve the de novo synthesis of sphingolipid, the LCB2 gene, encoding a subunit of serine palmitoyltransferase, which catalyzes the first committed step of sphingolipid synthesis, was cloned from P. ciferrii and overexpressed under the control of the P. ciferrii glyceraldehyde-3-phosphate dehydrogenase promoter. After transformation of an LCB2 gene expression cassette, several transformants that contained approximately five to seven copies of transforming DNA in the chromosome and exhibited about 50-fold increase in LCB2 mRNA relative to the wild type were identified. These transformants were observed to produce approximately two times more TAPS than the wild type.

  14. Construction of a Pichia pastoris cell-surface display system using Flo1p anchor system.

    Science.gov (United States)

    Tanino, Takanori; Fukuda, Hideki; Kondo, Akihiko

    2006-01-01

    A Pichia pastoris cell-surface display system was constructed using a Flo1p anchor system, which was developed in Saccharomyces cerevisiae. The lipase from Rhizopus oryzae with a pro sequence (ProROL) was used as the model protein and was genetically fused to the anchor consisting of amino acids 1-1099 of Flo1p (FS anchor). The resulting fusion protein FSProROL was expressed under the control of the alcohol oxidase 1 promoter (pAOX1). The fluorescence microscopy of immunolabeled P. pastoris cells revealed that ProROL was displayed on the cell surface, and Western blot analysis revealed that the fusion protein FSProROL was noncovalently attached to the cell wall and highly glycosylated. The lipase activity of P. pastoris cells was affected by the methanol concentration for the induction phase. Surprisingly, the activity of lipase displayed on the cells incubated at 60 degrees C was not only stable but also increased to about 6.5 times the initial value after 4 h incubation.

  15. Low pH D-xylonate production with Pichia kudriavzevii.

    Science.gov (United States)

    Toivari, Mervi; Vehkomäki, Maija-Leena; Nygård, Yvonne; Penttilä, Merja; Ruohonen, Laura; Wiebe, Marilyn G

    2013-04-01

    D-xylonic acid is one of the top 30 most desirable chemicals to be derived from biomass sugars identified by the US Department of Energy, being applicable as a non-food substitute for D-gluconic acid and as a platform chemical. We engineered the non-conventional yeast Pichia kudriavzevii VTT C-79090T to express a D-xylose dehydrogenase coding gene from Caulobacter crescentus. With this single modification the recombinant P. kudriavzevii strain produced up to 171 g L(-1) of D-xylonate from 171 g L(-1) D-xylose at a rate of 1.4 g L(-1) h(-1) and yield of 1.0 g [g substrate consumed](-1), which was comparable with D-xylonate production by Gluconobacter oxydans or Pseudomonas sp. The productivity of the strain was also remarkable at low pH, producing 146 g L(-1) D-xylonate at 1.2 g L(-1) h(-1) at pH 3.0. This is the best low pH production reported for D-xylonate. These results encourage further development towards industrial scale production.

  16. Modulation of cadmium bioaccumulation and enhancing cadmium tolerance in Pichia kudriavzevii by sodium chloride preincubation.

    Science.gov (United States)

    Ma, Ning; Li, Chunsheng; Zhang, Dandan; Yu, Jinzhi; Xu, Ying

    2016-07-01

    Application of growing microorganisms for cadmium removal is limited by the sensitivity of living cells to cadmium. The effects of sodium chloride (NaCl) preincubation on the cadmium bioaccumulation and tolerance of Pichia kudriavzevii and Saccharomyces cerevisiae were investigated in this study. NaCl preincubation significantly reduced the intracellular and cell-surface cadmium bioaccumulation of P. kudriavzevii at both 6 and 20 mg L(-1) cadmium, while no obvious effect was observed in S. cerevisiae except that the intracellular cadmium bioaccumulation at 20 mg L(-1) cadmium was reduced obviously by 20-60 g L(-1)  NaCl. For both yeasts, the improved contents of protein and proline after NaCl preincubation contributed to the cadmium tolerance. The thiol contents in P. kudriavzevii under cadmium stress were alleviated by NaCl preincubation, which might be due to the decrease of intracellular cadmium bioaccumulation. NaCl preincubation enhanced the contents of glycerol and trehalose in P. kudriavzevii under cadmium stress, while no acceleration was observed in S. cerevisiae. The results suggested that NaCl preincubation could be applied in cadmium removal by growing P. kudriavzevii to increase the cadmium tolerance of the yeast.

  17. High-level expression of Candida parapsilosis lipase/acyltransferase in Pichia pastoris.

    Science.gov (United States)

    Brunel, Laetitia; Neugnot, Virginie; Landucci, Laure; Boze, Hélène; Moulin, Guy; Bigey, Frédéric; Dubreucq, Eric

    2004-07-01

    Candida parapsilosis has been previously shown to produce a lipase/acyltransferase (EC 3.1.1.3) that preferentially catalyses transfer reactions such as alcoholysis over hydrolysis in the presence of suitable nucleophiles other than water, even in aqueous media (aw > 0.9 ). This enzyme has been shown to belong to a new family of lipases. The present work describes the cloning of the gene coding for this lipase/acyltransferase in the yeast Pichia pastoris and the heterologous high-level expression of the recombinant enzyme. The lipase/acyltransferase gene, in which the sequence encoding the signal peptide was replaced by that of the alpha-factor of Saccharomyces cerevisiae, was placed under the control of the methanol inducible promoter of the alcohol oxidase 1 gene (AOX1). A transformed P. pastoris clone, containing five copies of the lipase/acyltransferase gene, was selected for the production of recombinant enzyme. The fed-batch culture supernatant contained 5.8 gl(-1) (weighted) of almost pure recombinant lipase/acyltransferase displaying the same catalytic behavior as the original enzyme.

  18. Production of recombinant human bile salt-stimulated lipase in Pichia pastoris.

    Science.gov (United States)

    Murasugi, A; Asami, Y; Mera-Kikuchi, Y

    2001-11-01

    Recombinant human bile salt-stimulated lipase (rhBSSL) was efficiently expressed under the control of the AOX1 gene promoter in Pichia pastoris. Human BSSL has 16 successively repeated sequences in the carboxy terminal region. The sequence consists of 11 amino acid residues. The coding sequence for the middle 11 of the 16 repeats was removed from hBSSL cDNA to facilitate efficient secretory expression. The clone used for fermentation was a transformant of GS115 (his4) integrated with four copies of the expression cassette containing the modified hBSSL cDNA. Unique fermentation conditions were required for efficient expressions of rhBSSL in the high cell-density fermentation. A sufficient glycerol feed at 30 degrees C and pH 4 under an adequate concentration of dissolved oxygen in the growth phase make the cells active over a long induction period of approximately 15 days. On methanol induction, the concentration of dissolved oxygen should be maintained very low in the presence of sorbitol and skimmed milk at 20 degrees C and pH 5.7. Under these conditions, 0.8-1 g of rhBSSL was secreted in 1 liter of the medium. By immunoelectron microscopy, rhBSSL-tagged gold particles were located in secretion microbodies after the beginning of methanol induction. The secreted rhBSSL was efficiently captured and purified by expanded bed adsorption chromatography.

  19. Improved secretion of Candida antarctica lipase B with its native signal peptide in Pichia pastoris.

    Science.gov (United States)

    Vadhana, Ashok Kumar Prasanna; Samuel, Premsingh; Berin, Ronald M; Krishna, Jayachandran; Kamatchi, Kavitha; Meenakshisundaram, Sankaranarayanan

    2013-03-05

    Secretion efficiency of the 85-amino acid Sacchromyces cerevisiae alpha signal peptide and the 25-amino acid Candida antarctica lipase B signal (nsB) peptide were compared. Three reporter proteins used for the study are C. antarctica lipase A (CalA), lipase B (CalB) and hGMCSF. The copy number of recombinant α-CalB and nsB-CalB clones was determined by qPCR and clones with equivalent gene copies were used for comparative analysis. About threefold increased CalB production corresponding to an activity of 480 U ml(-1) was obtained with its native signal peptide, whereas with the alpha signal peptide the maximum activity was 160 U ml(-1). Also, CalB was secreted as a mature protein with native N-terminus when fused to its own signal peptide, while unprocessed CalB with N-terminal extension was detected with the alpha signal peptide. Real time PCR analysis of CalB strains indicated that the difference in protein expression was not at the transcriptional level. The nsB signal sequence was also effective in secreting CalA enzyme and its secretion efficiency was on par with the alpha signal sequence. Further, hGMCSF fused inframe with the nsB signal peptide was also efficiently secreted into the medium. These results indicate that the nsB signal peptide can be a better alternative to alpha signal peptide for heterologous protein expression in Pichia pastoris.

  20. Phosphate-responsive promoter of a Pichia pastoris sodium phosphate symporter.

    Science.gov (United States)

    Ahn, Jungoh; Hong, Jiyeon; Park, Myongsoo; Lee, Hyeokweon; Lee, Eungyo; Kim, Chunsuk; Lee, Joohwan; Choi, Eui-sung; Jung, Joon-ki; Lee, Hongweon

    2009-06-01

    To develop a functional phosphate-regulated promoter in Pichia pastoris, a phosphate-responsive gene, PHO89, which encodes a putative sodium (Na(+))-coupled phosphate symporter, was isolated. Sequencing analyses revealed a 1,731-bp open reading frame encoding a 576-amino-acid polypeptide with 12 putative transmembrane domains. The properties of the PHO89 promoter (P(PHO89)) were investigated using a bacterial lipase gene as a reporter in 5-liter jar fermentation experiments. P(PHO89) was tightly regulated by phosphate and was highly activated when the cells were grown in a phosphate-limited external environment. Compared to translation elongation factor 1alpha and the glyceraldehyde-3-phosphate dehydrogenase promoter, P(PHO89) exhibited strong transcriptional activity with higher specific productivity (amount of lipase produced/cell/h). Furthermore, a cost-effective and simple P(PHO89)-based fermentation process was developed for industrial application. These results demonstrate the potential for efficient use of P(PHO89) for controlled production of recombinant proteins in P. pastoris.

  1. Constitutive expression of human pancreatic lipase-related protein 1 in Pichia pastoris.

    Science.gov (United States)

    Aloulou, Ahmed; Grandval, Philippe; De Caro, Josiane; De Caro, Alain; Carrière, Frédéric

    2006-06-01

    High-level constitutive expression of the human pancreatic lipase-related protein 1 (HPLRP1) was achieved using the methylotrophic yeast Pichia pastoris. The HPLRP1 cDNA, including its original leader sequence, was subcloned into the pGAPZB vector and further integrated into the genome of P. pastoris X-33 under the control of the glyceraldehyde 3-phosphate dehydrogenase (GAP) constitutive promoter. A major protein with a molecular mass of 50 kDa was found to be secreted into the culture medium and was identified using anti-HPLRP1 polyclonal antibodies as HPLRP1 recombinant protein. The level of expression reached 100-120 mg of HPLRP1 per liter of culture medium after 40 h, as attested by specific and quantitative enzyme-linked immunosorbent assay. A single cation-exchange chromatography sufficed to obtain a highly purified recombinant HPLRP1 after direct batch adsorption onto S-Sepharose of the HPLRP1 present in the culture medium, at pH 5.5. N-terminal sequencing and mass spectrometry analysis were carried out to monitor the production of the mature protein and to confirm that its signal peptide was properly processed.

  2. Production of a novel cold-active lipase from Pichia lynferdii Y-7723.

    Science.gov (United States)

    Kim, Hak-Ryul; Kim, In-Hwan; Hou, Ching T; Kwon, Kwang-Il; Shin, Beom-Soo

    2010-01-27

    Lipase (triacylglycerol acylhydrolases, E.C. 3.1.1.3) is one of the most important enzymes applied to a broad range of industrial application fields. Especially, lipases with abnormal functionality such as thermostability and alkaline, acidic, and cold activities gain special attention because of their applicability in the restricted reaction conditions. In this study, 16 yeast strains prescreened for lipase induction were investigated for their actual lipase production, and we found a novel cold-active lipase produced from Pichia lynferdii Y-7723. The activity of lipase Y-7723 was retained by 74 and 70% at 20 and 10 degrees C, respectively, as compared to the maximum value at 35 degrees C. On the basis of an optimization study, the optimal lipase productivity was obtained at 96 h of incubation with 3% oil substrate in a medium composed of sucrose as a carbon source at pH 7.0. Among carbon sources tested, sucrose showed almost twice as high of a lipase production (184%) as the control, while the cell growth was similar (105%). Yeast extract and ammonium salts were effective as individual nitrogen sources for lipase production. This study demonstrated that the cold activity of lipase Y-7723 at 10 degrees C was highest among the cold-active lipases reported so far.

  3. Optimization of the high-level production of Rhizopus oryzae lipase in Pichia pastoris.

    Science.gov (United States)

    Minning, S; Serrano, A; Ferrer, P; Solá, C; Schmid, R D; Valero, F

    2001-03-09

    The lipases of the Rhizopus species family are important and versatile enzymes that are mainly used in fat and oil modification due to their strong 1,3-regiospecificity. Inexpensive synthetic medium was used for the production of Rhizopus oryzae lipase in the methylotrophic yeast Pichia pastoris. Methanol accumulation inside the bioreactor has previously been shown to negatively influence the production level. Three different methanol fed-batch strategies for maintaining the methanol concentration within optimal limits have been assayed in high-density cultures. One methanol feeding strategy, which is based on the monitoring of the methanol concentration by gas chromatography, resulted in a 2.5-fold higher productivity compared to an initial cultivation, where the feeding rate was adjusted according to the dissolved oxygen concentration (DO) in the supernatant. Finally, productivity could be further increased by introducing a transition phase that involved the simultaneous feeding of glycerol and methanol followed by a single methanol feed. This optimized strategy resulted in the highest productivity (12888 U l(-1) h(-1)), which is 13.6-fold higher than the DO-based strategy.

  4. High-level production of recombinant Geotrichum candidum lipases in yeast Pichia pastoris.

    Science.gov (United States)

    Holmquist, M; Tessier, D C; Cygler, M

    1997-10-01

    We describe the heterologous high-level expression of the two Geotrichum candidum lipase (GCL) isoenzymes from strain ATCC 34614 in the methylotrophic yeast Pichia pastoris. The lipase cDNAs were placed under the control of the methanol-inducible alcohol oxidase promoter. The lipases expressed in P. pastoris were fused to the alpha-factor secretion signal peptide of Saccharomyces cerevisiae and were secreted into the culture medium. Cultures of P. pastoris expressing lipase accumulated active recombinant enzyme in the supernatant to levels of approximately 60 mg/L virtually free from contaminating proteins. This yield exceeds that previously reported with S. cerevisiae by a factor of more than 60. Recombinant GCL I and GCL II had molecular masses of approximately 63 and approximately 66 kDa, respectively, as determined by SDS-PAGE. The result of endoglucosidase H digestion followed by Western blot analysis of the lipases suggested that the enzymes expressed in P. pastoris received N-linked high-mannose-type glycosylation to an extent, 6-8% (w/w), similar to that in G. candidum. The specific activities and substrate specificities of both recombinant lipases were determined and were found to agree with what has been reported for the enzymes isolated from the native source.

  5. Overexpression of Candida rugosa lipase Lip1 via combined strategies in Pichia pastoris.

    Science.gov (United States)

    Li, Xu; Liu, Zimin; Wang, Guilong; Pan, Dujie; Jiao, Liangcheng; Yan, Yunjun

    2016-01-01

    In this study, combined strategies were employed to heterologously overexpress Candida rugosa lipase Lip1 (CRL1) in a Pichia pastoris system. The LIP1 gene was systematically codon-optimized and synthesized in vitro. The Lip1 activity of a recombinant strain harboring three copies of the codon-optimized LIP1 gene reached 1200 U/mL in a shake flask culture. Higher lipase activity, 1450 U/mL, was obtained using a five copy number construct. Co-expressing one copy of the ERO1p and BiP chaperones with Lip1p, the CRL1 lipase yield further reached 1758 U/mL, which was significantly higher than that achieved by expressing Lip1p alone or only co-expressing one molecular chaperone. When cultivated in a 3 L fermenter under optimal conditions, the recombinant strain GS115/87-ZA-ERO1p-BiP #7, expressing the molecular chaperones Ero1p and BiP, produced 13,490 U/mL of lipase activity at 130 h, which was greater than the 11,400 U/mL of activity for the recombinant strain GS115/pAO815-α-mCRL1 #87, which did not express a molecular chaperone. This study indicates that a strategy of combining codon optimization with co-expression of molecular chaperones has great potential for the industrial-scale production of pure CRL1.

  6. Residue-specific global fluorination of Candida antarctica lipase B in Pichia pastoris.

    Science.gov (United States)

    Budisa, Nediljko; Wenger, Waltraud; Wiltschi, Birgit

    2010-09-01

    We report the in vivo fluorination of the tryptophan, tyrosine, and phenylalanine residues in a glycosylation-deficient mutant of Candida antarctica lipase B, CalB N74D, expressed in the methylotrophic yeast Pichia pastoris and subsequently segregated into the growth medium. To achieve this, a P. pastoris strain auxotrophic for all three aromatic amino acids was supplemented with 5-fluoro-L-tryptophan, meta-fluoro-(DL)-tyrosine, or para-fluoro-L-phenylalanine during expression of CalB N74D. The residue-specific replacement of the canonical amino acids by their fluorinated analogs was confirmed by mass analysis. Although global fluorination induced moderate changes in the secondary structure of CalB N74D, the fluorous variant proteins were still active lipases. However, their catalytic activity was lower than that of the non-fluorinated parent protein while their resistance to proteolytic degradation by proteinase K remained unchanged. Importantly, we observed that the global fluorination prolonged the shelf life of the lipase activity, which is an especially useful feature for the storage of, e.g., therapeutic proteins. Our study represents the first step on the road to the production of biotechnologically and pharmacologically relevant fluorous proteins in P. pastoris.

  7. Production of recombinant human bile salt stimulated lipase and its variant in Pichia pastoris.

    Science.gov (United States)

    Sahasrabudhe, A V; Solapure, S M; Khurana, R; Suryanarayan, V; Ravishankar, S; deSousa, S M; Das, G

    1998-12-01

    hBSSL and its truncated variant hBSSL-C cDNA clones were expressed in Pichia pastoris using two different signal peptides, native signal peptide and invertase signal peptide, respectively, to facilitate secretion of the recombinant proteins into the culture medium. Both recombinant proteins were secreted into the culture medium to a level of 45-50 mg/liter in shake flask cultures. Native signal peptide of hBSSL was recognized in P. pastoris and was cleaved at the same site as in humans. The level of expression of the hBSSL gene was found to be dependent on the number of its copies integrated into the host chromosome. The multicopy transformant clone was found to be very stable. When grown and induced in a fermentor, the level of accumulation of the recombinant hBSSL in the culture medium improved from 50 mg/liter in shake flask cultures to 300 mg/liter. The recombinant hBSSL purified from the culture supernatant was found to be similar to the native hBSSL in its biochemical properties except for the lectin-binding profile.

  8. Recombinant production of bacterial toxins and their derivatives in the methylotrophic yeast Pichia pastoris

    Directory of Open Access Journals (Sweden)

    Gurkan Cemal

    2005-12-01

    Full Text Available Abstract The methylotrophic yeast Pichia pastoris is a popular heterologous expression host for the recombinant production of a variety of prokaryotic and eukaryotic proteins. The rapid emergence of P. pastoris as a robust heterologous expression host was facilitated by the ease with which it can be manipulated and propagated, which is comparable to that of Escherichia coli and Saccharomyces cerevisiae. P. pastoris offers further advantages such as the tightly-regulated alcohol oxidase promoter that is particularly suitable for heterologous expression of foreign genes. While recombinant production of bacterial toxins and their derivatives is highly desirable, attempts at their heterologous expression using the traditional E. coli expression system can be problematic due to the formation of inclusion bodies that often severely limit the final yields of biologically active products. However, recent literature now suggests that P. pastoris may be an attractive alternative host for the heterologous production of bacterial toxins, such as those from the genera Bacillus, Clostridium, and Corynebacterium, as well as their more complex derivatives. Here, we review the recombinant production of bacterial toxins and their derivatives in P. pastoris with special emphasis on their potential clinical applications. Considering that de novo design and construction of synthetic toxin genes have often been necessary to achieve optimal heterologous expression in P. pastoris, we also present general guidelines to this end based on our experience with the P. pastoris expression of the Bacillus thuringiensis Cyt2Aa1 toxin.

  9. Characteristics of glycosylated streptokinase secreted from Pichia pastoris: enhanced resistance of SK to proteolysis by glycosylation.

    Science.gov (United States)

    Pratap, J; Rajamohan, G; Dikshit, K L

    2000-04-01

    Degradation of streptokinase (SK) has been frequently observed during large-scale protein production. An enhanced susceptibility of SK to degradation has been correlated with its existence in a partially unfolded state. The influence of the carbohydrate moiety on the stability and functional characteristics of SK has been examined by obtaining the glycoform of SK following its secretion through the methylotrophic yeast Pichia pastoris. Secretion of the protein product was achieved by replacing the native secretion signal codons of SK with those from alpha-factor leader peptide and expressing the fusion construct under the control of the methanol-inducible alcohol oxidase (ox) promoter of P. pastoris after its integration into the host chromosome. Western blot and zymographic analysis of proteins secreted from the recombinant P. pastoris indicated that SK was glycosylated by the host cells, which resulted in the appearance of a SK species migrating slowly, corresponding to a 55-kDa protein product as compared to the 47-kDa native SK. The glycosylated SK retained a plasminogen activation capability identical to that of its unglycosylated counterpart. Glycoform SK exhibited an enhanced stability profile at 25 degrees C and 37 degrees C and improved resistance towards protease treatment compared to unglycosylated SK secreted through P. pastoris after tunicamycin treatment or that secreted from the recombinant Escherichia coli. The results presented thus illustrate that N-linked glycosylation of SK results in 30-40% enhancement of the protein stability and resistance towards degradation but does not interfere with its fibrinolytic function.

  10. Expression and Purification of Enterovirus Type 71 Polyprotein P1 using Pichia pastoris system

    Institute of Scientific and Technical Information of China (English)

    Xue Han; Xiaoling Ying; Hao Huang; Shili Zhou; Qi Huang

    2012-01-01

    Enterovirus type 71(EV71) causes severe hand-foot-and-mouth disease(HFMD) resulting in hundreds of deaths of children every year; However,currently,there is no effective treatment for EV71.In this study,the EV71 poly-protein(EV71-P1 protein) gene was processed and cloned into the eukaryotic expression vector pPIC9k and then expressed in Pichia pastoris strain GS115.The EV71 P1 pretein with a molecular weight of 100 kD was produced and secreted into the medium.The soluble EV71 P1 protein was purified by column chromatography with a recovery efficiency of 70%.The result of the immunological analysis showed that the EV71 P1 protein had excellent immunogenicity and could stimulate the production of EV71-VP1 IgG antibody in injected rabbits.We suggest that EV71-P1 protein is an ideal candidate for an EV71 vaccine to prevent EV71 infection.

  11. Transcriptional control of ADH genes in the xylose-fermenting yeast Pichia stipitis

    Energy Technology Data Exchange (ETDEWEB)

    Cho, J.Y.; Jeffries, T.W. [Forest Service, Madison, WI (United States). Forest Products Lab.]|[Univ. of Wisconsin, Madison, WI (United States). Dept. of Bacteriology

    1999-06-01

    The authors studied the expression of the genes encoding group 1 alcohol dehydrogenases (PsADH1 and PsADH2) in the xylose-fermenting yeast Pichia stipitis CBS 6054. The cells expressed PsADH1 approximately 10 times higher under oxygen-limited conditions than under fully aerobic conditions when cultivated on xylose. Transcripts of PsADH2 were not detectable under either aeration condition. The authors used a PsADH1::lacZ fusion to monitor PsADH1 expression and found that expression increased as oxygen decreased. The level of PsADH1 transcript was expressed about 10-fold in cells grown in the presence of heme under oxygen-limited conditions. Concomitantly with the induction of PsADH1, PsCYC1 expression was regressed. These results indicate that oxygen availability regulates PsADH1 expression and that regulation may be mediated by heme. The regulation of PsADH2 expression was also examined in other genetic backgrounds. Disruption of PsADH1 dramatically increased PsADH2 expression on nonfermentable carbon sources under fully aerobic conditions, indicating that the expression of PsADH2 is subject to feedback regulation under these conditions.

  12. Transcriptional Control of ADH Genes in the Xylose-Fermenting Yeast Pichia stipitis

    Science.gov (United States)

    Cho, Jae-Yong; Jeffries, Thomas W.

    1999-01-01

    We studied the expression of the genes encoding group I alcohol dehydrogenases (PsADH1 and PsADH2) in the xylose-fermenting yeast Pichia stipitis CBS 6054. The cells expressed PsADH1 approximately 10 times higher under oxygen-limited conditions than under fully aerobic conditions when cultivated on xylose. Transcripts of PsADH2 were not detectable under either aeration condition. We used a PsADH1::lacZ fusion to monitor PsADH1 expression and found that expression increased as oxygen decreased. The level of PsADH1 transcript was repressed about 10-fold in cells grown in the presence of heme under oxygen-limited conditions. Concomitantly with the induction of PsADH1, PsCYC1 expression was repressed. These results indicate that oxygen availability regulates PsADH1 expression and that regulation may be mediated by heme. The regulation of PsADH2 expression was also examined in other genetic backgrounds. Disruption of PsADH1 dramatically increased PsADH2 expression on nonfermentable carbon sources under fully aerobic conditions, indicating that the expression of PsADH2 is subject to feedback regulation under these conditions. PMID:10347014

  13. Effect of inhibitors formed during wheat straw pretreatment on ethanol fermentation by Pichia stipitis.

    Science.gov (United States)

    Bellido, Carolina; Bolado, Silvia; Coca, Mónica; Lucas, Susana; González-Benito, Gerardo; García-Cubero, María Teresa

    2011-12-01

    The inhibitory effect of the main inhibitors (acetic acid, furfural and 5-hydroxymethylfurfural) formed during steam explosion of wheat straw was studied through ethanol fermentations of model substrates and hydrolysates from wheat straw by Pichia stipitis. Experimental results showed that an increase in acetic acid concentration led to a reduction in ethanol productivity and complete inhibition was observed at 3.5 g/L. Furfural produced a delay on sugar consumption rates with increasing concentration and HMF did not exert a significant effect. Fermentations of the whole slurry from steam exploded wheat straw were completely inhibited by a synergistic effect due to the presence of 1.5 g/L acetic acid, 0.15 g/L furfural and 0.05 g/L HMF together with solid fraction. When using only the solid fraction from steam explosion, hydrolysates presented 0.5 g/L of acetic acid, whose fermentations have submitted promising results, providing an ethanol yield of 0.45 g ethanol/g sugars and the final ethanol concentration reached was 12.2 g/L (10.9 g ethanol/100 g DM).

  14. Expression of Leishmania major LmSTI1 in Yeast Pichia Pastoris

    Directory of Open Access Journals (Sweden)

    Mehdi Shokri

    2017-01-01

    Full Text Available Background: Leishmania major LmSTI1 is a conserved protein among different species of leishmania, and expressed in both amastigote and promastigote forms of L. major life cycle. It has previously been expressed in bacterial systems.Materials and Methods: To express LmSTI1 in the methylotrophic yeast         Pichia pastoris (P. pastoris, the shuttle vector pPICZA containing gene lmsti1 was constructed under the control of the AOX1 promoter. The recombinant vector was electro-transformed into P. pastoris, and induced by 0.5% methanol in the buffered medium. The expression of the LmSTI1 protein was visualized in the total soluble protein of P. pastoris by 12% SDS-PAGE, and further confirmed by Western blotting with L.major-infected mouse sera and HRP-conjugated goat anti-mouse IgG as the first and secondary antibodies, respectively.Results: The expression level was 0.2% of total soluble proteins.Conclusion: It might be possible to use this formulation as a whole yeast candidate vaccine against cutaneous leishmanization.

  15. Isolation and characterization of the plasma membrane from the yeast Pichia pastoris.

    Science.gov (United States)

    Grillitsch, Karlheinz; Tarazona, Pablo; Klug, Lisa; Wriessnegger, Tamara; Zellnig, Günther; Leitner, Erich; Feussner, Ivo; Daum, Günther

    2014-07-01

    Despite similarities of cellular membranes in all eukaryotes, every compartment displays characteristic and often unique features which are important for the functions of the specific organelles. In the present study, we biochemically characterized the plasma membrane of the methylotrophic yeast Pichia pastoris with emphasis on the lipids which form the matrix of this compartment. Prerequisite for this effort was the design of a standardized and reliable isolation protocol of the plasma membrane at high purity. Analysis of isolated plasma membrane samples from P. pastoris revealed an increase of phosphatidylserine and a decrease of phosphatidylcholine compared to bulk membranes. The amount of saturated fatty acids in the plasma membrane was higher than in total cell extracts. Ergosterol, the final product of the yeast sterol biosynthetic pathway, was found to be enriched in plasma membrane fractions, although markedly lower than in Saccharomyces cerevisiae. A further characteristic feature of the plasma membrane from P. pastoris was the enrichment of inositol phosphorylceramides over neutral sphingolipids, which accumulated in internal membranes. The detailed analysis of the P. pastoris plasma membrane is discussed in the light of cell biological features of this microorganism especially as a microbial cell factory for heterologous protein production.

  16. Optimization of Media for Production of an Effective Yeast Biocontrol Agent Pichia membranefaciens

    Institute of Scientific and Technical Information of China (English)

    WAN Ya-kun; TIAN Shi-ping

    2004-01-01

    The growth of Pichia membranefaciens was studied using different nitrogen and carbonsources as substrates. Among nitrogen sources tested, soya peptone, yeast extract power,beef extract and polypeptone were relatively favorable to the growth of yeast. Thedensity of the yeast showed to be directly proportional to carbon sources supplementation.Glucose and fructose were good carbon sources for the yeast growth. However, lactoseshowed poor performance for the cell growth of the yeast. In this study, beef extractpresented a good synergic effect on the yeast growth with different carbonhydrates. Themedium for P.membranefaciens used glucose and beef extract as substrates. The higherconcentration of glucose and beef extract, the better growth of P.membranefaciens. Theaddition of chlorella growth factor (CGF) stimulated markedly the growth of P.membranefaci-ens. The increased concentration of CGF from 0.5 to 1% did not enhance the numbers ofP.membranefaciens. This result will help design a better strategy for scale-up produc-tion of P.membranefaciens.

  17. Cloning and expression of a functional core streptavidin in Pichia pastoris: strategies to increase yield.

    Science.gov (United States)

    Casteluber, Marisa C F; Damasceno, Leonardo M; da Silveira, Wendel B; Diniz, Raphael H S; Passos, Frederico J V; Passos, Flávia M L

    2012-01-01

    Streptavidin is widely used as an analytical tool and affinity tag together with biotinylated surfaces or molecules. We report for the first time a simple strategy that yields high biomass of a Pichia pastoris strain containing a methanol induced core streptavidin (cStp) gene. Three factors were evaluated for biomass production: glycerol concentration, aeration, and feed flow rates in a bioreactor. Recycling of recombinant cells, either free or immobilized, was investigated during induction. Concentration of 2.0 M glycerol, feeding flow rate of 0.11 mL min(-1) , and aeration by air injection dispersed with a porous stone combined with agitation at 500 rpm were the set of conditions resulting into maximum biomass yield (150 g L(-1) ). These parameters yielded 4.0 g L(-1) of cStp, after 96 h of induction. Recombinant biomass was recycled twice before being discarded, which can reduce production costs and simplify the process. Immobilized P. pastoris biomass produced 2.94 and 1.70 g L(-1) of cStp in the first and second induction cycle, respectively. Immobilization and recycling of recombinant P. pastoris biomass opens new possibilities as a potential strategy to improve volumetric productivity for heterologous protein expression. Copyright © 2012 American Institute of Chemical Engineers (AIChE).

  18. Process Control and Optimization for Heterologous Protein Production by Methylotrophic Pichia pastoris

    Institute of Scientific and Technical Information of China (English)

    GAO Minjie; SHI Zhongping

    2013-01-01

    The methylotrophic yeast Pichia pastoris is a highly successful system for production of a variety of heterologous proteins due to its unique features/abilities for effective protein expression,and tremendous efforts have been made to increase heterologous protein productivity by P.pastoris in recent years.When new engineered yeast strains are constructed and are ready to use for industrial protein production,process control and optimization techniques should be applied to improve the fermentation performance in the following aspects:(1)increase recombinant cell concentrations in fermentor to high density during growth phase;(2)effectively induce heterologous proteins by enhancing/stabilizing titers or concentrations of the proteins during induction phase;(3)decrease operation costs by relieving the working loads of heat-exchange and oxygen supply.This article reviews and discusses the key and commonly used techniques in heterologous protein production by P.pastoris,with the focus on optimizations of fermentation media and basic operation conditions,development of optimal glycerol feeding strategies for achieving high density cultivation of P.pastoris and effective heterologous protein induction methods by regulating specific growth rate,methanol concentration,temperatures,mixture ratio of multi-carbon substrates,etc.Metabolic analysis for recombinant protein production by P.pastoris is also introduced to interpret the mechanism of sub-optimal heterologous protein production and to explore further optimal expression methods.

  19. Characteristics and Applicability of Phytase of the Yeast Pichia anomala in Synthesizing Haloperoxidase.

    Science.gov (United States)

    Joshi, Swati; Satyanarayana, Tulasi

    2015-07-01

    The phytase of the yeast Pichia anomala is a histidine acid phosphatase based on signature sequences and catalytic amino acids identified by site-directed mutagenesis. Among modulators, N-bromosuccinimide and butanedione inhibit phytase, while Ca(2+) and Ni(2+) stimulate slightly. Vanadate exhibits competitive inhibition of phytase, making it bifunctional to act as haloperoxidase. Molecular docking supports vanadate to share its binding site with phytate. The T 1/2, activation energy (E a ), temperature quotient (Q 10), activation energy of thermal inactivation (Ed), and enthalpy (ΔH d (0) ) of the enzyme are 4.0 min (80 °C), 27.72 kJ mol(-1), 2.1, 410.62 kJ mol(-1), and ∼407.8 kJ mol(-1) (65-80 °C), respectively. The free energy of the process (ΔG d (o) ) increases from 49.56 to 71.58 kJ mol(-1) with rise in temperature, while entropy of inactivation (ΔS d (0) ) remains constant at ∼1.36 kJ mol(-1) K(-1). The supplementation of whole wheat dough with rPPHY resulted in 72.5 % reduction in phytic acid content of bread. These characteristics confirm that the phytase has adequate thermostability for its applicability as a food and feed additive.

  20. Dissociation and purification of the endogenous membrane-bound Vo complex from Pichia pastoris.

    Science.gov (United States)

    Li, Sumei; Hong, Tao; Wang, Kun; Lu, Yinghong; Zhou, Min

    2017-10-01

    Most proteins occur and function in complexes rather than as isolated entities in membranes. In most cases macromolecules with multiple subunits are purified from endogenous sources. In this study, an endogenous membrane-protein complex was obtained from Pichia pastoris, which can be grown at high densities to significantly improve the membrane protein yield. We successfully isolated the membrane-bound Vo complex of V-ATPase from P. pastoris using a fusion FLAG tag attached to the C-terminus of subunit a to generate the vph-tag strain, which was used for dissociation and purification. After FLAG affinity and size exclusion chromatography purification, the production quantity and purity of the membrane-bound Vo complex was 20 μg l(-1) and >98%, respectively. The subunits of the endogenous membrane-bound Vo complex observed in P. pastoris were similar to those obtained from S. cerevisiae, as demonstrated by liquid chromatography-tandem mass spectrometry (LC-MS-MS). Therefore, successful dissociation and purification of the membrane-bound Vo complex at a high purity and sufficient quantity was achieved via a rapid and simple procedure that can be used to obtain the endogenous membrane-protein complexes from P. pastoris. Copyright © 2017 Elsevier Inc. All rights reserved.