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https://www.readbyqxmd.com/read/27913218/methylotrophic-yeast-pichia-pastoris-as-a-chassis-organism-for-polyketide-synthesis-via-the-full-citrinin-biosynthetic-pathway
#1
Ying Xue, Chuixing Kong, Wei Shen, Chenxiao Bai, Yanna Ren, Xiangshan Zhou, Yuanxing Zhang, Menghao Cai
With the rapid development of synthetic biology, exploring various chassis organisms has become necessary to improve the heterologous biosynthesis of natural products and pharmaceuticals. In this study, we tested the potential of the industrial methylotrophic yeast strain Pichia pastoris for the heterologous synthesis of polyketides. A recombinant P. pastoris GS-pksCT-npgA carrying the Monascus purpureus citrinin polyketide synthase gene pksCT and the Aspergillus nidulans phosphopantetheinyl transferase gene npgA was constructed...
November 29, 2016: Journal of Biotechnology
https://www.readbyqxmd.com/read/27907132/a-simplified-and-efficient-process-for-insulin-production-in-pichia-pastoris
#2
Sulena Polez, Domenico Origi, Sotir Zahariev, Corrado Guarnaccia, Sergio G Tisminetzky, Nataša Skoko, Marco Baralle
A significant barrier to insulin is affordability. In this manuscript we describe improvements to key steps in the insulin production process in Pichia pastoris that reduce cost and time. The strategy for recovery and processing of human insulin precursor has been streamlined to two steps from bioreactor to the transpeptidation reaction. In the first step the insulin precursor secreted during the methanol induction phase is recovered directly from the culture broth using Tangential Flow Filtration with a Prostak™ module eliminating the laborious and time-consuming multi-step clarification, including centrifugation...
2016: PloS One
https://www.readbyqxmd.com/read/27905091/recent-advances-of-molecular-toolbox-construction-expand-pichia-pastoris-in-synthetic-biology-applications
#3
REVIEW
Zhen Kang, Hao Huang, Yunfeng Zhang, Guocheng Du, Jian Chen
Pichia pastoris: (reclassified as Komagataella phaffii), a methylotrophic yeast strain has been widely used for heterologous protein production because of its unique advantages, such as readily achievable high-density fermentation, tractable genetic modifications and typical eukaryotic post-translational modifications. More recently, P. pastoris as a metabolic pathway engineering platform has also gained much attention. In this mini-review, we addressed recent advances of molecular toolboxes, including synthetic promoters, signal peptides, and genome engineering tools that established for P...
January 2017: World Journal of Microbiology & Biotechnology
https://www.readbyqxmd.com/read/27904401/comparison-of-cultivable-acetic-acid-bacterial-microbiota-in-organic-and-conventional-apple-cider-vinegar
#4
Aleksandra Štornik, Barbara Skok, Janja Trček
Organic apple cider vinegar is produced from apples that go through very restricted treatment in orchard. During the first stage of the process, the sugars from apples are fermented by yeasts to cider. The produced ethanol is used as a substrate by acetic acid bacteria in a second separated bioprocess. In both, the organic and conventional apple cider vinegars the ethanol oxidation to acetic acid is initiated by native microbiota that survived alcohol fermentation. We compared the cultivable acetic acid bacterial microbiota in the production of organic and conventional apple cider vinegars from a smoothly running oxidation cycle of a submerged industrial process...
March 2016: Food Technology and Biotechnology
https://www.readbyqxmd.com/read/27897254/high-level-expression-of-improved-thermo-stable-alkaline-xylanase-variant-in-pichia-pastoris-through-codon-optimization-multiple-gene-insertion-and-high-density-fermentation
#5
Yihong Lu, Cheng Fang, Qinhong Wang, Yuling Zhou, Guimin Zhang, Yanhe Ma
In paper industry, xylanases are used to increase the pulp properties in bleaching process as its eco-friendly nature. The xylanases activity is hindered by high temperature and alkaline conditions with high enzyme production cost in the paper industry. Here, XynHB, an alkaline stable xylanase from Bacillus pumilus HBP8 was mutated at N188A to XynHBN188A. Expressed mutant in E. coli showed 1.5-fold higher xylanase activity than XynHB at 60 °C. The mutant expressed in Pichia pastoris was glycosylated, remained stable for 30 min at 60 °C...
November 29, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27890597/identification-eukaryotic-expression-and-structure-function-characterizations-of-%C3%AE-defensin-like-homologues-from-pelodiscus-sinensis
#6
Haining Yu, Xuelian Liu, Hui Wang, Lan Feng, Xue Qiao, Shasha Cai, Nannan Shi, Yipeng Wang
Defensins are a group of host defense peptides that play a central role in host innate immune responses. Here, 26 genes encoding β-defensin-like peptides have been identified for the first time in Pelodiscus sinensis using database mining approach. Phylogenetic study confirmed that β-defensins are fast evolving genes with high rates of sequence substitutions. The expression level of several selected genes in different tissues was examined by RT-PCR. Ps-BDs mainly adopt β-strands and/or α-helix conformations homology modeled by Rosetta program...
November 24, 2016: Developmental and Comparative Immunology
https://www.readbyqxmd.com/read/27889145/potential-probiotic-pichia-kudriavzevii-strains-and-their-ability-to-enhance-folate-content-of-traditional-cereal-based-african-fermented-food
#7
Anna Greppi, Fabien Saubade, Cristian Botta, Christèle Humblot, Jean-Pierre Guyot, Luca Cocolin
With the aim of selecting starter cultures with interesting probiotic potential and with the ability to produce folate in a food matrix, yeast strains isolated from fermented cereal-based African foods were investigated. A total of 93 yeast strains were screened for their tolerance to pH 2 and 0.3% of bile salts. Pichia kudriavzevii isolates gave the best results. Selected P. kudriavzevii strains were tested for survival to the simulated human digestion and for adhesion to Caco-2 cells. Moreover, presence of folate biosynthesis genes was verified and production of extra and intra-cellular folate determined during growth in culture medium...
April 2017: Food Microbiology
https://www.readbyqxmd.com/read/27889126/identification-of-the-major-yeasts-isolated-from-high-moisture-corn-and-corn-silages-in-the-united-states-using-genetic-and-biochemical-methods
#8
M C Santos, C Golt, R D Joerger, G D Mechor, Gerson B Mourão, L Kung
The objective of this study was to identify species of yeasts in samples of high moisture corn (HMC) and corn silage (CS) collected from farms throughout the United States. Samples were plated and colonies were isolated for identification using DNA analysis. Randomly selected colonies were also identified by fatty acid methyl esters (FAME) and by physiological substrate profiling (ID 32C). For CS, Candida ethanolica, Saccharomyces bulderi, Pichia anomala, Kazachstania unispora, and Saccharomyces cerevisiae were the predominant yeasts...
November 23, 2016: Journal of Dairy Science
https://www.readbyqxmd.com/read/27876784/expression-and-characterization-of-a-talaromyces-marneffei-active-phospholipase-b-expressed-in-a-pichia-pastoris-expression-system
#9
Yan He, Linghua Li, Fengyu Hu, Wanshan Chen, Huali Lei, Xiejie Chen, Weiping Cai, Xiaoping Tang
Phospholipase B is a virulence factor for several clinically important pathogenic fungi, including Candida albicans, Cryptococcus neoformans and Aspergillus fumigatus, but its role in the thermally dimorphic fungus Talaromyces marneffei remains unclear. Here, we provide the first report of the expression of a novel phospholipase gene, designated TmPlb1, from T. marneffei in the eukaryotic expression system of Pichia pastoris GS115. Sensitive real-time quantitative reverse-transcription PCR (qRT-PCR) demonstrated that the expression of TmPlb1 increased 1...
November 23, 2016: Emerging Microbes & Infections
https://www.readbyqxmd.com/read/27871374/catalytic-efficiency-and-thermostability-improvement-of-suc2-invertase-through-rational-site-directed-mutagenesis
#10
Nooshin Mohandesi, Kamahldin Haghbeen, Omid Ranaei, Seyed Shahriar Arab, Sorour Hassani
Engineering of invertases has come to attention because of increasing demand for possible applications of invertases in various industrial processes. Due to the known physicochemical properties, invertases from micro-organisms such as Saccharomyces cerevisiae carrying SUC2 gene are considered as primary models. To improve thermostability and catalytic efficiency of SUC2 invertase (SInv), six influential residues with Relative Solvent Accessibility<5% were selected through multiple-sequence alignments, molecular modelling, structural and computational analyses...
January 2017: Enzyme and Microbial Technology
https://www.readbyqxmd.com/read/27868164/efficient-production-of-recombinant-glycoprotein-d-of-herpes-simplex-virus-type-2-in-pichia-pastoris-and-its-protective-efficacy-against-viral-challenge-in-mice
#11
Man Wang, Shuai Jiang, Li Zhou, Chaoqun Wang, Ruifeng Mao, Murugavel Ponnusamy
Herpes simplex virus type 2 (HSV-2) infection is the leading cause of genital ulcer disease and a significant public health concern. However, there are no approved vaccines available to prevent HSV-2 infection. The glycoprotein D (gD) of HSV-2 is the most important candidate antigen for vaccine development. In this study, a truncated form of gD (codons 1-340, gD1-340) was produced as a secretory protein in the methylotrophic yeast Pichia pastoris. The recombinant gD1-340 with a His6 tag was purified to homogeneity by one-step affinity chromatography...
November 21, 2016: Archives of Virology
https://www.readbyqxmd.com/read/27867835/combined-13-c-assisted-metabolomics-and-metabolic-flux-analysis-reveals-the-impacts-of-glutamate-on-the-central-metabolism-of-high-%C3%AE-galactosidase-producing-pichia-pastoris
#12
Ping Liu, Mingzhi Huang, Menglei Guo, Jiangchao Qian, Weilu Lin, Ju Chu, Yingping Zhuang, Siliang Zhang
BACKGROUND: Pichia pastoris is a popular recombinant protein expression system for its accessibility of efficient gene manipulation and high protein production. Sufficient supply of precursors, energy, and redox cofactors is crucial for high recombinant protein production. In our present work, we found that the addition of glutamate improved the recombinant β-galactosidase (β-gal) production by P. pastoris G1HL. METHODS: To elucidate the impacts of glutamate on the central metabolism in detail, a combined (13)C-assisted metabolomics and (13)C metabolic flux analysis was conducted based on LC-MS/MS and GC-MS data...
2016: Bioresources and Bioprocessing
https://www.readbyqxmd.com/read/27860461/heterologous-expression-of-the-plant-cysteine-protease-bromelain-and-its-inhibitor-in-pichia-pastoris
#13
Nora Luniak, Peter Meiser, Sonja Burkart, Rolf Müller
Expression of proteases in heterologous hosts remains an ambitious challenge due to severe problems associated with digestion of host proteins. On the other hand, proteases are broadly used in industrial applications and resemble promising drug candidates. Bromelain is an herbal drug that is medicinally used for treatment of oedematous swellings and inflammatory conditions and consists in large part of proteolytic enzymes. Even though various experiments underline the requirement of active cysteine proteases for biological activity, so far no investigation succeeded to clearly clarify the pharmacological mode of action of bromelain...
November 14, 2016: Biotechnology Progress
https://www.readbyqxmd.com/read/27858135/expression-in-pichia-pastoris-and-characterization-of-two-novel-dirigent-proteins-for-atropselective-formation-of-gossypol
#14
Isabelle Effenberger, Michael Harport, Jens Pfannstiel, Iris Klaiber, Andreas Schaller
We established an efficient fed-batch fermentation process for two novel dirigent proteins from cotton plants, GbDIR2 from Gossypium barbadense and GhDIR3 from G. hirsutum, using the engineered Pichia pastoris GlycoSwitch® SuperMan5 strain to prevent hyperglycosylation. The two (His)6-tagged proteins were purified by metal-chelate affinity chromatography and obtained in quantities of 12 and 15 mg L(-1) of culture volume, respectively. Glycosylation sites were identified for the native and for the enzymatically deglycosylated proteins by mass spectrometry, confirming five to six of the seven predicted glycosylation sites in the NxS/T sequence context...
November 17, 2016: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/27857705/isolation-identification-and-characterization-of-yeasts-from-fermented-goat-milk-of-the-yaghnob-valley-in-tajikistan
#15
Linnea A Qvirist, Carlotta De Filippo, Francesco Strati, Irene Stefanini, Maddalena Sordo, Thomas Andlid, Giovanna E Felis, Paola Mattarelli, Duccio Cavalieri
The geographically isolated region of the Yaghnob Valley, Tajikistan, has allowed its inhabitants to maintain a unique culture and lifestyle. Their fermented goat milk constitutes one of the staple foods for the Yaghnob population, and is produced by backslopping, i.e., using the previous fermentation batch to inoculate the new one. This study addresses the yeast composition of the fermented milk, assessing genotypic, and phenotypic properties. The 52 isolates included in this study revealed small species diversity, belonging to Kluyveromyces marxianus, Pichia fermentans, Saccharomyces cerevisiae, and one Kazachstania unispora...
2016: Frontiers in Microbiology
https://www.readbyqxmd.com/read/27856231/bioprospecting-thermotolerant-ethanologenic-yeasts-for-simultaneous-saccharification-and-fermentation-from-diverse-environments
#16
Jairam Choudhary, Surender Singh, Lata Nain
Lignocellulosic biomass, a promising renewable energy source, can be used for the production of second generation bioethanol. Simultaneous saccharification and fermentation (SSF), the process which alleviates the problem of separate hydrolysis and fermentation (SHF), requires thermotolerant ethanologenic yeast for bioethanol production. Therefore, ten yeast strains isolated from diverse sources, belonging to various genera like Saccharomyces, Candida, Pichia and Wickerhamomyces were evaluated for their thermotolerance, sugar utilization pattern, inhibitor tolerance and ethanol production potential with glucose, xylose and alkali pretreated paddy straw...
November 14, 2016: Journal of Bioscience and Bioengineering
https://www.readbyqxmd.com/read/27854040/expression-and-characterization-of-a-novel-antifungal-exo-%C3%AE-1-3-glucanase-from-chaetomium-cupreum
#17
Cheng Jiang, Jinzhu Song, Hua Cong, Junzheng Zhang, Qian Yang
A novel β-1,3-glucanase gene, designated Ccglu17A, was cloned from the biological control fungus Chaetomium cupreum Ame. Its 1626-bp open reading frame encoded 541 amino acids. The corresponding amino acid sequence showed highest identity (67 %) with a glycoside hydrolase family 17 β-1,3-glucanase from Chaetomium globosum. The recombinant protein Ccglu17A was successfully expressed in Pichia pastoris, and the enzyme was purified to homogeneity with 10.1-fold purification and 47.8 % recovery yield. The protein's molecular mass was approximately 65 kDa, and its maximum activity appeared at pH 5...
November 16, 2016: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/27847501/chicken-igy-fc-linked-to-bordetella-avium-ompa-and-taishan-pinus-massoniana-pollen-polysaccharide-adjuvant-enhances-macrophage-function-and-specific-immune-responses
#18
Wenwen Dong, Hao Zhang, He Huang, Jianbo Zhou, Liping Hu, Ailing Lian, Lijun Zhu, Ningning Ma, Pingping Yang, Kai Wei, Ruiliang Zhu
Fc-fusion technologies, in which immunoglobulin Fc is genetically fused to an antigenic protein, have been developed to confer antibody-like properties to proteins and peptides. Mammalian IgG Fc fusion exhibits improved antigen-induced immune responses by providing aggregates with high avidity for the IgG Fc receptor and salvaging the antigenic portion from endosomal degradation. However, whether the linked chicken IgY Fc fragment shares similar characteristics to mammalian IgG Fc remains unclear. In this study, we linked the chicken IgY Fc gene to the outer membrane protein A (ompA) of Bordetella avium through overlapping PCR...
2016: Frontiers in Microbiology
https://www.readbyqxmd.com/read/27844110/matchout-deuterium-labelling-of-proteins-for-small-angle-neutron-scattering-studies-using-prokaryotic-and-eukaryotic-expression-systems-and-high-cell-density-cultures
#19
O Dunne, M Weidenhaupt, P Callow, A Martel, M Moulin, S J Perkins, M Haertlein, V T Forsyth
Small-angle neutron scattering (SANS) is a powerful technique for the characterisation of macromolecular structures and interactions. Its main advantage over other solution state approaches is the ability to use D2O/H2O solvent contrast variation to selectively match out specific parts of a multi-component system. While proteins, nucleic acids, and lipids are readily distinguished in this way, it is not possible to locate different parts of a protein-protein system without the introduction of additional contrast by selective deuteration...
November 14, 2016: European Biophysics Journal: EBJ
https://www.readbyqxmd.com/read/27842609/a-recombinant-multi-antigen-vaccine-formulation-containing-babesia-bovis-merozoite-surface-antigens-msa-2a1-msa-2b-and-msa-2c-elicits-invasion-inhibitory-antibodies-and-ifn-%C3%AE-producing-cells
#20
Alba Marina Gimenez, Katia S Françoso, Jonatan Ersching, Marcelo Y Icimoto, Vitor Oliveira, Anabel E Rodriguez, Leonhard Schnittger, Monica Florin-Christensen, Mauricio M Rodrigues, Irene S Soares
BACKGROUND: Babesia bovis is a tick-transmitted protozoan hemoparasite and the causative agent of bovine babesiosis, a potential risk to more than 500 million cattle worldwide. The vaccines currently available are based on attenuated parasites, which are difficult to produce, and are only recommended for use in bovines under one year of age. When used in older animals, these vaccines may cause life-threatening clinical symptoms and eventually death. The development of a multi-subunit recombinant vaccine against B...
November 14, 2016: Parasites & Vectors
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