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https://www.readbyqxmd.com/read/28653743/genetic-diversity-and-pectinolytic-activity-of-epiphytic-yeasts-from-grape-carposphere
#1
M Cilião Filho, M B D Bertéli, J S Valle, L D Paccola-Meirelles, G A Linde, F G Barcellos, N B Colauto
The genetic diversity of epiphytic yeasts from grape carposphere is susceptible to environmental variations that determine the predominant carposphere microbiota. Understanding the diversity of yeasts that inhabit grape carposphere in different environments and their pectinolytic activity is a way to understand the biotechnological potential that surrounds us and help improve winemaking. Therefore, this study aimed to evaluate the pectinolytic activity and characterize the genetic diversity of isolated epiphytic yeasts from grape carposphere...
June 20, 2017: Genetics and Molecular Research: GMR
https://www.readbyqxmd.com/read/28651974/variants-of-ppulcc-a-multi-dye-decolorizing-laccase-from-pleurotus-pulmonarius-expressed-in-pichia-pastoris
#2
Christoph J Behrens, Diana Linke, Aldrige B Allister, Katerina Zelena, Ralf G Berger
A laccase of the basidiomycete Pleurotus pulmonarius (PpuLcc) possessed strong decolorizing abilities towards artificial and natural dyes. The PpuLcc was purified from the culture supernatant via FPLC, and the corresponding gene cloned and expressed in Pichia pastoris GS115. To examine the impact of the C-terminal tail region and the signal peptide on the recombinant expression of PpuLcc, a non-modified version or different truncations (2, -5, -13 AA) of the target protein were combined with different secretion signals...
June 23, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28651356/heterologous-expression-in-pichia-pastoris-and-biochemical-characterization-of-the-unmodified-sulfatase-from-fusarium-proliferatum-le1
#3
Svetlana A Korban, Kirill S Bobrov, Maria A Maynskova, Stanislav N Naryzhny, Olga L Vlasova, Elena V Eneyskaya, Anna A Kulmisnkaya
Sulfatases are a family of enzymes (sulfuric ester hydrolases, EC 3.1.6.-) that catalyze the hydrolysis of a wide array of sulfate esters. To date, despite the discovery of many sulfatase genes and the accumulation of data on numerous sulfated molecules, the number of characterized enzymes that are key players in sulfur metabolism remains extremely limited. While mammalian sulfatases are well studied due to their involvement in a wide range of normal and pathological biological processes, lower eukaryotic sulfatases, especially fungal sulfatases, have not been thoroughly investigated at the biochemical and structural level...
June 23, 2017: Protein Engineering, Design & Selection: PEDS
https://www.readbyqxmd.com/read/28650069/transcriptional-engineering-of-the-glyceraldehyde-3-phosphate-dehydrogenase-promoter-for-improved-heterologous-protein-production-in-pichia-pastoris
#4
Özge Ata, Roland Prielhofer, Brigitte Gasser, Diethard Mattanovich, Pınar Çalık
The constitutive glyceraldehyde-3-phosphate dehydrogenase promoter (PGAP ), which is one of the benchmark promoters of Pichia pastoris, was analyzed in terms of putative transcription factor binding sites. We constructed a synthetic library with distinct regulatory properties through deletion and duplication of these putative transcription factor binding sites and selected transcription factor (TF) genes were overexpressed or deleted to understand their roles on heterologous protein production. Using enhanced green fluorescent protein, an expression strength in a range between 0...
June 26, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28641563/pentapeptide-prosequence-enhances-expression-and-structure-folding-of-recombinant-thermomyces-lanuginosus-lipase-in-pichia-pastoris
#5
Haiying Cai, Minjie Zhao, Yang Li, Jianwei Mao, Chenggang Cai, Fengqin Feng
Propeptides have been demonstrated to influence many properties of their mature proteins as intramolecular chaperones. In our study, the role of pentapeptide prosequence of Thermomyces lanuginosus lipase (TLL) was determined through its effect on the recombinant expression of TLL in P. pastoris. The result showed that the average lipase activity of recombinant strains GS-pTL reached 434.32 U/mL, higher than that of GS-TL 377.71 U/mL, indicating that the propeptide "SPIRR-" sequence is beneficial for enhancing TLL expression...
June 21, 2017: Protein and Peptide Letters
https://www.readbyqxmd.com/read/28638135/temperature-dependence-of-protein-water-interactions-in-a-gated-yeast-aquaporin
#6
Camilo Aponte-Santamaría, Gerhard Fischer, Petra Båth, Richard Neutze, Bert L de Groot
Regulation of aquaporins is a key process of living organisms to counteract sudden osmotic changes. Aqy1, which is a water transporting aquaporin of the yeast Pichia pastoris, is suggested to be gated by chemo-mechanical stimuli as a protective regulatory-response against rapid freezing. Here, we tested the influence of temperature by determining the X-ray structure of Aqy1 at room temperature (RT) at 1.3 Å resolution, and by exploring the structural dynamics of Aqy1 during freezing through molecular dynamics simulations...
June 21, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28637873/disease-linked-mutations-in-factor-h-reveal-pivotal-role-of-cofactor-activity-in-self-surface-selective-regulation-of-complement-activation
#7
Heather Kerr, Edwin Wong, Elisavet Makou, Yi Yang, Kevin Marchbank, David Kavanagh, Anna Richards, Andrew P Herbert, Paul N Barlow
Spontaneous activation enables the complement system to respond very rapidly to diverse threats. Activation is efficiently suppressed by complement factor H (CFH) on self surfaces but not foreign surfaces. The surface selectivity of CFH, a soluble protein containing 20 CCP modules (CCPs 1-20), may be compromised by disease-linked mutations. Which of the several functions of CFH drives its self-surface selectivity remains unknown. We made human CFH mutants in Pichia pastoris. In benchmark studies of CCP-1 variants, recombinant I62-CFH (protective against age-related macular degeneration) and V62-CFH functioned equivalently, matching or outperforming plasma-derived CFH while R53H-CFH, linked to atypical haemolytic uraemic syndrome (aHUS), was defective in C3bBb decay-accelerating activity (DAA) and factor I-cofactor activity (CA)...
June 21, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28633643/enhancement-of-thermoalkaliphilic-xylanase-production-by-pichia-pastoris-through-novel-fed-batch-strategy-in-high-cell-density-fermentation
#8
Tingting Shang, Dayong Si, Dongyan Zhang, Xuhui Liu, Longmei Zhao, Cong Hu, Yu Fu, Rijun Zhang
BACKGROUND: Xylanase degrades xylan into monomers of various sizes by catalyzing the endohydrolysis of the 1,4-β-D-xylosidic linkage randomly, possessing potential in wide industrial applications. Most of xylanases are susceptible to be inactive when suffering high temperature and high alkaline process. Therefore, it is necessary to develop a high amount of effective thermoalkaliphilic xylanases. 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...
June 21, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28632332/microscale-disruption-of-microorganisms-for-parallelized-process-development
#9
REVIEW
Cornelia Walther, Astrid Dürauer
Escherichia coli, Saccharomyces cerevisiae, and Pichia pastoris are the standard platforms for biopharmaceutical production with 40% of all between 2010 to 2014 approved protein drugs produced in those microbial hosts. Typically, products overexpressed E. coli and S. cerevisiae remain in the cytosol or are secreted into the periplasm. Consequently, efficient cell disruption is essential for high product recovery during microbial production. Process development platforms at microscale are essential to shorten time to market...
June 20, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28623735/biochemical-and-structural-characterization-of-penicillium-purpurogenum-%C3%AE-d-galactosidase-binding-of-galactose-to-an-alternative-pocket-may-explain-enzyme-inhibition
#10
Luis Morales-Quintana, Carolina Faúndez, Raúl Herrera, Vasni Zavaleta, María Cristina Ravanal, Jaime Eyzaguirre, María Alejandra Moya-León
The fungus Penicillium purpurogenum degrades plant cell walls by the action of cellulolytic, xylanolytic and pectinolytic enzymes. The α-D-galactosidase is one of the enzymes which may act on pectin degradation. This enzyme has several biotechnological and medical applications. The aim of this work was to better understand the molecular mechanism of α-D-galactosidase from P. purpurogenum (GALP1). For this purpose, a gene coding for the enzyme was identified from the fungal genome and heterologously expressed in Pichia pastoris...
June 9, 2017: Carbohydrate Research
https://www.readbyqxmd.com/read/28622854/linear-dna-vaccine-prepared-by-large-scale-pcr-provides-protective-immunity-against-h1n1-influenza-virus-infection-in-mice
#11
Fei Wang, Quanjiao Chen, Shuntang Li, Chenyao Zhang, Shanshan Li, Min Liu, Kun Mei, Chunhua Li, Lixin Ma, Xiaolan Yu
Linear DNA vaccines provide effective vaccination. However, their application is limited by high cost and small scale of the conventional polymerase chain reaction (PCR) generally used to obtain sufficient amounts of DNA effective against epidemic diseases. In this study, a two-step, large-scale PCR was established using a low-cost DNA polymerase, RKOD, expressed in Pichia pastoris. Two linear DNA vaccines encoding influenza H1N1 hemagglutinin (HA) 1, LEC-HA, and PTO-LEC-HA (with phosphorothioate-modified primers), were produced by the two-step PCR...
June 2017: Veterinary Microbiology
https://www.readbyqxmd.com/read/28619526/auto-induction-of-pichia-pastoris-aox1-promoter-for-membrane-protein-expression
#12
Jonas Y Lee, Hao Chen, Alan Liu, Benjamin M Alba, Ai Ching Lim
Pichia pastoris is a highly successful recombinant protein expression system due to its ability to quickly generate large quantities of recombinant proteins in simple media. P. pastoris has been used to successfully generate milligram quantities of many important human membrane proteins, including G-protein coupled receptors, ion channels, and transporters, which are becoming increasingly important therapeutic targets. Despite these successes, protein expression in P. pastoris is still cumbersome due to a need to change growth media from glycerol media to methanol induction media, which minimizes inhibition of the AOX1 promoter by residual glycerol...
June 13, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28613409/molecular-cloning-and-characterizations-of-xylanase-inhibitor-protein-from-wheat-triticum-aestivum
#13
Xinyu Liu, Yakun Zhang, Zhaohui Wei, Hongge Chen, Xincheng Jia
Xylanase inhibitor proteins (XIPs) were regarded to inhibit the activity of xylanases during baking and gluten-starch separation processes. To avoid the inhibition to xylanases, it is necessary to define the conditions under which the inhibition takes place. In this study, we cloned the XIP gene from 2 different variety of Triticum aestivum, that is, Zhengmai 9023 and Zhengmai 366, and investigated the properties of XIP protein expressed by Pichia pastoris. The results showed that the 2 XIP genes (xip-9023 and xip-366) were highly homologous with only 3 nucleotide differences...
June 14, 2017: Journal of Food Science
https://www.readbyqxmd.com/read/28611759/energy-charge-as-an-indicator-of-pexophagy-in-pichia-pastoris
#14
Jianguo Zhang, Taiyu Liu
Pichia pastoris is a good model for pexophagy research owing to its diverse pexophagy modes (macropexophagy and micropexophagy) exhibited during carbon-source shift from methanol to other carbon sources. The critical condition that triggers activation of macropexophagy and micropexophagy is important for clarifying the P. pastoris pexophagy mechanism and human peroxisomal disorders. In this study, the pexophagy modes of P. pastoris were confirmed by green fluorescent protein expression and alcohol oxidase and formate dehydrogenase activities...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28608761/biomedical-applications-of-yeast-a-patent-view-part-one-yeasts-as-workhorses-for-the-production-of-therapeutics-and-vaccines
#15
Farzin Roohvand, Mehdi Shokri, Meghdad Abdollahpour-Alitappeh, Parastoo Ehsani
Yeasts, as Eukaryotes, offer unique features for ease of growth and genetic manipulation possibilities, making it an exceptional microbial host. Areas covered: This review provides general and patent-oriented insights into production of biopharmaceuticals by yeasts. Patents, wherever possible, were correlated to the original or review articles. The review describes applications of major GRAS (generally regarded as safe) yeasts for the production of therapeutic proteins and subunit vaccines; additionally, immunomodulatory properties of yeast cell wall components were reviewed for use of whole yeast cells as a new vaccine platform...
June 22, 2017: Expert Opinion on Therapeutic Patents
https://www.readbyqxmd.com/read/28608428/fusion-of-plectasin-derivative-nz2114-with-hydrophilic-random-coil-polypeptide-recombinant-production-in-pichia-pastoris-and-antimicrobial-activity-against-clinical-strain-mrsa
#16
L A Lima, R de Vries, L S Biswaro, I M Vasconcelos, O L Franco, S C Dias
One of the roadblocks towards the practical use of antimicrobial peptides for medical use is their relatively high cost when synthesized chemically. Effective recombinant production has only been successful in some cases, such as the previously reported production in Pichia pastoris of the antimicrobial plectasin derivative peptide NZ2114. The same production host has also been used extensively to produce so-called protein-polymers: sequences that consist of repetitions of simple amino acid motifs found in structural proteins such as collagen and elastin, and that can be designed to self-assemble in micelles, fibers and hydrogels...
June 12, 2017: Biopolymers
https://www.readbyqxmd.com/read/28606662/comparison-of-cerato-platanin-family-protein-bcspl1-produced-in-pichia-pastoris-and-escherichia-coli
#17
Yi Zhang, Yingbo Liang, Dewen Qiu, Jingjing Yuan, Xiufen Yang
The Botrytis cinerea BcSpl1 protein is a member of the cerato-platanin family, and consists of 137 amino acids and two disulfide bridges. This protein induces the onset of necrosis in infiltrated plant hosts. Recombinant BcSpl1 proteins produced in Pichia pastoris (pBcSpl1) and Escherichia coli (eBcSpl1) were initially compared regarding their abilities to induce necrosis and systemic acquired resistance (SAR). The pBcSpl1 and eBcSpl1 treatments led to the development of necrotic lesions on tomato leaves, and provided tomato plants with SAR to B...
June 9, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28603986/str3-and-cys3-contribute-to-2-furfurylthiol-biosynthesis-in-chinese-sesame-flavored-baijiu-yeast
#18
Musu Zha, Sheng Yin, Baoguo Sun, Xin Wang, Chengtao Wang
2-Furfurylthiol is an important aroma compound with characteristic sesame flavor. It was recently identified as the representative aroma compound of Chinese sesame-flavored Baijiu. But its formation mechanism is unclear. In this study, the dominant yeast species Saccharomyces cerevisiae, Pichia kudriavzevii, and Wickerhamomyces anomalus were isolated from samples of Chinese sesame-flavored fermentation and were screened for the capacity of 2-furfurylthiol biosynthesis. Some isolates were found to be capable of generating 2-furfurylthiol using furfural and L-cysteine as precursors, among which S...
June 12, 2017: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/28603806/atp-sulfurylase-is-essential-for-the-utilization-of-sulfamate-as-a-sulfur-source-in-the-yeast-komagataella-pastoris-syn-pichia-pastoris
#19
Tomas Linder
The methylotrophic yeast Komagataella pastoris (syn. Pichia pastoris) is one of the few known yeasts that can utilize sulfamate ([Formula: see text]) as a sulfur source. The biochemical pathway responsible for the catabolism of sulfamate has yet to be identified. The present study sought to investigate whether sulfamate catabolism proceeds through either of the inorganic sulfur intermediates sulfate ([Formula: see text]) or sulfite ([Formula: see text]) before its assimilation and subsequent incorporation into sulfur-containing amino acids and their derivatives...
June 12, 2017: Current Microbiology
https://www.readbyqxmd.com/read/28602730/recombinant-expression-of-a-laccase-from-coriolopsis-gallica-in-pichia-pastoris-using-a-modified-%C3%AE-factor-preproleader
#20
Mayra Avelar, Clarita Olvera, Denise Aceves-Zamudio, Jorge Luis Folch, Marcela Ayala
In this work we communicate the heterologous expression of a laccase from Coriolopsis gallica in Pichia pastoris. This enzyme has been reported to efficiently degrade a variety of pollutants such as industrial dyes. The expression strategy included using a previously reported modified α-factor preproleader for enhanced secretion and pAOX1, a methanol-responsive promoter. Methanol concentration, copper salts concentration and temperature were varied in order to enhance laccase expression in this heterologous system...
June 5, 2017: Protein Expression and Purification
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