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https://www.readbyqxmd.com/read/28105485/biochemical-characterization-of-thermostable-%C3%AE-1-4-mannanase-belonging-to-the-glycoside-hydrolase-family-134-from-aspergillus-oryzae
#1
Kiyota Sakai, Mai Mochizuki, Miyuki Yamada, Yuta Shinzawa, Miho Minezawa, Saran Kimoto, Shunsuke Murata, Yuhei Kaneko, Saaya Ishihara, Sadanari Jindou, Tetsuo Kobayashi, Masashi Kato, Motoyuki Shimizu
A β-1,4-mannanase, termed AoMan134A, that belongs to the GH 134 family was identified in the filamentous fungus Aspergillus oryzae. Recombinant AoMan134A was expressed in Pichia pastoris, and the purified enzyme produced mannobiose, mannotriose, mannotetraose, and mannopentaose from galactose-free β-mannan, with mannotriose being the predominant reaction product. The catalytic efficiency (k cat/K m ) of AoMan134A was 6.8-fold higher toward galactomannan from locust bean gum, than toward galactomannan from guar gum, but similar toward galactomannan from locust bean gum and glucomannan from konjac flour...
January 19, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28101807/direct-bioethanol-production-from-wheat-straw-using-xylose-glucose-co-fermentation-by-co-culture-of-two-recombinant-yeasts
#2
Yuanyuan Zhang, Caiyun Wang, Lulu Wang, Ruoxin Yang, Peilei Hou, Junhong Liu
To achieve a cost-effective bioconversion of lignocellulosic materials, a novel xylose/glucose co-fermentation process by co-culture of cellulose-utilizing recombinant Saccharomyces cerevisiae (S. cerevisiae) and xylan-utilizing recombinant Pichia pastoris (P. pastoris) was developed, in which ethanol was produced directly from wheat straw without additional hydrolytic enzymes. Recombinant S. cerevisiae coexpressing three types of cellulase and recombinant P. pastoris coexpressing two types of xylanase were constructed, respectively...
January 18, 2017: Journal of Industrial Microbiology & Biotechnology
https://www.readbyqxmd.com/read/28101787/a-recombinant-highly-thermostable-%C3%AE-mannanase-retman26-from-thermophilic-bacillus-subtilis-tbs2-expressed-in-pichia-pastoris-and-its-ph-and-temperature-stability
#3
Zhangcai Luo, Jing Miao, Guoying Li, Yao Du, Xiaobin Yu
A gene encoding a highly thermostable β-mannanase from a thermophilic Bacillus subtilis (TBS2) was successfully expressed in Pichia pastoris. The maximum activity of the recombinant thermostable β-mannanase (ReTMan26) was 5435 U/mL, which was obtained by high-density, fed-batch cultivation after 168-h induction with methanol in a 50-L bioreactor. The protein yield reached 3.29 mg/mL, and the protein had a molecular weight of ~42 kDa. After fermentation, ReTMan26 was purified using a 10-kDa cut-off membrane and Sephadex G-75 column...
January 18, 2017: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/28088197/production-of-human-pro-relaxin-h2-in-the-yeast-pichia-pastoris
#4
D Cimini, K Della Corte, R Finamore, L Andreozzi, A Stellavato, A V A Pirozzi, F Ferrara, R Formisano, M De Rosa, M Chino, L Lista, A Lombardi, V Pavone, C Schiraldi
BACKGROUND: 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...
January 14, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28086806/age-related-macular-degeneration-associated-polymorphism-rs10490924-in-arms2-results-in-deficiency-of-a-complement-activator
#5
Sven Micklisch, Yuchen Lin, Saskia Jacob, Marcus Karlstetter, Katharina Dannhausen, Prasad Dasari, Monika von der Heide, Hans-Martin Dahse, Lisa Schmölz, Felix Grassmann, Medhanie Alene, Sascha Fauser, Harald Neumann, Stefan Lorkowski, Diana Pauly, Bernhard H Weber, Antonia M Joussen, Thomas Langmann, Peter F Zipfel, Christine Skerka
BACKGROUND: Age-related macular degeneration (AMD) is the leading cause of blindness in developed countries. The polymorphism rs10490924 in the ARMS2 gene is highly associated with AMD and linked to an indel mutation (del443ins54), the latter inducing mRNA instability. At present, the function of the ARMS2 protein, the exact cellular sources in the retina and the biological consequences of the rs10490924 polymorphism are unclear. METHODS: Recombinant ARMS2 was expressed in Pichia pastoris, and protein functions were studied regarding cell surface binding and complement activation in human serum using fluoresence-activated cell sorting (FACS) as well as laser scanning microscopy (LSM)...
January 5, 2017: Journal of Neuroinflammation
https://www.readbyqxmd.com/read/28067041/utility-of-thermostable-xylanases-of-mycothermus-thermophilus-in-generating-prebiotic-xylooligosaccharides
#6
Rui Ma, Yingguo Bai, Huoqing Huang, Huiying Luo, Sanfeng Chen, Yunliu Fan, Lei Cai, Bin Yao
Xylooligosaccharides as emerging prebiotics are able to promote the growth of probiotic bacteria. In the present study, four neutral, thermostable xylanases (MtXyn11A, MtXyn11At, MtXyn11B and MtXyn11C) from compost fungus Mycothermus thermophilus CGMCC3.18119 were overexpressed in Pichia pastoris GS115 and used to produce xylooligosaccharides from beechwood xylan. The enzymes showed similar enzymatic properties (maximal activities at pH 6.06.5 and 65 C) but varied in catalytic efficiency and cleaving actions...
January 9, 2017: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/28061345/bioaugmentation-as-a-strategy-for-the-remediation-of-pesticide-polluted-soil-a-review
#7
REVIEW
Mariusz Cycoń, Agnieszka Mrozik, Zofia Piotrowska-Seget
Bioaugmentation, a green technology, is defined as the improvement of the degradative capacity of contaminated areas by introducing specific microorganisms, has emerged as the most advantageous method for cleaning-up soil contaminated with pesticides. The present review discusses the selection of pesticide-utilising microorganisms from various sources, their potential for the degradation of pesticides from different chemical classes in liquid media as well as soil-related case studies in a laboratory, a greenhouse and field conditions...
December 26, 2016: Chemosphere
https://www.readbyqxmd.com/read/28056814/competition-assays-and-physiological-experiments-of-soil-and-phyllosphere-yeasts-identify-candida-subhashii-as-a-novel-antagonist-of-filamentous-fungi
#8
Maja Hilber-Bodmer, Michael Schmid, Christian H Ahrens, Florian M Freimoser
BACKGROUND: While recent advances in next generation sequencing technologies have enabled researchers to readily identify countless microbial species in soil, rhizosphere, and phyllosphere microbiomes, the biological functions of the majority of these species are unknown. Functional studies are therefore urgently needed in order to characterize the plethora of microorganisms that are being identified and to point out species that may be used for biotechnology or plant protection. Here, we used a dual culture assay and growth analyses to characterise yeasts (40 different isolates) and their antagonistic effect on 16 filamentous fungi; comprising plant pathogens, antagonists, and saprophytes...
January 5, 2017: BMC Microbiology
https://www.readbyqxmd.com/read/28050843/evaluating-the-probiotic-and-therapeutic-potentials-of-saccharomyces-cerevisiae-strain-obs2-isolated-from-fermented-nectar-of-toddy-palm
#9
Banoth Srinivas, Ganapathiwar Swarupa Rani, Bhukya Kiran Kumar, Banoth Chandrasekhar, Kommalapati Vamsi Krishna, Tangutur Anjana Devi, Bhukya Bhima
The purpose of this study is to evaluate the probiotic characteristics of 15 yeast strains isolated from nectar of toddy palm. Initially, the collected samples were inoculated on yeast extract peptone dextrose agar plates and the colonies so obtained were culturally and morphologically characterized. Commercial probiotic yeast, Saccharomyces boulardii served as the control in these experiments. Of the 15 yeast strains, the isolates that were resistant to antibiotics and worked synergistically with other cultures were considered for further evaluation...
December 2017: AMB Express
https://www.readbyqxmd.com/read/28039507/production-and-characteristics-of-the-recombinant-extracellular-bifunctional-endoglucanase-of-the-polyextremophilic-bacterium-bacillus-halodurans-and-its-applicability-in-saccharifying-agro-residues
#10
Ranjitha R Prabhu, Deepak Parashar, T Satyanarayana
The recombinant alkalistable and moderately thermostable bifunctional endoglucanase gene (BhCell-Xyl) of polyextremophilic bacterium Bacillus halodurans TSLV1 has been expressed in Pichia pastoris under constitutive GAP as well as inducible AOX promoters. A higher titre of recombinant BhCell-Xyl was attained after induction (4.8 U mL(-1)) as compared to that of the constitutive production (2.1 U mL(-1)). The recombinant P. pastoris strains integrated two copies of BhCell-Xyl under AOX and GAP promoters...
December 30, 2016: Bioprocess and Biosystems Engineering
https://www.readbyqxmd.com/read/28039426/yeast-an-overlooked-component-of-bactrocera-tryoni-diptera-tephritidae-larval-gut-microbiota
#11
Ania T Deutscher, Olivia L Reynolds, Toni A Chapman
Yeasts, often in hydrolyzed form, are key ingredients in the larval and adult diets of tephritid fruit fly colonies. However, very little is known about the presence or role of yeasts in the diets of tephritid fruit flies in nature. Previous studies have identified bacteria but not detected yeasts in the gut of Queensland fruit fly, Bactrocera tryoni (Froggatt), one of Australia's most economically damaging insect pests of horticultural crops and of significant biosecurity concern domestically and internationally...
December 29, 2016: Journal of Economic Entomology
https://www.readbyqxmd.com/read/28036287/n-glycosylation-converts-non-glycoproteins-into-mannose-receptor-ligands-and-reveals-antigen-specific-t-cell-responses-in-vivo
#12
Christoph Kreer, Janina M Kuepper, Matthias Zehner, Thomas Quast, Waldemar Kolanus, Beatrix Schumak, Sven Burgdorf
N-glycosylation is generally accepted to enhance the immunogenicity of antigens because of two main reasons. First, the attachment of glycans enables recognition by endocytic receptors like the mannose receptor (MR) and hence increased uptake by dendritic cells (DCs). Second, foreign glycans are postulated to be immunostimulatory and their recognition could induce DC activation. However, a direct comparison between the immunogenicity of N-glycosylated vs. de-glycosylated proteins in vivo and a direct effect of N-glycosylated antigens on the intrinsic capacity of DCs to activate T cells have not been assessed so far...
December 28, 2016: Oncotarget
https://www.readbyqxmd.com/read/28035480/a-comparative-study-of-glycerol-and-sorbitol-as-co-substrates-in-methanol-induced-cultures-of-pichia-pastoris-temperature-effect-and-scale-up-simulation
#13
Julio Berrios, María-Olga Flores, Alvaro Díaz-Barrera, Claudia Altamirano, Irene Martínez, Zaida Cabrera
The production of recombinant proteins by Pichia pastoris under AOX1 promoter is usually performed using methanol together with either glycerol or sorbitol as co-substrate. Although both co-substrates have been widely used, comparative studies are scarce. In addition, these comparisons have been performed at different specific growth rate (µ) that it is well known that has an important effect on productivity. Thus, the effect of using these co-substrates on the production of Rhyzopus oryzae lipase (ROL) by P...
December 29, 2016: Journal of Industrial Microbiology & Biotechnology
https://www.readbyqxmd.com/read/28032192/biotechnological-exploitation-of-tetrapisispora-phaffii-killer-toxin-heterologous-production-in-komagataella-phaffii-pichia-pastoris
#14
Rossella Chessa, Sara Landolfo, Maurizio Ciani, Marilena Budroni, Severino Zara, Murat Ustun, Zeynep Petek Cakar, Ilaria Mannazzu
The use of natural antimicrobials from plants, animals and microorganisms to inhibit the growth of pathogenic and spoilage microorganisms is becoming more frequent. This parallels the increased consumer interest towards consumption of minimally processed food and 'greener' food and beverage additives. Among the natural antimicrobials of microbial origin, the killer toxin produced by the yeast Tetrapisispora phaffii, known as Kpkt, appears to be a promising natural antimicrobial agent. Kpkt is a glycoprotein with β-1,3-glucanase and killer activity, which induces ultrastructural modifications to the cell wall of yeast of the genera Kloeckera/Hanseniaspora and Zygosaccharomyces...
December 28, 2016: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28013188/a-capture-method-based-on-the-vc1-domain-reveals-new-binding-properties-of-the-human-receptor-for-advanced-glycation-end-products-rage
#15
Genny Degani, Alessandra A Altomare, Mara Colzani, Caterina Martino, Angelica Mazzolari, Guenter Fritz, Giulio Vistoli, Laura Popolo, Giancarlo Aldini
The Advanced Glycation and Lipoxidation End products (AGEs and ALEs) are a heterogeneous class of compounds derived from the non-enzymatic glycation or protein adduction by lipoxidation break-down products. The receptor for AGEs (RAGE) is involved in the progression of chronic diseases based on persistent inflammatory state and oxidative stress. RAGE is a pattern recognition receptor (PRR) and the inhibition of the interaction with its ligands or of the ligand accumulation have a potential therapeutic effect...
December 18, 2016: Redox Biology
https://www.readbyqxmd.com/read/28012703/physicochemical-characterization-and-immunological-properties-of-pichia-pastoris-based-hpv16l1-and-18l1-virus-like-particles
#16
Gaurav Gupta, Reinhard Glueck, Narayan Rishi
There continues to be an urgent need for cost-effective prophylaxis for HPV-associated cancers in socio-economically underdeveloped nations. Presently HPV vaccines, which are commercially available, are adjuvanted virus-like particles (VLPs) expressed from various recombinant expression systems. They have been characterized by different methods as safe, pure, and potent HPV vaccine antigens. We cloned and expressed L1 proteins of HPV16 & 18 in Pichia pastoris and tested their immunogenicity. We observed that HPVL1 proteins (16L1 and 18L1) are expressed in Pichia pastoris at high levels...
December 21, 2016: Biologicals: Journal of the International Association of Biological Standardization
https://www.readbyqxmd.com/read/28010776/rational-design-for-the-stability-improvement-of-armillariella-tabescens-%C3%AE-mannanase-man47-based-on-n-glycosylation-modification
#17
Weixiong Hu, Xiaoyun Liu, Yufeng Li, Daling Liu, Zhihe Kuang, Chuiwen Qian, Dongsheng Yao
β-Mannanase has been widely used in industries such as food and feed processing and thus has been a target enzyme for biotechnological development. In this study, we sought to improve the stability and protease resistance of a recombinant β-mannanase, MAN47 from Armillariella tabescens, through rationally designed N-glycosylation. Based on homology modeling, molecular docking, secondary structure analysis and glycosylation feasibility analysis, an enhanced aromatic sequon sequence was introduced into specific MAN47 loop regions to facilitate N-glycosylation...
February 2017: Enzyme and Microbial Technology
https://www.readbyqxmd.com/read/28010766/heterologous-expression-in-pichia-pastoris-and-characterization-of-a-%C3%AE-glucosidase-from-the-xylophagous-cockroach-panesthia-angustipennis-spadica-displaying-high-specific-activity-for-cellobiose
#18
Yihai Li, Gaku Arakawa, Gaku Tokuda, Hirofumi Watanabe, Manabu Arioka
A β-glucosidase (BG), PaBG1b, from the xylophagous cockroach Panesthia angustipennis spadica was heterologously expressed in the methylotrophic yeast Pichia pastoris, purified, and biochemically characterized. Post-translational modification and N-terminal sequencing analysis demonstrated that the expression product was comprised of two polypeptides with different N-terminal sequences, presumably due to the presence of lysine-arginine (KR) sequence in the putative mature region. Substrate specificity analysis showed that PaBG1b hydrolyzed a broad range of substrates including cellohexaose, with the preference for aryl β-d-fucosyl linkage and laminaribiose...
February 2017: Enzyme and Microbial Technology
https://www.readbyqxmd.com/read/28007217/gene-cloning-heterologous-expression-and-characterization-of-a-coprinopsis-cinerea-endo-%C3%AE-1-3-4-glucanase
#19
Jun Wang, Liqin Kang, Zhonghua Liu, Sheng Yuan
A gene coding endo-β-1,3(4)-glucanase (ENG16A) was cloned from Coprinopsis cinerea and heterologously expressed in Pichia pastoris. ENG16A only acts on substrates containing β-1,3 glycosidic bonds but not on substrates containing only β-1,4- or β-1,6-glycosidic bonds. Interestingly, compared to the activity of this enzyme towards carboxymethyl (CM)-pachyman containing only β-1,3-glycosidic bonds, its activity towards barley β-glucan containing both β-1,3-glycosidic and β-1,4-glycosidic bonds was increased by 64...
January 2017: Fungal Biology
https://www.readbyqxmd.com/read/27998673/production-of-recombinant-human-epidermal-growth-factor-in-pichia-pastoris
#20
Samira Eissazadeh, Hassan Moeini, Maryam Ghandizadeh Dezfouli, Somayyeh Heidary, Rubina Nelofer, Mohd Puad Abdullah
This study was carried out to express human epidermal growth factor (hEGF) in Pichia pastoris GS115. For this aim, the hEGF gene was cloned into the pPIC9K expression vector, and then integrated into P. pastoris by electroporation. ELISA-based assay showed that the amount of hEGF secreted into the medium can be affected by the fermentation conditions especially by culture medium, pH and temperature. The best medium for the optimal hEGF production was BMMY buffered at a pH range of 6.0 and 7.0. The highest amount of hEGF with an average yield of 2...
December 17, 2016: Brazilian Journal of Microbiology: [publication of the Brazilian Society for Microbiology]
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