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https://www.readbyqxmd.com/read/29660471/efficient-production-of-human-goose-type-lysozyme-2-in-the-methylotrophic-yeast-pichia-pastoris
#1
Peng Huang, Qingwen Sun, Weijun Shi, Wangchun Du, Xue Li, Ning Zhang
Infectious diseases caused by antibiotic multidrug-resistant microorganisms are major causes of morbidity and mortality in humans. Hence, there is an urgent need to search for new antimicrobial agents. Initially known as a defensive effector in the innate immunity of certain organs of the human body, human goose-type lysozyme 2 (hLysG2) has been shown to possess therapeutically useful potential against multidrug-resistant microorganisms. Developing a novel strategy for large-scale production that provides high yields of this protein with high purity, quality, and potency is critical for pharmaceutical applications...
April 13, 2018: Journal of Biotechnology
https://www.readbyqxmd.com/read/29651700/engineering-pichia-pastoris-for-efficient-production-of-a-novel-bifunctional-strongylocentrotus-purpuratus-invertebrate-type-lysozyme
#2
Peng Huang, Jinlei Shi, Qingwen Sun, Xianping Dong, Ning Zhang
Lysozymes are known as ubiquitously distributed immune effectors with hydrolytic activity against peptidoglycan, the major bacterial cell wall polymer, to trigger cell lysis. In the present study, the full-length cDNA sequence of a novel sea urchin Strongylocentrotus purpuratus invertebrate-type lysozyme (sp-iLys) was synthesized according to the codon usage bias of Pichia pastoris and was cloned into a constitutive expression plasmid pPIC9K. The resulting plasmid, pPIC9K-sp-iLys, was integrated into the genome of P...
April 13, 2018: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/29648454/effect-of-n-linked-glycosylation-of-recombinant-windmill-palm-tree-peroxidase-on-its-activity-and-stability
#3
Bo Fu, Margaret R Baker, Qing X Li
Plant secretory peroxidases are valuable commercial enzymes. The windmill palm tree Trachycarpus fortunei produces one of the most stable and fastest peroxidases (WPTP) characterized to date, however, an economical source is needed. Pichia pastoris has been used as an expression system for WPTP and other peroxidases. However, yeast and plants synthesize different types of N-linked glycan structures and may differ the level of glycosylation at each site. Such non-native glycosylation can have unwanted consequences...
April 12, 2018: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/29642729/safety-evaluation-of-soy-leghemoglobin-protein-preparation-derived-from-pichia-pastoris-intended-for-use-as-a-flavor-catalyst-in-plant-based-meat
#4
Rachel Z Fraser, Mithila Shitut, Puja Agrawal, Odete Mendes, Sue Klapholz
The leghemoglobin protein (LegH) from soy ( Glycine max) expressed in Pichia pastoris (LegH preparation, LegH Prep) imparts a meat-like flavor profile onto plant-based food products. The safety of LegH Prep was evaluated through a series of in vitro and in vivo tests. The genotoxic potential of LegH Prep was assessed using the bacterial reverse mutation assay (Ames test) and the in vitro chromosome aberration test. LegH Prep was nonmutagenic and nonclastogenic in each test, respectively. Systemic toxicity was assessed in a 28-day dietary study in male and female Sprague Dawley rats...
January 1, 2018: International Journal of Toxicology
https://www.readbyqxmd.com/read/29627478/a-novel-fucolectin-from-apostichopus-japonicus-with-broad-pamp-recognition-pattern
#5
Ying Wang, Zhuang Xue, Qilin Yi, Hui Wang, Lingling Wang, Guangxia Lu, Yu Liu, Chen Qu, Yannan Li, Linsheng Song
F-type lectin (also known as fucolectin) is a newly identified family of fucose binding lectins with the sequence characters of a fucose binding motif and a unique lectin fold (the "F-type" fold). In the present study, a fucolectin was identified from sea cucumber Apostichopus japonicus (designated AjFL-1). The open reading frame (ORF) of AjFL-1 was of 546 bp, encoding a polypeptide of 181 amino acids with a predicted molecular mass of about 20 kDa. The deduced amino acid sequence of AjFL-1 shared 30%-40% similarity with the fucolectins from other animals...
April 5, 2018: Fish & Shellfish Immunology
https://www.readbyqxmd.com/read/29626731/microbial-composition-of-spoiled-industrial-scale-sichuan-paocai-and-characteristics-of-the-microorganisms-responsible-for-paocai-spoilage
#6
Feng Zhang, Yao Tang, Yao Ren, Kai Yao, Qiang He, Yuping Wan, Yuanlong Chi
The microorganisms of spoiled industrial-scale Sichuan paocai (ISSP) were isolated using six types of media, and 16S rRNA and 26S rRNA gene sequence analyses were used to identify the isolates. Meanwhile, the microbial composition was investigated using a culture-independent method through 16S rRNA and ITS sequencing on an Illumina MiSeq platform. The results obtained by these two methods were compared. Furthermore, characteristics of the isolated microorganisms responsible for ISSP spoilage were studied. Sixty-two strains belonging to twenty-three species, including three ammonia-producing genera, two gas-producing genera, two pectinase-producing genera, two cellulase-producing genera, three film-producing genera and one slime-producing genus, were isolated...
April 3, 2018: International Journal of Food Microbiology
https://www.readbyqxmd.com/read/29625371/purification-of-human-erythropoietin-by-affinity-chromatography-using-cyclic-peptide-ligands
#7
William S Kish, Matthew K Roach, Hiroyuki Sachi, Amith D Naik, Stefano Menegatti, Ruben G Carbonell
Prior work described the identification and characterization of erythropoietin-binding cyclic peptides SLFFLH, VVFFVH, FSLLHH and FSLLSH (all of the form cyclo[(Nα -Ac)Dap(A)-X1 -X6 -AE], wherein X1 -X6 is the listed sequences). In this work, the peptide ligands were synthesized on Toyopearl chromatographic resins and utilized for purifying recombinant human erythropoietin (rHuEPO) from complex sources. Elution buffer pH and composition were optimized to maximize the recovery of standard rHuEPO from the peptide resins...
March 27, 2018: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
https://www.readbyqxmd.com/read/29611040/cloning-and-expression-of-a-novel-%C3%AE-1-3-arabinofuranosidase-from-penicillium-oxalicum-sp-68
#8
Yanbo Hu, Xuecui Yan, Han Zhang, Jiaqi Liu, Feng Luo, Yingying Cui, Weiyang Wang, Yifa Zhou
The discovery and creation of biocatalysts for plant biomass conversion are essential for industrial demand and scientific research of the plant cell wall. α-1,2 and α-1,3-L-arabinofuranosidases are debranching enzymes that catalyzing hydrolytic release of α-L-arabinofuranosyl residues in plant cell wall. Gene database analyses shows that GH62 family only contains specific α-L-arabinofuranosidases that play an important role in the degradation and structure of the plant cell wall. At present, there are only 22 enzymes in this group has been characterized...
April 2, 2018: AMB Express
https://www.readbyqxmd.com/read/29609030/a-prokineticin-pk-like-cytokine-from-chinese-mitten-crab-eriocheir-sinensis-promotes-the-production-of-hemocytes-via-reactive-oxygen-species
#9
Zhihao Jia, Mengqiang Wang, Xiudan Wang, Jiachao Xu, Lingling Wang, Huan Zhang, Linsheng Song
Astakine is a cytokine-like factor containing a prokineticin domain, which directly participates in hematopoiesis and blood cell differentiation. In the present study, a novel Astakine gene was identified from Chinese mitten crab Eriocheir sinensis (designated as EsAst). The full-length cDNA of EsAst was of 1163 bp, consisting of a 5' untranslated region (UTR) of 120 bp, a 3' UTR of 656 bp, and an open reading frame (ORF) of 387 bp encoding a polypeptide of 128 amino acids. There were a signal peptide and a prokineticin domain with nine conserved cysteine residues in the deduced amino acid sequence of EsAst...
March 30, 2018: Fish & Shellfish Immunology
https://www.readbyqxmd.com/read/29602635/yeasts-and-filamentous-fungi-associated-with-some-dairy-products-in-egypt
#10
A-A H Moubasher, M A Abdel-Sater, Z S M Soliman
This work aimed to identify and evaluate the diversity of yeasts and filamentous fungi that contaminate some dairy products (fresh buffalo milk, plain yoghurt, butter and Kareisk cheese) in Assiut city, Egypt. The identification was based on phenotypic characteristics, and genotypically in case of yeasts. The pHs of all dairy products lie within the acidic range, but yoghurt registered the highest value. A total of 41 genera and 89 species+3 varieties were recovered from all dairy products investigated on DRBC (26 genera representing 59 species+1 variety), DG18 (32 genera, 56 species+2 varieties) and MY50G (31 genera, 64 species+2 varieties)...
March 27, 2018: Journal de Mycologie Médicale
https://www.readbyqxmd.com/read/29602390/the-effect-of-a-lytic-polysaccharide-monooxygenase-and-a-xylanase-from-gloeophyllum-trabeum-on-the-enzymatic-hydrolysis-of-lignocellulosic-residues-using-a-commercial-cellulase
#11
Cynthia Sanhueza, Gonzalo Carvajal, Javiera Soto-Aguilar, Maria Elena Lienqueo, Oriana Salazar
Hydrolysis of lignocellulosic biomass depends on the concerted actions of cellulases and accessory proteins. In this work we examined the combined action of two auxiliary proteins from the brown rot fungus Gloeophyllum trabeum: a family AA9 lytic polysaccharide monooxygenase (GtLPMO) and a GH10 xylanase (GtXyn10A). The enzymes were produced in the heterologous host Pichia pastoris. In the presence of an electron source, GtLPMO increased the activity of a commercial cellulase on filter paper, and the xylanase activity of GtXyn10A on beechwood xylan...
June 2018: Enzyme and Microbial Technology
https://www.readbyqxmd.com/read/29601964/a-systematic-analysis-of-vrc01-antibody-expression-in-pichia-pastoris-through-signal-peptide-optimization
#12
Rochelle Aw, Paul F McKay, Robin J Shattock, Karen M Polizzi
Pichia (syn. Komagataella) pastoris has been used for recombinant protein production for over 30 years with over 5000 proteins reported to date. However, yields of antibody are generally low. We have evaluated the effect of secretion signal peptides on the production of a broadly neutralizing antibody (VRC01) to increase yield. Eleven different signal peptides, including the murine IgG1 signal peptide, were combinatorially evaluated for their effect on antibody titer. Strains using different combinations of signal peptides were identified that secreted approximately 2-7 fold higher levels of VRC01 than the previous best secretor, with the highest yield of 6...
March 27, 2018: Protein Expression and Purification
https://www.readbyqxmd.com/read/29596989/high-level-expression-purification-and-enzymatic-characterization-of-a-recombinant-aspergillus-sojae-alkaline-protease-in-pichia-pastoris
#13
Ye Ke, XiaoMei Yuan, JiaSheng Li, Wei Zhou, XiaoHui Huang, Tao Wang
An alkaline protease (Ap) was cloned from Aspergillus sojae GIM3.33 via RT-PCR technique. A truncated Ap without the signal peptide was successfully expressed in the Pichia pastoris KM71 strain. The following describes the optimal process conditions for the recombinant engineering of a strain expressing a recombinant Ap (rAp) in a triangular flask: inoculum concentration OD600 value 20.0 in 40 mL working volume (in 500 mL flasks), methanol addition (1.0%; volume ratio), 0.02% biotin solution (60 μL), and YNB primary concentration (13...
March 26, 2018: Protein Expression and Purification
https://www.readbyqxmd.com/read/29595107/gene-and-process-level-modulation-to-overcome-the-bottlenecks-of-recombinant-proteins-expression-in-pichia-pastoris
#14
Ashish A Prabhu, Bibari Boro, Biju Bharali, Shuchishloka Chakraborty, V Venkata Dasu
BACKGROUND: Process development involving system metabolic engineering and bioprocess engineering has become one of the major thrust for the development of therapeutic proteins or enzymes. Pichia pastoris has emerged as a prominent host for the production of therapeutic protein or enzymes. Despite of producing high protein titers, various cellular and process level bottlenecks hinders the expression of recombinant proteins in P. pastoris. RESULT AND CONCLUSION: In the present review, we have summarized the recent developments in the expression of foreign proteins in P...
March 28, 2018: Current Pharmaceutical Biotechnology
https://www.readbyqxmd.com/read/29588473/spatial-temporal-profiling-of-prodiginines-and-serratamolides-produced-by-endophytic-serratia-marcescens-harbored-in-maytenus-serrata
#15
Dennis Eckelmann, Michael Spiteller, Souvik Kusari
An endophytic bacterium, Serratia marcescens MSRBB2, isolated from inner bark of a Cameroonian Maytenus serrata plant, was subjected to the OSMAC (One Strain Many Compounds) approach and metabolic profiling using HPLC-HRMSn . We identified 7 prodiginines along with 26 serratamolides. Their biosynthetic pathways were elucidated by feeding with labeled precursors in combination with HRMSn . Dual-culture confrontation/restriction assays of the bacterial endophyte were devised with coexisting fungal endophytes (Pestalotiopsis virgatula, Aspergillus caesiellus and Pichia spp...
March 27, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29577692/-cloning-of-oryza-sativa-n-glycanase-gene-ospngase-a-and-its-expression-in-pichia-pastoris
#16
Yuan Wang, Peng Jia, Xuejun Li, Yuhong Li, Peng Chen
N-glycanase is a class of deglycosylation enzymes, widely used in the study of N-glycosylation modification of glycoprotein. In this study, an N-glycanase gene (OsPNGase A, XM_015775832) with high GC content (69.48%) was cloned from rice and then the yeast secretory expression vector pPICZ(α)A-OsPNGase A was constructed for the purpose of transformation to Pichia pastoris. After induction in Pichia pastoris SMD1168H, the target protein was purified by DEAE Sepharose and HisTrap HP chromatography, with a yield of 12...
March 25, 2018: Sheng Wu Gong Cheng Xue Bao, Chinese Journal of Biotechnology
https://www.readbyqxmd.com/read/29577311/analysis-of-the-cocobiota-and-metabolites-of-moniliophthora-perniciosa-resistant-theobroma-cacao-beans-during-spontaneous-fermentation-in-southern-brazil
#17
Valdeci S Bastos, Maria F S Santos, Laidson P Gomes, Analy M O Leite, Vânia M Flosi Paschoalin, Eduardo M Del Aguila
BACKGROUND: Cocoa bean fermentation is a spontaneous process involving a succession of microbial activities; yeasts, lactic acid and acetic acid bacteria. The spontaneous fermentation of cocoa beans by Theobroma cacao TSH565 clonal variety, a highly productive hybrid resistant to Moniliophthora perniciosa and Phytophthora spp. was investigated. The natural cocobiota involved in the spontaneous fermentation of this hybrid, in Southern Brazil, was investigated by using both a culture-dependent microbiological and a molecular analysis...
March 25, 2018: Journal of the Science of Food and Agriculture
https://www.readbyqxmd.com/read/29572665/improved-fermentation-performance-to-produce-bioethanol-from-gelidium-amansii-using-pichia-stipitis-adapted-to-galactose
#18
Pailin Sukwong, Chae Hun Ra, In Yung Sunwoo, Sumate Tantratian, Gwi-Taek Jeong, Sung-Koo Kim
This study employed a statistical method to obtain optimal hyper thermal acid hydrolysis conditions using Gelidium amansii (red seaweed) as a source of biomass. The optimal hyper thermal acid hydrolysis using G. amansii as biomass was determined as 12% (w/v) slurry content, 358.3 mM H2 SO4 , and temperature of 142.6 °C for 11 min. After hyper thermal acid hydrolysis, enzymatic saccharification was carried out. The total monosaccharide concentration was 45.1 g/L, 72.2% of the theoretical value of the total fermentable monosaccharides of 62...
March 23, 2018: Bioprocess and Biosystems Engineering
https://www.readbyqxmd.com/read/29572144/locust-bean-gum-galactomannan-hydrolyzed-by-thermostable-%C3%AE-d-mannanase-may-reduce-the-secretion-of-pro-inflammatory-factors-and-the-release-of-granule-constituents
#19
Wei-Lin Chen, Huan-Ling Chen, Gui-Wen Guo, Yu-Chun Huang, Cheng-Yu Chen, Yu Tsai, Keh-Feng Huang, Chao-Hsun Yang
Locust bean gum (LBG) galactomannan has been claimed to have applications in the biopharmaceutical field. However, the effects of LBG galactomannan on immunomodulatory aspects are not yet clear. The purpose of this study was to over-express thermostable β-d-mannanase from the thermophilic actinomycete Thermobifida fusca BCRC 19214 using a Pichia pastoris expression system. The maximum intracellular β-d-mannanase activity obtained from the cell-free extract was approximately 40.0 U/mL after 72 h of cultivating a P...
March 20, 2018: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/29570735/microbial-diversity-in-two-traditional-bacterial-douchi-from-gansu-province-in-northwest-china-using-illumina-sequencing
#20
Weibing Zhang, Qiaoqiao Luo, Yan Zhu, Jiang Ma, Lei Cao, Min Yang, Pencheng Wen, Zhongmin Zhang, Xiaoling He
Douchi has been consumed as a flavoring ingredient for centuries. During production of douchi, numerous microorganisms play important roles in the hydrolysis and conversion of proteins and starch, which are related to the quality and flavor of the end product. Therefore, in the present study, the microbial diversity in two types of home-made traditional bacterial douchi from Gansu province in northwest China was studied by high-throughput sequencing, and a corresponding analysis of the bacterial and fungal communities were conducted...
2018: PloS One
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