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Lanfang Tian, Siyuan Chen, Haiyan Liu, Mingzhang Guo, Wentao Xu, Xiaoyun He, Yunbo Luo, Xiaozhe Qi, Hongxia Luo, Kunlun Huang
Hepcidin, one kind of antimicrobial peptides, is one of the promising alternatives to antibiotics with broad spectrum of antimicrobial activity. Hepcidins cloned from different kinds of fishes have been produced using exogenous expression systems, and their in vitro antimicrobial effects have been verified. However their in vivo effects on gut microbiota and gut health of hosts remain unclear. Here we performed a safety study of hepcidin so that it can be used to reduce microbial contaminations in the food and feed...
2016: PloS One
Natalia Eliza Domeradzka, Marc W T Werten, Frits A de Wolf, Renko de Vries
Previously, we developed triblock protein polymers that form fibrillar hydrogels at low protein polymer concentrations (denoted C2-SH48-C2). We here demonstrate that the structure of these hydrogels can be tuned via heterodimeric coiled coils that cross-link and bundle the self-assembled protein-polymer fibrils. We fused well-characterized, 47 amino acids-long heterodimeric coiled coil "linkers" (DA or DB) to the C-terminus of the triblock polymer. The resulting C2-SH48-C2-DA and C2-SH48-C2-DB polymers, were successfully produced as secreted proteins in Pichia pastoris, with titers of purified protein in the order of g L-1 of clarified broth...
October 21, 2016: Biomacromolecules
(no author information available yet)
[This corrects the article DOI: 10.1371/journal.pone.0161529.].
2016: PloS One
Su Jin Kang, Ji Yeon Seo, Kye Man Cho, Chang Kwon Lee, Jeong Hwan Kim, Jong-Sang Kim
A new type of doenjang was manufactured by mixing soaked soybean, koji (Rhizopus oryzae), cheonggukjang (Bacillus amyloliquefaciens MJ1-4 and B. amyloliquefaciens EMD17), and Pichia farinosa SY80 as a yeast, salt, and water, followed by fermentation with koji that was made by fermenting whole wheat with R. oryzae. The mixed culture doenjang was designed to have a more palatable flavor and stronger biological activities than the conventional product. The extract of mixed culture doenjang showed higher antioxidant activity than the commercial doenjang as evaluated by the ferric reducing antioxidant power assay although it was not significantly different from the commercial product in 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) radical scavenging activities...
September 2016: Preventive Nutrition and Food Science
Raida Jallouli, Madiha Bouali, Youssef Gargouri, Sofiane Bezzine
Two lipases from Fusarium solani, FSL and FSL2, were efficiently expressed in Pichia pastoris. To check the influence of the expression on interfacial properties of FSL and to study kinetic properties of FSL2, interfacial parameters of FSL2, native FSL, untagged recombinant and tagged recombinant forms of FSL were compared using the monomolecular film technique. Kinetic study on the dependence of the stereoselectivity of these lipases on the surface pressure was performed using three dicaprin isomers spread in the form of monomolecular films at the air-water interface...
October 13, 2016: International Journal of Biological Macromolecules
De-Mei Meng, Jing-Fang Zhao, Xiao Ling, Hong-Xia Dai, Ya-Jun Guo, Xiao-Fang Gao, Bin Dong, Zi-Qi Zhang, Xin Meng, Zhen-Chuan Fan
The antimicrobial peptide PaDef was isolated from Mexican avocado fruit and was reported to inhibit the growth of Escherichia coli and Staphylococcus aureus in 2013. In this study, an N-terminal 6 × His tagged recombinant PaDef (rPaDef) with a molecular weight of 7.5 KDa, for the first time, was expressed as a secreted peptide in Pichia pastoris. The optimal culture condition for rPaDef expression was determined to be incubation with 1.5% methanol for 72 h at 28 °C under pH 6.0. Under this condition, the amount of the rPaDef accumulation reached as high as 79...
October 11, 2016: Protein Expression and Purification
Liqiang Zhang, Rongqing Zhou, Ruiying Cui, Jun Huang, Chongde Wu
Present study was to characterize the physiochemical properties, free amino acids (FAAs), volatiles and microbial communities of various moromi, respectively sampled from different stages of high-salt dilute-state (HSDS) and low-salt solid-state (LSSS) fermentation, using multiphase analyzing methods. Phospholipid fatty acids (PLFA) analysis indicated that Gram-positive bacteria were dominant bacteria and fungi were principal microbes. For DGGE analysis, dominant microbes in moromi were mainly fell into Weissella, Tetragenococcus, Candida, Pichia, and Zygosaccharomyces...
October 14, 2016: Journal of Food Science
Eun Jae Kim, Jun Hyuck Lee, Sung Gu Lee, Se Jong Han
To survive in a sub-zero environment, polar organisms produce ice-binding proteins (IBPs). These IBPs prevent the formation of large intracellular ice crystals, which may be fatal to the organism. Recently, a recombinant FfIBP (an IBP from Flavobacterium frigoris PS1) was cloned and produced in Pichia pastoris using fed-batch fermentation with methanol feeding. In this study, we demonstrate that FfIBP produced by P. pastoris has a glycosylation site, which diminishes the thermal hysteresis (TH) activity of FfIBP...
October 13, 2016: Preparative Biochemistry & Biotechnology
Xiangfeng Zheng, Qiya Yang, Hongyin Zhang, Jing Cao, Xiaoyun Zhang, Maurice Tibiru Apaliya
In this work, we examined the mechanisms involved in the degradation of patulin by Pichia caribbica. Our results indicate that cell-free filtrate of P. caribbica reduced patutlin content. The heat-killed cells could not degrade patulin. However, the live cells significantly reduced the concentration of the patulin. In furtherance to this, it was observed that patulin was not detected in the broken yeast cells and cell wall. The addition of cycloheximide to the P. caribbica cells decreased the capacity of degradation of patulin...
October 9, 2016: Toxins
Tingjing Zhang, Jianqiang Liang, Panxue Wang, Ying Xu, Yutang Wang, Xinyuan Wei, Mingtao Fan
Phloretin-2'-O-glycosyltransferase (P2'GT) catalyzes the last glycosylation step in the biosynthesis of phloridzin that contributes to the flavor, color and health benefits of apples and processed apple products. In this work, a novel P2'GT of Malus x domestica (MdP2'GT) with a specific activity of 46.82 μkat/Kg protein toward phloretin and uridine diphosphate glucose (UDPG) at an optimal temperature of 30 °C and pH 8.0 was purified from the engineered Pichia pastoris broth to homogeneity by anion exchange chromatography, His-Trap affinity chromatography and gel filtration...
October 12, 2016: Scientific Reports
Xiaoyu Wang, Huoqing Huang, Xiangming Xie, Rui Ma, Yingguo Bai, Fei Zheng, Shuai You, Bingyu Zhang, Huifang Xie, Bin Yao, Huiying Luo
A xylanase gene of GH 10, Tlxyn10A, was cloned from Talaromyces leycettanus JCM12802 and expressed in Pichia pastoris. Purified recombinant TlXyn10A was acidic and hyperthermophilic, and retained stable over the pH range of 2.0-6.0 and at 90°C. Sequence analysis of TlXyn10A identified seven residues probably involved in substrate contacting. Three mutants (TlXyn10A_P, _N and _C) were then constructed by substituting some or all of the residues with corresponding ones of hyperthermal Xyl10C from Bispora sp...
October 3, 2016: Bioresource Technology
Joanna Berłowska, Katarzyna Pielech-Przybylska, Maria Balcerek, Urszula Dziekońska-Kubczak, Piotr Patelski, Piotr Dziugan, Dorota Kręgiel
Sugar beet pulp, a byproduct of sugar beet processing, can be used as a feedstock in second-generation ethanol production. The objective of this study was to investigate the effects of pretreatment, of the dosage of cellulase and hemicellulase enzyme preparations used, and of aeration on the release of fermentable sugars and ethanol yield during simultaneous saccharification and fermentation (SSF) of sugar beet pulp-based worts. Pressure-thermal pretreatment was applied to sugar beet pulp suspended in 2% w/w sulphuric acid solution at a ratio providing 12% dry matter...
2016: BioMed Research International
Manuel Nieto-Domínguez, Alicia Prieto, Beatriz Fernández de Toro, Francisco Javier Cañada, Jorge Barriuso, Zach Armstrong, Stephen G Withers, Laura I de Eugenio, María Jesús Martínez
BACKGROUND: Glycosides are compounds displaying crucial biological roles and plenty of applications. Traditionally, these molecules have been chemically obtained, but its efficient production is limited by the lack of regio- and stereo-selectivity of the chemical synthesis. As an interesting alternative, glycosidases are able to catalyze the formation of glycosides in a process considered green and highly selective. In this study, we report the expression and characterization of a fungal β-xylosidase in Pichia pastoris...
October 4, 2016: Microbial Cell Factories
Fabián Rueda, Brigitte Gasser, Alejandro Sánchez-Chardi, Mònica Roldán, Sandra Villegas, Verena Puxbaum, Neus Ferrer-Miralles, Ugutz Unzueta, Esther Vázquez, Elena Garcia-Fruitós, Diethard Mattanovich, Antonio Villaverde
BACKGROUND: 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...
October 1, 2016: Microbial Cell Factories
Cun-Duo Tang, Hong-Ling Shi, Qing-Hai Tang, Jun-Shi Zhou, Lun-Guang Yao, Zhu-Jin Jiao, Yun-Chao Kan
Two novel glycosyl hydrolase family 5 (GH5) β-mannanases (AoMan5A and AoMan5B) were identified from Aspergillus oryzae RIB40 by genome mining. The AoMan5A contains a predicted family 1 carbohydrate binding module (CBM-1), located at its N-terminal. The AoMan5A, AoMan5B and truncated mutant AoMan5AΔCL (truncating the N-terminal CBM and linker of AoMan5A) were expressed retaining the N-terminus of the native protein in Pichia pastoris GS115 by pPIC9K(M). The specific enzyme activity of the purified reAoMan5A, reAoMan5B and reAoMan5AΔCL towards locust bean gum at pH 3...
November 2016: Enzyme and Microbial Technology
Kentaro Inokuma, Tomohisa Hasunuma, Akihiko Kondo
N-acetyl-D-glucosamine (GlcNAc) is the building block of chitin, which is one of the most abundant renewable resources in nature after cellulose. Therefore, a microorganism that can utilize GlcNAc is necessary for chitin-based biorefinery. In this study, we report on the screening and characterization of yeast strains for bioethanol production from GlcNAc. We demonstrate that Scheffersomyces (Pichia) stipitis strains can use GlcNAc as the sole carbon source and produce ethanol. S. stipitis NBRC1687, 10007, and 10063 strains consumed most of the 50 g/L GlcNAc provided, and produced 14...
December 2016: AMB Express
Li-Wei Xing, Shi-Xun Tian, Wei Gao, Na Yang, Pei Qu, Di Liu, Jian Jiao, Jue Wang, Xing-Jun Feng
Fowlicidins are a group of cathelicidin antimicrobial peptides that were initially identified in chickens. Fowlicidin-2, which is composed of 31 amino acids, is widely expressed in the majority of tissues in chickens and has an important role in innate immunity. In the present study, a recombinant expression system for fowlicidin-2 was successfully constructed using Pichia pastoris X-33 and the expression vector pPICZα-A. Under the optimized fermentation conditions, 85.6 mg fowlicidin-2 with >95% purity was obtained from 1 liter culture medium following purification by ion exchange chromatography and reversed phase high performance liquid chromatography...
October 2016: Experimental and Therapeutic Medicine
Anay Kumar Maurya, Deepak Parashar, T Satyanarayana
Thermophilc mold Sporotrichum thermophile secretes an acidstable and thermostable phytase, which finds application as a food and feed additive because of its adequate thermostability, acid stability, protease insensitivity and broad substrate spectrum. Low extracellular phytase production by the mold is a major bottleneck for its application on a commercial scale. We have successfully overcome this problem by constitutive secretary expression of codon optimized rStPhy under glyceraldehyde phosphate dehydrogenase (GAP) promoter in Pichia pastoris...
September 30, 2016: International Journal of Biological Macromolecules
Takashi Kikuma, Takayuki Tadokoro, Jun-Ichi Maruyama, Katsuhiko Kitamoto
Autophagy is a conserved process in eukaryotic cells for degradation of cellular proteins and organelles. In filamentous fungi, autophagic degradation of organelles such as peroxisomes, mitochondria, and nuclei occurs in basal cells after the prolonged culture, but its mechanism is not well understood. Here, we functionally analyzed the filamentous fungus Aspergillus oryzae AoAtg26, an ortholog of the sterol glucosyltransferase PpAtg26 involved in pexophagy in the yeast Pichia pastoris. Deletion of Aoatg26 caused a severe decrease in conidiation and aerial hyphae formation, which is typically observed in the autophagy-deficient A...
October 4, 2016: Bioscience, Biotechnology, and Biochemistry
Bart W Faber, Stephan Hellwig, Sophie Houard, Nicolas Havelange, Jürgen Drossard, Hubert Mertens, Alexander Croon, Robin Kastilan, Richard Byrne, Nicole van der Werff, Marjolein van der Eijk, Alan W Thomas, Clemens H M Kocken, Edmond J Remarque
Plasmodium falciparum apical membrane antigen 1 (PfAMA1) is a leading asexual blood stage vaccine candidate for malaria. In preparation for clinical trials, three Diversity Covering (DiCo) PfAMA1 ectodomain proteins, designed to overcome the intrinsic polymorphism that is present in PfAMA1, were produced under Good Manufacturing Practice (GMP) in Pichia pastoris. Using identical methodology, the 3 strains were cultivated in 70-L scale fed-batch fermentations and PfAMA1-DiCos were purified by two chromatography steps, an ultrafiltration/diafiltration procedure and size exclusion chromatography, resulting in highly pure (>95%) PfAMA1-DiCo1, PfAMA1 DiCo2 and PfAMA1 DiCo3, with final yields of 1...
2016: PloS One
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