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Jianhua Yang, Anna Joëlle Ruff, Stefanie Nicole Hamer, Feng Cheng, Ulrich Schwaneberg
Escherichia coli is a common host for recombinant protein production in which production titers are highly dependent on the employed expression system. Promoters are thereby a key element to control gene expression levels. In this study, a novel PLICable promoter toolbox was developed which enables in a single cloning step and after a screening experiment to identify out of ten IPTG-inducible promoters (T7, A3, lpp, tac, pac, Sp6, lac, npr, trc and syn) the most suitable one for high level protein production...
October 18, 2016: Biotechnology Journal
Jie Zhou, Jia Ouyang, Qianqian Xu, Zhaojuan Zheng
The main barriers to cost-effective lactic acid production from lignocellulose are the high cost of enzymes and the ineffective utilization of the xylose within the hydrolysate. In the present study, the thermophilic Bacillus coagulans strain CC17 was used for the simultaneous saccharification and fermentation (SSF) of bagasse sulfite pulp (BSP) to produce l-lactic acid. Unexpectedly, SSF by CC17 required approximately 33.33% less fungal cellulase than did separate hydrolysis and fermentation (SHF). More interestingly, CC17 can co-ferment cellobiose and xylose without any exogenous β-glucosidase in SSF...
October 1, 2016: Bioresource Technology
Carolyn N Haarmeyer, Matthew D Smith, Shishir Chundawat, Deanne Sammond, Timothy A Whitehead
Biological-mediated conversion of pretreated lignocellulosic biomass to biofuels and biochemicals is a promising avenue towards energy sustainability. However, a critical impediment to the commercialization of cellulosic biofuel production is the high cost of cellulase enzymes needed to deconstruct biomass into fermentable sugars. One major factor driving cost is cellulase adsorption and inactivation in the presence of lignin, yet we currently have a poor understanding of the protein structure-function relationships driving this adsorption...
October 17, 2016: Biotechnology and Bioengineering
Suvi Arola, Markus B Linder
The interaction between cellulase enzymes and their substrates is of central importance to several technological and scientific challenges. Here we report that the binding of cellulose binding modules (CBM) from Trichoderma reesei cellulases Cel6A and Cel7A show a major difference in how they interact with substrates originating from wood compared to bacterial cellulose. We found that the CBM from TrCel7A recognizes the two substrates differently and as a consequence shows an unexpected way of binding. We show that the substrate has a large impact on the exchange rate of the studied CBM, and moreover, CBM-TrCel7A seems to have an additional mode of binding on wood derived cellulose but not on cellulose originating from bacterial source...
October 17, 2016: Scientific Reports
Toshio Mori, Hiroko Kako, Tomoki Sumiya, Hirokazu Kawagishi, Hirofumi Hirai
A lactic acid (LA)-producing strain of the hyper-lignin-degrading fungus Phanerochaete sordida YK-624 with the lactate dehydrogenase-encoding gene from Bifidobacterium longum (Blldh) was constructed. When the endogenous pyruvate decarboxylase gene-knocked down and Blldh-expressing transformant was cultured with beech wood meal, the transformant was able to successively delignify and ferment the substrate. Supplementation of calcium carbonate into the culture medium, significantly increased the level of LA accumulation...
October 13, 2016: Journal of Biotechnology
Neha Basotra, Baljit Kaur, Marcos Di Falco, Adrian Tsang, Bhupinder Singh Chadha
Mycothermus thermophilus (Syn. Scytalidium thermophilum/Humicola insolens), a thermophilic fungus, is being reported to produce appreciable titers of cellulases and hemicellulases during shake flask culturing on cellulose/wheat-bran/rice straw based production medium. The sequential and differential expression profile of endoglucanases, β-glucosidases, cellobiohydrolases and xylanases using zymography was studied. Mass spectrometry analysis of secretome (Q-TOF LC/MS) revealed a total of 240 proteins with 92 CAZymes of which 62 glycosyl hydrolases belonging to 30 different families were present...
October 8, 2016: Bioresource Technology
Waroon Khota, Suradej Pholsen, David Higgs, Yimin Cai
Natural lactic acid bacteria (LAB) populations in tropical grasses and their fermentation characteristics on silage prepared with cellulase enzyme and LAB inoculants were studied. A commercial inoculant Lactobacillus plantarum Chikuso 1 (CH), a local selected strain Lactobacillus casei TH14 (TH14), and 2 cellulases, Acremonium cellulase (AC) and Maicelase (MC; Meiji Seika Pharma Co. Ltd., Tokyo, Japan), were used as additives to silage preparation with fresh and wilted (6 h) Guinea grass and Napier grass. Silage was prepared using a laboratory-scale fermentation system...
October 13, 2016: Journal of Dairy Science
Christian Hernández, Anne-Marie Farnet Da Silva, Fabio Ziarelli, Isabelle Perraud-Gaime, Beatriz Gutiérrez-Rivera, José Antonio García-Pérez, Enrique Alarcón
The use of synthetic dyes for laccase induction in vivo has been scarcely explored. We characterized the effect of adding different synthetic dyes to liquid cultures of Pycnoporus sanguineus on laccase production. We found that carminic acid (CA) can induce 722 % and alizarin yellow 317 % more laccase than control does, and they promoted better fungal biomass development in liquid cultures. Aniline blue and crystal violet did not show such positive effect. CA and alizarin yellow were degraded up to 95 % during P...
October 14, 2016: Applied Microbiology and Biotechnology
Sen Wang, Dong Zhao, Xinfeng Bai, Weican Zhang, Xuemei Lu
: Cytophaga hutchinsonii is a gram-negative bacterium which can efficiently degrade crystalline cellulose by a unique mechanism different from free cellulases or cellulosomes strategy. In this study, chu_3220, encoding a hypothetical protein CHU_3220 (205 kDa), was identified by insertional mutation and gene deletion to be the first gene essential for degradation of crystalline region but not amorphous region of cellulose by C. hutchinsonii chu_3220-deletion mutant was defective in degradation of crystalline cellulose and increased the degree of crystallinity of Avicel PH101, but could still degrade amorphous cellulose completely...
October 14, 2016: Applied and Environmental Microbiology
Katrin Niegelhell, Michael Suessenbacher, Katrin Jammernegg, Thomas Ganner, Daniel Schwendenwein, Helmut Schwab, Franz Stelzer, Harald Plank, Stefan Spirk
The creation of nano- and micropatterned polymer films is a crucial step for innumerous applications in science and technology. However, there are several problems associated with environmental aspects concerning the polymer synthesis itself, crosslinkers to induce the patterns as well as toxic solvents used for the preparation and even more important development of the films (e.g. chlorobenzene). In this paper, we present a facile method to produce micro- and nanopatterned biopolymer thin films using enzymes as so called biodevelopers...
October 13, 2016: Biomacromolecules
Xiujun Zhang, Yinbo Qu, Yuqi Qin
BACKGROUND: Heterochromatin protein 1 (HP1, homologue HepA in Penicillium oxalicum) binding is associated with a highly compact chromatin state accompanied by gene silencing or repression. HP1 loss leads to the derepression of gene expression. We investigated HepA roles in regulating cellulolytic enzyme gene expression, as an increasingly number of studies have suggested that cellulolytic enzyme gene expression is not only regulated by transcription factors, but is also affected by the chromatin status...
2016: Biotechnology for Biofuels
Xia Zhao, Canhuang Chen, Xingyu Jiang, Wei Shen, Guangtao Huang, Shuai Le, Shuguang Lu, Lingyun Zou, Qingshan Ni, Ming Li, Yan Zhao, Jing Wang, Xiancai Rao, Fuquan Hu, Yinling Tan
The impact of phage infection on the host cell is severe. In order to take over the cellular machinery, some phage proteins were produced to shut off the host biosynthesis early in the phage infection. The discovery and identification of these phage-derived inhibitors have a significant prospect of application in antibacterial treatment. This work presented a phage protein, gp70.1, with non-specific inhibitory effects on Pseudomonas aeruginosa and Escherichia coli. Gp70.1 was encoded by early gene - orf 70...
2016: Frontiers in Microbiology
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
Yonglan Tian, Huayong Zhang, Yang Chai, Lijun Wang, Xueyue Mi, Luyi Zhang, Maxwell Adam Ware
The importance of nickel (added as NiCl2) on mesophilic anaerobic fermentation of Phragmites australis straw and cow dung was demonstrated by investigating the biogas properties, pH values, organic matter degradation [chemical oxygen demand (COD)] and enzyme activities (cellulase, protease and dehydrogenase) during the fermentation process. The results showed that Ni(2+) addition increased the cumulative biogas yields by >18 % by improving the efficiency of first peak stage and bringing forward the second peak stage...
October 7, 2016: Biodegradation
Kai Zhao, Jiwei Duan, Xueling Ma, Yue Zhang, Xiaohua Wang
Termites are destructive to agriculture, forestry and buildings, but they can also promote agro-ecosystem balance through the degradation of lignocellulose. Termite-triggered cellulose digestion may be clarified through microbial metabolism of cellulose products. In the present study, we characterized the activities of cellulase and its three components synthesized by the cellulase-producing fungal strain HDZK-BYTF620 isolated from the gut of Odontotermes formosanus. The protein components of cellulase were synthesized by strain HDZK-BYTF620, which were isolated and characterized using polyacrylamide gel electrophoresis, and the expression of cellulose was studied at the proteome level...
October 6, 2016: Journal of Microbiology and Biotechnology
Jianfeng Zhang, Hongyan Hou, Guang Chen, Shusheng Wang, Jiejing Zhang
The straw can be degraded efficiently into humus by powerful enzymes from microorganisms, resulting in the accelerated circulation of N,P,K and other effective elements in ecological system. We isolated a strain through screening the straw degradation strains from natural humic straw in the low temperature area in northeast of china, which can produce cellulase efficiently. The strain was identified as Pseudomonas mendocina by using morphological, physiological, biochemical test, and molecular biological test, with the functional clarification on producing cellulase for Pseudomonas mendocina for the first time...
September 2, 2016: Bioengineered
Thaila Fernanda Dos Reis, Pollyne Borborema Almeida de Lima, Nádia Skorupa Parachin, Fabiana Bombonato Mingossi, Juliana Velasco de Castro Oliveira, Laure Nicolas Annick Ries, Gustavo Henrique Goldman
BACKGROUND: The conversion of lignocellulosic biomass to biofuels (second-generation biofuel production) is an environmentally friendlier alternative to petroleum-based energy sources. Enzymatic deconstruction of lignocellulose, catalyzed by filamentous fungi such as Aspergillus nidulans, releases a mixture of mono- and polysaccharides, including hexose (glucose) and pentose (xylose) sugars, cellodextrins (cellobiose), and xylooligosaccharides (xylobiose). These sugars can subsequently be fermented by yeast cells to ethanol...
2016: Biotechnology for Biofuels
Yihui Chen, Junzheng Sun, Hetong Lin, Yen-Con Hung, Shen Zhang, Yifen Lin, Ting Lin
BACKGROUND: Huanghua pear will loss its firmness quickly during postharvest storage at ambient temperature, and hence with limited storage and marketing potential. In this study, Huanghua pears treated with paper containing 0 (control) or 0.9 μL L(-1) 1-methylcyclopropene (1-MCP) for 12 h, and then stored at (25 ± 1) °C for 30 days were investigated on the effect of a paper-based 1-MCP treatment on fruit firmness, cell wall composition and activities of cell wall-degrading enzymes...
October 5, 2016: Journal of the Science of Food and Agriculture
A F N Volpini, T Thomazine, S H Umeo, G A Pereira, G A Linde, J S Valle, N B Colauto, F G Barcellos, S G H Souza
Enzymes produced by basidiomycetes that are involved in the cellulose degradation process, and their respective codifying genes, must be identified to facilitate the development of novel biotechnological strategies and applications in the agro-industry. The objective of this study was to identify prospective cellulase-producing genes and characterize their cellulolytic activity, in order to elucidate the potential biotechnological applications (with respect to vegetal residues) of basidiomycetes. The basidiomycete strains Lentinula edodes U8-1, Lentinus crinitus U9-1, and Schizophyllum commune U6-7 were analyzed in this study...
September 16, 2016: Genetics and Molecular Research: GMR
Meiling Hou, Gentu Ge, Tingyu Liu, Yushan Jia, Yimin Cai
Silage preparation and fermentation quality of natural grasses in meadow steppe (MS) and typical steppe (TS) were studied. MS and TS contained thirty-three and nine species of natural grasses, respectively. Stipa Baicalensis in MS and Stipa grandi in TS were the dominant grasses with the highest dry matter (DM) yield. The crude protein (CP), neutral detergent fiber (NDF) and water-soluble carbohydrate (WSC) of the mixed natural grasses in both steppes were 8.02 to 9.03, 66.75 to 69.47 and 2.02 to 2.20% on a DM basis, respectively...
September 27, 2016: Asian-Australasian Journal of Animal Sciences
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