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Hanjun He, Mei Bai, Panpan Tong, Yanting Hu, Ming Yang, Hong Wu
Cellulases, hemicellulases and pectinases play important roles in fruit development and maturation, but mutants with defects in the fruit have not been reported for cellulase or hemicellulase genes. Here we report the functional characterization of cellulase gene CEL6 and hemicellulase gene MAN7 in silique development and dehiscence in Arabidopsis. These genes were found to be expressed in vegetative and reproductive organs, and their expression in the silique partially depended on the IND and ALC transcriptional factors...
January 18, 2018: Plant Physiology
Veeresh Juturu, Jin Chuan Wu
Pichia pastoris is a well-known platform strain for heterologous protein expression. Over the past five years, different strategies to improve the efficiency of recombinant protein expression by this yeast strain have been developed; these include a patent-free protein expression kit, construction of the P. pastoris CBS7435Ku70 platform strain with its high efficiency in site-specific recombination of plasmid DNA into the genomic DNA, the design of synthetic promoters and their variants by combining different core promoters with multiple putative transcription factors, the generation of mutant GAP promoter variants with various promoter strengths, codon optimization, engineering the α-factor signal sequence by replacing the native glutamic acid at the Kex2 cleavage site with the other 19 natural amino acids and the addition of mammalian signal sequence to the yeast signal sequence, and the co-expression of single chaperones, multiple chaperones or helper proteins that aid in recombinant protein folding...
December 13, 2017: Chembiochem: a European Journal of Chemical Biology
Rafaela I S Ladeira Ázar, Túlio Morgan, Antonio Carlos Freitas Dos Santos, Eduardo de Aquino Ximenes, Michael R Ladisch, Valéria M Guimarães
Cellulase and hemicellulase activities in a 1:1 ratio of enzymes extracted from Chrysoporthe cubensis and Penicillium pinophilum were evaluated in the presence of known monocomponent phenolic inhibitors and also with phenol mixtures derived from alkali pretreated sugarcane bagasse. The cellulolytic activities from C. cubensis:P. pinophilum displayed a much higher tolerance to phenolic inhibitors than equivalent enzyme activities obtained from Trichoderma reesei and Aspergillus niger. Enzymes from T. reesei and A...
February 2018: Enzyme and Microbial Technology
Chuannan Long, Yijin Cheng, Jingjing Cui, Jian Liu, Lihui Gan, Bin Zeng, Minnan Long
The role of the transcription factor creA-mediating carbon catabolite repression in Trichoderma orientalis EU7-22 was investigated for cellulase and hemicellulase production. The binary vector pUR5750G/creA::hph was constructed to knock out creA by homologous integration, generating the ΔcreA mutant Trichoderma orientalis CF1D. For strain CF1D, the filter paper activities (FPA), endoglucanase activities (CMC), cellobiohydrolase activity(CBH), β-glucosidase activity (BG), xylanase activity (XYN), and extracellular protein concentration were 1...
December 6, 2017: Molecular Biotechnology
Tomoko Maehara, Haruka Yagi, Tomoko Sato, Mayumi Ohnishi-Kameyama, Zui Fujimoto, Kei Kamino, Yoshiaki Kitamura, Franz St John, Katsuro Yaoi, Satoshi Kaneko
Endoxylanases are important enzymes in bioenergy research because they specifically hydrolyze xylan, the predominant polysaccharide in the hemicellulose fraction of lignocellulosic biomass. For effective biomass utilization, it is important to understand the mechanism of substrate recognition by these enzymes. Recent studies have shown that the substrate specificity of bacterial and fungal endoxylanases classified into glycoside hydrolase family 30 (GH30) were quite different. While the functional differences have been described the mechanism of substrate recognition is still unknown...
November 27, 2017: Applied and Environmental Microbiology
Yanfen Cheng, Ying Wang, Yuanfei Li, Yipeng Zhang, Tianyi Liu, Yu Wang, Thomas J Sharpton, Weiyun Zhu
The aim of this study was to improve the utilization of rice straw as forage in ruminants by investigating the degradation pattern of rice straw in the dairy cow rumen. Ground up rice straw was incubated in situ in the rumens of three Holstein cows over a period of 72 h. The rumen fluid at 0 h and the rice straw at 0.5, 1, 2, 4, 6, 12, 24, 48, and 72 h were collected for analysis of the bacterial community and the degradation of the rice straw. The bacterial community and the carbohydrate-active enzymes in the rumen fluid were analyzed by metagenomics...
2017: Frontiers in Microbiology
Silvia Hüttner, Thanh Thuy Nguyen, Zoraide Granchi, Thomas Chin-A-Woeng, Dag Ahrén, Johan Larsbrink, Vu Nguyen Thanh, Lisbeth Olsson
Background: Genome and transcriptome sequencing has greatly facilitated the understanding of biomass-degrading mechanisms in a number of fungal species. The information obtained enables the investigation and discovery of genes encoding proteins involved in plant cell wall degradation, which are crucial for saccharification of lignocellulosic biomass in second-generation biorefinery applications. The thermophilic fungus Malbranchea cinnamomea is an efficient producer of many industrially relevant enzymes and a detailed analysis of its genomic content will considerably enhance our understanding of its lignocellulolytic system and promote the discovery of novel proteins...
2017: Biotechnology for Biofuels
Bareket Dassa, Ilya Borovok, Vincent Lombard, Bernard Henrissat, Raphael Lamed, Edward A Bayer, Sarah Moraïs
The bacterial cellulosome is an extracellular, multi-enzyme machinery, which efficiently depolymerizes plant biomass by degrading plant cell wall polysaccharides. Several cellulolytic bacteria have evolved various elaborate modular architectures of active cellulosomes. We present here a genome-wide analysis of a dozen mesophilic clostridia species, including both well-studied and yet-undescribed cellulosome-producing bacteria. We first report here, the presence of cellulosomal elements, thus expanding our knowledge regarding the prevalence of the cellulosomal paradigm in nature...
November 18, 2017: Microorganisms
Lara Hassan, Manfred J Reppke, Nils Thieme, Steffen A Schweizer, Carsten W Mueller, J Philipp Benz
Background: The industrial applications of cellulases are mostly limited by the costs associated with their production. Optimized production pathways are therefore desirable. Based on their enzyme inducing capacity, celluloses are commonly used in fermentation media. However, the influence of their physiochemical characteristics on the production process is not well understood. In this study, we examined how physical, structural and chemical properties of celluloses influence cellulase and hemicellulase production in an industrially-optimized and a non-engineered filamentous fungus: Trichoderma reesei RUT-C30 and Neurospora crassa...
2017: Fungal Biology and Biotechnology
Daniel P Kiesenhofer, Robert L Mach, Astrid R Mach-Aigner
Trichoderma reesei can produce up to 100 g/l of extracellular proteins. The major and industrially relevant products are the cellobiohydrolase CBHI and the hemicellulase XYNI. Both encoding genes are transcriptionally activated by the regulatory protein Xyr1. The first 850 nucleotides of the cbh1 promoter contain fourteen Xyr1-binding sites (XBS), in the xyn1 promoter are eight XBS present. Some of these XBS are arranged in tandem, others as inverted repeats. One such cis element, an inverted repeat, plays a crucial role for the inducibility of the xyn1 promoter...
October 27, 2017: Applied and Environmental Microbiology
Yujia Jiang, Tianpeng Chen, Weiliang Dong, Min Zhang, Wenming Zhang, Hao Wu, Jiangfeng Ma, Min Jiang, Fengxue Xin
A wild type solventogenic Clostridium beijerinckii NJP7 capable of converting polysaccharides, such as hemicellulose, into butanol and isopropanol via a unique acetone-isopropanol-butanol (AIB) pathway was isolated and characterized. This represents the first wild type isopropanol-butanol generating bacterium which could achieve butanol production directly from lignocellulose through consolidated bioprocessing (CBP). Strain NJP7 was isolated from decomposite soil from Laoshan Nature Park, China, and its genome shows 98...
October 24, 2017: Current Microbiology
Ronglin He, Xue Bai, Penglin Cai, Cheng Sun, Dongyuan Zhang, Shulin Chen
Many species of Penicillium have exhibited great potential for lignocellulose hydrolysis. The filamentous fungus Talaromyces piceus 9-3 (anamorph: Penicillium piceum), which was isolated from compost wastes in China, was sequenced in this study. Compared with the cellulase producer T. reesei, T. piceus 9-3 processes a lignocellulolytic enzyme system comprising more diverse enzymatic components, especially hemicellulases. This report will facilitate the use of this strain for biomass degradation.
December 2017: 3 Biotech
Chengcheng Li, Fengming Lin, Le Zhou, Lei Qin, Bingzhi Li, Zhihua Zhou, Mingjie Jin, Zhan Chen
BACKGROUND: The induction of cellulase production by insoluble carbon source cellulose was a common and efficient strategy, but has some drawbacks, such as difficult fermentation operation, substantial cellulase loss, long fermentation time, and high energy-consumption, resulting in high cost of cellulase production in industry. These drawbacks can be overcome if soluble carbon sources are utilized as the inducers for cellulase production. However, until now the induction efficiency of most soluble carbon sources, especially lactose and glucose, is still inferior to cellulose despite extensive efforts have been made by either optimizing the fermentation process or constructing the recombinant strains...
2017: Biotechnology for Biofuels
Virginia D Dimaki, Gregoris Iatrou, Fotini N Lamari
A number of beneficial medicinal properties are attributed to the extract and essential oil of the aerial parts of Sideritis species (Lamiaceae). Hydrodistillation of the aerial parts of wild Sideritis clandestina ssp. peloponnesiaca (an endemic taxon in northern Peloponnesus, Greece) gave a low essential oil yield (<0.12%); about 65 components, mainly α-pinene, β-caryophyllene, β-pinene, globulol, caryophyllene oxide, were identified via GC-MS. Internal and external standards were used for quantification...
October 6, 2017: Journal of Chromatography. A
A Gallo, G Giuberti, S Bruschi, P Fortunati, F Masoero
The study was performed on forages ( = 8), nonforage fibrous feeds ( = 10), and crop residues ( = 2). Samples were characterized for in situ NDF degradability (NDFD) at 6, 12, 18, 24, 30, 36, 48, 72, 96, 120, and 240 h of ruminal incubation. Then, samples were characterized for enzymatic NDFD by adopting a multistep enzymatic method consisting of a preincubation (PreInc) phase followed by enzymatic incubation (EnzInc) steps. In the PreInc phase, samples were incubated in a NaOH solution for 0, 30, 60, or 90 min...
September 2017: Journal of Animal Science
Hirak Saxena, Bryan Hsu, Marc de Asis, Mirko Zierke, Lyann Sim, Stephen G Withers, Warren Wakarchuk
Bacteria in the genus Cellulomonas are well known as secretors of a variety of mesophilic carbohydrate degrading enzymes (e.g. cellulases and hemicellulases), active against plant cell wall polysaccharides. Recent proteomic analysis of the mesophilic bacterium <i>Cellulomonas fimi</i> ATCC484 revealed uncharacterized enzymes for the hydrolysis of plant cell wall biomass. Celf_1230 (CfCel6C), a secreted protein of Cellulomonas fimi ATCC484, is a novel member of the GH6 family of cellulases which could be successfully expressed in Escherichia coli...
October 5, 2017: Biochemistry and Cell Biology, Biochimie et Biologie Cellulaire
Damhan S Scully, Amit K Jaiswal, Nissreen Abu-Ghannam
Conventional coffee brewing techniques generate vast quantities of spent espresso grounds (SEGs) rich in lignocellulose and valuable bioactives. These bioactive compounds can be exploited as a nutraceutical or used in a range of food products, while breakdown of lignocellulose generates metabolizable sugars that can be used for the production of various high-value products such as biofuels, amino acids and enzymes. Response surface methodology (RSM) was used to optimize the enzymatic saccharification of lignocellulose in SEGs following a hydrothermal pretreatment...
November 21, 2016: Bioengineering
Huyen Thi Do, Khoa Trong Dao, Viet Khanh Hoang Nguyen, Giang Ngoc Le, Phuong Thi Mai Nguyen, Lam Tung Le, Nguyet Thu Phung, Nico van Straalen, Dick Roelofs, Hai Nam Truong
Objective: In a previous study, analysis of Illumina sequenced metagenomic DNA data of bacteria in Vietnamese goats' rumen showed a high diversity of putative lignocellulolytic genes. In this study, taxonomy speculation of microbial community and lignocellulolytic bacteria population in the rumen was conducted to elucidate a role of bacterial structure for effective degradation of plant materials. Methods: The metagenomic data had been subjected into Basic Local Alignment Search Tool (BLASTX) algorithm and the National Center for Biotechnology Information (NCBI) non-redundant (NR) sequence database...
September 18, 2017: Asian-Australasian Journal of Animal Sciences
Christa Ivanova, Jonas Ramoni, Thiziri Aouam, Alexa Frischmann, Bernhard Seiboth, Scott E Baker, Stéphane Le Crom, Sophie Lemoine, Antoine Margeot, Frédérique Bidard
BACKGROUND: The hydrolysis of biomass to simple sugars used for the production of biofuels in biorefineries requires the action of cellulolytic enzyme mixtures. During the last 50 years, the ascomycete Trichoderma reesei, the main source of industrial cellulase and hemicellulase cocktails, has been subjected to several rounds of classical mutagenesis with the aim to obtain higher production levels. During these random genetic events, strains unable to produce cellulases were generated...
2017: Biotechnology for Biofuels
Chunliang Xie, Wenbing Gong, Li Yan, Zuohua Zhu, Zhenxiu Hu, Yuande Peng
In the textile industry, ramie stalk is byproducts with a low economic value. The potential use of this leftover as a substrate ingredient for Flammulina velutipes (F. velutipe) cultivation was evaluated. The degradation and utilization of ramie stalk by F. velutipes was evaluated through mushroom production, lignocelluloses degradation and lignocellulolytic enzymes activity. The best substrate mixture for F. velutipes cultivation comprised 50% ramie stalk, 20% cottonseed hulls, 25% wheat bran, 4% cornstarch and 2% CaCO3...
September 12, 2017: AMB Express
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