Profiling and production of hemicellulases by thermophilic fungus Malbranchea flava and the role of xylanases in improved bioconversion of pretreated lignocellulosics to ethanol

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• 作者：Manju Sharma ; Chhavi Mahajan ; Manpreet S. Bhatti ; Bhupinder Singh Chadha
• 关键词：Xylanases ; Xylan ; debranching accessory enzymes ; Secretome analysis ; Response surface methodology (RSM) ; Saccharification of lignocellulosics
• 刊名：3 Biotech
• 出版年：2016
• 出版时间：December 2016
• 年：2016
• 卷：6
• 期：1
• 全文大小：1,393 KB
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• 作者单位：Manju Sharma (1)
  Chhavi Mahajan (1)
  Manpreet S. Bhatti (2)
  Bhupinder Singh Chadha (1)
  
  1. Department of Microbiology, Guru Nanak Dev University, Amritsar, Punjab, 143005, India
  2. Department of Botanical and Environmental Sciences, Guru Nanak Dev University, Amritsar, Punjab, India
  
• 刊物主题：Biotechnology; Agriculture; Cancer Research; Bioinformatics; Stem Cells; Biomaterials;
• 出版者：Springer Berlin Heidelberg
• ISSN：2190-5738

文摘

This study reports thermophilic fungus Malbranchea flava as a potent source of xylanase and xylan-debranching accessory enzymes. M. flava produced high levels of xylanase on sorghum straw containing solidified culture medium. The optimization of culture conditions for production of hemicellulases was carried out using one factor at a time approach and Box–Behnken design of experiments with casein (%), inoculum age (h) and inoculum level (ml) as process variables and xylanase, β-xylosidase, acetyl esterases and arabinofuranosidase as response variables. The results showed that casein concentration between 3.0 and 3.5 %, inoculum age (56–60 h) and inoculum level (2–2.5 ml) resulted in production of 16,978, 10.0, 67.7 and 3.8 (U/gds) of xylanase, β-xylosidase, acetyl esterase and α-l-arabinofuranosidase, respectively. Under optimized conditions M. flava produced eight functionally diverse xylanases with distinct substrate specificity against different xylan types. The peptide mass fingerprinting of 2-D gel electrophoresis resolved proteins indicated to the presence of cellobiose dehydrogenase and glycosyl hydrolases suggesting the potential of this strain in oxidative and classical cellulase-mediated hydrolysis of lignocellulosics. Addition of xylanase (300 U/g substrate) during saccharification (at 15 % substrate loading) of different pretreated (acid/alkali) substrates (cotton stalks, wheat straw, rice straw, carrot grass) by commercial cellulase (NS28066) resulted in 9–36 % increase in saccharification and subsequent fermentation to ethanol when compared to experiment with commercial enzyme only. High ethanol level 46 (g/l) was achieved with acid pretreated cotton stalk when M. flava xylanase was supplemented as compared to 39 (g/l) with xylanase without xylanase addition. Keywords Xylanases Xylan-debranching accessory enzymes Secretome analysis Response surface methodology (RSM) Saccharification of lignocellulosics