Highly thermostable GH51 α-arabinofuranosidase from Hungateiclostridium clariflavum DSM 19732

Autor:	Hongcheng Wang, Jianzhong Sun, Jian Wu, Rongrong Xie, Xia Li, Alei Geng, Yanfang Wu, Fuxiang Chang
Rok vydání:	2019
Předmět:	Hot Temperature Glycoside Hydrolases Stereochemistry medicine.disease_cause Applied Microbiology and Biotechnology Substrate Specificity 03 medical and health sciences chemistry.chemical_compound Hydrolysis Bacterial Proteins Enzyme Stability Arabinoxylan medicine Enzyme kinetics Catalytic efficiency Escherichia coli Incubation 030304 developmental biology Thermostability Clostridiales 0303 health sciences 030306 microbiology Chemistry Substrate (chemistry) General Medicine Hydrogen-Ion Concentration Arabinose Kinetics Xylans Biotechnology
Zdroj:	Applied Microbiology and Biotechnology. 103:3783-3793
ISSN:	1432-0614 0175-7598
DOI:	10.1007/s00253-019-09753-8
Popis:	Arabinofuranosidase plays an essential role in the process of hydrolysis of arabinoxylan (AX). Thermostable, versatile, and efficient arabinofuranosidase is thus of great interest for the biorefinery industry. A GH51 arabinofuranosidase, Abf51, from Hungateiclostridium clariflavum DSM 19732 was heterogeneously expressed in Escherichia coli. Abf51 was found to have an optimal pH and temperature of 6.5 and 60 °C, respectively, with very high thermostability. At the optimal working temperature (60 °C), Abf51 retained over 90% activity after a 2-day incubation and over 60% activity after a 6-day incubation. Abf51 could effectively remove the arabinofuranosyls from three kinds of AX oligosaccharides [23-α-L-arabinofuranosyl-xylotriose (A2XX), 32-α-L-arabinofuranosyl-xylobiose (A3X), and 2333-di-α-L-arabinofuranosyl-xylotriose (A2 + 3XX)], which characterized as either single substitution or double substitution by arabinofuranosyls on terminal xylopyranosyl units. The maximal catalytic efficiency (Kcat/Km) was observed using p-nitrophenyl-α-L-arabinofuranoside (pNPAF) as a substrate (205.0 s-1 mM-1), followed by using A3X (22.8 s-1 mM-1), A2XX (6.9 s-1 mM-1), and A2 + 3XX (0.5 s-1 mM-1) as substrates. Moreover, the presence of Abf51 significantly stimulated the saccharification level of AX (18.5 g L-1) up to six times along with a β-xylanase as well as a β-xylosidase. Interestingly, in our survey of top thermostable arabinofuranosidases, most members were found from GH51, probably due to their owning of (β/α)8-barrel architectures. Our results suggested the great importance of GH51s as candidates for thermostable, versatile, and efficient arabinofuranosidases toward industry application.
Databáze:	OpenAIRE
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