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J Agric Food Chem. 2018 Sep 12;66(36):9465-9472. doi: 10.1021/acs.jafc.8b03327. Epub 2018 Aug 30.

Glycoside Hydrolase Family 39 β-Xylosidase of Sphingomonas Showing Salt/Ethanol/Trypsin Tolerance, Low-pH/Low-Temperature Activity, and Transxylosylation Activity.

Li N1,2,3, Han X1,2,3, Xu S1,2,3, Li C1,2,3, Wei X4, Liu Y1,2,3, Zhang R1,2,3, Tang X1,2,3, Zhou J1,2,3, Huang Z1,2,3.

Author information

1
Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education , Yunnan Normal University , Kunming , 650500 , People's Republic of China.
2
College of Life Sciences , Yunnan Normal University , Kunming , 650500 , People's Republic of China.
3
Key Laboratory of Yunnan for Biomass Energy and Biotechnology of Environment , Yunnan, Kunming , 650500 , People's Republic of China.
4
State Key Laboratory of Phytochemistry and Plant Resources in West China , Kunming Institute of Botany, Chinese Academy of Sciences , Kunming 650201 , People's Republic of China.

Abstract

Mining for novel enzymes from new microorganisms is a way to obtain β-xylosidases with promising applications. A Sphingomonas β-xylosidase was expressed in Escherichia coli. The purified recombinant enzyme (rJB13GH39) was most active at pH 4.5 and 50 °C, retaining 10%-50% of its maximum activity at 0-20 °C. Most salts and chemical reagents including 3.0%-20.0% (w/v) NaCl showed little or no effect on the enzymatic activity. rJB13GH39 exhibited 71.9% and 55.2% activity in 10.0% and 15.0% (v/v) ethanol, respectively. rJB13GH39 was stable below 60 °C in 3.0%-30.0% (w/v) NaCl, 3.0%-20.0% (v/v) ethanol, and 2.2-87.0 mg/mL trypsin. The enzyme transferred one xylosyl moiety to certain sugars and alcohols. The salt/ethanol tolerance and low-temperature activity of the enzyme may be attributed to its high structural flexibility caused by high proportions of small amino acids ACDGNSTV and random coils.

KEYWORDS:

Sphingomonas sp; ethanol tolerance; glycoside hydrolase family 39; low-temperature activity; salt tolerance; transxylosylation; trypsin tolerance; β-xylosidase

PMID:
30132665
DOI:
10.1021/acs.jafc.8b03327
[Indexed for MEDLINE]

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