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Identification and characterization of an unusual glycosyltransferase-like enzyme with β-galactosidase activity from a soil metagenomic library

Paper ID Volume ID Publish Year Pages File Format Full-Text
17058 42637 2014 10 PDF Available
Title
Identification and characterization of an unusual glycosyltransferase-like enzyme with β-galactosidase activity from a soil metagenomic library
Abstract

•A novel glycosyltransferase (Glyt110) was obtained from a metagenomic library.•It is an unusual enzyme with β-galactosidase activity but grouped into family GT4.•The specific activities toward ONPG and lactose were 314 ± 18.3 and 32 ± 2.7 U/mg, respectively.•A GOS yield of 47.2% (w/w) was achieved by Glyt110 using lactose as substrate.•Glyt110 may be a potential candidate for industrial production of GOS.

Glycosyltransferases and glycoside hydrolases are two diversified groups of carbohydrate-active enzymes (CAZymes) in existence, they serve to build and break down the glycosidic bonds, respectively, and both categories have formed many sequence-based families. In this study, a novel gene (glyt110) conferring β-galactosidase activity was obtained from a metagenomic library of Turpan Basin soil. Sequence analysis revealed that glyt110 encoded a protein of 369 amino acids that, rather than belonging to a family typically known for β-galactosidase activity, belonged to glycosyltransferase family 4. Because of this unusual sequence information, the novel gene glyt110 was subsequently expressed in Escherichia coli BL21(DE3), and the recombinant enzyme (Glyt110) was purified and characterized. Biochemical characterization revealed that the β-galactosidase activity of Glyt110 toward o-nitrophenyl-β-d-galactopyranoside (ONPG) and lactose were identified to be 314 ± 18.3 and 32 ± 2.7 U/mg, correspondingly. In addition, Glyt110 can synthesize galacto-oligosaccharides (GOS) using lactose as substrate. A GOS yield of 47.2% (w/w) was achieved from 30% lactose solution at 50 °С, pH 8.0 after 10 h reaction. However, Glyt110 was unable to glycosylate either N-acetylated saccharides or lactose and galactose using UDP-gal as sugar donor, and its glycosyltransferase activity needs further investigation. These results indicated that Glyt110 is an unusual enzyme with β-galactosidase activity but phylogenetically related to glycosyltransferase. Our findings may provide opportunities to improve the insight into the relationship between glycosyltransferases and glycoside hydrolases and the sequence-based classification.

Keywords
β-Galactosidase; Glycosyltransferase; Metagenomic library; Sequence-based classification; Enzyme characterization
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Identification and characterization of an unusual glycosyltransferase-like enzyme with β-galactosidase activity from a soil metagenomic library
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 57, 10 April 2014, Pages 26–35
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us