Cloning and enzymatic characterization of a xylanase gene from a soil-derived metagenomic library with an efficient approach

Applied Microbiology and Biotechnology
Yong HuLi Xin Ma

Abstract

Screening interesting biocatalysts directly from soil samples is a more convenient and applicable approach than conventional cultivation-dependent ones. In our present work, a soil-derived metagenomic library containing 24,000 transformants was constructed with an efficient strategy for cloning xylanase genes. A gene encoding the enzyme (XynH) able to hydrolyze xylan was obtained. Similarity analysis revealed that this enzyme is a new member in the family 10 of xylanases. The molecular mass of XynH purified from Escherichia coli was estimated to be 39 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis. It was found to display the maximal activity at lower temperature, under weakly alkaline conditions, different from most of xylanases. The K(m) and V(max) values of XynH with birchwood xylan as substrate are 7.5 mg/ml and 190 micromol min(-1 )mg(-1), respectively. It is greatly interesting to note that the activity of XynH was not reduced significantly by Mn(2+), Zn(2+), Co(2+), Ag(+), and Cu(2+), even at the concentration of 5 mM, which strongly inhibits most of the other xylanases studied previously.

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Citations

Apr 16, 2010·Applied Microbiology and Biotechnology·Guozeng WangBin Yao
Jun 23, 2010·Applied Microbiology and Biotechnology·Xin-chun MoJia-xun Feng
Feb 22, 2011·Applied Microbiology and Biotechnology·Chengzhang FuLixin Zhang
Apr 16, 2011·Applied Microbiology and Biotechnology·Julie NeveuMichael S DuBow
Jul 19, 2012·World Journal of Microbiology & Biotechnology·Mahjabeen SaleemMuhammad Ibrahim Rajoka
Jan 5, 2012·Applied Biochemistry and Biotechnology·Yu Seok JeongHoon Kim
Aug 17, 2010·Applied and Environmental Microbiology·Shengguo ZhaoZhongtang Yu
May 25, 2010·Biotechnology Advances·In Taek HwangNo-Joong Park
Oct 7, 2009·Biotechnology and Bioengineering·Min Hye ShinKyoung Heon Kim
Oct 15, 2014·Journal of Food Science·T A Sathya, Mahejibin Khan
Aug 6, 2014·Protein Expression and Purification·La XiangYanhe Ma
Mar 13, 2015·Applied Biochemistry and Biotechnology·Rosmeriana Afnis Marioto GarciaEliana Gertrudes de Macedo Lemos
Oct 24, 2009·Current Opinion in Biotechnology·Taku Uchiyama, Kentaro Miyazaki
Jun 13, 2017·Critical Reviews in Food Science and Nutrition·Mahejibin Khan, T A Sathya
Aug 9, 2012·Animal Biotechnology·Jiakun WangJianxin Liu
Aug 18, 2017·Applied Biochemistry and Biotechnology·Aravind MadhavanAshok Pandey

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