Cloning, over-expression and enzymatic characterization of two xylanase?-encoding genes from Streptomyces sp. J103 in Escherichia coli

Title
Cloning, over-expression and enzymatic characterization of two xylanase?-encoding genes from Streptomyces sp. J103 in Escherichia coli
Author(s)
권영경; 이지현; 이영득; 이수진; 박건후; 허수진; 강도형; 오철홍
KIOST Author(s)
Lee, Youngdeuk(이영득)Heo, Soo Jin(허수진)Kang, Do-Hyung(강도형)Oh, Chulhong(오철홍)
Publication Year
2014-10-17
Abstract
Xylan, a component of plant cell wall, is the most abundant hemicellulosic polysaccharide in nature. Xylanases are glycosidases that catalyze the endohydrolysis of 1, 4-β-D-xylosidic linkages in xylan into short xylooligosaccharides. Xylanases are used various industries including bioethanol production, bio-bleaching of pulp, textile, and production of animal feed. A xylanase-producing bacterium, Streptomyces sp. J103, identified by selection on an agar palate containing xylan as carbon source followed by 16S rDNA sequence analysis. The genome of Streptomyces sp. J103 was sequenced by Genome Sequencer-FLX system (GS-FLX), a next generation sequencing (NGS) technology. The predicted xylanase coding genes were identified by the NCBI BLAST algorithm (designated as J103-x2 and J103-x7). The nucleotide sequence of J103-x2 contains a 693 bp ORF, which encodes 230 amino acid (aa) protein with a predicted molecular mass of 24 kDa and theoretical isoelectric point (pI) of 8.4. J103-x7 consists of 1011 bp ORF coding for 693 aa. The predicted molecular mass and isoelectric point of J103-x7 were 35 kDa and 8.8, respectively. The deduced amino acid sequence of the J103-x2 and J103-x7 showed 79.8 and 66.4% similarity with xylanase from Streptomyces sp. e14 and Streptomyces sp. TN119, respectively. The xylanase genes were cloned and overexpressed in E. coli expression system. The recombinant enzymes were purified using ion-exchangases are used various industries including bioethanol production, bio-bleaching of pulp, textile, and production of animal feed. A xylanase-producing bacterium, Streptomyces sp. J103, identified by selection on an agar palate containing xylan as carbon source followed by 16S rDNA sequence analysis. The genome of Streptomyces sp. J103 was sequenced by Genome Sequencer-FLX system (GS-FLX), a next generation sequencing (NGS) technology. The predicted xylanase coding genes were identified by the NCBI BLAST algorithm (designated as J103-x2 and J103-x7). The nucleotide sequence of J103-x2 contains a 693 bp ORF, which encodes 230 amino acid (aa) protein with a predicted molecular mass of 24 kDa and theoretical isoelectric point (pI) of 8.4. J103-x7 consists of 1011 bp ORF coding for 693 aa. The predicted molecular mass and isoelectric point of J103-x7 were 35 kDa and 8.8, respectively. The deduced amino acid sequence of the J103-x2 and J103-x7 showed 79.8 and 66.4% similarity with xylanase from Streptomyces sp. e14 and Streptomyces sp. TN119, respectively. The xylanase genes were cloned and overexpressed in E. coli expression system. The recombinant enzymes were purified using ion-exchang
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/25948
Bibliographic Citation
2014 한국해양바이오학회 학술대회, pp.60, 2014
Publisher
한국해양바이오학회
Type
Conference
Language
English
Publisher
한국해양바이오학회
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