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Title of Journal: World J Microbiol Biotechnol

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Abbravation: World Journal of Microbiology and Biotechnology

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Springer Netherlands

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DOI

10.1007/s10573-010-0052-2

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1573-0972

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Molecular cloning and heterologous expression of a

Authors: Liangwei Mao Po Meng Cheng Zhou Lixin Ma Guimin Zhang Yanhe Ma
Publish Date: 2011/10/26
Volume: 28, Issue: 3, Pages: 777-784
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Abstract

A new xylanase gene named xyn186 was cloned by the genomewalking PCR method from the Alternaria sp HB186 The sequence of xyn186 contains a 748 bp open reading frame separated by one intron with the size of 52 bp The cDNA was obtained by DpnImediated intron deletion The cDNA was cloned into pHBM905A and transformed into Pichia pastoris GS115 to screen xylanasesecreting transformants on RBBxylan plates The molecular mass of the enzyme was estimated to be 23 kDa on SDSPAGE The optimal pH and temperature of the purified enzyme is 6 and 50°C respectively The K m and V max valued for birchwood xylan are 1404 mg ml−1 and 02748 mmol min−1 mg−1 respectively The inhibitory effects of various metal ions were investigated Cu2+ and Hg2+ ions inhibited most of the enzyme activity The gene copy number of xyn186 in the genome of P pastoris was estimated as two by the Realtime PCR To date xyn186 gene is the first xylanase gene cloned from the genus AlternariaThis study was supported by the National Nature Science Foundation 30600014 Hubei Province Nature Science Foundation Key Project Knowledge Innovative Program of the Chinese Academy of Sciences KSCX2EWG8 and the Ministry of Sciences and Technology of China 973 programs 2012CB721000

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