Cloning and Expression of ywtD Gene from B.subtilis NX-2 and the Enzymatic Degradation of γ-Polyglutamic Acid

China Biotechnology

2007, Vol. 27

Issue (5): 34-38 DOI:

Cloning and Expression of ywtD Gene from B.subtilis NX-2 and the Enzymatic Degradation of γ-Polyglutamic Acid

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Abstract

The B.subtilis ywtD gene, encoding a γ-polyglutamic acid (γ-PGA) depolymerase, was amplified from the genome of B.subtilis NX-2 by PCR.The comparability between the cloned ywtD gene sequence to the reported sequence is high to 99.0%. Only one of the substituted nucleotide base caused the change to the amino acid sequence. The recombinant plasmid pET-15b-ywtD was then transformed into E. coli Rosetta (DE3) and the ywtD gene product could be expressed with the induction of 0.5mmol/L IPTG. The YwtD protein exhibited a remarkable activity in γ-polyglutamic acid degradation. The molecular weight of γ-PGA could be reduced from 700kDa to 20kDa after 72h through the enzymatic hydrolysis and consequently trended to be constant.

Key words： depolymerase Bacillus subtilis molecular weight γ-polyglutamic acid

Received: 15 December 2006 Published: 25 May 2007

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. Cloning and Expression of ywtD Gene from B.subtilis NX-2 and the Enzymatic Degradation of γ-Polyglutamic Acid. China Biotechnology, 2007, 27(5): 34-38.

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https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2007/V27/I5/34

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