Cloning and Expression of ywtD Gene from B. subtilis NX-2 and the Enzymatic Degradation of ?-Polyglutamic Acid
- VernacularTitle:枯草杆菌NX-2聚谷氨酸解聚酶的克隆表达及其降解性质研究
- Author:
Jing JIN
;
Jun YAO
;
Hong XU
;
Lin XU
;
- Publication Type:Journal Article
- Keywords:
?-polyglutamic acid Depolymerase Bacillus subtilis Molecular weight
- From:
China Biotechnology
2006;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
The B.subtilis ywtD gene,encoding a ?-polyglutamic acid(?-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 ?-polyglutamic acid degradation.The molecular weight of ?-PGA could be reduced from 700kDa to 20kDa after 72h through the enzymatic hydrolysis and consequently trended to be constant.
