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Pak J Biol Sci ; 13(10): 504-8, 2010 May 15.
Article in English | MEDLINE | ID: mdl-21848075

ABSTRACT

The aim of this study is over-expression of Meso-diaminopimelate decarboxylase enzyme (EC 4.1.1.20) and enhancement of L-lysine production rate. The C. glutamicum LysA gene which encodes a Meso-diaminopimelate decarboxylase was cloned in E. coli. The cloned gene was sequenced; it encodes a 445 amino acids protein with molecular weight of 47 kD. Expression of the LysA gene in E. coli resulted in an increase in Meso-diaminopimelate decarboxylase activity, correlated with the presence in sodium dodecyl sulfate-polyacrylamid gels of a clear protein band that corresponds to this enzyme. The induction of cloned gene by IPTG has been shown to have an inhibitory effect on cell growth due to over-expression of the cloned gene. A two fold increase in lysine production rate was observed after introduction of the cloned gene into E. coli.


Subject(s)
Biotechnology/methods , Carboxy-Lyases/biosynthesis , Corynebacterium glutamicum/enzymology , Escherichia coli/enzymology , Lysine/metabolism , Amino Acids/chemistry , Bacterial Proteins/genetics , Carboxy-Lyases/genetics , Carboxy-Lyases/metabolism , Cloning, Molecular , Corynebacterium glutamicum/genetics , DNA/chemistry , Electrophoresis, Polyacrylamide Gel , Escherichia coli/genetics , Gene Expression Regulation, Bacterial , Molecular Weight , Plasmids/metabolism , Polymerase Chain Reaction/methods , Time Factors
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