Sequencing of the L-glutaminase gene isolated from Streptomyces of sea water and cloning in Escherichia coli origami for clinical and industrial usage

Background and Aim : Streptomyces are a gram-positive bacteria and have a unique role in the production of secondary metabolites and biological active substances.L-glutaminase is an enzyme is used for industrial applications such as pharmaceutical and food industries for increasing the flavor and taste of food. Because of the importance of using of this enzyme,the aim of this study is sequencing of the L-glutaminase gene isolated from Streptomyces of sea water and cloning in Escherichia coli origami bacteria for clinical, industrial and food utilization .Methods : After sampling from the Persian Gulf waters,Streptomyces were isolated and DNA was extracted.Then by the PCR test the strains of Streptomyces which had a L-glutaminase gene were identified.The L-glutaminase gene positive strains were cloned through the vector into the E.coli and cloned by TA cloning technique.Finally,with the Real Time PCR technique measured the expression of these genes in E.coli origami.Results : After screening of sea water samples,۱۲ isolates of Streptomyces were isolated totally, of these, ۷ isolates had the L-glutaminase gene.In order to confirm the cloning results,the DNA was extracted from the suspected colonies and were evaluated with the PCR and finally,the Real time PCR product confirmed the expression of L-glutaminase gene in E.coli origami.Conclusion : The results of this study showed that this procedure can be a suitable candidates for design and manufacture of drugs and antimicrobial compounds. In the present study, the production of glutaminase enzyme was performed by recombinant plasmid and TA cloning,but subsequent studies could be used to optimize different conditions for producing this enzyme.

کلیدواژه ها:

Sea Streptomyces ، L-glutaminase ، Cloning