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| Cloning and Function Study of amtS Gene from Saccharopolyspora spinosa |
| TAO Wen-na, XIA Li-qiu, DING Xue-zhi, TANG Ying |
| College of Life Science, Hunan Normal University, Hunan Provincial Key Laboratory of Microbial Molecular Biology-State Key Laboratory Breeding Base of Microbial Molecular Biology, Changsha 410081, China |
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Abstract Objective: Ammonium ion is one of the precursors for biosynthesis of nucleic acids, amino acids and cofactors. The transportation of NH4+ through the cell plasma membrane is mediated by ammonium carrier protein. The ammonium carrier protein encoding gene of Saccharopolyspora spinosa was cloned, and its effects on the production of secondary metabolites of Streptomyces coelicolor and Streptomyces lividans were evaluated. Methods:The amtS gene of S. spinosa was cloned into E. coli-Streptomyces shuttle vector pMF and transferred into S. coelicolor M145 and S. lividans TK24 by conjugation. The phenotypes of the recombinants were compared with the parental strains, and the effects of amtS overexpression on the production of secondary metabolites were analyzed. Results:The yields of actinorhodin and undecylprodigiosin in both M145/pMF-amtS and TK24/pMF-amtS were greatly improved. The yields of actinorhodin were elevated by 2.85 and 30.02 times, respectively. Conclusion:Heterologous expression of amtS from Saccharopolyspora spinosa could improve the production of secondary metabolites, which provided the important basis for its function study in Saccharopolyspora spinosa.
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Received: 03 December 2014
Published: 25 February 2015
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