Development and application of a T7 RNA polymerase-dependent expression system for antibiotic production improvement in Streptomyces
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Abstract
Objective
To develop a reliable and easy to use expression system for antibiotic production improvement of Streptomyces.
Results
A two-compound T7 RNA polymerase-dependent gene expression system was developed to fulfill this demand. In this system, the T7 RNA polymerase coding sequence was optimized based on the codon usage of Streptomyces coelicolor. To evaluate the functionality of this system, we constructed an activator gene overexpression strain for enhancement of actinorhodin production. By overexpression of the positive regulator actII-ORF4 with this system, the maximum actinorhodin yield of engineered strain was 15-fold higher and the fermentation time was decreased by 48 h.
Conclusion
The modified two-compound T7 expression system improves both antibiotic production and accelerates the fermentation process in Streptomyces. This provides a general and useful strategy for strain improvement of important antibiotic producing Streptomyces strains.
Keywords
Antibiotics Production improvement Streptomyces T7 RNA polymeraseNotes
Acknowledgements
This work was financially supported by grants from National Natural Science Foundation of China (Grant Nos. 31470250 and 31601678), Natural Science Foundation of Chongqing (Grant No. 2015jcyjys80001) and Fundamental Research Funds for the Central Universities (Grant No. XDJK2016C012). We want to thank Dr. Guojian Liao (College of Pharmaceutical Sciences, Southwest University, Chongqing, China) for kindly providing Streptomyces coelicolor M145, ET12567/pUZ8002 and pSET152 in this study.
Supporting information
Supplementary Table 1—Primers used in this study.
Supplementary Sequence 1—Sequence of modified T7 RNA polymerase.
Supplementary Fig. 1—bar graph showing actinorhodin production profiles of all strains.
Compliance with ethical standards
Conflict of interest
The authors declare no conflict of interest.
Supplementary material
References
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