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journal article Aug 23, 2017

Combination of traditional mutation and metabolic engineering to enhance ansamitocin P‐3 production in Actinosynnema pretiosum

Zhi‐Qiang Du Yuan Zhang ORCID Zhi‐Gang Qian Han Xiao ORCID Jian‐Jiang Zhong ORCID
Biotechnology and Bioengineering Vol. 114 No. 12 pp. 2794-2806 · Wiley
View at Publisher Save 10.1002/bit.26396
Abstract
AbstractAnsamitocin P‐3 (AP‐3) is a maytansinoid with its most compelling antitumor activity, however, the low production titer of AP‐3 greatly restricts its wide commercial application. In this work, a combinatorial approach including random mutation and metabolic engineering was conducted to enhance AP‐3 biosynthesis in Actinosynnema pretiosum. First, a mutant strain M was isolated by N‐methyl‐N'‐nitro‐N‐nitrosoguanidine mutation, which could produce AP‐3 almost threefold that of wild type (WT) in 48 deep‐well plates. Then, by overexpressing key biosynthetic genes asmUdpg and asm13‐17 in the M strain, a further 60% increase of AP‐3 production in 250‐ml shake flasks was achieved in the engineered strain M‐asmUdpg:asm13‐17 compared to the M strain, and its maximum AP‐3 production reached 582.7 mg/L, which is the highest as ever reported. Both the gene transcription levels and intracellular intermediate concentrations in AP‐3 biosynthesis pathway were significantly increased in the M and M‐asmUdpg:asm13‐17 during fermentation compared to the WT. The good fermentation performance of the engineered strain was also confirmed in a lab‐scale bioreactor. This work demonstrated that combination of random mutation and metabolic engineering could promote AP‐3 biosynthesis and might be helpful for increasing the production of other industrially important secondary metabolites.
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References
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Metrics
25
Citations
36
References
Details
Published
Aug 23, 2017
Vol/Issue
114(12)
Pages
2794-2806
License
View
Authors
Z
Zhi‐Qiang Du

State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic and Developmental Sciences, and Laboratory of Molecular Biochemical Engineering and Advanced Fermentation Technology, Department of Bioengineering, School of Life Sciences and Biotechnology Shanghai Jiao Tong University Shanghai China

Y
Yuan Zhang

State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic and Developmental Sciences, and Laboratory of Molecular Biochemical Engineering and Advanced Fermentation Technology, Department of Bioengineering, School of Life Sciences and Biotechnology Shanghai Jiao Tong University Shanghai China

Z
Zhi‐Gang Qian

State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic and Developmental Sciences, and Laboratory of Molecular Biochemical Engineering and Advanced Fermentation Technology, Department of Bioengineering, School of Life Sciences and Biotechnology Shanghai Jiao Tong University Shanghai China

H
Han Xiao

State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic and Developmental Sciences, and Laboratory of Molecular Biochemical Engineering and Advanced Fermentation Technology, Department of Bioengineering, School of Life Sciences and Biotechnology Shanghai Jiao Tong University Shanghai China

J
Jian‐Jiang Zhong

State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic and Developmental Sciences, and Laboratory of Molecular Biochemical Engineering and Advanced Fermentation Technology, Department of Bioengineering, School of Life Sciences and Biotechnology Shanghai Jiao Tong University Shanghai China

Funding
National Natural Science Foundation of China Award: 31370083
Cite This Article
Zhi‐Qiang Du, Yuan Zhang, Zhi‐Gang Qian, et al. (2017). Combination of traditional mutation and metabolic engineering to enhance ansamitocin P‐3 production in Actinosynnema pretiosum. Biotechnology and Bioengineering, 114(12), 2794-2806. https://doi.org/10.1002/bit.26396
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