Reverse biological engineering of hrdB to enhance the production of avermectins in an industrial strain of Streptomyces avermitilis

Ying Zhuo, Wenquan Zhang, Difei Chen, Hong Gao, Jun Tao, Mei Liu, Zhongxuan Gou, Xianlong Zhou, Bang-Ce Ye, Qing Zhang, Siliang Zhang, Li-Xin Zhang

Research output: Contribution to journalArticle

Abstract

Avermectin and its analogues are produced by the actinomycete Streptomyces avermitilis and are widely used in the field of animal health, agriculture, and human health. Here we have adopted a practical approach to successfully improve avermectin production in an industrial overproducer. Transcriptional levels of the wild-type strain and industrial overproducer in production cultures were monitored using microarray analysis. The avermectin biosynthetic genes, especially the pathway-specific regulatory gene, aveR, were up-regulated in the high-producing strain. The upstream promoter region of aveR was predicted and proved to be directly recognized by sigma(hrdB) in vitro. A mutant library of hrdB gene was constructed by error-prone PCR and selected by high-throughput screening. As a result of evolved hrdB expressed in the modified avermectin high-producing strain, 6.38 g/L of avermectin B1a was produced with over 50% yield improvement, in which the transcription level of aveR was significantly increased. The relevant residues were identified to center in the conserved regions. Engineering of the hrdB gene can not only elicit the overexpression of aveR but also allows for simultaneous transcription of many other genes. The results indicate that manipulating the key genes revealed by reverse engineering can effectively improve the yield of the target metabolites, providing a route to optimize production in these complex regulatory systems.

Original languageEnglish
Pages (from-to)11250-4
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume107
Issue number25
DOIs
Publication statusPublished - 22 Jun 2010

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Bioengineering
Streptomyces
Genes
Actinobacteria
Health
Regulator Genes
Microarray Analysis
Agriculture
Gene Library
Genetic Promoter Regions
Polymerase Chain Reaction
avermectin

Cite this

Zhuo, Ying ; Zhang, Wenquan ; Chen, Difei ; Gao, Hong ; Tao, Jun ; Liu, Mei ; Gou, Zhongxuan ; Zhou, Xianlong ; Ye, Bang-Ce ; Zhang, Qing ; Zhang, Siliang ; Zhang, Li-Xin. / Reverse biological engineering of hrdB to enhance the production of avermectins in an industrial strain of Streptomyces avermitilis. In: Proceedings of the National Academy of Sciences of the United States of America. 2010 ; Vol. 107, No. 25. pp. 11250-4.
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Reverse biological engineering of hrdB to enhance the production of avermectins in an industrial strain of Streptomyces avermitilis. / Zhuo, Ying; Zhang, Wenquan; Chen, Difei; Gao, Hong; Tao, Jun; Liu, Mei; Gou, Zhongxuan; Zhou, Xianlong; Ye, Bang-Ce; Zhang, Qing; Zhang, Siliang; Zhang, Li-Xin.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 107, No. 25, 22.06.2010, p. 11250-4.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Reverse biological engineering of hrdB to enhance the production of avermectins in an industrial strain of Streptomyces avermitilis

AU - Zhuo, Ying

AU - Zhang, Wenquan

AU - Chen, Difei

AU - Gao, Hong

AU - Tao, Jun

AU - Liu, Mei

AU - Gou, Zhongxuan

AU - Zhou, Xianlong

AU - Ye, Bang-Ce

AU - Zhang, Qing

AU - Zhang, Siliang

AU - Zhang, Li-Xin

PY - 2010/6/22

Y1 - 2010/6/22

N2 - Avermectin and its analogues are produced by the actinomycete Streptomyces avermitilis and are widely used in the field of animal health, agriculture, and human health. Here we have adopted a practical approach to successfully improve avermectin production in an industrial overproducer. Transcriptional levels of the wild-type strain and industrial overproducer in production cultures were monitored using microarray analysis. The avermectin biosynthetic genes, especially the pathway-specific regulatory gene, aveR, were up-regulated in the high-producing strain. The upstream promoter region of aveR was predicted and proved to be directly recognized by sigma(hrdB) in vitro. A mutant library of hrdB gene was constructed by error-prone PCR and selected by high-throughput screening. As a result of evolved hrdB expressed in the modified avermectin high-producing strain, 6.38 g/L of avermectin B1a was produced with over 50% yield improvement, in which the transcription level of aveR was significantly increased. The relevant residues were identified to center in the conserved regions. Engineering of the hrdB gene can not only elicit the overexpression of aveR but also allows for simultaneous transcription of many other genes. The results indicate that manipulating the key genes revealed by reverse engineering can effectively improve the yield of the target metabolites, providing a route to optimize production in these complex regulatory systems.

AB - Avermectin and its analogues are produced by the actinomycete Streptomyces avermitilis and are widely used in the field of animal health, agriculture, and human health. Here we have adopted a practical approach to successfully improve avermectin production in an industrial overproducer. Transcriptional levels of the wild-type strain and industrial overproducer in production cultures were monitored using microarray analysis. The avermectin biosynthetic genes, especially the pathway-specific regulatory gene, aveR, were up-regulated in the high-producing strain. The upstream promoter region of aveR was predicted and proved to be directly recognized by sigma(hrdB) in vitro. A mutant library of hrdB gene was constructed by error-prone PCR and selected by high-throughput screening. As a result of evolved hrdB expressed in the modified avermectin high-producing strain, 6.38 g/L of avermectin B1a was produced with over 50% yield improvement, in which the transcription level of aveR was significantly increased. The relevant residues were identified to center in the conserved regions. Engineering of the hrdB gene can not only elicit the overexpression of aveR but also allows for simultaneous transcription of many other genes. The results indicate that manipulating the key genes revealed by reverse engineering can effectively improve the yield of the target metabolites, providing a route to optimize production in these complex regulatory systems.

U2 - 10.1073/pnas.1006085107

DO - 10.1073/pnas.1006085107

M3 - Article

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JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 25

ER -