Designed biosynthesis of 25-methyl and 25-ethyl ivermectin with enhanced insecticidal activity by domain swap of avermectin polyketide synthase

Microbial Cell Factories

Research

Sheng-Xiong Huang¹  Xiang-Jing Wang²  Jing An²  Ji Zhang²  Yi-Jun Yan³  Wen-Sheng Xiang⁴ 

[1] Kunming Institute of Botany, Chinese Academy of Sciences, No. 132 Lanhei Road, Panlong District, 650201, Kunming, China;School of Life Science, Northeast Agricultural University, No. 59 Mucai Street, Xiangfang District, 150030, Harbin, China;School of Life Science, Northeast Agricultural University, No. 59 Mucai Street, Xiangfang District, 150030, Harbin, China;Kunming Institute of Botany, Chinese Academy of Sciences, No. 132 Lanhei Road, Panlong District, 650201, Kunming, China;School of Life Science, Northeast Agricultural University, No. 59 Mucai Street, Xiangfang District, 150030, Harbin, China;State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, No. 2 Yuanmingyuan West Road, Haidian District, 100193, Beijing, China;

关键词: 25-Ethyl ivermectin;    25-Methyl ivermectin;    Domain swap;    Insecticidal activity;   

DOI  :  10.1186/s12934-015-0337-y

 received in 2015-06-30, accepted in 2015-09-10,  发布年份 2015

来源: Springer

【 摘 要 】

BackgroundAvermectin and milbemycin are important 16-membered macrolides that have been widely used as pesticides in agriculture. However, the wide use of these pesticides inevitably causes serious drug resistance, it is therefore imperative to develop new avermectin and milbemycin analogs. The biosynthetic gene clusters of avermectin and milbemycin have been identified and the biosynthetic pathways have been elucidated. Combinatorial biosynthesis by domain swap provides an efficient strategy to generate chemical diversity according to the module polyketide synthase (PKS) assembly line.ResultsThe substitution of aveDH2-KR2 located in avermectin biosynthetic gene cluster in the industrial avermectin-producing strain Streptomyces avermitilis NA-108 with the DNA regions milDH2-ER2-KR2 located in milbemycin biosynthetic gene cluster in Streptomyces bingchenggensis led to S. avermitilis AVE-T27, which produced ivermectin B1a with high yield of 3450 ± 65 μg/ml. The subsequent replacement of aveLAT-ACP encoding the loading module of avermectin PKS with milLAT-ACP encoding the loading module of milbemycin PKS led to strain S. avermitilis AVE-H39, which produced two new avermectin derivatives 25-ethyl and 25-methyl ivermectin (1 and 2) with yields of 951 ± 46 and 2093 ± 61 μg/ml, respectively. Compared to commercial insecticide ivermectin, the mixture of 25-methyl and 25-ethyl ivermectin (2:1 = 3:7) exhibited 4.6-fold increase in insecticidal activity against Caenorhabditis elegans. Moreover, the insecticidal activity of the mixture of 25-methyl and 25-ethyl ivermectin was 2.5-fold and 5.7-fold higher than that of milbemycin A3/A4 against C. elegans and the second-instar larva of Mythimna separate, respectively.ConclusionsTwo new avermectin derivatives 25-methyl and 25-ethyl ivermectin were generated by the domain swap of avermectin PKS. The enhanced insecticidal activity of 25-methyl and 25-ethyl ivermectin implied the potential use as insecticide in agriculture. Furthermore, the high yield and genetic stability of the engineered strains S. avermitilis AVE-T27 and AVE-H39 suggested the enormous potential in industrial production of the commercial insecticide ivermectin and 25-methyl/25-ethyl ivermectins, respectively.

【 授权许可】

CC BY   
© Zhang et al. 2015