Identification and Heterologous Reconstitution of a 5-Alk(en)ylresorcinol Synthase from Endophytic Fungus Shiraia sp. Slf14

Authors

Huiwen Yan, Utah State University
Lei Sun, Utah State UniversityFollow
Jinge Huang, Utah State UniversityFollow
Yixing Qiu, Utah State University
Fuchao Xu, Utah State UniversityFollow
Riming Yan, Utah State University
Du Zhu, Jiangxi Normal University
Wei Wang, Hunan University of Chinese Medicine
Jixun Zhan, Utah State UniversityFollow

Document Type

Article

Journal/Book Title

Journal of Microbiology

Publication Date

10-24-2018

Publisher

The Microbiological Society of Korea

Volume

56

Issue

11

Abstract

A new type III polyketide synthase gene (Ssars) was discovered from the genome of Shiraia sp. Slf14, an endophytic fungal strain from Huperzia serrata. The intron-free gene was cloned from the cDNA and ligated to two expression vectors pET28a and YEpADH2p-URA3 for expression in Escherichia coli BL21(DE3) and Saccharomyces cerevisiae BJ5464, respectively. SsARS was efficiently expressed in E. coli BL21(DE3), leading to the synthesis of a series of polyketide products. Six major products were isolated from the engineered E. coli and characterized as 1,3-dihydroxyphenyl-5-undecane, 1,3-dihydroxyphenyl-5-cis-6'-tridecene,1,3-dihydroxyphenyl-5-tridecane, 1,3-dihydroxyphenyl-5-cis-8'-pentadecene, 1,3-dihydroxyphenyl-5-pentadecane and 1,3-dihydroxyphenyl-5-cis-10'-heptadecene, respectively, based on the spectral data and biosynthetic origin. Expression of SsARS in the yeast also led to the synthesis of the same polyketide products, indicating that this enzyme can be reconstituted in both heterologous hosts. Supplementation of soybean oil into the culture of E. coli BL21(DE3)/SsARS increased the production titers of 1-6 and led to the synthesis of an additional product, which was identified as 5-(8'Z,11'Z-heptadecadienyl)resorcinol. This work thus allowed the identification of SsARS as a 5-alk(en)ylresorcinol synthase with flexible substrate specificity toward endogenous and exogenous fatty acids. Desired resorcinol derivatives may be synthesized by supplying corresponding fatty acids into the culture medium.

First Page

1

Last Page

29

Comments

This is a post-peer-review, pre-copyedit version of an article published in Journal of Microbiology. The final authenticated version is available online at: http://dx.doi.org/10.1007/s12275-018-8278-x

Recommended Citation

Yan, H., Sun, L., Huang, J. et al. J Microbiol. (2018) 56: 805. https://doi.org/10.1007/s12275-018-8278-x