title: Engineering validamycin production by tandem deletion of γ-butyrolactone receptor genes in Streptomyces hygroscopicus 5008.
abstract: Paired homologs of γ-butyrolactone (GBL) biosynthesis gene afsA and GBL receptor gene arpA are located at different positions in genome of Streptomyces hygroscopicus 5008. Inactivation of afsA homologs dramatically decreased biosynthesis of validamycin, an important anti-fungal antibiotic and a critical substrate for antidiabetic drug synthesis, and the deletion of arpA homologs increased validamycin production by 26% (ΔshbR1) and 20% (ΔshbR3). By double deletion, the ΔshbR1/R3 mutant showed higher transcriptional levels of adpA-H (the S. hygroscopicus ortholog of the global regulatory gene adpA) and validamycin biosynthetic genes, and validamycin production increased by 55%. Furthermore, by engineering a high-producing industrial strain via tandem deletion of GBL receptor genes, validamycin production and productivity were enhanced from 19 to 24 g/L (by 26%) and from 6.7 to 9.7 g/L(-1) d(-1) (by 45%), respectively, which was the highest ever reported. The strategy demonstrated here may be useful to engineering other Streptomyces spp. with multiple pairs of afsA-arpA homologs.
authors.affiliation: State Key Laboratory of Microbial Metabolism, and School of Life Sciences & Biotechnology, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240, China; Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, School of Pharmaceutical Sciences, Wuhan University, Wuhan 430071, China.
authors.forename: Gao-Yi
authors.initials: GY
authors.lastname: Tan
authors.affiliation: State Key Laboratory of Microbial Metabolism, and School of Life Sciences & Biotechnology, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240, China.
authors.forename: Yao
authors.initials: Y
authors.lastname: Peng
authors.affiliation: State Key Laboratory of Microbial Metabolism, and School of Life Sciences & Biotechnology, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240, China.
authors.forename: Chenyang
authors.initials: C
authors.lastname: Lu
authors.affiliation: State Key Laboratory of Microbial Metabolism, and School of Life Sciences & Biotechnology, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240, China. Electronic address: bailq@sjtu.edu.cn.
authors.forename: Linquan
authors.initials: L
authors.lastname: Bai
authors.affiliation: State Key Laboratory of Microbial Metabolism, and School of Life Sciences & Biotechnology, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240, China; Shanghai Collaborative Innovation Center for Biomanufacturing Technology, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, China. Electronic address: jjzhong@sjtu.edu.cn.
authors.forename: Jian-Jiang
authors.initials: JJ
authors.lastname: Zhong
mesh_terms: D000935:Antifungal Agents / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics*; D017353:Gene Deletion; D005798:Genes, Bacterial; D007294:Inositol / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000235:genetics; D011963:Receptors, GABA-A / Q000235:genetics*; D013302:Streptomyces / Q000235:genetics / Q000378:metabolism
chemical_list: C037481:4-butyrolactone receptor; D000935:Antifungal Agents; D001426:Bacterial Proteins; D011963:Receptors, GABA-A; D007294:Inositol; C003749:validamycins
keywords: Antibiotic fermentation; Engineering; Secondary metabolism regulation; Streptomyces; Tandem deletion; γ-butyrolactone regulatory system