title: In vivo investigation to the macrolide-glycosylating enzyme pair DesVII/DesVIII in Saccharopolyspora erythraea.
abstract: Glycosyltransferase DesVII and its auxiliary partner DesVIII from Streptomyces venezulae, homologs of EryCIII and EryCII in Saccharopolyspora erythraea, have previously been demonstrated to be flexible on their substrates in vitro. Herein, we investigated their in vivo function by interspecies complementation in the mutant strains of Sac. erythraea A226. As desVII and desVIII were concomitantly expressed in the ΔeryCIII mutant, the erythromycin A (Er-A) production was restored. Interestingly, co-expression of desVII and desVIII in the ΔeryBV mutant exhibited an increased Er-A yield by 15 % in comparison to A226. Hence, DesVII/DesVIII not only replaced EryCIII to upload D-desosamine to C5 position of 3-O-mycarosyl erythronolide B (MEB) but also in vivo attached L-mycarose, not D-desosamine to C3 position of erythronolide B (EB) with a higher activity than EryBV. Furthermore, expression of desVII in ΔeryCIII and ΔeryBV-CIII partially restored the Er-A production; however, no Er-A was detected while desVII was expressed in ΔeryBV. It was implicated that DesVII coupled with EryCII to form the DesVII/EryCII complex for attaching above two deoxysugars in the absence of EryCIII in Sac. erythraea. In addition, when desVII and desVIII were co-expressed in ΔeryBV-CII, Er-A was recovered with a lower yield than ΔeryBV-CIII. Our study presents an opportunity with Sac. erythraea as a cell factory for macrolide glycodiversification.
authors.affiliation: Institute of Health Sciences, School of Life Sciences, Anhui University, Hefei, 230601, China.
authors.forename: Hang
authors.initials: H
authors.lastname: Wu
authors.affiliation: Institute of Health Sciences, School of Life Sciences, Anhui University, Hefei, 230601, China.
authors.forename: Weiwei
authors.initials: W
authors.lastname: Li
authors.affiliation: Institute of Health Sciences, School of Life Sciences, Anhui University, Hefei, 230601, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Xin
authors.affiliation: Institute of Health Sciences, School of Life Sciences, Anhui University, Hefei, 230601, China.
authors.forename: Congming
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Institute of Health Sciences, School of Life Sciences, Anhui University, Hefei, 230601, China.
authors.forename: Yansheng
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Institute of Health Sciences, School of Life Sciences, Anhui University, Hefei, 230601, China.
authors.forename: Shaohua
authors.initials: S
authors.lastname: Ren
authors.affiliation: Institute of Health Sciences, School of Life Sciences, Anhui University, Hefei, 230601, China.
authors.forename: Min
authors.initials: M
authors.lastname: Ren
authors.affiliation: Institute of Health Sciences, School of Life Sciences, Anhui University, Hefei, 230601, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhao
authors.affiliation: Institute of Health Sciences, School of Life Sciences, Anhui University, Hefei, 230601, China.
authors.forename: Li
authors.initials: L
authors.lastname: Yuan
authors.affiliation: School of Life Sciences, Hefei Normal University, Hefei, 230601, China.
authors.forename: Zhongdong
authors.initials: Z
authors.lastname: Xu
authors.affiliation: School of Life Sciences, Hefei Normal University, Hefei, 230601, China.
authors.forename: Hualing
authors.initials: H
authors.lastname: Yuan
authors.affiliation: School of Life Sciences, Hefei Normal University, Hefei, 230601, China.
authors.forename: Ming
authors.initials: M
authors.lastname: Geng
authors.affiliation: Institute of Health Sciences, School of Life Sciences, Anhui University, Hefei, 230601, China. lzhang03@gmail.com.|CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China. lzhang03@gmail.com.
authors.forename: Lixin
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Institute of Health Sciences, School of Life Sciences, Anhui University, Hefei, 230601, China. David.weaver.t@gmail.com.
authors.forename: David T
authors.initials: DT
authors.lastname: Weaver
authors.affiliation: Institute of Health Sciences, School of Life Sciences, Anhui University, Hefei, 230601, China. zhbc@ahu.edu.cn.
authors.forename: Buchang
authors.initials: B
authors.lastname: Zhang
mesh_terms: D000900:Anti-Bacterial Agents / Q000378:metabolism*; D004917:Erythromycin / Q000378:metabolism*; D055786:Gene Knockout Techniques; D005816:Genetic Complementation Test; D016695:Glycosyltransferases / Q000235:genetics / Q000378:metabolism*; D016930:Saccharopolyspora / Q000201:enzymology* / Q000378:metabolism*; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism*
chemical_list: D000900:Anti-Bacterial Agents; D004917:Erythromycin; D016695:Glycosyltransferases
keywords: DesVII/DesVIII; EryBV; EryCII/EryCIII; Glycosyltransferase; Saccharopolyspora erythraea