title: [Effect of ribosome engineering on butenyl-spinosyns synthesis of Saccharopolyspora pogona].
abstract: Through introducing mutations into ribosomes by obtaining spontaneous drug resistance of microorganisms, ribosome engineering technology is an effective approach to develop mutant strains that overproduce secondary metabolites. In this study, ribosome engineering was used to improve the yield of butenyl-spinosyns produced by Saccharopolyspora pogona by screening streptomycin resistant mutants. The yields of butenyl-spinosyns were then analyzed and compared with the parent strain. Among the mutants, S13 displayed the greatest increase in the yield of butenyl-spinosyns, which was 1.79 fold higher than that in the parent strain. Further analysis of the metabolite profile of S13 by mass spectrometry lead to the discovery of Spinosyn α1, which was absent from the parent strain. DNA sequencing showed that there existed two point mutations in the conserved regions of rpsL gene which encodes ribosomal protein S12 in S13. The mutations occurred a C to A and a C to T transversion mutations occurred at nucleotide pair 314 and 320 respectively, which resulted in the mutations of Proline (105) to Gultamine and Alanine (107) to Valine. It also demonstrated that S13 exhibited genetic stability even after five passages.
authors.forename: Lin'gen
authors.initials: L
authors.lastname: Luo
authors.forename: Yan
authors.initials: Y
authors.lastname: Yang
authors.forename: Hui
authors.initials: H
authors.lastname: Wei
authors.forename: Jie
authors.initials: J
authors.lastname: Rang
authors.forename: Qiong
authors.initials: Q
authors.lastname: Tang
authors.forename: Shengbiao
authors.initials: S
authors.lastname: Hu
authors.forename: Yunjun
authors.initials: Y
authors.lastname: Sun
authors.forename: Ziquan
authors.initials: Z
authors.lastname: Yu
authors.forename: Xuezhi
authors.initials: X
authors.lastname: Ding
authors.forename: Liqiu
authors.initials: L
authors.lastname: Xia
mesh_terms: D005818:Genetic Engineering; D018942:Macrolides / Q000378:metabolism*; D017354:Point Mutation; D012269:Ribosomal Proteins / Q000235:genetics; D012270:Ribosomes / Q000378:metabolism*; D016930:Saccharopolyspora / Q000378:metabolism*
chemical_list: D018942:Macrolides; D012269:Ribosomal Proteins; C034212:ribosomal protein S12