title: A Novel Approach for Gene Expression Optimization through Native Promoter and 5' UTR Combinations Based on RNA-seq, Ribo-seq, and TSS-seq of Streptomyces coelicolor.
abstract: Streptomycetes are Gram-positive mycelial bacteria, which synthesize a wide range of natural products including over two-thirds of the currently available antibiotics. However, metabolic engineering in Streptomyces species to overproduce a vast of natural products are hampered by a limited number of genetic tools. Here, two promoters and four 5' UTR sequences showing constant strengths were selected based upon multiomics data sets from Streptomyces coelicolor M145, including RNA-seq, Ribo-seq, and TSS-seq, for controllable transcription and translation. A total eight sets of promoter/5' UTR combinations, with minimal interferences of promoters on translation, were constructed using the transcription start site information, and evaluated with the GusA system. Expression of GusA could be controlled to various strengths in three different media, in a range of 0.03- to 2.4-fold, compared to that of the control, ermE*P/Shine-Dalgarno sequence. This method was applied to engineer three previously reported promoters to enhance gene expressions. The expressions of ActII-ORF4 and MetK were also tuned for actinorhodin overproductions in S. coelicolor as examples. In summary, we provide a novel approach and tool for optimizations of gene expressions in Streptomyces coelicolor.
authors.forename: Jeong Sang
authors.initials: JS
authors.lastname: Yi
authors.forename: Min Woo
authors.initials: MW
authors.lastname: Kim
authors.forename: Minsuk
authors.initials: M
authors.lastname: Kim
authors.affiliation: Department of Biological Sciences and KI for the BioCentury, Korea Advanced Institute of Science and Technology , Daejeon 34141, Republic of Korea.
authors.forename: Yujin
authors.initials: Y
authors.lastname: Jeong
authors.forename: Eun-Jung
authors.initials: EJ
authors.lastname: Kim
authors.affiliation: Department of Biological Sciences and KI for the BioCentury, Korea Advanced Institute of Science and Technology , Daejeon 34141, Republic of Korea.|Intelligent Synthetic Biology Center, Daejeon 34141, Republic of Korea.
authors.forename: Byung-Kwan
authors.initials: BK
authors.lastname: Cho
authors.forename: Byung-Gee
authors.identifier: 0000-0002-3776-1001
authors.initials: BG
authors.lastname: Kim
mesh_terms: D020121:5' Untranslated Regions / Q000235:genetics*; D000880:Anthraquinones / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000096:biosynthesis; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics*; D001483:Base Sequence / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics*; D060847:Metabolic Engineering / Q000379:methods; D011401:Promoter Regions, Genetic / Q000235:genetics*; D014176:Protein Biosynthesis / Q000235:genetics; D012270:Ribosomes / Q000235:genetics; D048372:Streptomyces coelicolor / Q000235:genetics* / Q000378:metabolism; D024363:Transcription Initiation Site / Q000502:physiology; D014158:Transcription, Genetic / Q000235:genetics*; D000095622:Benzoisochromanequinones
chemical_list: D020121:5' Untranslated Regions; D000880:Anthraquinones; D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; C013390:actinorhodin; D000095622:Benzoisochromanequinones
keywords: Streptomyces coelicolor; antibiotics; metabolic engineering; promoter engineering; ribosome binding site