title: Re-engineering of genetic circuit for 2-deoxystreptamine (2-DOS) biosynthesis in Escherichia coli BL21 (DE3).
abstract: Various approaches for monocistronic constructions of genetic circuits have been designed for metabolite production but there has been no attempt to apply such methodology for aminoglycosides biosynthesis. Here, a simple and commercially available bio-part, despite the current trend focusing on the standardized BioBricks bio-parts available in the registry, is used. A 181-bp nucleotide fragment was designed for the efficient construction of an expression vector for monocistronic assembly of genes. Furthermore, a single vector with multi-monocistronic assembled genes for 2-deoxystreptamine (2-DOS) synthesis was constructed for production in engineered Escherichia coli. The working efficiency of model vector was concluded by reporter assay whereas the expressions of biosynthesis genes were confirmed by RT-PCR and SDS-PAGE. Production of 2-DOS was confirmed by TLC, LC-ELSD, and ESI-MS/MS.
authors.affiliation: Institute of Biomolecule Reconstruction, SunMoon University, 100 Kalsan-ri, Tangjeonmyun, Chungnam, Asansi, Republic of Korea.
authors.forename: Amit Kumar
authors.initials: AK
authors.lastname: Chaudhary
authors.forename: Je Won
authors.initials: JW
authors.lastname: Park
authors.forename: Yeo Joon
authors.initials: YJ
authors.lastname: Yoon
authors.forename: Byung-Gee
authors.initials: BG
authors.lastname: Kim
authors.forename: Jae Kyung
authors.initials: JK
authors.lastname: Sohng
mesh_terms: D002855:Chromatography, Thin Layer; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors; D006595:Hexosamines / Q000096:biosynthesis; D013058:Mass Spectrometry; D060847:Metabolic Engineering; D053858:Metabolic Networks and Pathways / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D006595:Hexosamines; C007265:2-deoxystreptamine