title: Design and application of a novel high-throughput screening technique for 1-deoxynojirimycin.
abstract: High-throughput screening techniques for small molecules can find intensive applications in the studies of biosynthesis of these molecules. A sensitive, rapid and cost-effective technique that allows high-throughput screening of endogenous production of the natural iminosugar 1-deoxynojirimycin (1-DNJ), an α-glucosidase inhibitor relevant to the pharmaceutical industry, was developed in this study, based on the inhibitory effects of 1-DNJ on the activity of the β-glycosidase LacS from Sulfolobus solfataricus. This technique has been demonstrated effective in engineering both the key enzyme and the expression levels of enzymes in the 1-DNJ biosynthetic pathway from Bacillus atrophaeus cloned in E. coli. Higher biosynthetic efficiency was achieved using directed evolution strategies.
authors.affiliation: CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Peixia
authors.initials: P
authors.lastname: Jiang
authors.affiliation: 1] CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China [2] University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Shanshan
authors.initials: S
authors.lastname: Mu
authors.affiliation: 1] CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China [2] University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Heng
authors.initials: H
authors.lastname: Li
authors.affiliation: CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Youhai
authors.initials: Y
authors.lastname: Li
authors.affiliation: CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Congmin
authors.initials: C
authors.lastname: Feng
authors.affiliation: Beijing Key Laboratory of Plant Resources Research and Development, Beijing Technology and Business University, Beijing 100048, China.
authors.forename: Jian-Ming
authors.initials: JM
authors.lastname: Jin
authors.affiliation: CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Shuang-Yan
authors.initials: SY
authors.lastname: Tang
mesh_terms: D017485:1-Deoxynojirimycin / Q000737:chemistry* / Q000378:metabolism; D001407:Bacillus / Q000201:enzymology; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D005959:Glucosidases / Q000037:antagonists & inhibitors / Q000378:metabolism; D057166:High-Throughput Screening Assays; D007700:Kinetics; D005810:Multigene Family; D016296:Mutagenesis; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D048229:Sulfolobus solfataricus / Q000201:enzymology
chemical_list: D001426:Bacterial Proteins; D004791:Enzyme Inhibitors; D017485:1-Deoxynojirimycin; D010088:Oxidoreductases; D005959:Glucosidases; C104851:Sulfolobus solfataricus beta-glycosidase