title: Genetic modification of Bacillus subtilis for production of D-chiro-inositol, an investigational drug candidate for treatment of type 2 diabetes and polycystic ovary syndrome.
abstract: D-chiro-inositol (DCI) is a drug candidate for the treatment of type 2 diabetes and polycystic ovary syndrome, since it improves the efficiency with which the body uses insulin and also promotes ovulation. Here, we report genetic modification of Bacillus subtilis for production of DCI from myo-inositol (MI). The B. subtilis iolABCDEFGHIJ operon encodes enzymes for the multiple steps of the MI catabolic pathway. In the first and second steps, MI is converted to 2-keto-MI (2KMI) by IolG and then to 3D-(3,5/4)-trihydroxycyclohexane-1,2-dione by IolE. In this study, we identified iolI encoding inosose isomerase, which converts 2KMI to 1-keto-D-chiro-inositol (1KDCI), and found that IolG reduces 1KDCI to DCI. Inactivation of iolE in a mutant constitutively expressing the iol operon blocked the MI catabolic pathway to accumulate 2KMI, which was converted to DCI via the activity of IolI and IolG. The mutant was able to convert at least 6% of input MI in the culture medium to DCI.
authors.affiliation: Department of Biofunctional Chemistry, Faculty of Agriculture, Kobe University, 1-1 Rokkodai, Kobe, Hyogo 657-8501, Japan. kenyoshi@kobe-u.ac.jp
authors.forename: Ken-ichi
authors.initials: K
authors.lastname: Yoshida
authors.forename: Masanori
authors.initials: M
authors.lastname: Yamaguchi
authors.forename: Tetsuro
authors.initials: T
authors.lastname: Morinaga
authors.forename: Maya
authors.initials: M
authors.lastname: Ikeuchi
authors.forename: Masaki
authors.initials: M
authors.lastname: Kinehara
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Ashida
mesh_terms: D001412:Bacillus subtilis / Q000235:genetics* / Q000378:metabolism*; D001483:Base Sequence; D001711:Biotransformation; D002851:Chromatography, High Pressure Liquid; D004269:DNA, Bacterial / Q000235:genetics; D003924:Diabetes Mellitus, Type 2 / Q000188:drug therapy; D015507:Drugs, Investigational; D005260:Female; D005798:Genes, Bacterial; D006801:Humans; D007294:Inositol / Q000096:biosynthesis* / Q000737:chemistry / Q000494:pharmacology; D009154:Mutation; D009876:Operon; D011085:Polycystic Ovary Syndrome / Q000188:drug therapy; D013237:Stereoisomerism; D035703:Therapies, Investigational
chemical_list: D004269:DNA, Bacterial; D015507:Drugs, Investigational; D007294:Inositol