title: Establishment of a novel gene expression method, BICES (biomass-inducible chromosome-based expression system), and its application to the production of 2,3-butanediol and acetoin.
abstract: In this study, we describe a novel method for producing valuable chemicals from glucose and xylose in Escherichia coli. The notable features in our method are avoidance of plasmids and expensive inducers for foreign gene expression to reduce production costs; foreign genes are knocked into the chromosome, and their expression is induced with xylose that is present in most biomass feedstock. As loci for the gene knock-in, lacZYA and some pseudogenes are chosen to minimize unexpected effects of the knock-in on cell physiology. The promoter of xylF is inducible with xylose and is combined with the T7 RNA polymerase-T7 promoter system to ensure strong gene expression. This expression system was named BICES (biomass-inducible chromosome-based expression system). As examples of BICES application, 2,3-butanediol and acetoin were successfully produced from glucose and xylose, and the maximal concentrations reached 54gL(-1) [99.6% in (R,S)-form] and 31gL(-1), respectively. 2,3-Butanediol and acetoin are industrially important chemicals that are, at present, produced primarily through petrochemical processes. To demonstrate usability of BICES in practical situations, we produced these chemicals from a saccharified cedar solution. From these results, we can conclude that BICES is suitable for practical production of valuable chemicals from biomass.
authors.affiliation: Bioproduction Research Institute, National Institute of Advanced Industrial Sciences and Technology (AIST), 2-17-2-1 Tsukisamu-Higashi, Toyohira-ku, Sapporo 062-8517, Japan; Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 2-12-1-M6-5 Ookayama, Meguro-ku, Tokyo 152-8550, Japan. Electronic address: nnakashima@bio.titech.ac.jp.
authors.forename: Nobutaka
authors.initials: N
authors.lastname: Nakashima
authors.affiliation: Biomass Refinery Research Center, National Institute of Advanced Industrial Sciences and Technology (AIST), 3-11-32 Kagamiyama, Higashi-Hiroshima, Hiroshima 739-0046, Japan.
authors.forename: Hironaga
authors.initials: H
authors.lastname: Akita
authors.affiliation: Bioproduction Research Institute, National Institute of Advanced Industrial Sciences and Technology (AIST), 2-17-2-1 Tsukisamu-Higashi, Toyohira-ku, Sapporo 062-8517, Japan; Biomass Refinery Research Center, National Institute of Advanced Industrial Sciences and Technology (AIST), 3-11-32 Kagamiyama, Higashi-Hiroshima, Hiroshima 739-0046, Japan.
authors.forename: Tamotsu
authors.initials: T
authors.lastname: Hoshino
mesh_terms: D000093:Acetoin / Q000378:metabolism*; D002072:Butylene Glycols / Q000378:metabolism*; D002876:Chromosomes, Bacterial / Q000235:genetics*; D004926:Escherichia coli / Q000502:physiology*; D029968:Escherichia coli Proteins / Q000502:physiology*; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D024861:Genetic Enhancement / Q000379:methods; D060847:Metabolic Engineering / Q000379:methods*; D015202:Protein Engineering / Q000379:methods*; D014994:Xylose / Q000378:metabolism
chemical_list: D002072:Butylene Glycols; D029968:Escherichia coli Proteins; C026978:2,3-butylene glycol; D014994:Xylose; D000093:Acetoin
keywords: 2,3-Butanediol; Acetoin; Butanediol dehydrogenase; Diacetyl acetoin reductase; T7 RNA polymerase; xylF