title: Construction of heterologous gene expression cassettes for the development of recombinant Clostridium beijerinckii.
abstract: Gene-expression cassettes for the construction of recombinant Clostridium beijerinckii were developed as potential tools for metabolic engineering of C. beijerinckii. Gene expression cassettes containing ColE1 origin and pAMB origin along with the erythromycin resistance gene were constructed, in which promoters from Escherichia coli, Lactococcus lactis, Ralstonia eutropha, C. acetobutylicum, and C. beijerinckii are examined as potential promoters in C. beijerinckii. Zymogram analysis of the cell extracts and comparison of lipase activities of the recombinant C. beijerinckii strains expressing Pseudomonas fluorescens tliA gene suggested that the tliA gene was functionally expressed by all the examined promoters with different expression level. Also, recombinant C. beijerinckii expressing C. beijerinckii secondary alcohol dehydrogenase by the constructed expression cassettes successfully produced 2-propanol from glucose. The best promoter for TliA expression was the R. eutropha phaP promoter while that for 2-propanol production was the putative C. beijerinckii pta promoter. Gene expression cassettes developed in this study may be useful tools for the construction of recombinant C. beijerinckii strains as host strains for the valuable chemicals and fuels from renewable resources.
authors.affiliation: Center for Bio-based Chemistry, Division of Convergence Chemistry, Korea Research Institute of Chemical Technology, 141 Gajeong-ro, Yuseong-gu, P.O. Box 107, Taejon, 305-600, Republic of Korea.
authors.forename: Young Hoon
authors.initials: YH
authors.lastname: Oh
authors.affiliation: Center for Industrial Chemical Biotechnology, Ulsan Research and Business Development Division, Korea Research Institute of Chemical Technology, 45 Jongga-ro, Jung-gu, Ulsan, 681-802, Republic of Korea.
authors.forename: Gyeong Tae
authors.initials: GT
authors.lastname: Eom
authors.affiliation: Center for Bio-based Chemistry, Division of Convergence Chemistry, Korea Research Institute of Chemical Technology, 141 Gajeong-ro, Yuseong-gu, P.O. Box 107, Taejon, 305-600, Republic of Korea.
authors.forename: Kyoung Hee
authors.initials: KH
authors.lastname: Kang
authors.affiliation: Center for Bio-based Chemistry, Division of Convergence Chemistry, Korea Research Institute of Chemical Technology, 141 Gajeong-ro, Yuseong-gu, P.O. Box 107, Taejon, 305-600, Republic of Korea.
authors.forename: Jeong Chan
authors.initials: JC
authors.lastname: Joo
authors.affiliation: Center for Bio-based Chemistry, Division of Convergence Chemistry, Korea Research Institute of Chemical Technology, 141 Gajeong-ro, Yuseong-gu, P.O. Box 107, Taejon, 305-600, Republic of Korea.
authors.forename: Young-Ah
authors.initials: YA
authors.lastname: Jang
authors.affiliation: Center for Bio-based Chemistry, Division of Convergence Chemistry, Korea Research Institute of Chemical Technology, 141 Gajeong-ro, Yuseong-gu, P.O. Box 107, Taejon, 305-600, Republic of Korea.|Department of Biological and Chemical Engineering, Hongik University, Chungnam, 339-701, Republic of Korea.
authors.forename: Jae Woo
authors.initials: JW
authors.lastname: Choi
authors.affiliation: Center for Bio-based Chemistry, Division of Convergence Chemistry, Korea Research Institute of Chemical Technology, 141 Gajeong-ro, Yuseong-gu, P.O. Box 107, Taejon, 305-600, Republic of Korea.
authors.forename: Bong Keun
authors.initials: BK
authors.lastname: Song
authors.affiliation: Department of Biotechnology and Bioengineering, College of Engineering, Chonnam National University, 77 Yongbong-ro, Buk-gu, Kwangju, 500-757, Republic of Korea. leesh@chonnam.ac.kr.
authors.forename: Seung Hwan
authors.initials: SH
authors.lastname: Lee
authors.affiliation: Department of Environmental Engineering and Energy, Myongji University, 116 Myongji-ro, Cheoin-gu, Yongin, Gyeonggi-do, 449-728, Republic of Korea. parksj93@mju.ac.kr.
authors.forename: Si Jae
authors.initials: SJ
authors.lastname: Park
mesh_terms: D046970:Clostridium beijerinckii / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D010957:Plasmids / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D011994:Recombinant Proteins
keywords: 2-Propanol; Clostridium beijerinckii; E. coli-Clostridia shuttle vector; Gene-expression cassettes; Lipase