title: Change in the plasmid copy number in acetic acid bacteria in response to growth phase and acetic acid concentration.
abstract: Plasmids pGE1 (2.5 kb), pGE2 (7.2 kb), and pGE3 (5.5 kb) were isolated from Gluconacetobacter europaeus KGMA0119, and sequence analyses revealed they harbored 3, 8, and 4 genes, respectively. Plasmid copy numbers (PCNs) were determined by real-time quantitative PCR at different stages of bacterial growth. When KGMA0119 was cultured in medium containing 0.4% ethanol and 0.5% acetic acid, PCN of pGE1 increased from 7 copies/genome in the logarithmic phase to a maximum of 12 copies/genome at the beginning of the stationary phase, before decreasing to 4 copies/genome in the late stationary phase. PCNs for pGE2 and pGE3 were maintained at 1-3 copies/genome during all phases of growth. Under a higher concentration of ethanol (3.2%) the PCN for pGE1 was slightly lower in all the growth stages, and those of pGE2 and pGE3 were unchanged. In the presence of 1.0% acetic acid, PCNs were higher for pGE1 (10 copies/genome) and pGE3 (6 copies/genome) during the logarithmic phase. Numbers for pGE2 did not change, indicating that pGE1 and pGE3 increase their PCNs in response to acetic acid. Plasmids pBE2 and pBE3 were constructed by ligating linearized pGE2 and pGE3 into pBR322. Both plasmids were replicable in Escherichia coli, Acetobacter pasteurianus and G. europaeus, highlighting their suitability as vectors for acetic acid bacteria.
authors.affiliation: Institute of Applied Microbiology, Marukan Vinegar Co. Ltd., 5-6 Koyo-cho West, Higashinada-ku, Kobe, Hyogo 658-0033, Japan.
authors.forename: Naoki
authors.initials: N
authors.lastname: Akasaka
authors.affiliation: Department of Bioscience, Graduate School of Science and Technology, Kwansei Gakuin University, 2-1 Gakuen, Sanda, Hyogo 669-1337, Japan.
authors.forename: Wiwik
authors.initials: W
authors.lastname: Astuti
authors.affiliation: Department of Bioscience, Graduate School of Science and Technology, Kwansei Gakuin University, 2-1 Gakuen, Sanda, Hyogo 669-1337, Japan.
authors.forename: Yuri
authors.initials: Y
authors.lastname: Ishii
authors.affiliation: Research Center for Environmental Bioscience, Graduate School of Science and Technology, Kwansei-Gakuin University, Sanda, Hyogo 669-1337, Japan.
authors.forename: Ryota
authors.initials: R
authors.lastname: Hidese
authors.affiliation: Institute of Applied Microbiology, Marukan Vinegar Co. Ltd., 5-6 Koyo-cho West, Higashinada-ku, Kobe, Hyogo 658-0033, Japan.
authors.forename: Hisao
authors.initials: H
authors.lastname: Sakoda
authors.affiliation: Department of Bioscience, Graduate School of Science and Technology, Kwansei Gakuin University, 2-1 Gakuen, Sanda, Hyogo 669-1337, Japan; Research Center for Environmental Bioscience, Graduate School of Science and Technology, Kwansei-Gakuin University, Sanda, Hyogo 669-1337, Japan. Electronic address: fujiwara-s@kwansei.ac.jp.
authors.forename: Shinsuke
authors.initials: S
authors.lastname: Fujiwara
mesh_terms: D019342:Acetic Acid / Q000378:metabolism* / Q000494:pharmacology; D000091:Acetobacter / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D018628:Gene Dosage / Q000187:drug effects; D005822:Genetic Vectors / Q000096:biosynthesis / Q000235:genetics; D041821:Gluconacetobacter / Q000187:drug effects / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*; D010957:Plasmids / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D019342:Acetic Acid
keywords: Acetic acid bacteria; Acetobacter pasteurianus; Gluconacetobacter europaeus; Plasmid copy number; Real-time quantitative PCR