Marukan Vinegar Co.

Kōbe-shi, Japan

Marukan Vinegar Co.

Kōbe-shi, Japan
SEARCH FILTERS
Time filter
Source Type

Ishii Y.,Kwansei Gakuin University | Akasaka N.,Marukan Vinegar Co. | Goda I.,Kwansei Gakuin University | Sakoda H.,Marukan Vinegar Co. | Fujiwara S.,Kwansei Gakuin University
Applied and Environmental Microbiology | Year: 2015

Acetoin in vinegar is an attractant to fruit flies when combined with acetic acid. To make vinegar more effective in attracting fruit flies with increased acetoin production, Komagataeibacter europaeus KGMA0119 was modified by specific gene disruption of the acetohydroxyacid isomeroreductase gene (ilvC). A previously constructed mutant lacking the putative ligand-sensing region in the leucine-responsive regulatory protein (KeLrp, encoded by Kelrp) was also used. The ilvC and Kelrp disruptants (KGMA5511 and KGMA7203, respectively) produced greater amounts of acetoin (KGMA5511, 0.11%; KGMA7203, 0.13%) than the wild-type strain KGMA0119 (0.069%). KGMA7203 produced a trace amount of isobutyric acid (0.007%), but the other strains did not. These strains produced approximately equal amounts of acetic acid (0.7%). The efficiency of fruit fly attraction was investigated with cultured Drosophila melanogaster. D. melanogaster flies (approximately 1,500) were released inside a cage (2.5mby 2.5mby 1.5 m) and were trapped with a device containing vinegar and a sticky sheet. The flies trapped on the sticky sheet were counted. The cell-free supernatant from KGMA7203 culture captured significantly more flies (19.36 to 36.96% of released flies) than did KGMA0119 (3.25 to 11.40%) and KGMA5511 (6.87 to 21.50%) cultures. Contrastingly, a 0.7% acetic acid solution containing acetoin (0.13%) and isobutyric acid (0.007%), which mimicked the KGMA7203 supernatant, captured significantly fewer flies (0.88 to 4.57%). Furthermore, the KGMA0119 supernatant with additional acetoin (0.13%) and isobutyric acid (0.007%) captured slightly more flies than the original KGMA0119 supernatant but fewer than the KGMA7203 supernatant, suggesting that the synergistic effects of acetic acid, acetoin, isobutyric acid, and unidentified metabolites achieved the efficient fly trapping of the KGMA7203 supernatant. © 2015, American Society for Microbiology.


Akasaka N.,Marukan Vinegar Co. | Ishii Y.,Kwansei Gakuin University | Hidese R.,Kwansei Gakuin University | Sakoda H.,Marukan Vinegar Co. | Fujiwara S.,Kwansei Gakuin University
Journal of Bioscience and Bioengineering | Year: 2014

Vinegar with increased amounts of branched-chain amino acids (BCAAs; valine, leucine and isoleucine) is favorable for human health as BCAAs decrease diet-induced obesity and hyperglycemia. To construct Gluconacetobacter europaeus which produces BCAAs, leucine responsive regulator (. GeLrp) is focused and two Gelrp mutants were constructed. Wild-type KGMA0119 didn't produce significant amount of valine (0.13mM) and leucine (0mM) and strain KGMA7110 which lacks complete Gelrp accumulated valine (0.48mM) and leucine (0.11mM) but showed impaired growth, and it was fully restored in the presence of essential amino acids. Strain KGMA7203 was then constructed with a nonsense mutation at codon Trp132 in the Gelrp, which leads a specific deletion at an estimated ligand-sensing region in the C-terminal domain. KGMA7203 produced greater quantities of valine (0.80mM) and leucine (0.26mM) and showed the same growth characteristics as KGMA0119. mRNA levels of BCAAs biosynthesis genes (. ilvI and ilvC) and probable BCAAs efflux pump (. leuE) were determined by quantitative reverse-transcription PCR. Expression rates of ilvI and ilvC in the two Gelrp disruptants were greater than those in KGMA0119. leuE was highly expressed in KGMA7110 only, suggesting that the accumulation in KGMA7110 culture was caused by increased expression of the biosynthesis genes and abnormal enhanced export of amino acids resulting in impaired cell growth. In contrast, KGMA7203 would achieve the high level production through enhanced expression of the biosynthesis genes without enhancing that for the efflux pump. KGMA7203 was considered advantageous for production of vinegar with higher amounts of valine and leucine. © 2014 The Society for Biotechnology, Japan.


Akasaka N.,Marukan Vinegar Co. | Sakoda H.,Marukan Vinegar Co. | Fujiwara S.,Kwansei Gakuin University
Seibutsu-kogaku Kaishi | Year: 2012

Comparative genomics suggested that Gluconacetobacter europaeus possesses a biosynthetic pathway for the production of branched-chain amino acids (BCAAs) similar to that in the well-studied BCAAproducing bacteria Corynebacterium glutamicum and Serratia marcescens. To obtain BCAA-producing Ga. europaeus, we attempted to design a screening system using Ga. europaeus M0119 isolated from rice vinegar. Alpha-amino butyric acid (ABA) is a well-known valine analogue. It inhibits growth by inducing feedback inhibition during BCAA biosynthesis. Ga. europaeus M0119 was treated with N-methyl-N'- nitro-N-nitrosoguanidine, and ABA resistant (ABAr) mutants were isolated. Almost all the ABAr mutants showed valine and leucine accumulation in the culture medium, whereas the wild-type strain did not. Among the mutants that were resistant to 50 mM ABA, strain ABAr1-56 showed the highest BCAA production. ABAr1-56 showed valine accumulation in the logarithmic phase (111 mg/l) and leucine accumulation in the stationary phase (20 mg/l). Addition of sodium lactate to the medium promoted BCAA production by ABAr1-56. These results suggest that Ga. europaeus is applicable as a BCAA producing bacterium.


Akasaka N.,Marukan Vinegar Co. | Sakoda H.,Marukan Vinegar Co. | Hidese R.,Kwansei Gakuin University | Ishii Y.,Kwansei Gakuin University | Fujiwara S.,Kwansei Gakuin University
Applied and Environmental Microbiology | Year: 2013

Gluconacetobacter europaeus, one of the microorganisms most commonly used for vinegar production, produces the unfavorable flavor compound acetoin. Since acetoin reduction is important for rice vinegar production, a genetic approach was attempted to reduce acetoin produced by G. europaeus KGMA0119 using specific gene knockout without introducing exogenous antibiotic resistance genes. A uracil-auxotrophic mutant with deletion of the orotate phosphoribosyltransferase gene (pyrE) was first isolated by positive selection using 5-fluoroorotic acid. The pyrE disruptant designated KGMA0704 (ΔpyrE) showed 5-fluoroorotic acid resistance. KGMA0704 and the pyrE gene were used for further gene disruption experiments as a host cell and a selectable marker, respectively. Targeted disruption of aldC or als, which encodes α-acetolactate decarboxylase or α-acetolactate synthase, was attempted in KGMA0704. The disruption of these genes was expected to result in a decrease in acetoin levels. A disruption vector harboring the pyrE marker within the targeted gene was constructed for double-crossover recombination. The cells of KGMA0704 were transformed with the exogenous DNA using electroporation, and genotypic analyses of the transformants revealed the unique occurrence of targeted aldC or als gene disruption. The aldC disruptant KGMA4004 and the als disruptant KGMA5315 were cultivated, and the amount of acetoin was monitored. The acetoin level in KGMA4004 culture was significantly reduced to 0.009% (wt/vol) compared with KGMA0119 (0.042% [wt/vol]), whereas that of KGMA5315 was not affected (0.037% [wt/vol]). This indicates that aldC disruption is critical for acetoin reduction. G. europaeus KGMA4004 has clear application potential in the production of rice vinegar with less unfavorable flavor. © 2013, American Society for Microbiology.


Akasaka N.,Marukan Vinegar Co. | Astuti W.,Kwansei Gakuin University | Ishii Y.,Kwansei Gakuin University | Hidese R.,Kwansei Gakuin University | And 2 more authors.
Journal of Bioscience and Bioengineering | Year: 2015

Plasmids pGE1 (2.5kb), pGE2 (7.2kb), and pGE3 (5.5kb) 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. © 2014 The Society for Biotechnology, Japan.


PubMed | Marukan Vinegar Co.. and Kwansei Gakuin University
Type: Journal Article | Journal: Journal of bioscience and bioengineering | Year: 2016

Vinegar produced by acetic acid bacteria is used as an attractant for fruit flies. Apple cider vinegar (ACV) and brown rice vinegar (BRV) are used as lures to detect Drosophila suzukii (also known as the spotted wing drosophila [SWD], a newly emerging invasive pest of soft-skinned fruits) and to capture Drosophila melanogaster, respectively. In the present study, we evaluated the attractiveness of BRV and ACV to SWD in laboratory trapping experiments using an upturned microcentrifuge tube with a pipette tip as a trap. We transferred SWD (approximately 20, 7-10 days old) to a glass vial containing a trap baited with BRV or ACV and counted the captured flies. BRV attracted more flies (52.889.75%) than ACV (35.787.47%) in 6h. Based on high-performance liquid chromatography, we found that BRV contained greater amounts of putrescine (12.360.44M) and spermidine (35.084.34M) than ACV (putrescine, 0.310.067M; spermidine, not detected). The attractiveness of ACV supplemented with putrescine (12M) and spermidine (35M) (68.564.69%) was significantly higher than that of ACV, indicating that the enhanced attractiveness of BRV to SWD was accomplished by the additive effects of polyamines and other known attractive volatiles, such as acetic acid and acetoin. BRV is expected to be a powerful tool for the efficient management of SWD.


PubMed | Marukan Vinegar Co. and Kwansei Gakuin University
Type: Journal Article | Journal: Journal of bioscience and bioengineering | Year: 2014

Vinegar with increased amounts of branched-chain amino acids (BCAAs; valine, leucine and isoleucine) is favorable for human health as BCAAs decrease diet-induced obesity and hyperglycemia. To construct Gluconacetobacter europaeus which produces BCAAs, leucine responsive regulator (GeLrp) is focused and two Gelrp mutants were constructed. Wild-type KGMA0119 didnt produce significant amount of valine (0.13 mM) and leucine (0 mM) and strain KGMA7110 which lacks complete Gelrp accumulated valine (0.48 mM) and leucine (0.11 mM) but showed impaired growth, and it was fully restored in the presence of essential amino acids. Strain KGMA7203 was then constructed with a nonsense mutation at codon Trp132 in the Gelrp, which leads a specific deletion at an estimated ligand-sensing region in the C-terminal domain. KGMA7203 produced greater quantities of valine (0.80 mM) and leucine (0.26 mM) and showed the same growth characteristics as KGMA0119. mRNA levels of BCAAs biosynthesis genes (ilvI and ilvC) and probable BCAAs efflux pump (leuE) were determined by quantitative reverse-transcription PCR. Expression rates of ilvI and ilvC in the two Gelrp disruptants were greater than those in KGMA0119. leuE was highly expressed in KGMA7110 only, suggesting that the accumulation in KGMA7110 culture was caused by increased expression of the biosynthesis genes and abnormal enhanced export of amino acids resulting in impaired cell growth. In contrast, KGMA7203 would achieve the high level production through enhanced expression of the biosynthesis genes without enhancing that for the efflux pump. KGMA7203 was considered advantageous for production of vinegar with higher amounts of valine and leucine.


PubMed | Marukan Vinegar Co.
Type: Journal Article | Journal: Applied and environmental microbiology | Year: 2013

Gluconacetobacter europaeus, one of the microorganisms most commonly used for vinegar production, produces the unfavorable flavor compound acetoin. Since acetoin reduction is important for rice vinegar production, a genetic approach was attempted to reduce acetoin produced by G. europaeus KGMA0119 using specific gene knockout without introducing exogenous antibiotic resistance genes. A uracil-auxotrophic mutant with deletion of the orotate phosphoribosyltransferase gene (pyrE) was first isolated by positive selection using 5-fluoroorotic acid. The pyrE disruptant designated KGMA0704 (pyrE) showed 5-fluoroorotic acid resistance. KGMA0704 and the pyrE gene were used for further gene disruption experiments as a host cell and a selectable marker, respectively. Targeted disruption of aldC or als, which encodes -acetolactate decarboxylase or -acetolactate synthase, was attempted in KGMA0704. The disruption of these genes was expected to result in a decrease in acetoin levels. A disruption vector harboring the pyrE marker within the targeted gene was constructed for double-crossover recombination. The cells of KGMA0704 were transformed with the exogenous DNA using electroporation, and genotypic analyses of the transformants revealed the unique occurrence of targeted aldC or als gene disruption. The aldC disruptant KGMA4004 and the als disruptant KGMA5315 were cultivated, and the amount of acetoin was monitored. The acetoin level in KGMA4004 culture was significantly reduced to 0.009% (wt/vol) compared with KGMA0119 (0.042% [wt/vol]), whereas that of KGMA5315 was not affected (0.037% [wt/vol]). This indicates that aldC disruption is critical for acetoin reduction. G. europaeus KGMA4004 has clear application potential in the production of rice vinegar with less unfavorable flavor.

Loading Marukan Vinegar Co. collaborators
Loading Marukan Vinegar Co. collaborators