Thermal adaptation of acetic acid bacteria for practical high-temperature vinegar fermentation.

Thermotolerant microorganisms are useful for high-temperature fermentation. Several thermally adapted strains were previously obtained from Acetobacter pasteurianus in a nutrient-rich culture medium, while these adapted strains could not grow well at high temperature in the nutrient-poor practical culture medium, "rice moromi." In this study, A. pasteurianus K-1034 originally capable of performing acetic acid fermentation in rice moromi was thermally adapted by experimental evolution using a "pseudo" rice moromi culture. The adapted strains thus obtained were confirmed to grow well in such the nutrient-poor media in flask or jar-fermentor culture up to 40 or 39 °C; the mutation sites of the strains were also determined. The high-temperature fermentation ability was also shown to be comparable with a low-nutrient adapted strain previously obtained. Using the practical fermentation system, "Acetofermenter," acetic acid production was compared in the moromi culture; the results showed that the adapted strains efficiently perform practical vinegar production under high-temperature conditions.

Mutated fabG gene encoding oxidoreductase enhances the cost-effective fermentation of jasmine rice vinegar in the adapted strain of Acetobacter pasteurianus SKU1108.

A low-nutrient adapted strain, Acetobacter pasteurianus G-40, was successfully obtained by repetitive cultivation of A. pasteurianus 7E-13 under selective pressure. The adapted strain could grow well and produce 3.45-fold higher amounts of acetic acid than 7E-13 in jasmine rice wine containing 6% ethanol at 37 °C in a shaking flask. The G-40 strain also exhibited higher amounts of acetic acid (5.16%) in 2-L jar fermentor compared with 7E-13, where the bio-conversion yield to acetic acid from ethanol was 71% and 55.5% in the adapted strain and parental strain, respectively. In addition, genome sequence analysis of G-40 revealed that the strain has mutations in the 6 genes, of which the fabG gene encoding oxidoreductase is largely mutated by the partial recombination with a highly homologous fabG homolog present in the large plasmid of the strain. Over-expression of the mutated fabG gene and also the replacement of the original fabG gene in the chromosome with the mutated one obviously enhanced growth and acetic acid production of 7E-13 in the rice wine without any nutrient supplementation, indicating that the mutation in the fabG gene is mainly involved in higher fermentation ability under low-nutrient conditions. Thus, the results suggest that the adapted G-40 strain has proven useful for the cost-effective fermentation of rice vinegar.