Phathanathavorn Theerisara, Naloka Kallayanee, Matsutani Minenosuke, Yakushi Toshiharu, Matsushita Kazunobu, Theeragool Gunjana
Department of Microbiology, Faculty of Science, Kasetsart University, Bangkok 10900, Thailand.
Graduate School of Science and Technology for Innovation, Yamaguchi University, Yamaguchi 753-8515, Japan.
J Biosci Bioeng. 2019 Jun;127(6):690-697. doi: 10.1016/j.jbiosc.2018.12.006. Epub 2019 Jan 22.
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.
通过在选择性压力下对巴氏醋酸杆菌7E-13进行重复培养,成功获得了一种低营养适应性菌株——巴氏醋酸杆菌G-40。在37℃的摇瓶中,该适应性菌株在含6%乙醇的茉莉米酒中能良好生长,产酸量比7E-13高3.45倍。与7E-13相比,G-40菌株在2-L罐式发酵罐中也表现出更高的醋酸含量(5.16%),其中适应性菌株和亲本菌株从乙醇到醋酸的生物转化率分别为71%和55.5%。此外,G-40的基因组序列分析表明,该菌株在6个基因中存在突变,其中编码氧化还原酶的fabG基因因与该菌株大质粒中存在的高度同源fabG同源物部分重组而发生大量突变。在米酒中不添加任何营养物质的情况下,突变型fabG基因的过表达以及用突变型fabG基因替换染色体上的原始fabG基因,明显增强了7E-13的生长和醋酸产量,这表明fabG基因的突变主要参与了低营养条件下较高的发酵能力。因此,结果表明适应性G-40菌株已被证明可用于米醋的经济高效发酵。