Lim Hee Seon, Cha In-Tae, Roh Seong Woon, Shin Hae-Hun, Seo Myung-Ji
Department of Life Sciences, Graduate School of Incheon National University, Incheon 22012, Republic of Korea.
Division of Bioengineering, Incheon National University, Incheon 22012, Republic of Korea.
J Microbiol Biotechnol. 2017 Mar 28;27(3):450-459. doi: 10.4014/jmb.1610.10008.
This study evaluated the effects of culture conditions, including carbon and nitrogen sources, L-monosodium glutamate (MSG), and initial pH, on gamma-aminobutyric acid (GABA) production by HYE1 isolated from kimchi, a Korean traditional fermented food. HYE1 was screened by the production analysis of GABA and genetic analysis of the glutamate decarboxylase gene, resulting in 14.64 mM GABA after 48 h of cultivation in MRS medium containing 1% (w/v) MSG. In order to increase GABA production by HYE1, the effects of carbon and nitrogen sources on GABA production were preliminarily investigated via one-factor-at-a-time optimization strategy. As the results, 2% maltose and 3% tryptone were determined to produce 17.93 mM GABA in modified MRS medium with 1% (w/v) MSG. In addition, the optimal MSG concentration and initial pH were determined to be 1% and 5.0, respectively, resulting in production of 18.97 mM GABA. Thereafter, response surface methodology (RSM) was applied to determine the optimal conditions of the above four factors. The results indicate that pH was the most significant factor for GABA production. The optimal culture conditions for maximum GABA production were also determined to be 2.14% (w/v) maltose, 4.01% (w/v) tryptone, 2.38% (w/v) MSG, and an initial pH of 4.74. In these conditions, GABA production by HYE1 was predicted to be 21.44 mM using the RSM model. The experiment was performed under these optimized conditions, resulting in GABA production of 18.76 mM. These results show that the predicted and experimental values of GABA production are in good agreement.
本研究评估了培养条件,包括碳源、氮源、L-谷氨酸钠(味精)和初始pH值,对从韩国传统发酵食品泡菜中分离出的HYE1产γ-氨基丁酸(GABA)的影响。通过GABA产量分析和谷氨酸脱羧酶基因的遗传分析筛选出HYE1,在含有1%(w/v)味精的MRS培养基中培养48小时后,GABA产量为14.64 mM。为了提高HYE1的GABA产量,通过单因素优化策略初步研究了碳源和氮源对GABA产量的影响。结果表明,在含有1%(w/v)味精的改良MRS培养基中,2%的麦芽糖和3%的胰蛋白胨可产生17.93 mM的GABA。此外,确定最佳味精浓度和初始pH值分别为1%和5.0,GABA产量为18.97 mM。此后,应用响应面法(RSM)确定上述四个因素的最佳条件。结果表明,pH值是影响GABA产量的最显著因素。最大GABA产量的最佳培养条件还确定为2.14%(w/v)麦芽糖、4.01%(w/v)胰蛋白胨、2.38%(w/v)味精和初始pH值4.74。在这些条件下,使用RSM模型预测HYE1的GABA产量为21.44 mM。在这些优化条件下进行实验,GABA产量为18.76 mM。这些结果表明,GABA产量的预测值与实验值吻合良好。