Singhvi Mamata, Zendo Takeshi, Iida Hiroshi, Gokhale Digambar, Sonomoto Kenji
Laboratory of Microbial Technology, Division of Systems Bioengineering, Department of Bioscience and Biotechnology, Faculty of Agriculture, Graduate School, Kyushu University, 6-10-1 Hakozaki, Higashi-Ku, Fukuoka 812-8581, Japan.
Laboratory of Zoology, Division of Agricultural Bioresource Sciences, Department of Bioresource Sciences, Faculty of Agriculture, Graduate School, Kyushu University, Fukuoka 812-8581, Japan.
J Biosci Bioeng. 2017 Dec;124(6):674-679. doi: 10.1016/j.jbiosc.2017.07.009. Epub 2017 Aug 8.
The present study revealed the effect of nitrogen sources on lactic acid production and stimulation of d- and l-lactate dehydrogenases (LDH) of parent Lactobacillus lactis NCIM 2368 and its mutant RM2-24 generated after UV mutagenesis. Both the parent and mutant strains were evaluated for d-lactic acid production in control and modified media. The modified media did not show remarkable effect on lactic acid production in case of parent whereas mutant exhibited significant enhancement in d-lactic acid production along with the appearance of l-lactic acid in the broth. Both LDH activities and specific activities were found to be higher in mutant than the parent strain. These results suggested that the diammonium hydrogen phosphate in modified media triggered the expression of LDH genes leading to enhanced lactic acid production. This observation has been proved by studying the expression levels of d- and l-LDH genes of parent and mutant in control and modified media using quantitative RT-PCR technique. In case of mutant, the transcriptional levels of d-LDH and l-LDH increased ∼17 fold and ∼1.38 fold respectively in modified medium compared to the values obtained with control medium. In case of parent, no significant change in transcriptional levels of d- and l-LDH was found when the cells were grown in either control medium or modified medium. This study suggested that the mutant, RM2-24 has l-LDH gene which is expressed in presence of (NH)HPO resulting in l-lactic acid production. Co-production of l-lactic acid in d-lactic acid fermentation may be detrimental in the PLA production.
本研究揭示了氮源对乳酸乳球菌NCIM 2368亲本菌株及其经紫外线诱变产生的突变体RM2-24的乳酸生成以及d-和l-乳酸脱氢酶(LDH)活性的影响。对亲本菌株和突变菌株在对照培养基和改良培养基中进行d-乳酸生成的评估。改良培养基对亲本菌株的乳酸生成未显示出显著影响,而突变菌株在肉汤中d-乳酸生成显著增加,同时出现了l-乳酸。发现突变体的LDH活性和比活性均高于亲本菌株。这些结果表明,改良培养基中的磷酸氢二铵触发了LDH基因的表达,从而导致乳酸生成增加。通过使用定量RT-PCR技术研究亲本菌株和突变菌株在对照培养基和改良培养基中d-和l-LDH基因的表达水平,证实了这一观察结果。对于突变体,与对照培养基相比,改良培养基中d-LDH和l-LDH的转录水平分别增加了约17倍和约1.38倍。对于亲本菌株,当细胞在对照培养基或改良培养基中生长时,d-和l-LDH的转录水平未发现显著变化。本研究表明,突变体RM2-24具有l-LDH基因,该基因在(NH₄)₂HPO₄存在的情况下表达,导致l-乳酸的产生。在d-乳酸发酵过程中共同产生l-乳酸可能对聚乳酸(PLA)的生产不利。