Thakur Kiran, Tomar Sudhir K, Wei Zhao-Jun
Dairy Microbiology Division, Indian Council of Agricultural Research - National Dairy Research InstituteKarnal, India; School of Food Science and Engineering, Hefei University of TechnologyHefei, China.
Dairy Microbiology Division, Indian Council of Agricultural Research - National Dairy Research Institute Karnal, India.
Front Microbiol. 2017 Mar 17;8:427. doi: 10.3389/fmicb.2017.00427. eCollection 2017.
With the aim to bioprospect potent riboflavin producing lactobacilli, the present study was carried out to evaluate the relative mRNA expression of riboflavin biosynthesis genes namely 1, 2, 3, and 4 from potent riboflavin producers obtained from our previous studies. All the four genes were successfully cloned and sequenced for further analysis by procedures. As studied by non-denaturing Polyacrylamide gel electrophoresis, no difference in size of all the four genes among those of various lactobacilli was observed. The relative fold increase in mRNA expression in 1, 2, 3, and 4 genes has been observed to be 10-, 1-, 0.7-, and 8.5-fold, respectively. Due to increase in relative mRNA expression for all the genes as well as phenotypic production attribute, KTLF1 strain was used further for expression studies in milk and whey. The fold increase in mRNA expression for all the four genes was higher at 12 and 18 h in milk and whey respectively. After exposure to roseoflavin, resistant variant of KTLF1 showed considerable increase in expression of all the targets genes. This is the first ever study to compare the mRNA expression of riboflavin biosynthesis pathway genes in lactobacilli and it also under lines the effect of media and harvesting time which significantly affect the expression of genes. The use of roseoflavin-resistant strains capable of synthesizing riboflavin in milk and whey paves a way for an exciting and economically viable biotechnological approach to develop novel riboflavin bio-enriched functional foods.
为了对产核黄素的潜在乳酸菌进行生物勘探,本研究旨在评估从我们之前的研究中获得的产核黄素能力较强的乳酸菌中核黄素生物合成基因1、2、3和4的相对mRNA表达。通过相关程序成功克隆并测序了所有这四个基因,以便进行进一步分析。通过非变性聚丙烯酰胺凝胶电泳研究发现,不同乳酸菌的这四个基因在大小上没有差异。观察到基因1、2、3和4的mRNA表达相对增加倍数分别为10倍、1倍、0.7倍和8.5倍。由于所有这些基因的相对mRNA表达以及表型生产属性都有所增加,KTLF1菌株被进一步用于牛奶和乳清中的表达研究。这四个基因的mRNA表达增加倍数在牛奶中于12小时时更高,在乳清中于18小时时更高。在接触玫瑰红菌素后,KTLF1的抗性变体显示所有目标基因的表达有显著增加。这是首次比较乳酸菌中核黄素生物合成途径基因的mRNA表达的研究,它还强调了培养基和收获时间对基因表达有显著影响。使用能够在牛奶和乳清中合成核黄素的玫瑰红菌素抗性菌株为开发新型核黄素生物强化功能食品开辟了一条令人兴奋且经济可行的生物技术途径。