Tehri Nimisha, Kumar Naresh, Raghu H V, Thakur Geetika, Kumar Sharma Pradip
Microbial Biosensors and Food Safety Laboratory, Dairy Microbiology Division, ICAR-National Dairy Research Institute, Karnal, Haryana 132001 India.
3 Biotech. 2017 Aug;7(4):259. doi: 10.1007/s13205-017-0897-0. Epub 2017 Jul 24.
The present study was undertaken with the objective to assess the effect of distinct stereoisomeric forms of nutrient germinants (selected sugars and amino acids) on the process of germination onset in dormant spores of MTCC 2949. In this respect, epimers of glucose and enantiomers of alanine were employed in current work. When supplemented with these stereoisomers, spores were found germinated only with d-glucose and d-mannose among epimers of glucose and only with l-alanine among enantiomers of alanine. Interestingly, germination in spores was observed to negligible extent with d-galactose and d-alanine. These findings were obtained on the basis of four type of germination assays, namely reduction in absorbance measured at 600 nm (≤5 to ≥30%), refractility examination (phase bright and dark), esterase assay [fluorescence units 0.455-94.62 (×10)] and fluorescent staining (fluorescent/non-fluorescent signals). Understanding of spores germination process and efficacy of different forms of germinants to trigger germination is of immense importance. It aids in development of sensing and sterilization indicating tools employing chiefly spores as biorecognition elements and in uncovering the mechanism of diseases, food contamination and spoilages resulting from the germination of spores. The findings of current work support the possibility to explore such germination mechanism by significantly giving the clue for potential existence of stereospecific receptor sites on the surface of spores. Perhaps, these sites can specifically differentiate and recognize stereoisomerically diverse forms of germinants for induction of germination.
本研究旨在评估营养萌发剂(选定的糖类和氨基酸)的不同立体异构形式对MTCC 2949休眠孢子萌发起始过程的影响。在这方面,本研究采用了葡萄糖的差向异构体和丙氨酸的对映体。当添加这些立体异构体时,发现孢子仅在葡萄糖差向异构体中与D-葡萄糖和D-甘露糖一起萌发,并且仅在丙氨酸对映体中与L-丙氨酸一起萌发。有趣的是,观察到孢子与D-半乳糖和D-丙氨酸的萌发程度可忽略不计。这些发现是基于四种类型的萌发试验获得的,即测量600nm处吸光度的降低(≤5%至≥30%)、折射检查(亮相和暗相)、酯酶测定[荧光单位0.455-94.62(×10)]和荧光染色(荧光/非荧光信号)。了解孢子萌发过程以及不同形式的萌发剂触发萌发的功效非常重要。它有助于开发主要以孢子作为生物识别元件的传感和杀菌指示工具,并有助于揭示由孢子萌发导致的疾病、食品污染和变质的机制。当前工作的发现支持了通过显著提供孢子表面立体特异性受体位点潜在存在的线索来探索这种萌发机制的可能性。也许,这些位点可以特异性地区分和识别立体异构形式不同的萌发剂以诱导萌发。