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首页> 外文期刊>African Journal of Biotechnology >Comparison of three methods for determination of protein concentration in lactic acid bacteria for proteomics studies
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Comparison of three methods for determination of protein concentration in lactic acid bacteria for proteomics studies

机译：蛋白质组学研究中三种测定乳酸菌蛋白质浓度的方法的比较

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Finding the best method of cell lysis?and extraction of protein from the lysed cells?is the key step in detection and identification of?extra- and intra-cellular proteins in all applications of proteomics.?To develop an optimized?protein extraction protocol,Enterococcus?faecalis?V583,?Lactococcus?lactis?NIZO 0900 and?Pediococcus pentosaceus?OZF strains, respectively, belonging to each genus of?Enterococcus, Lactococcus?and?Pediococcus?were used as a representative cells in a study of lactic acid bacteria (LAB).?This report covers the use?and comparison?of threedifferent protein extraction methods including sonication, centrifugation and rupture by glass beads (FastPrep) to get a better understanding about which methods give better extract quality and higher amount of proteins when applied to one dimensional (1D) sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and for subsequent analysis by two dimensional (2D)-PAGE. The resultsclearly showed that,?all methods can be used to lyse LAB strains. However, a six fold greater amount of protein was obtained when FastPrep was applied to lyse LAB cells.?Our results also indicate that, this fast and easy extraction method allows more spot-abundant polyacrylamide gels. More?clear and consistent strips were detected by SDS-PAGE when proteins were extracted by FastPrep.?These results testify to the suitability of FastPrep protein extraction protocols for 2D proteomicstudies of representative strains of LAB.
机译：在蛋白质组学的所有应用中，寻找最佳的细胞裂解方法和从裂解细胞中提取蛋白质的方法是检测和鉴定细胞外和细胞内蛋白质的关键步骤。要开发优化的蛋白质提取方案，乳酸菌研究中，分别属于肠球菌，乳球菌和戊二球菌各属的粪肠球菌V583，乳酸乳球菌，NIZO 0900和戊糖小球菌OZF菌株分别作为代表性细胞。 （LAB）。本报告涵盖了三种不同的蛋白质提取方法的使用和比较，包括超声，离心和玻璃珠破裂（FastPrep），以更好地了解哪种方法在使用时可提供更好的提取质量和更多的蛋白质进行一维（1D）十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE），然后通过二维（2D）-PAGE分析。结果清楚地表明，所有方法均可用于裂解LAB菌株。但是，将FastPrep应用于裂解LAB细胞后，蛋白质的含量增加了六倍。我们的结果还表明，这种快速简便的提取方法可实现更多斑点丰富的聚丙烯酰胺凝胶。用FastPrep提取蛋白质时，通过SDS-PAGE检测出更清晰，一致的条带。这些结果证明FastPrep蛋白质提取方案适用于LAB代表性菌株的2D蛋白质组学研究。

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