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Evaluation of the site-unspecified peptide identification method for proteolytic peptide mapping

机译：评价蛋白水解肽测绘的位点未指定的肽鉴定方法

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摘要

Proteases are widely used in the food industry to hydrolyze proteins and prepare bioactive peptides. Peptide mapping identification supports the application of proteases in the food industry. The site-specified peptide identification method, which was developed for site-specific proteases like trypsin, is relatively mature and reliable but cannot be applied using most industrial proteases with weak site specificity. To address this issue, the performance and reliability of the site-unspecified peptide identification method should be investigated and evaluated. In this study, tryptic hydrolysates of a single protein and a protein mixture were used to evaluate the site-unspecified identification method. The species origin of the hydrolyzed proteins was not specified in a database search, meaning that millions of protein sequences were included for calculating and matching. At least 98% of the tryptic peptides were successfully identified via the site-unspecified method, demonstrating that the site-unspecified method shows promising reliability. Moreover, the site-unspecified method identified more peptides than the site-specified method, including those from the low-frequency site-unspecific hydrolysis of trypsin, suggesting that the method has strong capabilities for peptide mapping. The results indicate the applicability of the site-unspecified peptide identification method in the study of site-unspecific industrial proteases.
机译：蛋白酶广泛用于食品工业中，以水解蛋白质并制备生物活性肽。肽测绘鉴定支持蛋白酶在食品工业中的应用。为胰蛋白酶等现场特异性蛋白酶开发的位点指定的肽鉴定方法是相对成熟和可靠的，但不能使用具有弱部位特异性的工业蛋白酶施用。为了解决这个问题，应研究和评估网站未指定的肽鉴定方法的性能和可靠性。在该研究中，使用单一蛋白质和蛋白质混合物的胰蛋白水解产物来评估位点未指定的鉴定方法。水解蛋白质的物种原产地未在数据库搜索中规定，这意味着包含数百万蛋白序列用于计算和匹配。通过现场未指定的方法成功鉴定了至少98％的胰蛋白肽，证明了站点未指定的方法显示有前途的可靠性。此外，该部位未指定的方法鉴定了比现场指定方法更多的肽，包括来自胰蛋白酶的低频部位 - 未特异性水解的方法，表明该方法具有肽映射的强能力。结果表明，位点未指定的肽鉴定方法在现场 - 非特异性产业蛋白酶研究中的适用性。

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