Hong Hui, Chaplot Shreyak, Chalamaiah Meram, Roy Bimol C, Bruce Heather L, Wu Jianping
Department of Agricultural, Food and Nutritional Science, University of Alberta , Edmonton, Alberta T6G 2P5, Canada.
J Agric Food Chem. 2017 Aug 30;65(34):7491-7499. doi: 10.1021/acs.jafc.7b02319. Epub 2017 Aug 15.
The low-molecular-weight (LMW) peptides derived from collagen have shown a potential for various nutritional and pharmaceutical applications. However, production of LMW peptides from vertebrate collagen remains a challenge. Herein, we report a new method to produce LMW collagen peptides using pepsin pretreatment that removed cross-linked telopeptides in collagen molecules. After the pretreatment, the proportion of LMW collagen peptides (<1.4 kDa) that were obtained from pepsin-soluble collagen increased to 32.59% compared to heat-soluble collagen peptides (16.10%). Fourier transform infrared spectroscopy results indicated that telopeptide cleavage retained the triple-helical conformation of collagen. Liquid chromatography-tandem mass spectrometry analysis suggested that Gly-X-Y (X is often proline, while Y is either hydroxyproline or hydroxylysine) repeats were not the main factors that hindered the enzymatic hydrolysis of collagen molecules. However, cross-link quantification demonstrated that trivalent cross-links that included pyridinolines and pyrroles were the primary obstacles to producing small peptides from collagen of spent hens. This study demonstrated for the first time that removing cross-linked telopeptides could enhance the production of LMW peptides from spent hen collagen, which is also of interest to manufacturers who produce LMW collagen peptides from other vertebrate animals, such as bovids and porcids.
源自胶原蛋白的低分子量（LMW）肽已显示出在各种营养和制药应用中的潜力。然而，从脊椎动物胶原蛋白生产LMW肽仍然是一项挑战。在此，我们报告了一种使用胃蛋白酶预处理来生产LMW胶原蛋白肽的新方法，该预处理去除了胶原蛋白分子中的交联端肽。预处理后，与热溶性胶原蛋白肽（16.10%）相比，从胃蛋白酶可溶性胶原蛋白中获得的LMW胶原蛋白肽（<1.4 kDa）的比例增加到32.59%。傅里叶变换红外光谱结果表明，端肽切割保留了胶原蛋白的三螺旋构象。液相色谱-串联质谱分析表明，Gly-X-Y（X通常是脯氨酸，而Y是羟脯氨酸或羟赖氨酸）重复序列不是阻碍胶原蛋白分子酶解的主要因素。然而，交联定量表明，包括吡啶啉和吡咯的三价交联是从蛋鸡胶原蛋白生产小肽的主要障碍。这项研究首次证明，去除交联端肽可以提高从蛋鸡胶原蛋白中生产LMW肽的产量，这对于从其他脊椎动物如牛科动物和猪科动物生产LMW胶原蛋白肽的制造商来说也是有意义的。
