Study of peptides generated during milk fermentation of Lactobacilli and protease related to the peptide formation

乳酸杆菌乳发酵过程中产生的肽及与肽形成相关的蛋白酶的研究

• 批准号: 08660334
• 负责人: IGOSHI Keiji
• 金额: $ 1.54万
• 依托单位: Kyushu Tokai University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08660334/
• 关键词: yogurt; peptide; Lactobacillus delbruekii subsp bulgaricus; protease; fermented milk; ヨーグルト; カゼイン

The peptides generated during the skim milk fermentation of Lactobacillus delbrueckii subsp. bulgaricus(Lb. bulgaricus) and the protease which was related to the peptide formation were studied. 1, The peptides in the yogurt were examined by using the high performance liquid chromatography(HPLC). The primary structure of 22 peptides which were found in the yogurt was determined after the purification by HPLC.Most of determined peptides were from beta casein and some peptides were from kappa -casein. 2, The peptides generated during the skim milk fermentation of Lb bulgaricus were examined with the HPLC.The primary structure of the main peaks, which were found during fermentation, were cleared by automatic peptide sequencer. Most of them were from beta-casein and were located in two region of beta-casein, beta -casein(47-91) and beta-casein(193-207). 3, Protease were highly purified from the skim milk culture medium of Lb.bulgaricus. The protease was purified about 275 fold with a yield of 6.5 % using various chromatogray. The enzyme was most activity at pH 6.0, and had a temperature optimum at 35ﾟC.The enzyme was strongly inhibited by DlFP, and also inhibited by EDTA.The enzyme could degradedbeta-casein, but had no effect on as1- and k-casein. The many peptides originated from the beta- and alpha s2-casein were found as a result of action of casein by the enzyme.

德氏乳杆菌亚种脱脂奶发酵过程中产生的肽。对保加利亚乳杆菌（Lb. bulgaricus）以及与肽形成相关的蛋白酶进行了研究。 1、采用高效液相色谱法（HPLC）对酸奶中的肽进行检测。经HPLC纯化后，对酸奶中发现的22种肽进行了一级结构测定。测定的肽大部分来自β酪蛋白，部分肽来自κ-酪蛋白。 2、采用HPLC对保加利亚乳杆菌脱脂乳发酵过程中产生的肽进行检测，通过自动肽测序仪对发酵过程中发现的主峰进行一级结构分析。它们大多数来自β-酪蛋白，并且位于β-酪蛋白的两个区域：β-酪蛋白(47-91)和β-酪蛋白(193-207)。 3、蛋白酶是从保加利亚乳杆菌脱脂乳培养基中高度纯化的。使用各种色谱法将蛋白酶纯化约 275 倍，产率为 6.5%。该酶在pH 6.0时活性最强，最适温度为35℃。该酶受DlFP强烈抑制，也被EDTA抑制。该酶能降解β-酪蛋白，但对as1-和k-酪蛋白无影响。许多源自β-和α-s2-酪蛋白的肽是由于酶作用于酪蛋白而被发现的。