School of Biomedical and Health Sciences, Victoria University, Werribee Campus, PO Box 14428, Melbourne, Victoria 8001, Australia.
J Dairy Sci. 2013 Jan;96(1):40-9. doi: 10.3168/jds.2012-5761. Epub 2012 Oct 22.
The aim of this study was to investigate the effect of substitution of NaCl with KCl at different pH levels and salt concentrations on proteinase activity of cell-free extract and cell-free supernatant of the probiotics Lactobacillus acidophilus and Lactobacillus casei. de Man, Rogosa, and Sharpe broth aliquots were mixed with 2 pure salts (NaCl and KCl) and 2 salt concentrations at 2 concentration levels (5 and 10%), inoculated with Lactobacillus acidophilus or Lactobacillus casei, and incubated aerobically at 37°C for 22 h. The cultures were then centrifuged at 4,000×g for 30 min, and the collected cell pellets were used to prepare cell-wall proteinases and the supernatants used as a source of supernatant (extracellular) proteinases. The proteolytic activity and protein content of both portions were determined. After incubation of both portions with 3 milk caseins (α-, β-, κ-casein), the supernatants were individually subjected to analysis of angiotensin-converting enzyme (ACE)-inhibitory activity and proteolytic activity using the o-phthalaldehyde method. Significant differences were observed in ACE-inhibitory and proteolytic activities between salt substitution treatments of cell-free extract and cell-free supernatant from both probiotic strains at the same salt concentration and pH level.
本研究旨在探讨在不同 pH 值和盐浓度下,用 KCl 替代 NaCl 对嗜酸乳杆菌和干酪乳杆菌无细胞提取物和无细胞上清液中蛋白酶活性的影响。将 de Man、Rogosa 和 Sharpe 肉汤等分试样与 2 种纯盐(NaCl 和 KCl)和 2 种盐浓度(5%和 10%)混合,接种嗜酸乳杆菌或干酪乳杆菌,在 37°C 下有氧孵育 22 小时。然后将培养物在 4000×g 下离心 30 分钟,收集的细胞沉淀用于制备细胞壁蛋白酶,上清液用作上清液(细胞外)蛋白酶的来源。测定两部分的蛋白水解活性和蛋白含量。用 3 种牛奶酪蛋白(α-、β-、κ-酪蛋白)孵育两部分后,分别用邻苯二甲醛法分析上清液的血管紧张素转换酶(ACE)抑制活性和蛋白水解活性。在相同盐浓度和 pH 值下,两种益生菌菌株的无细胞提取物和无细胞上清液的盐替代处理之间,ACE 抑制活性和蛋白水解活性存在显著差异。