Shukla Pratik, Sakure Amar, Pipaliya Rinkal, Basaiawmoit Bethsheba, Maurya Ruchika, Bishnoi Mahendra, Kondepudi Kanthi Kiran, Hati Subrota
Dairy Microbiology Department, SMC College of Dairy Science, Anand, Gujarat, India.
Department of Plant Biotechnology, B.A College of Agriculture, Anand, Gujarat, India.
J Food Biochem. 2022 Dec;46(12):e14449. doi: 10.1111/jfbc.14449. Epub 2022 Oct 7.
The goal of this investigation was to find antidiabetic peptides and inhibit angiotensin converting enzyme (ACE) in Lacticaseibacillus paracasei (M11) fermented dromedary camel milk (Camelus dromedaries). According to the findings, the rate of antidiabetic activity increased along with the incubation periods and reached its peak after 48 hr of fermentation. The inhibitions of α-amylase, α-glucosidase, and lipase were 80.75, 59.62, and 65.46%, respectively. The inhibitory activity of ACE was 78.33%, and the proteolytic activity was 8.90 mg/mL. M11 at 0.25 mg/mL effectively suppressed LPS-induced pro-inflammatory cytokines and their mediators such as NO, TNF-α, IL-6, and IL-1β in RAW 264.7 cells. The rate of inoculum in the optimization phase was 1.5-2.5%, and the greatest proteolytic activity was observed after 48 hr of fermentation. The investigation of the above property in the ultrafiltered fermented milk exhibited the highest antidiabetic and ACE inhibition activities in the 3 kDa than 10 kDa fractions. The molecular weight was determined employing SDS-PAGE, and the six-peptide sequences were identified using 2D gel electrophoresis. Due to its high proteolytic activity, the L. paracasei strain has been reported to be useful in the production of ACE-inhibitory and antidiabetic peptides. Amino acid sequences such from ɑ1, ɑ2, and β-caseins have been identified within fermented camel milk by searching on online databases, including BIOPEP (for antidiabetic peptides) and AHTPDB (for hypertension peptides) to validate the antidiabetic and ACE-inhibitory actions of several peptides. PRACTICAL APPLICATIONS: The study aims to identify antidiabetic peptides and inhibit ACE in dromedary camel milk fermented with Lacticaseibacillus paracasei M11. Maximum antidiabetic and ACE-inhibitory actions of the fermented camel milk were observed in 3 kDa permeate fractions. Fermented camel milk significantly reduced the excessive TNF-α, IL-6, and IL-1β production in LPS-activated RAW 264.7 cells. RP-LC/MS was used to identify 6 bioactive peptides from dromedary fermented camel milk. This fermented camel milk could be used for the management of hypertension and diabetic related problems.
本研究的目的是在副干酪乳杆菌（M11）发酵的单峰驼奶（骆驼属单峰驼）中寻找抗糖尿病肽并抑制血管紧张素转换酶（ACE）。研究结果表明，抗糖尿病活性随着发酵时间的延长而增加，并在发酵48小时后达到峰值。α-淀粉酶、α-葡萄糖苷酶和脂肪酶的抑制率分别为80.75%、59.62%和65.46%。ACE的抑制活性为78.33%，蛋白水解活性为8.90mg/mL。0.25mg/mL的M11能有效抑制RAW 264.7细胞中LPS诱导的促炎细胞因子及其介质，如NO、TNF-α、IL-6和IL-1β。优化阶段的接种率为1.5-2.5%，发酵48小时后观察到最大的蛋白水解活性。对超滤发酵乳上述特性的研究表明，3kDa组分的抗糖尿病和ACE抑制活性高于10kDa组分。采用SDS-PAGE测定分子量，用二维凝胶电泳鉴定六个肽序列。据报道，由于其高蛋白水解活性，副干酪乳杆菌菌株可用于生产ACE抑制肽和抗糖尿病肽。通过在在线数据库（包括BIOPEP（用于抗糖尿病肽）和AHTPDB（用于高血压肽））中搜索，已在发酵驼奶中鉴定出来自α1、α2和β-酪蛋白的氨基酸序列，以验证几种肽的抗糖尿病和ACE抑制作用。实际应用：本研究旨在鉴定副干酪乳杆菌M11发酵的单峰驼奶中的抗糖尿病肽并抑制ACE。发酵驼奶的最大抗糖尿病和ACE抑制作用在3kDa渗透组分中观察到。发酵驼奶显著降低了LPS激活的RAW 264.7细胞中过量的TNF-α、IL-6和IL-1β的产生。采用RP-LC/MS从单峰驼发酵驼奶中鉴定出6种生物活性肽。这种发酵驼奶可用于治疗高血压和糖尿病相关问题。
