Zhang Xia, Zhao Wen-Xiao, Wang Yan, Cheng Jun-Hui, Bao Kai, He Jin, Chen Xiu-Lan
State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan, China.
State Key Laboratory of Microbial Technology, Marine Biotechnology Center, Shandong University, Qingdao, China.
J Sci Food Agric. 2023 Nov;103(14):7153-7163. doi: 10.1002/jsfa.12797. Epub 2023 Jul 18.
Marine bacteria secrete a variety of proteases, which are a good source to explore proteases with application value. However, only a few marine bacterial proteases with a potential in bioactive peptides preparation have been reported.
The metalloprotease A69 from the marine bacterium Anoxybacillus caldiproteolyticus 1A02591 was successfully expressed in the food safe bacterium Bacillus subtilis as a secreted enzyme. A technique to efficiently produce protease A69 in a 15-L bioreactor was established, with a production of 8988 U mL . Based on optimizing the hydrolysis parameters of A69 on soybean protein, a process for soybean protein peptides (SPs) preparation was set up, in which soybean protein was hydrolyzed by A69 at 4000 U g and 60 °C for 3 h. The prepared SPs had a high content (> 90%) of peptides with a molecular mass less than 3000 Da and contained 18 amino acids. The prepared SPs showed high angiotensin-converting enzyme (ACE)-inhibitory activity, with an IC value of 0.135 mg mL . Moreover, three ACE-inhibitory peptides, RPSYT, VLIVP and LAIPVNKP, were identified from the SPs using liquid chromatography-mass spectrometry analysis.
The marine bacterial metalloprotease A69 has a promising potential for preparing SPs with good nutritional and potential antihypertensive effects, laying a good foundation for its industrial production and application. © 2023 Society of Chemical Industry.
海洋细菌分泌多种蛋白酶，是探索具有应用价值蛋白酶的良好来源。然而，仅有少数具有制备生物活性肽潜力的海洋细菌蛋白酶被报道。
来自嗜热解蛋白嗜碱芽孢杆菌1A02591的金属蛋白酶A69在食品安全细菌枯草芽孢杆菌中成功作为分泌酶表达。建立了在15升生物反应器中高效生产蛋白酶A69的技术，产量为8988 U/mL。基于优化A69对大豆蛋白的水解参数，建立了大豆蛋白肽（SPs）的制备工艺，其中大豆蛋白在4000 U/g和60℃下用A69水解3小时。制备的SPs中分子量小于3000 Da的肽含量高（>90%），且含有18种氨基酸。制备的SPs显示出高的血管紧张素转换酶（ACE）抑制活性，IC值为0.135 mg/mL。此外，使用液相色谱-质谱分析从SPs中鉴定出三种ACE抑制肽，RPSYT、VLIVP和LAIPVNKP。
海洋细菌金属蛋白酶A69在制备具有良好营养和潜在降压作用的SPs方面具有广阔的潜力，为其工业化生产和应用奠定了良好基础。©2023化学工业协会。
