Laboratorio de Biología Molecular, Facultad de Farmacia y Bioquímica, Universidad Nacional Mayor de San Marcos, Lima 01, Peru.
Laboratory of Nanobiotechnology and Applied Microbiology, Department of Food Science, Federal University of Rio Grande do Sul, Porto Alegre, 91501-970, Brazil.
Arch Microbiol. 2024 Aug 14;206(9):377. doi: 10.1007/s00203-024-04109-x.
The high content and quality of protein in Andean legumes make them valuable for producing protein hydrolysates using proteases from bacteria isolated from extreme environments. This study aimed to carry out a single-step purification of a haloprotease from Micrococcus sp. PC7 isolated from Peru salterns. In addition, characterize and apply the enzyme for the production of bioactive protein hydrolysates from underutilized Andean legumes. The PC7 protease was fully purified using only tangential flow filtration (TFF) and exhibited maximum activity at pH 7.5 and 40 °C. It was characterized as a serine protease with an estimated molecular weight of 130 kDa. PC7 activity was enhanced by Cu (1.7-fold) and remained active in the presence of most surfactants and acetonitrile. Furthermore, it stayed completely active up to 6% NaCl and kept ̴ 60% of its activity up to 8%. The protease maintained over 50% of its activity at 25 °C and 40 °C and over 70% at pH from 6 to 10 for up to 24 h. The determined K and V were 0.1098 mg mL and 273.7 U mL, respectively. PC7 protease hydrolyzed 43%, 22% and 11% of the Lupinus mutabilis, Phaseolus lunatus and Erythrina edulis protein concentrates, respectively. Likewise, the hydrolysates from Lupinus mutabilis and Erythrina edulis presented the maximum antioxidant and antihypertensive activities, respectively. Our results demonstrated the feasibility of a simple purification step for the PC7 protease and its potential to be applied in industrial and biotechnological processes. Bioactive protein hydrolysates produced from Andean legumes may lead to the development of nutraceuticals and functional foods contributing to address some United Nations Sustainable Development Goals (SDGs).
安第斯豆类的高蛋白含量和质量使其成为使用从极端环境中分离的细菌蛋白酶生产蛋白质水解物的有价值原料。本研究旨在从秘鲁盐田分离的微球菌属 PC7 中进行单一纯化步骤。此外，对该酶进行表征并应用于生产未充分利用的安第斯豆类的生物活性蛋白水解物。仅使用切向流过滤（TFF）即可完全纯化 PC7 蛋白酶，其最大活性在 pH7.5 和 40°C。它被表征为一种丝氨酸蛋白酶，估计分子量为 130 kDa。PC7 活性被 Cu（提高 1.7 倍）增强，并且在存在大多数表面活性剂和乙腈的情况下仍保持活性。此外，它在高达 6%的 NaCl 中仍保持完全活性，并保持在 8%左右的活性约 60%。该蛋白酶在 25°C 和 40°C 下保持超过 50%的活性，在 pH6 到 10 范围内保持超过 70%的活性，持续 24 小时。确定的 K 和 V 分别为 0.1098 mg mL 和 273.7 U mL。PC7 蛋白酶分别水解 Lupinus mutabilis、Phaseolus lunatus 和 Erythrina edulis 蛋白浓缩物的 43%、22%和 11%。同样，Lupinus mutabilis 和 Erythrina edulis 的水解产物分别具有最大的抗氧化和降血压活性。我们的结果证明了 PC7 蛋白酶的简单纯化步骤的可行性，并且它可能适用于工业和生物技术过程。从安第斯豆类生产的生物活性蛋白水解物可能会导致开发营养保健品和功能性食品，有助于实现一些联合国可持续发展目标（SDG）。
