Sisa Alisson, Sotomayor Cristina, Buitrón Lucía, Gómez-Estaca Joaquín, Martínez-Alvarez Oscar, Mosquera Mauricio
Department of Food Science and Biotechnology, Escuela Politécnica Nacional, Quito, P.O. Box 17-01-2759, Ecuador.
Institute of Food Science, Technology and Nutrition (ICTAN-CSIC), 6 José Antonio Novais St., 28040, Madrid, Spain.
Heliyon. 2023 Oct 10;9(10):e20735. doi: 10.1016/j.heliyon.2023.e20735. eCollection 2023 Oct.
This study presents an approach that utilizes low-value agro-industrial by-products as culture media for producing high-value proteolytic enzymes. The objective was to assess the impact of six agro-industrial by-products as culture media on the production of proteolytic enzymes. Bacillus subtilis strains, confirmed through comprehensive biochemical, morphological, and molecular analyses, were isolated and identified. Enzymatic activity was evaluated using azocasein and casein substrates, and the molecular sizes of the purified extract components were determined. The results demonstrated that the isolated bacteria exhibited higher metabolic and enzymatic activity when cultured in media containing 1 % soybean oil cake or feather meal. Furthermore, higher concentrations of the culture media were found to hinder the production of protease. Optimal protease synthesis on soybean oil cake and feather meal media was achieved after 4 days, using both the azocasein and casein methods. Semi-purification of the enzymatic extract obtained from Bacillus subtilis in feather meal and soybean oil cake resulted in a significant increase in azocaseinolytic and caseinolytic activities. Gel electrophoresis analysis revealed multiple bands in the fractions with the highest enzymatic activity in soybean oil cake, indicating the presence of various enzymes with varying molecular sizes. These findings highlight the potential of utilizing low-value agro-industrial by-products as efficient culture media for the sustainable and economically viable production of proteolytic enzymes with promising applications in various industries.
本研究提出了一种利用低价值农业工业副产品作为培养基来生产高价值蛋白水解酶的方法。目的是评估六种农业工业副产品作为培养基对蛋白水解酶生产的影响。通过全面的生化、形态学和分子分析确认的枯草芽孢杆菌菌株被分离和鉴定。使用偶氮酪蛋白和酪蛋白底物评估酶活性，并测定纯化提取物成分的分子大小。结果表明，在含有1%大豆油饼或羽毛粉的培养基中培养时，分离出的细菌表现出更高的代谢和酶活性。此外，发现较高浓度的培养基会阻碍蛋白酶的产生。使用偶氮酪蛋白法和酪蛋白法，在大豆油饼和羽毛粉培养基上培养4天后实现了蛋白酶的最佳合成。对从羽毛粉和大豆油饼中的枯草芽孢杆菌获得的酶提取物进行半纯化，导致偶氮酪蛋白分解和酪蛋白分解活性显著增加。凝胶电泳分析显示，大豆油饼中酶活性最高的组分中有多条带，表明存在各种分子大小不同的酶。这些发现突出了利用低价值农业工业副产品作为高效培养基来可持续且经济可行地生产蛋白水解酶的潜力，这些酶在各个行业都有广阔的应用前景。
