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Optimization of media composition and growth conditions for production of milk-clotting protease (MCP) from Aspergillus oryzae DRDFS13 under solid-state fermentation
Brazilian Journal of Microbiology ( IF 2.2 ) Pub Date : 2020-03-24 , DOI: 10.1007/s42770-020-00243-y
Jermen Mamo 1, 2 , Martin Kangwa 3 , Hector Marcelo Fernandez-Lahore 3 , Fassil Assefa 1
Affiliation  

This study reports the optimization of milk-clotting protease production from Aspergillus oryzae DRDFS13 under solid-state fermentation (SSF) in both one-variable-at-a-time and response surface methodology (RSM). The production and optimization of milk-clotting protease obtained from Aspergillus oryzae DRDFS13 under solid-state fermentation (SSF) using different agro-industrial wastes as solid substrates were studied. The agro-industrial wastes used included wheat bran, rice bran, pea bran, and grass pea bran. The chemical composition of the best solid substrate was tested using standard methods. Others cultivation parameters were studied, and the results showed that the optimum fermentation medium composed of wheat bran, casein (1% w/w), and glucose (0.5% w/w) and the conditions for maximum milk-clotting protease production were at the moisture content of 55.0%, inoculum of 0.5*106 spores/mL, incubation temperature of 30 °C, pH of 6.0, and fermentation time of 5 days. The highest milk-clotting activity was obtained from the crude enzyme extracted using 0.1 M NaCl and partial purification of the crude enzyme using chilled acetone, and 80% (NH4)2SO4 increased the ratio of MCA/PA from 0.56 to 1.30 and 0.65, respectively. Moreover, the highest MCA (137.58 U/mL) was obtained at a casein concentration of 0.5%, pH 4.0, and 25 °C, using RSM. Thus, results from the present study showed that the optimization of milk-clotting protease production from A. oryzae DRDFS 13 under SSF by both one-variable-at-a-time and RSM significantly increased the milk-clotting activity. This is the first report from a fungus in the Ethiopian setting and a modest contribution to highlight the potential of harnessing microbial protease enzymes for industrial applications.

中文翻译:

米曲霉DRDFS13固态发酵生产凝乳蛋白酶(MCP)培养基组成和生长条件的优化

本研究报告了在固态发酵 (SSF) 下,米曲霉 DRDFS13 的凝乳蛋白酶生产在一次一个变量和响应面方法 (RSM) 中的优化。以不同农工业废弃物为固体底物,研究了米曲霉DRDFS13在固态发酵(SSF)下的凝乳蛋白酶的生产和优化。使用的农工业废弃物包括麦麸、米糠、豌豆麸和草豌豆麸。使用标准方法测试最佳固体基材的化学成分。对其他培养参数进行了研究,结果表明,最佳发酵培养基由麦麸、酪蛋白(1% w/w)和葡萄糖(0. 5% w/w),最大凝乳蛋白酶产量的条件是水分含量为 55.0%,接种量为 0.5*106 个孢子/mL,孵育温度为 30°C,pH 为 6.0,发酵时间为 5天。使用 0.1 M NaCl 提取的粗酶和使用冷冻丙酮对粗酶进行部分纯化获得了最高的凝乳活性,80% (NH4)2SO4 使 MCA/PA 的比率分别从 0.56 增加到 1.30 和 0.65 . 此外,使用 RSM 在酪蛋白浓度为 0.5%、pH 为 4.0 和 25°C 时获得了最高的 MCA (137.58 U/mL)。因此,本研究的结果表明,在 SSF 下,通过一次一个变量和 RSM 优化米曲霉 DRDFS 13 的凝乳蛋白酶生产显着提高了凝乳活性。
更新日期:2020-03-24
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