Optimization of Nutritional Parameters by One-Factor-At-A-Time Method for the Biosynthesis of Alkaline Protease from the Isolated Strain Alternaria alternata TUSGF1

Tapasi Polley, U. Ghosh
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Abstract

Alkaline proteases are essential enzymes that have several applications in our industry. The aim was optimization of nutritional parameters by one-factor-at-a-time (OFAT) method in solid-state fermentation. Production of protease by employing our laboratory's new isolate, Alternaria alternata TUSG1 (strain accession number- MF401426) under solid-state fermentation was optimized. The nutritional factors were investigated, and only one agricultural residue (cauliflower leaves) with different particle sizes was checked. Highest enzyme production was obtained with a medium particle size of cauliflower leaves (610 U/gds) followed by coarse waste (603U/gds) and fine waste (596 U/gds) using 106 spores/ml as inoculum at 30° C for 7 days. The organism utilized carbon sources 0.5% (w/w) dextrose, fructose, maltose, sucrose, lactose, and starch. Among them, maltose was found to be the best carbon source. A variety of inorganic and organic media components were investigated for nitrogen sources of 0.3% (w/w), and skim milk was turned out to the best. The maximum enzyme activity was obtained with 1% maltose, 0.5% skim milk and 0.05% MgSO4. With optimized media 1.53 fold increase in the protease production at agricultural residue cauliflower leaves was obtained.
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单因子单次法优化菌株TUSGF1合成碱性蛋白酶的营养条件
碱性蛋白酶是一种重要的酶,在工业上有多种用途。采用单因素法(OFAT)对固态发酵过程中的营养参数进行优化。利用本实验室新分离菌株alternnaria alternataTUSG1(菌株编号:MF401426)在固态发酵条件下优化蛋白酶的生产。研究了营养因子,只检测了一种不同粒径的农用残渣(花椰菜叶)。以中等粒径的菜叶(610 U/gds)为酶产率最高,其次是粗废(603U/gds)和细废(596 U/gds),接种量为106孢子/ml,在30°C下接种7天。该生物利用0.5% (w/w)葡萄糖、果糖、麦芽糖、蔗糖、乳糖和淀粉作为碳源。其中,麦芽糖是最佳碳源。以0.3% (w/w)为氮源,考察了多种无机和有机培养基组分,以脱脂乳为最佳。在1%麦芽糖、0.5%脱脂牛奶和0.05% MgSO4的条件下酶活性最高。优化后的培养基可使农用菜叶蛋白酶产量提高1.53倍。
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