红海沿岸Streptomyces sp.H1深层培养产碱性蛋白酶的条件优化

Hind A. A. Al-Zahrani
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引用次数: 1

摘要

从红海沿岸土壤中分离到一株产碱性蛋白酶的强效菌株Streptomyces sp. H1。该酶在细胞外的深层培养中产生,在固定期早期达到最高水平。碱性蛋白酶在培养时间、pH和接种量分别为3 d、9%和8%时活性最高。在不同碳源中,甜菜糖蜜的产量最大,其次是淀粉、蔗糖和果糖。以酪蛋白、蛋白胨、酵母浸膏和硫酸铵为主要原料,获得了较高的蛋白酶产量。以7g/l NaCl为最佳处理条件,可提高蛋白酶水平。优化工艺参数后,碱性蛋白酶的产量提高了约3.4倍。在50%的饱和度下,用硫酸铵进行分级沉淀,实现了粗酶的部分纯化。由于在较便宜的底物(如甜菜糖蜜)存在下蛋白酶的最大产量,在碱性pH值9和温度高达70℃下的稳定性以及耐盐性使该菌株及其酶在不同的工业应用中很有用。
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Optimization conditions of alkaline protease production by Streptomyces sp.H1 isolated from red sea coastal region in submerged culture
A potent alkaline protease producing strain characterized and identified as Streptomyces sp. H1 was isolated from soil around red sea shore. The enzyme was produced extracellulary in submerged culture revealed maximum level during early stationary phase. Alkaline protease showed the highest activity at incubation time, pH and inoculum size of 3 days, 9 and 8% respectively. Among different carbon sources beet molasses gave a maximum production followed by starch, sucrose and fructose. High yield of protease production was noticed with casein followed by peptone, yeast extract and ammonium sulphate. Furthermore, it was optimized with 7g/l NaCl resulted in higher level of protease. Optimization of the process parameters resulted in about 3.4 fold increase in the alkaline protease. Partial purification of the crude enzyme was achieved by fractional precipitation using ammonium sulfate at 50% saturation. Due to the maximum production of protease in the presence of cheaper substrate as beet molasses, stability at alkaline pH 9 and temperature up to 70 oC besides salt tolerance make the strain and its enzyme useful in different industrial applications.  
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