Production of Alkaline Protease Using Industrial Waste Effluent as Low-cost Fermentation Substrate

H. Zare, F. Meiguni, G. Najafpour
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Abstract

Alkaline proteases are the most important groups of commercial enzymes, which have been broadly used in industrial processes. In this study, Bacillus sp. PTCC 1538 was selected as a biological agent to produce alkaline protease. Enzyme production under submerge fermentation using industrial waste effluent was investigated. Since the costs of the raw material plays an important role in the cost of enzyme production, corn steep liquor (CSL) was selected as a low-cost substrate to reduce the cost of enzyme production. Various carbon sources were used as the auxiliary substrates to enhance enzyme production. Results showed that maximum enzyme activity was obtained when wheat bran was used as an auxiliary substrate. Optimal media composition and growth conditions for alkaline protease production were defined. The optimum conditions were found to be pH 8, incubation temperature of 37 °C, CSL inoculum size of 5 v/v %, yeast extract and wheat bran concentrations of 2 and 6 g/l, respectively. CaCl2 was used as an activator to enhance proteolytic activity of the enzyme. Under optimum condition, enzyme activity of 100.7 U/ml was obtained at CaCl2 concentration of 1.5 g/l.
利用工业废液作为低成本发酵底物生产碱性蛋白酶
碱性蛋白酶是最重要的一类商业酶,在工业生产中得到了广泛的应用。本研究选择芽孢杆菌PTCC 1538作为生产碱性蛋白酶的生物制剂。研究了利用工业废液进行水下发酵法产酶。由于原料成本在酶的生产成本中占有重要的地位,为了降低酶的生产成本,我们选择玉米浸泡液作为低成本的底物。利用多种碳源作为辅助底物促进酶的生成。结果表明,以麦麸为辅助底物时酶活性最高。确定了生产碱性蛋白酶的最佳培养基组成和生长条件。最佳条件为pH为8,培养温度为37℃,CSL接种量为5 v/v %,酵母膏和麦麸浓度分别为2和6 g/l。用CaCl2作为活化剂提高酶的蛋白水解活性。在最佳条件下,CaCl2浓度为1.5 g/l时,酶活为1000.7 U/ml。
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