花生壳制备黑曲霉SCSGAF0145单宁酶的纯化及部分特性研究

O. Olaleye, M. Omotayo, Rosemary Meekness Oni, Tawakalitu Jumoke Orisadare
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引用次数: 0

摘要

单宁酶在工业上广泛应用于真菌利用富含单宁的农业废弃物作为底物分泌。以黑曲霉SCSGAF0145为底物,以花生壳为底物进行深层发酵分泌单宁酶。分泌的粗丹宁酶用硫酸铵沉淀出来,在Sephadex G-150上纯化。以单宁酸为底物,测定了分泌单宁酶的动力学参数。发酵后粗酶的生物量、单宁浓度、没食子酸浓度和单宁酶活性分别为81.00±0.08 mg、1.31±0.01 mg/mL、0.67±0.03µg/mL和6.04 U/mL。纯化倍数和纯化后的酶收率分别为13.44%和47.86%。纯化后的酶以单宁酸为唯一碳源,在pH为5.5、温度为35℃时活性最高,Km和Vmax分别为0.43 mol /L和21.55 U/mL。综上所述,黑曲霉SCSGAF0145以花生壳为单宁酸来源,分泌出高活性的单宁酶,可用于多种工业用途。
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Purification and Partial Characterization of Tannase produced by Aspergillus niger SCSGAF0145 using Arachis hypogaea (Groundnut) Shell
Tannase have been widely secreted industrially by fungi using tannin rich agricultural waste as substrate. Aspergillus niger SCSGAF0145, was used for the secretion of tannase using groundnut shell as substrate under submerged fermentation. The crude tannase secreted was precipitated out using ammonium sulphate and purified on Sephadex G-150. Kinetic parameters of the secreted tannase were determined using tannic acid as substrate. The estimated biomass weight, tannin concentration, gallic acid concentration and tannase activity of the crude enzyme after fermentation was 81.00±0.08 mg, 1.31±0.01 mg/mL, 0.67±0.03 µg/mL and 6.04 U/mL respectively. The purification fold and enzyme yield after purification were 13.44 and 47.86%. The purified enzyme showed maximal activity at a pH of 5.5 and temperature of 35oC while producing Km and Vmax of 0.43 moles/L and Vmax is 21.55 U/mL respectively using tannic acid as its sole carbon source. In conclusion, Aspergillus niger SCSGAF0145 secreted tannase of high activity using groundnut shell as source of tannic acid which could be utilized for various industrial purposes.
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