局部黄热无氧杆菌TP-01嗜热果胶酶的部分纯化

Asni Putri, Galih Ayhusta Laras, Ismi Aulia Syafira, A. Baktir
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摘要

果胶酶是一种能水解果胶物质的酶。它在日常生活中有多种应用,例如在果汁厂,葡萄酒厂,纸浆和造纸厂,咖啡和茶厂使用发酵。果胶酶包括果胶裂解酶、果胶裂解酶和聚半乳糖醛酸酶。本研究的目的是用硫酸铵沉淀法和透析法对果胶酶蛋白进行部分纯化。利用晕区分析法对果胶酶活性进行初步测定。对硫酸铵进行了部分提纯,得到了40-90%硫酸铵组成的最佳工艺条件。采用DNS法测定果胶酶活性,Bradford法测定蛋白质浓度。黄热无氧杆菌TP-01的部分纯化率为70%。果胶酶活性降低是由于辅酶的缺失。综上所述,黄热无氧杆菌TP-01的果胶酶为聚半乳糖醛酸酶。
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Partial Purification of Thermophilic Pectinase from Local Isolate Anoxybacillus flavithermus TP-01
Pectinase is an enzyme that can hydrolyze substances of pectin. It has several applications in daily life, for example in juice factories, wine factories, pulp and paper factories, coffee and tea factories that use fermentation. Pectinase consists of pectate lyase, pectin lyase, and polygalacturonase. The aim of this research is to partially purify pectinase protein by ammonium sulfate precipitation and dialysis. A preliminary test of pectinase activity used halo zone analysis. The partial purification was needed to optimize ammonium sulfate and get the optimal condition in 40–90% of the ammonium sulfate composition. Pectinase activity was measured using DNS method, and the concentration of protein was measured using Bradford method. The result of this partial purification of Anoxybacillus flavithermus TP-01 was 70%. Decreasing od pectinase activity was resulted in this research due to the loss of cofactor. Based on the result, it can be concluded that the pectinase from Anoxybacillus flavithermus TP-01 was polygalacturonase.
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