活性炭吸附固定化粪产碱杆菌青霉素G酰化酶

Immobilized Penicillin G Acylase from Alcaligenes faecalis with Activated Charcoal

目的:以活性炭为载体固定化粪产碱杆菌来源的青霉素G酰化酶,考察固定化酶的性质。方法:对影响酶固定化的因素优化筛选,确定有显著影响的因素:pH、离子强度、酶量、固定化时间进行L934的正交实验,获得最佳固定化条件,并对固定化酶的最适反应温度、pH及批次稳定性进行研究。结果:最佳固定化条件为:载体0.3g,酶量5mL,总反应体系为12mL,离子强度1mol/L,温度4℃,pH 7.0,固定化40h;最高固定化酶活性为135.9U/g湿载体。固定化酶性最适反应温度为55℃,最适pH为10,重复使用12次后没有活性损失。结论:活性炭吸附固定化青霉素G酰化酶的活性高,批次反应稳定,具有工业应用潜力。

Objective： Penicillin G acylase from Alcaligenes faecalis was immobilized with activated charcoal,and then study its properties.Method： By the optimization of the immobilized conditions using single factor,and chose four factors（immobilized time,enzyme volume,pH,ionic strength） for the orthogonal experiment.Finally,the optimum temperature,pH and batch cycle numbers of immobilized enzyme were studied.Results： The optimum conditions for the enzyme immobilization were： the ratio of carrier and enzyme was 3：50（g/V）,temperature 4℃,pH 7.0,ionic strength 1 mol/L and for 40 h.The optimum temperature and pH of the immobilized enzyme were 55℃ and pH 10 respectively,and no activity was lost after 12 cycles in the conversion of penicillin G to 6-aminopenicillanic acid.Conclusion： The immobilized enzyme had high activity and better batch cycle numbers,which could be potential to be used as industrial biocatalysts.