岩藻聚糖硫酸酯酶提取方法比较研究及响应面优化
The study of extraction methods for fucoidanase and optimization using response surface methodology
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摘要: 为了研究海洋细菌Flavobacteriaceae sp.CZ1127产胞内岩藻聚糖硫酸酯酶的高效提取方法,本研究比较了超声破碎、化学试剂法、溶菌酶法、渗透压作用、反复冻融等几种方法对Flavobacteriaceae sp.CZ1127胞内岩藻聚糖硫酸酯酶的提取效果,确定超声破碎法为最优方法。进一步通过单因素实验及响应面实验考察了超声破碎各因素对提取效果的影响,建立了预测超声破碎法提取岩藻聚糖硫酸酯酶酶活的数学模型,并对各因素条件进行了优化。最终确立的超声破碎提取最佳条件为:破碎功率600 W,菌悬液浓度20 mg/m L,破碎总时长9 min,单次破碎时长为6 s,间歇时长4 s。利用优化方法可从单位质量(g)菌体中获得岩藻聚糖硫酸酯酶酶活(1640.47±84.81)m U/g,提取效果较优化前提高了56.80%。Abstract: In order to obtain high efficiency methods of extracting fucoidanase from Flavobacteriaceae sp.CZ1127,this study compared several kinds of methods,and ultrasonic method was selected as the best method. The single factor experiments and response surface methodogy were adopted to search optimalizing conditions of extraction fucoidanase. And a second order quadratic equation was established. Results indicated that the optimalizing conditions of ultrasonic were as follows:ultrasonic power 600 W,bacterial concentration 20 mg/m L,total ultrasonic time 9 min,single ultrasonic time 6 s and internal time 4 s. After optimization,the enzyme activity of fucoidanase by ultrasonic method was improved to(1640.47±84.81) m U/g,and the enzyme activity increased by 56.80%.
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