一株产柚苷酶菌株的ARTP诱变育种及培养基的优化

ARTP Mutain Breeding of Naringinase-Producing Strains and Optimization of Fermentation Conditions

DOI：10.3969/j.issn.1673-1689.2018.09.010

中文关键词: 柚苷酶 互隔交链孢 常温室压等离子体 摇瓶发酵

英文关键词: Alternaria alternate,naringinase,atmospheric and room temperature,fermentation medium

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中文摘要:

利用常温室压等离子体技术对一株产柚苷酶菌株互隔交链孢霉（A. alternate）SK.37001进行快速诱变处理，获得一株高产柚苷酶菌株 A. alternate SK.37002，与野生菌株相比，该菌株发酵酶活提高了208%。通过单因素实验确定发酵产柚苷酶的最适碳源为蔗糖，最适诱导物为柚皮苷，最适氮源为硝酸钠，利用Plackett-Burman试验筛选出影响液态摇瓶发酵酶产量的3个重要因素：硝酸钠、氯化钙、磷酸氢二钾，在此基础上运用最陡爬坡实验逼近最大响应值区域，再利用Box-Behnken设计及响应面分析法进行回归分析，获得最佳培养基配方为：1 g/L柚皮苷，5 g/L蔗糖，5 g/L硝酸钠，0.8 g/L磷酸氢二钾，0.7 g/L氯化钙，0.5 g/L氯化钾，0.5 g/L硫酸镁。采用优化后的培养基进行摇瓶发酵产柚苷酶酶活为624.73 U/mL，与模型预测接近，发酵产酶量比优化前提高了90%。

英文摘要:

Atmospheric and room temperature plasma（ARTP） was used to mutagenizeAlternaria alternatewhich is a naringinase-producing strain. The naringinase activity of the mutagenized stain SK37.002 reached 327.32 U/mL，which was 208% higher than that of original strain.The optimum carbon sources for naringinase-producing is sucrose. The most suitable inducer is naringin and nitrogen sources being NaNO3. According to Plackett-Burman design，NaNO3，CaCl2 and K2HPO4 were screened out of 8 factors as main affecting variables of naringinase production. Then steepest ascent method was used to approach themaximum response regions and it was followed by Box-Behnken and response surface analysis. As a result，the optimum medium formula（g/L） determined was composed of naringin 1，sucrose 5，NaNO3 5，K2HPO4 0.8，CaCl2 0.7，KCl 0.5，MgCl2 0.5. The resulting naringinase activity in fermentation broth reached up to 624.73±30.33 U/mL and was 90% higher than before which was consistent with the predicted maximum value.

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