一株产壳聚糖酶的噬纤维菌分离及产酶发酵条件优化

生物技术通报 ›› 2013, Vol. 0 ›› Issue (8): 150-154.

一株产壳聚糖酶的噬纤维菌分离及产酶发酵条件优化

孙玉英^{1, 2}, 张继泉², 邬世昂¹

（1.淮海工学院海洋学院，连云港 222005；2.中国科学院海洋研究所，青岛 266071）

收稿日期:2013-03-03 修回日期:2013-08-11 出版日期:2013-08-11 发布日期:2013-09-02
作者简介:孙玉英，女，博士研究生，副教授，研究方向：微生物酶技术；E-mail：sunyuying125@163.com
基金资助:
中国博士后科学基金项目（2011M501169），江苏省海洋生物技术重点实验室开放课题（2009HS01）

Screening of a Chitosanase-producing Strain Cellulophaga sp. and Optimization of the Fermentation Conditions

Sun Yuying^{1, 2}, Zhang Jiquan², Wu Shiang¹

（1. Marine College, Huaihai Institute of Techology, Lianyungang 222005；2. Institute Oceanology, Chinese Academy of Sciences, Qingdao 266071）

Received:2013-03-03 Revised:2013-08-11 Published:2013-08-11 Online:2013-09-02

摘要/Abstract

摘要： 从连云港海域中筛选出一株产壳聚糖酶活力较高的菌株M5。根据其形态特征、生理生化以及16S rDNA鉴定，初步认定该菌为噬纤维菌属，命名为Cellulophaga sp. M5。通过单因素优化结合正交试验初步确定菌株M5产酶的培养基（g/L）及培养条件：粉末壳聚糖10，酵母提取物3，硫酸铵5，葡萄糖1.0，K₂HPO₄?3H₂O 1.4，NaCl 5，MgSO₄ ?7H₂O 1.4，陈海水1 L；发酵起始pH为 6.5，250 mL三角瓶中装70 mL培养基，1%接种量，30℃ 180 r/min培养84 h。经过优化Cellulophaga sp. M5发酵产酶由2.67 U/mL提高到6.67 U/mL，产酶活力提高了1.5倍。

关键词: 壳聚糖酶, 筛选, 鉴定, 发酵优化, 噬纤维菌属

Abstract: A chitosanase-producing bacterium M5 was isolated from Lianyungang sea area. It was identified as Cellulophaga sp. based on its morphological, physiological and biochemical properties and 16S rDNA sequence analysis and named Cellulophaga sp. M5. By single factors combination with orthogonal experimental, the optimum medium composition for producing chitosanase by Cellulophaga sp. M5 was as follows（g/L）：chitosan powder 10, yeast extract 3, ammonium sulfate 5, glucose 1.0, NaCl 5, MgSO₄ ?7H₂O 1.4, K₂HPO₄ ?3H₂O 1.4, the initial pH6.5. The optimum cultivation condition was to incubate in 250 mL flask installed with 70 mL medium of 1% inoculum size at 30℃, 180 r/min for 84 h. The maximal chitosanase activity could reach 6.67 U/mL when Cellulophaga sp. M5 was incubated in flask under the optimum cultural conditions. The activity increased by 1.5 times in comparision with initial activity of 2.67 U/mL.

Key words: Chitosanase Screening, Identification, Fermentation optimization, Cellulophaga

孙玉英, 张继泉, 邬世昂. 一株产壳聚糖酶的噬纤维菌分离及产酶发酵条件优化[J]. 生物技术通报, 2013, 0(8): 150-154.

Sun Yuying, Zhang Jiquan, Wu Shiang. Screening of a Chitosanase-producing Strain Cellulophaga sp. and Optimization of the Fermentation Conditions[J]. Biotechnology Bulletin, 2013, 0(8): 150-154.

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