壳聚糖酶高产菌的筛选、鉴定与发酵产酶初探
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上海海洋大学博士启动基金项目(080219);上海海洋大学优秀青年教师基金(080236)


Screening, identification and fermentation of a chitosanaseproducing strain
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    摘要:

    壳聚糖酶(EC3.2.1.132)能催化壳聚糖分子中β1,4糖苷键的水解,以获得具有特殊生理活性的壳聚寡糖(聚氨基葡萄糖,聚合度2-10)。该酶在细菌、真菌等多种微生物中存在。采用透明圈法,以壳聚糖为唯一碳源,从11份虾蟹壳堆积的土壤中分离出51株能降解壳聚糖的菌株。经平板初筛、摇瓶发酵复筛以及产酶动力学研究,确定H2为产壳聚酶活力高、发酵时间短的优良菌株。根据其形态及培养特征、生理生化特性及16S rDNA序列分析,鉴定该菌株为浅玫瑰色链霉菌,并命名为(Streptomyces roseolus DH)。进一步的基本发酵条件研究显示:该菌株发酵最适温度为30 ℃,发酵液初始pH为7.2,最适碳源为1%胶体壳聚糖,最适氮源为0.5%蛋白胨。此条件下经60 h发酵,发酵液中壳聚糖酶活力可达(6.10±0.12) U/mL。此菌株产酶量高,发酵周期短,具有良好的应用潜力。

    Abstract:

    Chitosanase (EC3.2.1.132) catalyses the hydrolysis of the β-1,4 glycosidic bonds in chitosan to produce physiologically active chitooligosaccharides, polyglucosamines with 2 to 10 degrees of polymerization. It has been found in many microorganisms including bacteria and fungi. 51 chitosanaseproducing strains were screened and isolated from 11 soilsamples rich in shrimp and crab shells by transparent circles in the culture media containing chitosan as sole carbon source. After screening of flatpanel, flaskshaking cultivation and enzyme kinetics in fermentation, a strain (H2) with higher chitosanlytic activity was acquired. According to its morphological, cultural, biochemical and physiological characteristics and phylogenetic analysis of 16S rDNA, the strain was identified and named as Streptomyces roseolus DH. Further studies of fermentation conditions showed that the optimum temperature and initial pH of medium were 30 ℃ and 7.2, respectively. Moreover, the colloid chitosan (1.0%) and peptone (0.5%) were chosen as the best carbon and nitrogen sources among the tested ones. Under these conditions, the activity of chitosanase in media reached 6.10±0.12 U/mL after 60hour fermentation. This strain with higher yield of chitosanase and shorter fermentation period displayed good potential for future applications.

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魏福卫,蒋霞云,陈舜胜,陈道春,党培育.壳聚糖酶高产菌的筛选、鉴定与发酵产酶初探[J].上海海洋大学学报,2011,20(2):296-302.
WEI Fu-wei, JIANG Xia-yun, CHEN Shun-sheng, CHEN Dao-chun, DANG Pei-yu. Screening, identification and fermentation of a chitosanaseproducing strain[J]. Journal of Shanghai Ocean University,2011,20(2):296-302.

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