食品科学

• 生物工程 • 上一篇    下一篇

1 株产壳聚糖酶细菌的分离、鉴定和发酵条件优化

冯志彬1,薛 钰1,陈国忠1,张 娟2,罗 乐1,程仕伟1   

  1. 1.鲁东大学生命科学学院,山东 烟台 264025;2.鲁东大学农学院,山东 烟台 264025
  • 出版日期:2016-10-15 发布日期:2016-12-01

Isolation, Identification of Bacillus mojavensis amyP216 and Optimization of Its Fermentation Conditions for Enhanced Chitosanase Production

FENG Zhibin1, XUE Yu1, CHEN Guozhong1, ZHANG Juan2, LUO Le1, CHENG Shiwei1   

  1. 1. College of Life Science, Ludong University, Yantai 264025, China;
    2. College of Agriculture, Ludong University, Yantai 264025, China
  • Online:2016-10-15 Published:2016-12-01

摘要:

从烟台海岸带沙质土壤中分离筛选壳聚糖酶产生菌株,对其进行分类鉴定,优化其发酵条件,为酶法生产壳寡糖提供技术依据。通过透明圈法初步判断产酶能力,液体发酵复筛测定酶活力,筛选到酶活力达36.20 U/mL的菌株amyP216。通过菌体形态、菌落特征、生理生化及16S rDNA序列分析鉴定菌株amyP216为莫哈韦芽孢杆菌(Bacillus mojavensis)。通过单因素和正交试验确定最佳发酵条件为:胶体壳聚糖添加量20 g/L、酵母浸粉添加量12.5 g/L、吐温-80添加量1.0 g/L、初始发酵pH 6.0、发酵温度30 ℃。在最优条件下利用5 L自控发酵罐培养45 h壳聚糖酶活力可达到80.60 U/mL,较优化前(36.20 U/mL)提高了1.2 倍。莫哈韦芽孢杆菌amyP216是一株壳聚糖酶活力较高的生产菌株,具有潜在的研究和应用价值。

关键词: 莫哈韦芽孢杆菌, 壳聚糖酶, 壳寡糖, 分离鉴定, 发酵条件优化

Abstract:

Bacterial strains were isolated from coastal sandy soil collected in Yantai, Shandong province, and were initially
screened to obtain 5 chitosanase-producing strains by transparent circle method. Further secondary screening provided a
strain producing the highest chitosanase activity named amyP216. The strain amyP216 was identified as Bacillus mojavensis
based on morphological, physiological and biochemical characteristics and 16S rDNA sequence analysis. The fermentation
conditions of amyP216 for chitosanase production were optimized by one-factor-at-a-time and orthogonal array designs.
The results showed that the maximum chitosanase activity of 80.60 U/mL, which was increased by 2.2 folds as compared
to that before optimization (36.20 U/mL), was obtained when the fermentation was carried out at 30 ℃ for 45 h in a 5-L
auto-controlled fermentor using optimized culture medium containing 20 g/L colloid chitosan, 12.5 g/L yeast extract
powder and 1.0 g/L Tween-80 at initial pH 6.0. Therefore, the strain amyP216 has potential application in the industrial
production of chitosanase.

Key words: Bacillus mojavensis, chitosanase, chitosan oligosaccharide, isolation and identification, optimization of fermentation conditions

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