| 张子瑞,孙谧.一株壳聚糖酶产生菌的鉴定和产酶条件优化D张子瑞1,2,孙谧1*,郝建华1,刘均忠1[J].中国海洋药物,2016,35(6):63-71. |
| 一株壳聚糖酶产生菌的鉴定和产酶条件优化D张子瑞1,2,孙谧1*,郝建华1,刘均忠1 |
| Identification of a Chitosanase Producing Strain and Optimized of Its Fermentation Conditions |
| 投稿时间：2016-03-25 修订日期：2016-04-25 |
| DOI： |
| 中文关键词: 海洋芽孢杆菌,壳聚糖酶,鉴定,产酶条件优化,响应面 |
| English Keywords:marine bacillus, chitosanase, identification, optimization, response surface |
| Fund Project: |
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| 中文摘要: |
| 对一株产壳聚糖酶菌株Y116进行了形态学观察、生理生化特征鉴定和16S rDNA序列同源性分析,初步确定该菌株属于芽孢杆菌属(Bacillus)。采用单因素法和响应面法,对菌株Y116产酶的培养基成分和培养条件进行了优化。首先筛选碳源、氮源、初始pH、金属离子、NaCl浓度、酵母浸粉浓度、接种量、发酵温度和时间8个因素。研究结果显示：当酶活达到最高值时,最终确定各因素的值分别为：乳糖 35 g/L,豆饼粉30 g/L, Mg2+ 0.5 g/L,NaCl 5 g/L,初始pH 5.0,接种量5%,培养温度30 ℃和发酵时间96 h。再通过Plackett-Burman(PB)设计对影响壳聚糖酶产量的8个主要因素进行评价,确定了豆饼粉浓度、初始pH和Mg2+浓度是影响产酶量的3个主要因素；利用中心组合设计(CCD)及响应面分析,最终得到3个主要因素的最优值：豆饼粉浓度39.44 g/L,pH 5.12,Mg2+ 浓度5.12 g/L,酶活力比优化前提高了5倍。 |
| English Summary: |
| A chitosanase producing strain Y116 was subsequently identified as Bacillus based on the morphology characters, physiological and biochemical test and 16S rDNA sequences analysis.The culture medium and culture conditions of strain Y116 were optimized in order to improve the yield of chitosanase by one-single factor and response surface methodology. The single factor experiment: the nitrogen source, carbon source, initial pH, Mg2 , NaCl, Yeast extract powder, inoculation, fermentation temperature and fermentation time. The best result carried out of each single factor was as follows: lactose 35 g/L, bean cake powder 30 g/L, Mg2 0.5 g/L, NaCl 5 g/L, Yeast extract powder5 g/L, initial pH 5.0, inoculation 5%, the culture temperature at 30℃ and the culture time at 96h. Secondly, those main variables were valuated , Result of Plackett-Burman design showed, the significantly affecting variables were bean cake powder, initial pH and Mg2 . The optimal level of the three main factors were determine by the central composite design and the analysis.The most suitable variables were identified as follows: bean cake powder 39.44 g/L, pH 5.12, Mg2 5.12 g/L. Comparison with that before optimization, the productivity of chitosanase increased 5 times under the optimum conditions |
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