食品科学 ›› 2009, Vol. 30 ›› Issue (21): 262-266.doi: 10.7506/spkx1002-6300-200921061

• 生物工程 • 上一篇    下一篇

β-甘露聚糖酶产生菌HDYM-04的分离鉴定及产酶条件优化

葛菁萍,赵丹,宋刚,凌宏志,平文祥*   

  1. 微生物黑龙江省高校重点实验室,黑龙江大学生命科学学院
  • 收稿日期:2008-11-28 修回日期:2009-07-26 出版日期:2009-11-01 发布日期:2010-12-29
  • 通讯作者: 平文祥 E-mail:zhaodan4u@163.com;zhaodan4u@126.com
  • 基金资助:

    黑龙江省自然科学基金项目(C200505); 黑龙江省博士后启动基金项目(LBH-Q05113);
    哈尔滨市科技攻关计划项目(2007AA6CN032); 黑龙江省教育厅振兴老工业基地成果转化项目(1152GZH03);
    黑龙江大学校青年科学基金项目(QL200648); 黑龙江省人事厅留学回国择优资助项目

Isolation and Identification of Beta-Mannanase Producing Strain HDYM-04 and Optimization of Fermentation Conditions

GE Jing-ping,ZHAO Dan,SONG Gang,LING Hong-zhi,PING Wen-xiang*   

  1. Key Laboratory ofMicrobiology of Heilongjiang Province, Life Science College, Heilongjiang University, Harbin 150080, China
  • Received:2008-11-28 Revised:2009-07-26 Online:2009-11-01 Published:2010-12-29
  • Contact: PING Wen-xiang E-mail:zhaodan4u@163.com;zhaodan4u@126.com

摘要:

从实验室温水沤麻液中分离并筛选到了一株产β-甘露聚糖酶的细菌HDYM-04,经生理生化试验及16S rDNA序列分析鉴定为地衣芽孢杆菌(Bacillus licheniformis)。研究了HDYM-04菌株72h连续发酵过程中产酶及生长的动态变化,通过单因素及序贯正交试验优化得到其摇瓶发酵产酶的最佳培养基组分及条件:碳源60g/L魔芋粉,氮源30g/L蛋白胨,MgSO4•7H2O 0.2g/L,K2HPO4 5g/L,初始pH 8.0,装液量100ml/250ml三角瓶,接种量2%,37℃振荡培养48h。在研究条件下发酵液酶活力4890U/ml,比优化前提高了3.2倍。

关键词: β-甘露聚糖酶, 16S rDNA, 优化, 正交试验

Abstract:

A -mannanase producing bacterial strain HDYM-04 was isolated from laboratory-prepared flax retting liquor. HDYM-04 was identified as Bacillus licheniformis according to physiological and biochemical tests and phylogenetic analysis based on16S rDNA sequence. Single factor and orthogonal array design experiments were performed to obtain the optimum fermentation conditions for -mannanase production: culture with an inoculation size of 2% for 48 h at 37 in a 250 ml shaking flask filled with 100 ml liquid medium consisting of 60 g/L konjac powder utilized as carbon source, 30 g/L peptone utilized as nitrogen source, 0.2 g/L MgSO4 7H2O, 5 g/L K2HPO4 at initial pH 8.0. Under these optimum conditions, a -mannanase activity of 4890 U/ml fermentation broth was achieved, which was 3.2 times higher than before the optimization.

Key words: β--mannanase , 16S rDNA , optimization , orthogonal array design

中图分类号: