科微学术

微生物学通报

产壳聚糖酶菌株的筛选、鉴定及酶学特性分析
DOI:
CSTR:
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

基金项目:

福建省自然科学基金项目(No. 2010J05056); 福建省教育厅科技项目(No. JB10201)


Screening, identification and enzymatic properties of chitosanase-producing strain
Author:
Affiliation:

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    【目的】利用筛选培养基, 从福建沿海潮间带泥样中分离筛选产壳聚糖酶的菌株, 并研究菌株的产酶特性。【方法】通过形态学观察, 结合26S rDNA序列进行分类鉴定, 采用DNS法测定酶活力。【结果】筛选得到产壳聚糖酶的菌株KQ-1002与草酸青霉(Penicillium oxalicum)的同源性为99%, 并初步鉴定为青霉属的一种。发酵培养的最适温度为30 °C, 最适碳源为1.0%水溶性壳聚糖, 最适氮源为1.87% (NH4)2SO4, 最适pH为6.0。该菌株液体发酵培养72 h产壳聚糖酶活性最高, 经优化后最高产酶量为18 U/mL。纯化后的壳聚糖酶经SDS-PAGE分析其分子量约40 kD。酶促反应最适pH为5.0, 最适反应温度为55 °C, Km值为1.293 g/L。在离子浓度为1.0×10?3 mol/L时, 金属离子Cu2+、Hg2+、Ag+对酶的活性均有强烈的抑制作用。壳聚糖酶对不同底物及脱乙酰度的壳聚糖具有不同的降解作用。【结论】筛选获得产壳聚糖酶的真菌菌株KQ-1002的壳聚糖酶活力经优化后提高了约7倍, 是一株具有研究和应用潜力的产壳聚糖酶菌株。

    Abstract:

    [Objective] By screening of chitosanase strains in mud samples from the Fujian coastal intertidal with screening medium, studied the characteristics of the strains for enzyme production. [Methods] Through morphological observation, combined with 26S rDNA sequences were classified and identified, DNS method was use for the determination of enzyme activity. [Results] The conclusion showed a 99% similarity between the chitosanase strain KQ-1002 and Penicillium oxalicum. The strain was tentatively identified as a species of Penicillium fungus. The optimal fermentation temperature was 30 °C, the optimum carbon source was the 1.0% soluble chitosan, the optimal nitrogen source was 1.87% (NH4)2SO4, and the optimum pH was 6.0. Cultured for 72 h under liquid fermentation on the strains, the chitosanase-producing activity reached the maximum, and the maximum of enzyme production had been optimized to 18 U/mL. The molecular masses of purified enzymes was 40 kD which analyzed by SDS-PAGE. Enzymatic reactions’ optimum pH was 5.0, the optimum temperature was 55 °C, and Km was 1.293 g/L. When ion concentration reached 1.0×10?3 mol/L, metal ions of Cu2+, Hg2+, Ag+ would strongly inhibit the enzyme’ activity. There was a different degradation of chitosanase on various substrates and deacetylation of chitosan. [Conclusion] Through screening, the chitosanase activity of chitosanase-producing fungal strains KQ-1002 which had been optimized boosted by approximately 7 times, and it is a chitosanase-producing strains of research and potential applications.

    参考文献
    相似文献
    引证文献
引用本文

王艳君,卓少玲,陈盛,杨谦. 产壳聚糖酶菌株的筛选、鉴定及酶学特性分析[J]. 微生物学通报, 2012, 39(12): 1734-1745

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:
  • 最后修改日期:
  • 录用日期:
  • 在线发布日期: 2012-12-20
  • 出版日期:
文章二维码