动物营养学报 >

2016 , Vol. 28 >Issue 11: 3567 - 3575

DOI: https://doi.org/10.3969/j.issn.1006-267x.2016.11.025

饲料营养 Feed Science and Technology

植物角质降解菌的种属鉴定、发酵条件优化及酶学性质研究

  • 梁争文 ,
  • 张铁鹰 ,
  • 李爽 ,
  • 刘俊丽
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  • 1. 中国农业科学院北京畜牧兽医研究所, 动物营养学国家重点实验室, 北京 100193;
    2. 甘肃农业大学动物科学技术学院, 兰州 730070

收稿日期: 2016-05-01

  网络出版日期: 2016-11-18

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Identification, Fermentation Conditions Optimization and Enzymatic Properties of Cutin Degrading Bacteria

  • LIANG Zhengwen ,
  • ZHANG Tieying ,
  • LI Shuang ,
  • LIU Junli
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  • 1. State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Science, Beijing 100193, China;
    2. College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China

Received date: 2016-05-01

  Online published: 2016-11-18

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摘要

本试验旨在通过角质降解菌株X8P的种属鉴定、发酵条件优化和酶学性质研究，探讨降解植物表面角质层，进一步改善动物对植物纤维利用的可能新方案。试验通过形态观察和16S rDNA测序鉴定菌株X8P种属，并对其产酶所需碳源、氮源、发酵温度与时间进行优化，其发酵液经硫酸铵盐析沉淀获得其胞外蛋白粗酶，并对其粗酶催化的适宜pH和pH稳定性、温度和温度稳定性，以及有机溶剂、表面活性剂和金属离子对其活性的影响进行研究。结果表明：1）菌株X8P经形态观察和分子鉴定为东方醋杆菌（Acetobacter orientalis）。2）菌株X8P适宜产酶发酵条件为溶菌肉汤（LB）培养基中37℃发酵4 d，1%橄榄油和1%葡萄糖明显促进菌株产酶，而1%可溶性淀粉明显抑制菌株产酶。3）该菌株胞外粗酶催化适宜pH和温度分别为6.5和45℃，且表现出一定pH稳定性，但在有机溶剂中不稳定，仅甘油中可保留全部活性，在二甲基亚砜（50%）中活性可保留66%。吐温（Tween）-20（1 mmol/L）、Tween-80（1 mmol/L）和聚乙二醇辛基苯基醚（1和10 mmol/L）可使菌株X8P粗酶活性提高3%~35%。金属离子钾离子（K+）、锰离子（Mn2+）（1和10 mmol/L）可使菌株X8P粗酶活性提高2%~20%。由此可见，菌株X8P具有一定的产角质酶潜力和应用前景，可进一步深入研究。

关键词： 角质; 角质酶; 角质降解菌; 发酵条件优化; 酶学性质

本文引用格式

梁争文 , 张铁鹰 , 李爽 , 刘俊丽 . 植物角质降解菌的种属鉴定、发酵条件优化及酶学性质研究[J]. 动物营养学报, 2016 , 28(11) : 3567 -3575 . DOI: 10.3969/j.issn.1006-267x.2016.11.025

Abstract

The present study was designed to identify the cutin degrading strain X8P, their fermentation conditions and enzymatic properties were also explored, to investigate the degradation of plant cuticular surface, and to improve the possible new schemes for the utilization of plant fiber. The strain X8P was identified by morphology observation and 16S rDNA sequencing, and the optimal fermentation conditions of carbon source, nitrogen source, fermentation temperature and fermentation time for enzyme production were investigated. After precipitating crude extracellular enzyme protein by ammonium sulfate, we studied the optimum pH and pH stability and temperature and temperature stability of crude extracellular enzyme, and the effect of organic solvents, surfactants and metal ions on crude enzyme. The results showed as follows:1) the strain X8P was successfully identified as Acetobacter orientalis by colony morphological characteristic observing and 16S rDNA sequencing. 2) The optimal fermentation conditions of the strain X8P were temperature 37℃ and fermentation time for 4 days in the lysogeny broth (LB) medium. The LB medium supplemented with 1% olive oil and glucose was significantly promoted enzyme production, and supplemented with 1% soluble starch inhibited the enzyme production. 3) The optimum temperature and pH of the strain X8P's extracellular esterase with good pH stability were 45℃ and pH 6.5. In tested organic solvents, the enzyme exhibited part of cutinolytic esterase activity only in dimethyl sulfoxide (DMSO)(50%). Tween-20 (1 mmol/L), Tween-80 (1 mmol/L) and Triton-100 (1 and 10 mmol/L) could improve the extracellular enzyme activity as 3% to 35%. Metal ions such as K+, Mn2+ (1 and 10 mmol/L) could improve extracellular enzyme activity as 2% to 20%. Therefore, the strain X8P has certain enzyme potential and application prospects, worthy of further study.

Key words： cutin; cutinase; cutin degrading bacteria; fermentation conditions optimization; enzymatic properties

参考文献

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