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

LIANG Zhengwen; ZHANG Tieying; LI Shuang; LIU Junli

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

Chinese Journal of Animal Nutrition >

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

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

FEED SCIENCE AND TECHNOLOGY

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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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

Cite this article

LIANG Zhengwen , ZHANG Tieying , LI Shuang , LIU Junli . Identification, Fermentation Conditions Optimization and Enzymatic Properties of Cutin Degrading Bacteria[J]. Chinese Journal of Animal Nutrition, 2016 , 28(11) : 3567 -3575 . DOI: 10.3969/j.issn.1006-267x.2016.11.025

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