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Front. Microbiol. | doi: 10.3389/fmicb.2018.02848

Characterization of a new cyclohexylamine oxidase from Acinetobacter sp. YT-02

 Hui Zhou1, Zheng-gang Han1,  Ti Fang2, Yuan-yuan Chen1, Shang-bo Ning1, Ya-ting Gan1 and  Da-zhong Yan1*
  • 1College of Biology and Pharmaceutical Engineering, Wuhan Polytechnic University, China
  • 2Wuhan Institute Of Virology (CAS), China

Abstract
Cyclohexylamine (CHAM) is widely used in various industries, but it is harmful to human beings and the environment. Acinetobacter sp. YT-02 can degrade CHAM via cyclohexanone as an intermediate. In this study, the cyclohexylamine oxidase (CHAO) gene from Acinetobacter sp. YT-02 was cloned. Amino acid sequence alignment indicated that CHAOYT-02 was 48% identical to its homolog from Brevibacterium oxydans IH-35A. The enzyme was expressed in Escherichia coli BL21 (DE3), and purified to apparent homogeneity by Ni-affinity chromatography. The purified enzyme was proposed to be a dimer of molecular mass of approximately 91 kDa. The enzyme exhibited its maximum activity at 50℃ and at pH 7.0. The enzyme was thermolabile as demonstrated by loss of its important percentage of activity after 30 min incubation at 50℃. Metal ions Mg2+, Co2+, and K+ had certain inhibitory effect on the enzyme activity. The kinetic parameters Km and Vmax were 0.252 ± 0.0180 mM and 4.30 ± 0.0833 M min−1, respectively. The biochemical properties, substrate specificities, and three-dimensional structures of CHAOYT-02 and CHAOIH-35 were compared.

Keywords: Cyclohexylamine oxidase, Acinetobacter sp. YT-02, Cyclohexylamine, Cyclohexanone, Biodegradation

Received: 26 Aug 2018; Accepted: 06 Nov 2018.

Edited by:

Mariusz Cycoń, Medical University of Silesia, Poland

Reviewed by:

Jun-Jie Zhang, School of Medicine, Indiana University Bloomington, United States
Xinjiong Fan, Anhui Medical University, China
Chandradipa Ghosh, Vidyasagar University, India
Gabriela Briceño, Universidad de La Frontera, Chile  

Copyright: © 2018 Zhou, Han, Fang, Chen, Ning, Gan and Yan. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence: PhD. Da-zhong Yan, College of Biology and Pharmaceutical Engineering, Wuhan Polytechnic University, Wuhan, China, yandz6808@163.com