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Structural and functional analysis of a low-temperature-active alkaline esterase from South China Sea marine sediment microbial metagenomic library

  • Biocatalysis
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

A low-temperature-active alkaline esterase, Est12, from a marine sediment metagenomic fosmid library was identified. Est12 prefers short- and middle-chain p-nitrophenol esters as substrate with optimum temperature and pH value of 50 °C and 9.0, respectively, and nearly 50 % of maximum activity retained at 5 °C. The hydrolysis activity of Est12 was stable at 40 °C. Ca2+ especially activated the activity of Est12 to about 151 % of the control. DEPC and PMSF inhibited the activity of Est12 to 34 and 25 %, respectively. In addition, Est12 was more tolerable to methanol compared to other organic solvents tested. The crystal structure of Est12 at 1.39 Å resolution showed that the cap domain which is composed of an α-helix and a flexible region resulted in a relatively wide spectrum of substrate, with p-nitrophenol caproate as the preferred one. Furthermore, the flexible cap domain and the high percentage of Gly, Ser, and Met may play important roles in the adaptation of Est12 to low temperature.

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Acknowledgments

This work was supported by the National High Technology Research and Development Program of China (2007AA09Z443 and 2007AA021301), Knowledge Innovation Project of The Chinese Academy of Sciences (KSCX2-YW-G-022), the National Key Basic Research Program of China ‘973 Program’ (2011CBA00803), and the Hundred Talents Program of the Chinese Academy of Sciences (Grant No. A1097). We cordially thank the staff of beamline BL17U1 at the Shanghai Synchrotron Radiation Facility, People’s Republic of China for assistance in synchrotron X-ray data collection.

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    1. CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China

      Yongfei Hu, Jing Li, Na Lu & Baoli Zhu

    2. Beijing Key Laboratory of Microbial Drug Resistance and Resistome, Beijing, 100101, China

      Yongfei Hu, Jing Li, Na Lu & Baoli Zhu

    3. Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, 310006, China

      Yongfei Hu & Baoli Zhu

    4. Marine Bioengineering Group, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, China

      Yinghui Liu, Yanbin Feng & Song Xue

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    Y. Hu and Y. Liu are contributed equally to this work.

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    Hu, Y., Liu, Y., Li, J. et al. Structural and functional analysis of a low-temperature-active alkaline esterase from South China Sea marine sediment microbial metagenomic library. J Ind Microbiol Biotechnol 42, 1449–1461 (2015). https://doi.org/10.1007/s10295-015-1653-2

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