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Applied Microbiology and Biotechnology

, Volume 103, Issue 13, pp 5231–5241 | Cite as

A new cold-active and alkaline pectate lyase from Antarctic bacterium with high catalytic efficiency

  • Yumeng Tang
  • Pan Wu
  • Sijing Jiang
  • Jonathan Nimal Selvaraj
  • Shihui Yang
  • Guimin ZhangEmail author
Biotechnologically relevant enzymes and proteins
  • 161 Downloads

Abstract

Cold-active enzymes have become attractive biocatalysts in biotechnological applications for their ability to retain high catalytic activity below 30 °C, which allows energy reduction and cost saving. Here, a 1041 bp gene pel1 encoding a 34.7 KDa pectate lyase was cloned from a facultatively psychrophilic Antarctic bacterium Massilia eurypsychrophila and heterologously expressed in Escherichia coli. PEL1 presented the highest 66% identity to the reported mesophilic pectate lyase PLXc. The purified PEL1 exhibits the optimum temperature and pH of 30 °C and 10 toward polygalacturonic acid, respectively. PEL1 is a cold-active enzyme that can retain 60% and 25% relative activity at 10 °C and 0 °C, respectively, while it loses most of activity at 40 °C for 10 min. PEL1 has the highest specific activity (78.75 U mg−1) than all other reported cold-active pectinase, making it a better choice for use in industry. Based on the detailed sequence and structure comparison between PEL1 and PLXc and mutation analysis, more flexible structure and some loop regions may contribute to the cold activity and thermal instability of PEL1. Our investigations of the cold-active mechanism of PEL1 might guide the rational design of PEL1 and other related enzymes.

Keywords

Pectate lyase Enzymatic assay Cold-active Specific activity Mutation analysis 

Notes

Acknowledgment

We thank Prof. Peng Fang for providing M. eurypsychrophila for our research. We thank Dr. Zhao Jing (Tianjin Institute of Industrial Biotechnology) for helpful discussions.

Funding information

This work was supported by Technical innovation special fund of Hubei Province (2017ACA171), Hubei Provincial Natural Science Foundation (2018CFA042, 2018CFB319), and 2016 Wuhan Yellow Crane Talents (Science) Program.

Compliance with ethical standards

This article does not contain any studies with human participants or animals performed by any of the authors. All authors confirm that ethical principles have been followed in the research as well as in manuscript preparation, and approved this submission.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

253_2019_9803_MOESM1_ESM.pdf (1.3 mb)
ESM 1 (PDF 1355 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.State Key Laboratory of Biocatalysis and Enzyme Engineering, Hubei Collaborative Innovation Center for Green Transformation of Bio-Resources, School of Life SciencesHubei UniversityWuhanChina

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