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Archives of microbiology: screening of pectinase-producing bacteria from citrus peel and characterization of a recombinant pectate lyase with applied potential

Abstract

Pectinase is widely used in numerous industrial fields, including the food, wine, and paper industries. In this work, seven bacteria were isolated from orange peel and their pectinase production activity was assayed. One bacterium (OR-B2) identified as a Bacillus sp. showed the highest enzyme activity towards others. A gene encoding a pectate lyase designed as PelB-B2 in this work was amplified and heterogeneous expressed in E.coli. PelB-B2 was defined as a member of the PelB pectate lyase family after phylogenic tree analysis. 3D model of PelB-B2 was constructed by SWISS-MODEL and PelB-B2 showed conserved para structure. After inducing culture and purified by Ni-affinity chromatography, the properties of the purified PelB-B2 were assayed. Optimal pH and temperature for PelB-B2 was pH 8.0 and 50 °C, respectively. PelB-B2 showed excellent pH stability and thermostability. It was stable within pH range 3.0–11.0 and retained more than 51% activity after incubation at 40 °C, 50 °C, or 60 °C for 1 h. Furthermore, we determined that PelB-B2 was a Ca2+-dependent pectinase and the pectin extracted from citrus was the benefit substrate for PelB-B2. The Km and Vmax of PelB-B2 were 1.64 g/L and 232.56 mol/(L min), respectively. The OR-B2 can be a new resource for pectinase production and the PelB-B2 has potential for industrial application.

Graphic abstract

7 bacteria were isolated from orange peel, namely OR-B1 to OR-B7 and their pectinase activities were assayed. One pectate lyase belongs to PelB family was cloned from OR-B2 and heterogeneous expressed in E. coli. Purified PelB-B2 was further studied with its properties. Effects of pH, temperature, chemicals, substrate on the enzyme activity were assayed and the enzyme kinetic was also measured.

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Acknowledgements

The authors would like to thank Gale A. Kirking at English Editorial Services, s.r.o. for language correction and additional assistance in improving the text.

Funding

This work was supported by National Natural Science Foundation of China (31701844), Natural Science Foundation of Fujian province (2018J05057), the Foundation of Fujian Educational Bureau (JAT170078), Fuzhou University Testing Fund of precious apparatus (2019T018), and with institutional support RVO: 60077344 of the Biology Centre CAS, Institute of Entomology, Czech Republic.

Author information

YG, DW, CL, YZ, IG, and XY contributed equally to the study design, collection of data, development of the sampling, analyses, interpretation of results, and preparation of the paper. All authors read and approved the final manuscript.

Correspondence to Ivan Gelbic or Xiuyun Ye.

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Communicated by Erko Stackebrandt.

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Guan, Y., Wang, D., Lv, C. et al. Archives of microbiology: screening of pectinase-producing bacteria from citrus peel and characterization of a recombinant pectate lyase with applied potential. Arch Microbiol (2020). https://doi.org/10.1007/s00203-020-01807-0

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Keywords

  • Pectate lyase
  • PelB
  • Bacillus sp.
  • Heterologous expression
  • Enzyme properties