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Indian Journal of Microbiology

, Volume 59, Issue 1, pp 64–72 | Cite as

Cloning, Expression and Characterization of Two Beta Carbonic Anhydrases from a Newly Isolated CO2 Fixer, Serratia marcescens Wy064

  • Fanbing Chen
  • Wensong Jin
  • Huifang Gao
  • Zewang Guo
  • Hui Lin
  • Jiahuan Li
  • Kaihui Hu
  • Xiong Guan
  • Vipin C. Kalia
  • Jung-Kul LeeEmail author
  • Liaoyuan ZhangEmail author
  • Yongyu LiEmail author
Original Research Article
  • 43 Downloads

Abstract

Bacterial strains from karst landform soil were enriched via chemostat culture in the presence of sodium bicarbonate. Two chemolithotrophic strains were isolated and identified as Serratia marcescens Wy064 and Bacillus sp. Wy065. Both strains could grow using sodium bicarbonate as the sole carbon source. Furthermore, the supplement of the medium with three electron donors (Na2S, NaNO2, and Na2S2O3) improved the growth of both strains. The activities of carbonic anhydrase (CA) and ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) could be detected in the crude enzyme of strain Wy064, implying that the strain Wy064 might employ Calvin cycle to fix CO2. S. marcescens genome mining revealed four potential CA genes designated CA1CA4. The proteins encoded by genes CA13 were cloned and expressed in Escherichia coli. The purified recombinant enzymes of CA1 and CA3 exhibited CO2 hydration activities, whereas enzyme CA2 was expressed in inclusion bodies. A CO2 hydration assay demonstrated that the specific activity of CA3 was significantly higher than that of CA1. The maximum CO2 hydration activities for CA1 and CA3 were observed at pH 7.5 and 40 °C. The activities of CA1 and CA3 were significantly enhanced by several metal ions, especially Zn2+, which resulted in 21.1-fold and 26.1-fold increases of CO2 hydration activities, respectively. The apparent Km and Vmax for CO2 as substrate were 27 mM and 179 WAU/mg for CA1, and 14 mM and 247 WAU/mg for CA3, respectively. Structure modeling combined with sequence analysis indicated that CA1 and CA3 should belong to the Type II β-CA.

Keywords

Serratia marcescens CO2 fixation Carbonic anhydrase Expression Enzymatic properties 

Notes

Acknowledgements

This work was supported by the New Century Excellent Talents Supporting Plan of the Provincial Education Department of Fujian Province of China (No. K8015056A), the Project of Fujian-Taiwan Joint Center for Ecological Control of Crop Pests (Minjiaoke[2013]51), the Leading Talents of Fujian Province College (No. K8012012a), Science Fund of the Provincial Education Department of Fujian Province of China (No. KLA17041A). This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (NRF-2018H1D3A2001746, 2013M3A6A8073184).

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

12088_2018_773_MOESM1_ESM.docx (2.2 mb)
Supplementary material 1 (DOCX 2269 kb)

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Copyright information

© Association of Microbiologists of India 2018

Authors and Affiliations

  1. 1.College of Life Sciences, Gutian Edible Fungi Research InstituteFujian Agriculture and Forestry UniversityFuzhouPeople’s Republic of China
  2. 2.Department of Chemical EngineeringKonkuk UniversitySeoulRepublic of Korea
  3. 3.College of HorticultureFujian Agriculture and Forestry UniversityFuzhouPeople’s Republic of China

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