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Characterization and engineering of two new GH9 and GH48 cellulases from a Bacillus pumilus isolated from Lake Bogoria

Abstract

Objectives

To search for new alkaliphilic cellulases and to improve their efficiency on crystalline cellulose through molecular engineering

Results

Two novel cellulases, BpGH9 and BpGH48, from a Bacillus pumilus strain were identified, cloned and biochemically characterized. BpGH9 is a modular endocellulase belonging to the glycoside hydrolase 9 family (GH9), which contains a catalytic module (GH) and a carbohydrate-binding module belonging to class 3 and subclass c (CBM3c). This enzyme is extremely tolerant to high alkali pH and remains significantly active at pH 10. BpGH48 is an exocellulase, belonging to the glycoside hydrolase 48 family (GH48) and acts on the reducing end of oligo-β1,4 glucanes. A truncated form of BpGH9 and a chimeric fusion with an additional CBM3a module was constructed. The deletion of the CBM3c module results in a significant decline in the catalytic activity. However, fusion of CBM3a, although in a non native position, enhanced the activity of BpGH9 on crystalline cellulose.

Conclusions

A new alkaliphilic endocellulase BpGH9, was cloned and engineered as a fusion protein (CBM3a-BpGH9), which led to an improved activity on crystalline cellulose.

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Acknowledgements

LO was a recipient of a PhD scholarship (2014–2018) from the Embassy of France in Kenya, and of a stipendium (2015–2019), from the University of Nairobi. This work was supported by the Swedish International Development Cooperation Agency (Sida) sponsored Bioinnovate-Africa Program and UNEP GEF Agency Project No. 00876 on developing the microbial biotechnology industry from Kenya’s Soda Lakes in line with the Nagoya Protocol.

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Correspondence to Charles Tellier.

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Ogonda, L.A., Saumonneau, A., Dion, M. et al. Characterization and engineering of two new GH9 and GH48 cellulases from a Bacillus pumilus isolated from Lake Bogoria. Biotechnol Lett 43, 691–700 (2021). https://doi.org/10.1007/s10529-020-03056-z

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Keywords

  • Cellulose
  • Endoglucanase
  • Cellobiohydrolase
  • Alkaliphilic
  • CBM
  • Bacillus pumilus