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Engineering cellulase activity into Clostridium acetobutylicum.

Henri-Pierre Fierobe¹, Florence Mingardon, Angélique Chanal

¹Laboratoire de Chimie Bactérienne, UPR9043, CNRS IMM, Marseille, France.

Methods in Enzymology

|May 22, 2012

Summary

Engineered Clostridium acetobutylicum can produce biofuels via consolidated bioprocessing. Researchers improved secretion of key enzymes by modifying cellulase N-termini, enabling efficient biofuel production.

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Area of Science:

Biotechnology
Microbial Engineering
Biofuel Production

Background:

Clostridium acetobutylicum is a key organism for butanol production.
Engineered strains offer potential for consolidated bioprocessing (CBP) in biofuel applications.
Previous work demonstrated heterologous production of minicellulosomes in C. acetobutylicum.

Purpose of the Study:

To investigate the heterologous secretion of essential cellulosomal enzymes in Clostridium acetobutylicum.
To overcome limitations in enzyme translocation and secretion due to absent chaperones.
To enhance biofuel production efficiency through improved enzyme secretion.

Main Methods:

Engineering Clostridium acetobutylicum for cellulolytic capabilities.
Heterologous expression of cellulosomal enzymes, including GH48 and GH9 cellulases.
Utilizing scaffoldin modules fused to cellulases as cargo domains for secretion.

Main Results:

Achieved secretion yields for individual minicellulosomes ranging from 0.3 to 15 mg/L.
Identified the necessity of specific chaperones for certain cellulosomal enzymes (GH48, GH9).
Successfully secreted target cellulases by grafting scaffoldin modules to their N-termini.

Conclusions:

The engineered C. acetobutylicum strain shows promise for consolidated bioprocessing.
Grafting scaffoldin modules is an effective strategy to enable heterologous secretion of challenging cellulosomal enzymes.
This approach enhances the potential for efficient biofuel production from cellulosic biomass.