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Efficient whole-cell-catalyzing cellulose saccharification using engineered Clostridium thermocellum.

Jie Zhang1,2,3,4, Shiyue Liu1,2,3,4, Renmin Li1,2,3,4

  • 1Shandong Provincial Key Laboratory of Energy Genetics, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, 266101 People's Republic of China.

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Engineered Clostridium thermocellum overcomes cellobiose inhibition for efficient lignocellulose saccharification. This whole-cell catalyst strategy offers a cost-effective solution for industrial applications.

Keywords:
CelSCellulosomeFermentable sugarGenome editingLignocelluloseβ-Glucosidase

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Area of Science:

  • Biotechnology
  • Microbial Engineering
  • Biomass Conversion

Background:

  • Cost-efficient saccharification is a major hurdle in industrial lignocellulose conversion.
  • Clostridium thermocellum utilizes cellulosomes for efficient lignocellulose degradation.
  • Cellobiose inhibition of cellulosomes limits industrial application of C. thermocellum.

Purpose of the Study:

  • To engineer a Clostridium thermocellum strain that overcomes cellobiose feedback inhibition.
  • To develop an efficient whole-cell catalyst for enhanced cellulose saccharification.
  • To establish a cost-effective method for lignocellulose conversion.

Main Methods:

  • Constructed a recombinant C. thermocellum strain (∆pyrF::CaBglA) using seamless genome editing.
  • Integrated a heterologous beta-glucosidase (BglA) into the C. thermocellum genome.
  • Employed a two-stage saccharification process with Avicel as the carbon source.

Main Results:

  • The recombinant strain ∆pyrF::CaBglA showed over a twofold increase in saccharification compared to the parent strain.
  • Achieved a relative saccharification level of 79.4% and produced 490 mM reducing sugars.
  • Demonstrated enhanced sugar production and saccharification with increased cell density without external enzyme addition.

Conclusions:

  • The whole-cell catalysis strategy using engineered C. thermocellum is a promising approach for cellulose saccharification.
  • The C. thermocellum ∆pyrF::CaBglA strain shows potential as an efficient whole-cell catalyst for industrial cellulose saccharification.
  • Overcoming cellobiose inhibition is key to improving the efficiency of lignocellulose conversion.