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Area of Science:

  • Metabolic Engineering
  • Synthetic Biology
  • Biotechnology

Background:

  • Engineering novel carbon fixation pathways offers potential for higher yields than natural systems like the Calvin-Benson-Bassham (CBB) cycle.
  • Predicting the in vivo performance and optimal design criteria for these artificial pathways remains a challenge.

Purpose of the Study:

  • To computationally explore and compare aerobic carbon fixation pathways, including novel cycles, based on specific activity and yield.
  • To evaluate performance using C1 substrates (methanol, formate) and CO2/H2, considering reaction kinetics and thermodynamics.

Main Methods:

  • Utilized computational approaches to analyze existing and novel carbon fixation pathways.
  • Collected comprehensive kinetic data and employed the Parameter Balancing algorithm for missing data.
  • Applied the Enzyme Cost Minimization algorithm to assess kinetic and thermodynamic consistency and calculate pathway activities.

Main Results:

  • The reductive glycine pathway, CETCH cycle, and a new reductive citramalyl-CoA cycle showed predicted specific activities matching natural cycles with superior product-substrate yield.
  • The Calvin-Benson-Bassham (CBB) cycle demonstrated higher activity than previously assumed.
  • Novel pathways were found to be pareto-optimal for specific activity and product-substrate yield with C1 substrates and CO2/H2.

Conclusions:

  • Stoichiometric yield may not be the primary design criterion for the CBB cycle.
  • Engineered carbon fixation pathways hold significant potential for industrial biotechnology and synthetic biology applications.
  • Further research into alternative pathways is warranted for optimizing C1 substrate utilization.