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Related Experiment Videos

High-level ATP production by a genetically-engineered Candida yeast

Y Sakai1, T Rogi, T Yonehara

  • 1Department of Agricultural Chemistry, Faculty of Agriculture, Kyoto University, Japan.

Bio/Technology (Nature Publishing Company)
|March 1, 1994
PubMed
Summary

Enhancing adenylate kinase (ADK) activity in Candida boidinii yeast significantly boosted adenosine triphosphate (ATP) production. This genetic modification led to a 23-fold increase in ATP yield, demonstrating a viable method for industrial ATP synthesis.

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Area of Science:

  • Biotechnology
  • Enzymology
  • Microbial Engineering

Background:

  • Adenylate kinase (ADK) catalyzed AMP phosphorylation is a rate-limiting step in ATP production in Candida boidinii.
  • Improving ADK activity is a potential strategy to enhance cellular ATP productivity.

Purpose of the Study:

  • To investigate the impact of enhanced ADK activity on ATP production in Candida boidinii.
  • To determine if expressing Saccharomyces cerevisiae ADK1 gene in C. boidinii improves ATP yield.

Main Methods:

  • The Saccharomyces cerevisiae ADK1 gene was expressed in Candida boidinii under the C. boidinii AOD1 promoter.
  • Methanol induction was used to enhance ADK expression and activity.
  • ATP production was measured from adenosine in a pH-controlled reaction system with successive feeding.

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Main Results:

  • Methanol-induced transformants exhibited a 10,000-fold increase in ADK activity.
  • ATP production from adenosine was 23-fold higher in engineered C. boidinii compared to the parent strain.
  • A high ATP concentration of 230 mM (117 g/l) was achieved over 45 hours with a 78% overall yield.

Conclusions:

  • Enhancing ADK activity in Candida boidinii is an effective strategy for significantly improving ATP production.
  • Genetically engineered C. boidinii offers a robust platform for high-yield industrial ATP synthesis.
  • The developed process allows for efficient ATP purification with a high overall yield.