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Related Experiment Video

Updated: Jan 26, 2026

A Soluble Tetrazolium-Based Reduction Assay to Evaluate the Effect of Antibodies on Candida tropicalis Biofilms
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RNA accumulation in Candida tropicalis based on cofactor engineering.

Bingbing Li1,2,3, Ying Liu1,2,3, Lianzhe Wang1,2,3

  • 1School of Life Science and Engineering, Henan University of Urban Construction, Pingdingshan, 367036, Henan, P.R. China.

FEMS Yeast Research
|April 4, 2019
PubMed
Summary

Altering cellular redox cofactor balance by expressing specific enzymes impacts metabolic flux and RNA content. This research explores how manipulating NADH/NAD+ and NADPH/NADP+ ratios affects cellular growth and RNA biosynthesis.

Keywords:
Candida tropicalisNADH oxidaseRNAcofactor engineeringmetabolismtranshydrogenase

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Area of Science:

  • Metabolic Engineering
  • Biotechnology
  • Synthetic Biology

Background:

  • Redox cofactors are crucial for cellular energy transfer and biosynthetic pathways.
  • Understanding cofactor dynamics is key to optimizing metabolic processes and product yields.
  • Perturbing cofactor balance offers a strategy to study and manipulate cellular metabolism.

Purpose of the Study:

  • To investigate the effects of impaired intracellular cofactor balance on RNA accumulation and metabolism.
  • To analyze the impact of expressing water-forming NADH oxidase (NoxE) and transhydrogenase (PntAB) on cofactor ratios and metabolic flux in *Candida tropicalis*.
  • To elucidate the role of ATP in RNA biosynthesis under altered cofactor conditions.

Main Methods:

  • Genetically engineered *Candida tropicalis* no. 121 by expressing *Lactococcus lactis* NoxE and *Escherichia coli* PntAB.
  • Quantified intracellular NADH/NAD+ and NADPH/NADP+ ratios.
  • Assessed changes in biomass, RNA content, ATP concentration, and metabolic flux redistribution.

Main Results:

  • NoxE expression significantly decreased the NADH/NAD+ ratio, while PntAB increased the NADH pool and decreased the NADPH/NADP+ ratio.
  • Metabolic flux redistribution occurred in response to cofactor perturbation.
  • NoxE expression reduced biomass and RNA content, whereas PntAB increased RNA content without significantly affecting biomass.
  • ATP concentration and RNA yield decreased with NoxE and increased with PntAB.

Conclusions:

  • Intracellular cofactor balance significantly influences metabolic flux and cellular composition, including RNA content.
  • ATP plays a critical role in RNA biosynthesis, as evidenced by its correlation with RNA yield under varying cofactor conditions.
  • Targeted manipulation of redox cofactors presents a viable strategy for metabolic engineering to control cellular metabolism and product formation.