L-Lactate dehydrogenase from Cyanidioschyzon merolae shows high catalytic efficiency for pyruvate reduction and is inhibited by ATP

Plant Mol Biol. 2024 Sep 10;114(5):98. doi: 10.1007/s11103-024-01495-0.

Abstract

L-Lactate is a commodity chemical used in various fields. Microorganisms have produced L-lactate via lactic fermentation using saccharides derived from crops as carbon sources. Recently, L-lactate production using microalgae, whose carbon source is carbon dioxide, has been spotlighted because the prices of the crops have increased. A red alga Cyanidioschyzon merolae produce L-lactate via lactic fermentation under dark anaerobic conditions. The L-lactate titer of C. merolae is higher than those of other microalgae but lower than those of heterotrophic bacteria. Therefore, an increase in the L-lactate titer is required in C. merolae. L-Lactate dehydrogenase (L-LDH) catalyzes the reduction of pyruvate to L-lactate during lactic fermentation. C. merolae possesses five isozymes of L-LDH. The results of previous transcriptome analysis suggested that L-LDHs are the key enzymes in the lactic fermentation of C. merolae. However, their biochemical characteristics, such as catalytic efficiency and tolerance for metabolites, have not been revealed. We compared the amino acid sequences of C. merolae L-LDHs (CmLDHs) and characterized one of the isozymes, CmLDH1. BLAST analysis revealed that the sequence similarities of CmLDH1 and the other isozymes were above 99%. The catalytic efficiency of CmLDH1 under its optimum conditions was higher than those of L-LDHs of other organisms. ATP decreased the affinity and turnover number of CmLDH1 for NADH. These findings contribute to understanding the characteristics of L-LDHs of microalgae and the regulatory mechanisms of lactic fermentation in C. merolae.

Keywords: Cyanidioschyzon merolae; l-Lactate dehydrogenase; Catalytic efficiency; Lactic fermentation.

MeSH terms

  • Adenosine Triphosphate* / metabolism
  • Amino Acid Sequence
  • Catalysis
  • Fermentation
  • L-Lactate Dehydrogenase* / genetics
  • L-Lactate Dehydrogenase* / metabolism
  • Lactic Acid / metabolism
  • Microalgae / enzymology
  • Microalgae / genetics
  • Microalgae / metabolism
  • Pyruvic Acid* / metabolism
  • Rhodophyta* / enzymology
  • Rhodophyta* / genetics
  • Rhodophyta* / metabolism

Substances

  • L-Lactate Dehydrogenase
  • Pyruvic Acid
  • Adenosine Triphosphate
  • Lactic Acid