Acid-resistant yeast strain for efficient production of l-malic acid, and construction method therefor and use thereof

A method for preparing a genetically modified malate-producing yeast strain, comprising conferring on the strain the activities of or enhancing in the strain the activities of malate transport protein and NADPH-dependent malate dehydrogenase, optionally further comprising conferring or enhancing the activity of at least one of:

• (i) pyruvate carboxylase activity, (ii) phosphoenolpyruvate carboxykinase activity, (iii) phosphoenolpyruvate carboxylase activity, preferably Escherichia coli phosphoenolpyruvate carboxylase activity, and (iv) biotin transport protein activity,
• preferably, said malate transport protein is selected from the group consisting of SpMAE1 protein, C4T318 protein and AsDct protein;
• preferably, said pyruvate carboxylase is derived from Aspergillus oryzae or Pichia kudriavzevii,
• preferably, said NADPH-dependent malate dehydrogenase is derived from a plant, more preferably a C4 plant, more preferably a plant of the family Gramineae, Cyperaceae, Compositae, Euphorbiaceae, Chenopodiaceae, Portulacaceae or Amaranthaceae, or derived from the genus Euglena or Thermobacillus, more preferably from Sorghum bicolor, Zea mays, Saccharum officinarum, Pisum sativum, Cicer arietinum, Spinacia oleracea, Euglena gracilis or Methanothermobacter thermautotrophicus, more preferably, said NADPH-dependent malate dehydrogenase is a Sorghum bicolor-derived NADPH-dependent malate dehydrogenase.