The role of PaAAC1 encoding a mitochondrial ADP/ATP carrier in the biosynthesis of extracellular glycolipids, mannosylerythritol lipids, in the basidiomycetous yeast Pseudozyma antarctica.
Pseudozyma antarctica produces large amounts of the glycolipid biosurfactants known as mannosylerythritol lipids (MEL), which show not only excellent surface-active properties but also versatile biochemical actions. A gene homologous with a mitochondrial ADP/ATP carrier was dominantly expressed in P. antarctica under MEL-producing conditions on the basis of previous gene expression analysis. The gene encoding the mitochondrial ADP/ATP carrier of P. antarctica (PaAAC1) contained a putative open reading frame of 954 bp and encodes a polypeptide of 317 amino acids. The deduced translation product shared high identity of 66%, 70%, 69%, 74%, 75% and 52% with the mitochondrial ADP/ATP carrier of Saccharomyces cerevisiae (AAC1), S. cerevisiae (AAC2), S. cerevisiae (AAC3), Kluyveromyces lactis (KlAAC), Neurospora crassa (NcAAC) and human (ANT1), respectively, and conserved the consensus sequences of all ADP/ATP carrier proteins. The gene expression by introducing a plasmid pUXV1-PaAAC1 into the yeast cells increased the MEL production. In addition, the expression of PaAAC1 in which the conserved arginine and leucine required for ATP transport activity were replaced with isoleucine and serine, respectively, failed to increase MEL production. Accordingly, these results suggest that PaAAC1 encoding a mitochondrial ADP/ATP carrier should be involved in MEL biosynthesis in the yeast.
Morita
Tomotake
T
Research Institute for Innovations in Sustainable Chemistry, National Institute of Advanced Industrial Science and Technology (AIST), Central 5-2, Higashi 1-1, Tsukuba, Ibaraki 305-8565, Japan.
Ito
Emi
E
Fukuoka
Tokuma
T
Imura
Tomohiro
T
Kitamoto
Dai
D
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001483:Base Sequence; D003001:Cloning, Molecular; D017124:Conserved Sequence; D004271:DNA, Fungal / Q000235:genetics; D005656:Fungal Proteins / Q000378:metabolism*; D015870:Gene Expression; D005822:Genetic Vectors; D006017:Glycolipids / Q000096:biosynthesis*; D006801:Humans; D007716:Kluyveromyces / Q000201:enzymology / Q000235:genetics; D000226:Mitochondrial ADP, ATP Translocases / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D015394:Molecular Structure; D016297:Mutagenesis, Site-Directed; D009492:Neurospora crassa / Q000201:enzymology / Q000235:genetics; D016366:Open Reading Frames; D010957:Plasmids; D017433:Protein Structure, Secondary; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D014587:Ustilaginales / Q000201:enzymology* / Q000378:metabolism*
D004271:DNA, Fungal; D005656:Fungal Proteins; D006017:Glycolipids; C439671:mannosylerythritol lipid; D000226:Mitochondrial ADP, ATP Translocases