Cloning and functional characterization of the SUR2/SYR2 gene encoding sphinganine hydroxylase in Pichia ciferrii.
Saccharomyces cerevisiae sphinganine C4-hydroxylase encoded by the SUR2 gene catalyses the conversion of sphinganine to phytosphingosine. We isolated the SUR2 gene from Pichia ciferrii using nucleotide sequence homology to S. cerevisiae SUR2 to study hydroxylation of sphinganine in the sphingoid base overproducing yeast P. ciferrii. A positive clone was confirmed by nucleotide sequencing. A syringomycin-E resistance phenotype of a S. cerevisiae sur2-null mutant was complemented by expression of the cloned P. ciferrii SUR2 gene. Restoration of phytosphingosine production in the complemented strain was also confirmed, indicating that the cloned gene is a functional homologue of S. cerevisiae SUR2. .
Bae
Jung-Hoon
JH
Laboratory of Microbial Functions, Korea Research Institute of Bioscience and Biotechnology, Yusong, Taejon 305-333, Korea.
Sohn
Jung-Hoon
JH
Park
Chang-Seo
CS
Rhee
Joon-Shick
JS
Choi
Eui-Sung
ES
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000494:pharmacology; D001483:Base Sequence; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000235:genetics; D025141:Drug Resistance, Fungal / Q000235:genetics; D017353:Gene Deletion; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D005822:Genetic Vectors; D006899:Mixed Function Oxygenases / Q000235:genetics*; D008969:Molecular Sequence Data; D010843:Pichia / Q000201:enzymology* / Q000235:genetics*; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000201:enzymology / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D017386:Sequence Homology, Amino Acid; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000378:metabolism
D001426:Bacterial Proteins; D004271:DNA, Fungal; D029701:Saccharomyces cerevisiae Proteins; C032677:syringomycin; D006899:Mixed Function Oxygenases; C109313:SUR2 protein, S cerevisiae; D013110:Sphingosine; C005682:safingol