Molecular cloning and characterization of the ATP citrate lyase from carotenogenic yeast Phaffia rhodozyma
作者： Cipriano Chávez-Cabrera , Rodolfo Marsch , Yaneth Bartolo-Aguilar , Zoila Rosa Flores-Bustamante , María Eugenia Hidalgo-Lara
关键词: Molecular cloning 、 Biology 、 Gene 、 Biochemistry 、 Recombinant DNA 、 Yeast 、 Complementary DNA 、 Peptide sequence 、 ATP citrate lyase 、 Amino acid
摘要: ATP citrate lyase (ACL), is a key cytosolic source of acetyl-CoA for fatty acid and sterol biosynthesis appear to be involved in carotenoid yeasts. Three homologous DNA sequences encoding ACLs Phaffia rhodozyma were isolated i.e two genes one cDNA. The multi-intronic, with 3450-bp-coding both genes, as the cDNA, encoded identical 120.1-kDa polypeptides. Full-length amino these showed multidomains, PLN02235 PLN02522, which are necessary activity. 82-87% similarity putative from other basidiomycetes 71% human ACL. acl cDNA was used express heterologous ACL 6XHis-tagged identified using MALDI-TOF-MS. sequenced peptides 42.2% coverage 100% identity sequence generated silico. recombinant purified homogeneity an activity 2 U. This first study characterize carotenogenic yeast. present provides foundation future studies assess (a) possible occurrence alternative splicing, (b) identify promoter(s) sequence(s) (c) involvement differential regulation
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