title: Two acetyl-CoA synthetase isoenzymes are encoded by distinct genes in marine yeast Rhodosporidium diobovatum.
abstract: OBJECTIVE
  Two genes encoding two acetyl-CoA synthetase (ACS) isoenzymes have been identified in the marine yeast Rhodosporidium diobovatum MCCC 2A00023.
  
  
  RESULTS
  ACS1 encoded a polypeptide with a sequence of 578 amino acid residues, a predicted molecular weight of 63.73 kDa, and pI of 8.14, while the ACS2 encoded a polypeptide containing 676 amino acid residues with a deduced molecular mass of 75.61 kDa and a pI of 5.95. Biological activity of Acs1p and Acs2p was confirmed by heterologous expression in Escherichia coli. A 1.5-kb DNA fragment of the ACS1 gene and a 2.7-kb DNA fragment of the ACS2 gene were deleted using the RNA guide CRISPR-Cas9 system. The strain lacking ACS1 was unable to grow on acetate and ethanol media, while the ACS2 deletant was unable to grow on glucose medium. ACS1-ACS2 double mutants of R. diobovatum were non-viable.
  
  
  CONCLUSIONS
  ACS isoenzymes are essential to the yeast metabolism, and other sources of ACSs cannot compensate for the lack of ACSs encoded by the two genes.
authors.affiliation: Key Laboratory of Microbial Diversity Research and Application of Hebei Province, College of Life Sciences, Hebei University, Baoding, 071002, China.
authors.forename: Yuxuan
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Key Laboratory of Microbial Diversity Research and Application of Hebei Province, College of Life Sciences, Hebei University, Baoding, 071002, China.
authors.forename: Meiru
authors.initials: M
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Microbial Diversity Research and Application of Hebei Province, College of Life Sciences, Hebei University, Baoding, 071002, China.
authors.forename: Tianshi
authors.initials: T
authors.lastname: Wang
authors.affiliation: Key Laboratory of Microbial Diversity Research and Application of Hebei Province, College of Life Sciences, Hebei University, Baoding, 071002, China.
authors.forename: Xunxun
authors.initials: X
authors.lastname: Shi
authors.affiliation: Key Laboratory of Microbial Diversity Research and Application of Hebei Province, College of Life Sciences, Hebei University, Baoding, 071002, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Li
authors.affiliation: Key Laboratory of Microbial Diversity Research and Application of Hebei Province, College of Life Sciences, Hebei University, Baoding, 071002, China.
authors.forename: Lu
authors.initials: L
authors.lastname: Jia
authors.affiliation: Key Laboratory of Microbial Diversity Research and Application of Hebei Province, College of Life Sciences, Hebei University, Baoding, 071002, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Tang
authors.affiliation: Key Laboratory of Microbial Diversity Research and Application of Hebei Province, College of Life Sciences, Hebei University, Baoding, 071002, China. zhanglphbu@sohu.com.
authors.forename: Liping
authors.initials: L
authors.lastname: Zhang
mesh_terms: D000106:Acetate-CoA Ligase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D059001:Aquatic Organisms / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D003470:Culture Media / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D017353:Gene Deletion; D015870:Gene Expression; D007526:Isoelectric Point; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008970:Molecular Weight; D012248:Rhodotorula / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism
chemical_list: D003470:Culture Media; D007527:Isoenzymes; D000106:Acetate-CoA Ligase
keywords: Acetyl-CoA synthetase; CRISPR-Cas9; Isoenzymes; Rhodosporidium diobovatum