title: Recombinant expression, purification, and characterization of an acyl-CoA binding protein from Aspergillus oryzae.
abstract: OBJECTIVE
  To characterize biochemically the lipid metabolism-regulating acyl-CoA binding protein (ACBP) from the industrially-important fungus Aspergillus oryzae.
  
  
  RESULTS
  A full-length cDNA encoding a candidate ACBP from A. oryzae (AoACBP) was cloned and expressed in Escherichia coli as a maltose-binding protein (MBP) fusion protein. The MBP-AoACBP protein was purified by an amylose resin chromatography column. SDS-PAGE showed that MBP-AoACBP has an estimated molecular weight of 82 kDa. Microscale thermophoresis binding assay showed that the recombinant AoACBP displayed much greater affinity for palmitoyl-CoA (K d = 80 nM) than for myristoyl-CoA (K d = 510 nM), thus demonstrating the preference of AoACBP for long-chain acyl-CoA.
  
  
  CONCLUSIONS
  The data support the identification of AoACBP as a long-chain ACBP in A. oryzae.
authors.affiliation: Key Laboratory of Industrial Fermentation Microbiology, National Engineering Laboratory for Industrial Enzymes, College of Biotechnology, Ministry of Education, Tianjin University of Science & Technology, Tianjin, 300457, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Hao
authors.affiliation: College of Chemical Engineering and Materials Science, Tianjin University of Science & Technology, Tianjin, 300457, China.
authors.forename: Xiaoguang
authors.initials: X
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, Jiangsu, China. zgzjiangnan@163.com.
authors.forename: Guozhong
authors.initials: G
authors.lastname: Zhao
authors.affiliation: College of Life Science, Jiangxi Science & Technology Normal University, Nanchang, 330013, China.
authors.forename: Lu
authors.initials: L
authors.lastname: Jiang
authors.affiliation: Key Laboratory of Industrial Fermentation Microbiology, National Engineering Laboratory for Industrial Enzymes, College of Biotechnology, Ministry of Education, Tianjin University of Science & Technology, Tianjin, 300457, China.
authors.forename: Ming
authors.initials: M
authors.lastname: Li
authors.affiliation: College of Life Science, Jiangxi Science & Technology Normal University, Nanchang, 330013, China. Zengtx001@aliyun.com.
authors.forename: Bin
authors.initials: B
authors.lastname: Zeng
mesh_terms: D000595:Amino Acid Sequence; D001236:Aspergillus oryzae / Q000235:genetics / Q000378:metabolism*; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D026261:Diazepam Binding Inhibitor / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
chemical_list: D026261:Diazepam Binding Inhibitor; D011994:Recombinant Proteins
keywords: Acyl-CoA binding affinity; Acyl-CoA binding protein; Aspergillus oryzae; Microscale thermophoresis binding assay; Myristoyl-CoA; Palmitoyl-CoA