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Updated: Mar 28, 2026
A Facile Protocol to Generate Site-Specifically Acetylated Proteins in Escherichia Coli
Published on: December 9, 2017
Recombinant expression, purification, and characterization of an acyl-CoA binding protein from Aspergillus oryzae
Qing Hao1, Xiaoguang Liu2, Guozhong Zhao3
1Key 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.
Objectives:
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.
Conclusion:
The data support the identification of AoACBP as a long-chain ACBP in A. oryzae.
