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Recombinant expression, purification, and characterization of an acyl-CoA binding protein from Aspergillus oryzae

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Abstract

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.

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Acknowledgments

This study was financially supported by these projects in China (31171731, 31460447, 31401682, 2015T80498, 20142BDH80003, 2013-CXTD002, 3000035402, 00001384, 30000411, 2014QNBJRC0010, 31401682, BK20140146, “555 talent project” of Jiangxi Province) and Jiangxi Key Laboratory of Bioprocess Engineering.

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    Authors and Affiliations

    1. 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

      Qing Hao & Ming Li

    2. College of Chemical Engineering and Materials Science, Tianjin University of Science & Technology, Tianjin, 300457, China

      Xiaoguang Liu

    3. State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, Jiangsu, China

      Guozhong Zhao

    4. College of Life Science, Jiangxi Science & Technology Normal University, Nanchang, 330013, China

      Lu Jiang & Bin Zeng

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    1. Qing Hao
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    2. Xiaoguang Liu
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    3. Guozhong Zhao
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    4. Lu Jiang
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    5. Ming Li
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    Corresponding authors

    Correspondence to Guozhong Zhao or Bin Zeng.

    Additional information

    Qing Hao and Xiaoguang Liu contributed equally to this work.

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    Cite this article

    Hao, Q., Liu, X., Zhao, G. et al. Recombinant expression, purification, and characterization of an acyl-CoA binding protein from Aspergillus oryzae . Biotechnol Lett 38, 519–525 (2016). https://doi.org/10.1007/s10529-015-2003-1

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    • DOI: https://doi.org/10.1007/s10529-015-2003-1

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