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Application of an S-layer protein as a self-aggregating tag for cost-effective separation of recombinant human and yeast d-amino acid oxidases in the aqueous two-phase system

  • Minkyeong Ahn
  • Sangje Park
  • Junhyun Jeon
  • Jung Kyu Choi
  • Yongho KhangEmail author
Original Research Paper
  • 45 Downloads

Abstract

Objective

To evaluate whether the surface layer (S-layer) protein of Lactobacillus brevis serves as a self-aggregating protein tag for cost-effective separation of human and yeast d-amino acid oxidases (hDAAO and yDAAO) expressed in E. coli.

Results

In aqueous two-phase (PEG–phosphate) system, the S-layer:DAAO fusion proteins (shDAAO and syDAAO) were separated at the interface with a recovery of 82 ± 10.6% for shDAAO and 95 ± 1.9% for syDAAO. Some shDAAO proteins were separated as precipitates with a recovery of 41 ± 0.5% in phosphate (9%, w/w) using PEG 3000 and PEG 4000 (16%, w/w), while some syDAAO proteins were also isolated as precipitates with a recovery of 75 ± 17.5% in phosphate (9%, w/w) using PEG 4000 and PEG 8000 (16%, w/w).

Conclusions

The S-layer of L. brevis was applied to a self-assembled protein tag to enable cost-effective separation of human and yeast d-amino acid oxidases expressed in E. coli cells. Because of the self-assembling properties of S-layer proteins, human and yeast d-amino acid oxidases fused with S-layer proteins could be easily separated by aggregates at the interface and/or in a few conditions by precipitates to the bottom of the PEG–phosphate aqueous system.

Keywords

Aqueous two-phase system d-Amino acid oxidase Fusion tag Self-aggregating tag S-layer protein 

Notes

Acknowledgements

This work was supported in part by Yeungnam University.

Author contributions

All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by MA, SP, JJ, and JKC. The first draft of the manuscript was written by YK and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Department of BiotechnologyYeungnam UniversityGyeongsanRepublic of Korea

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