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Bioprocess and Biosystems Engineering

, Volume 42, Issue 1, pp 107–116 | Cite as

Towards efficient enzymatic conversion of d-galactose to d-tagatose: purification and characterization of l-arabinose isomerase from Lactobacillus brevis

  • Mengge Du
  • Dongying Zhao
  • Sisi Cheng
  • Di Sun
  • Ming Chen
  • Ziqing Gao
  • Chunzhi Zhang
Research Paper
  • 72 Downloads

Abstract

l-arabinose isomerase (l-AI) (EC 5. 3. 1. 4. l-AI) that mediates the isomerization of d-galactose to d-tagatose was isolated from Lactobacillus brevis (MF 465792), and was further purified and characterized. Pure enzyme with molecular weight of 60.1 kDa was successfully obtained after the purification using Native-PAGE gel extraction method, which was a monomer in solution. The l-AI was found to be stable at 45–75 °C, and at pH 7.0–9.0. Its optimum temperature and pH was determined as 65 °C and 7.0, respectively. Besides, we found that Ca2+, Cu2+, and Ba2+ ions inhibited the enzyme activity, whereas the enzyme activity was significantly enhanced in the presence of Mg2+, Mn2+, or Co2+ ions. The optimum concentration of Mn2+ and Co2+ was determined to be 1 mM. Furthermore, we characterized the kinetic parameters for l-AI and determined the Km (129 mM) and the Vmax (0.045 mM min− 1) values. Notably, L. brevisl-AI exhibited a high bioconversion yield of 43% from d-galactose to d-tagatose under the optimal condition, and appeared to be a more efficient catalyst compared with other l-AIs from various organisms.

Keywords

Purification Characterization l-arabinose isomerase Lactobacillus brevis d-tagatose 

Abbreviations

EDTA

Ethylenediaminetetraacetic acid

SDS

Sodium dodecyl sulfate

PAGE

Polyacrylamide gel electrophoresis

l-AI

l-arabinose isomerase

L. brevis

Lactobacillus brevis

Notes

Acknowledgements

This research was supported by the State Ethnic Affairs Commission & Ministry of Education, China. We thank Professor Shuxia Lv from Shenyang Agricultural University for helpful discussions on this manuscript.

Funding

This study was funded by the Doctoral Science Research Foundation of Liaoning Province of China (No. 20141111).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with animals performed by any of the authors.

Supplementary material

449_2018_2018_MOESM1_ESM.doc (54 kb)
Supplementary material 1 (DOC 54 KB)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Mengge Du
    • 1
  • Dongying Zhao
    • 2
  • Sisi Cheng
    • 1
  • Di Sun
    • 1
  • Ming Chen
    • 1
  • Ziqing Gao
    • 1
  • Chunzhi Zhang
    • 1
  1. 1.School of Biological EngineeringDalian Polytechnic UniversityDalianPeople’s Republic of China
  2. 2.School of Life Science and BiotechnologyDalian University of TechnologyDalianPeople’s Republic of China

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