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Journal of the American Oil Chemists' Society

, Volume 92, Issue 1, pp 37–45 | Cite as

Biochemical Characterization of Purified Esterase from Soybean (Glycine max) Seed

  • Márcio de BarrosEmail author
  • Gabriela Alves Macedo
Original Paper
  • 283 Downloads

Abstract

Alkaline esterase (carboxylic-ester hydrolases; EC 3.1.1.1) extracted from germinated soybean seeds (Glycine max) was purified approximately 3.6 times by chromatography in a DEAE-cellulose anion exchange column and filtration in Sephadex G100 gel. The molecular mass of the enzyme was estimated at 45 kDa by gel electrophoresis (SDS-PAGE). The purified enzyme showed a specific activity of 5.6 U mg−1 using p-nitrophenyl butyrate as substrate. The esterase showed optimal activity at 47 °C in moderately alkaline pH, low stability in temperatures higher than 50 °C, and high stability at pH values between 6 and 9.5. The Ca2+ and Co2+ ions proved to have a positive effect on enzyme activity; however, Hg2+ completely inhibited esterase activity. Using p-nitrophenyl butyrate as substrate, the enzyme showed a K m of 0.39 mM, V max of 31.5 mM mg−1 min−1 and k cat 7.60 × 10s−1. Regarding substrate affinity, the enzyme showed greater activity for substrates containing short-chain fatty acids, especially p-nitrophenyl acetate. Such characteristics give the enzyme great potential for application in the production of low molecular weight esters, in the food industry, and in chemical products. This enzyme is another new member of the family of lipases and esterases from vegetable seeds with high activity and stability in alkaline pH.

Keywords

Glycine max Esterase Purification Characterization 

Notes

Acknowledgments

This project was supported by the CNPq (Conselho Nacional de Pesquisa e Desenvolvimento).

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Copyright information

© AOCS 2014

Authors and Affiliations

  1. 1.Biochemistry and Biotechnology DepartmentState University of Londrina (UEL)LondrinaBrazil
  2. 2.Food Science Department, School of Food EngineeringState University of Campinas (UNICAMP)CampinasBrazil

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