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Induction, purification and characterization of malolactic enzyme from Oenococcus oeni SD-2a

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Abstract

The aim of this study was to purify a malolactic enzyme (MLE) from Oenococcus oeni (O. oeni) strain and determine its properties in detail. O. oeni SD-2a was cultivated in the ATB broth supplemented with 7 g/L l-malic acid for harvesting the cells. After harvest, the cells were washed and disrupted for purification of MLE. MLE was purified from the supernatant of the disrupted cells through protamine sulfate precipitation, anion exchange chromatography and gel filtration chromatography. The purified MLE was identified using mass spectrometry. The MLE was purified by 43-fold with a yield of 0.42 % and possessed a specific activity of 419.2 U/mg. The purified enzyme with a nominal molecular mass of 59 kDa and a theoretical pI of 4.76 exhibited a maximum enzyme activity at 35 °C and pH 6.0, which retained over 50 % of its initial activity in the presence of 14 % (v/v) ethanol. Mn2+ was proven to be the most effective divalent cation to promote enzyme activity. Under the conditions of temperature 30 °C and pH 6.0, the K m and V max of MLE on l-malic acid were 12.5 × 10−3 M and 43.86 μmol/(min × mg), respectively. Moreover, the purified enzyme exhibited a higher stability with 0.1 M NaCl in addition and had a half-life of 30 days at 4 °C.

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Acknowledgments

This work was financially supported by the National Production of Grape Industry (CRAS-30-gi-3).

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This article does not contain any studies with human or animal subjects.

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Authors and Affiliations

  1. College of Food Science and Engineering, Northwest A&F University, Yangling, 712100, Shaanxi, People’s Republic of China

    Panxue Wang, Aixia Li, Mei Dong & Mingtao Fan

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  1. Panxue Wang
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  2. Aixia Li
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  3. Mei Dong
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  4. Mingtao Fan
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Correspondence to Mingtao Fan.

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Wang, P., Li, A., Dong, M. et al. Induction, purification and characterization of malolactic enzyme from Oenococcus oeni SD-2a. Eur Food Res Technol 239, 827–835 (2014). https://doi.org/10.1007/s00217-014-2276-y

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