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Purification, Properties, and Application of a Novel Acid Urease from Enterobacter sp.

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Abstract

It has been demonstrated that acid urease is capable of decomposing urea in fermented beverage and foods. As urea is a precursor of ethylcarbamate, a potential carcinogenic compound, measures must be taken to control the level of urea. We herein describe the purification and characterization of a novel acid urease from Enterobacter sp. R-SYB082 and its application to the removal of urea in Chinese rice wine. The enzyme was purified to electrophoretic homogeneity using ethanol precipitation, Superdex 200 and Mono Q with a fold purification of 21.1 and a recovery of 49%. The molecular weight of the enzyme was 430,000 Da by gel filtration and 72,000 Da by sodium dodecyl sulfate polyacrylamide gel electrophoresis, suggesting that it was a hexamer. The activity of this purified enzyme was optimal at pH 4.5 and 35 °C. The temperature stability was under 55 °C, and the pH stability was 4.0~5.0. The enzyme exhibited an apparent K m of 19.5 μmol/l and a V max of 109 μmol urea/mg·min at 35 °C and pH 4.5. When incubating two different kinds of Chinese rice wine with the enzyme (0.08 U/ml) at 35 °C for 7 days, over 85% of urea was decomposed, and at 20 °C, above 78% was removed. The result showed that the enzyme is applicable to elimination of urea in Chinese rice wine.

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  1. The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, 214122, China

    Lu-qiang Yang, Song-hua Wang & Ya-ping Tian

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  1. Lu-qiang Yang
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  2. Song-hua Wang
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  3. Ya-ping Tian
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Correspondence to Ya-ping Tian.

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Yang, Lq., Wang, Sh. & Tian, Yp. Purification, Properties, and Application of a Novel Acid Urease from Enterobacter sp.. Appl Biochem Biotechnol 160, 303–313 (2010). https://doi.org/10.1007/s12010-008-8159-6

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  • DOI: https://doi.org/10.1007/s12010-008-8159-6

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