Effects of Polyethylene Glycol on Bovine Intestine Alkaline Phosphatase Activity and Stability
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概要
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In this study, we evaluated the effects of polyethylene glycol (PEG) on bovine intestine alkaline phosphatase (BIALP) activity and stability. In the hydrolysis of p-nitrophenylphosphate (pNPP) at pH 9.8 at 20 °C, the kcat⁄Km values of BIALP plus 5–15% w/v free PEG with molecular masses of 1, 2, 6, and 20 kDa (PEG1000, PEG2000, PEG6000, and PEG20000 respectively) were 120–140%, 180–300%, 130–170%, and 110–140% respectively of that of BIALP without free PEG (1.8 μM−1 s−1), indicating that activation by PEG2000 was the highest. Unmodified BIALP plus 5% PEG2000 and BIALP pegylated with 2,4-bis(O-methoxypolyethylene glycol)-6-chloro-s-triazine exhibited 1.3-fold higher activity on average than that of BIALP without free PEG under various conditions, including pH 7.0–10.0 and 20–65 °C. The temperatures reducing initial activity by 50% in 30-min incubation of unmodified BIALP plus 5% PEG2000 and pegylated BIALP were 51 and 47 °C respectively, similar to that of BIALP without free PEG (49 °C). These results indicate that the addition of PEG2000 and pegylation increase BIALP activity without affecting its stability, suggesting that they can be used in enzyme immunoassay with BIALP to increase sensitivity and rapidity.
- 2011-11-23
著者
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Inouye Kuniyo
Div. Of Food Sci. And Biotechnology Graduate School Of Agriculture Kyoto Univ. Sakyo-ku Kyoto 606-85
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Yasukawa Kiyoshi
Div. Of Food Sci. And Biotechnology Graduate School Of Agriculture Kyoto Univ.
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Yasukawa Kiyoshi
Division Of Food Science And Biotechnology Graduate School Of Agriculture Kyoto University
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Sekiguchi Satoshi
Division Of Advanced Surgical Science And Technology Graduate School Of Medicine Tohoku University
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Inouye Kuniyo
Division If Applied Life Sciences Graduate School Of Agriculture Kyoto University
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Sekiguchi Satoshi
Division Of Food Science And Biotechnology Graduate School Of Agriculture Kyoto University
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