Kinetic studies on the oxidation of nitrite by horseradish peroxidase and lactoperoxidase.

Kinetic studies on the oxidation of nitrite by horseradish peroxidase and lactoperoxidase.
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辣根过氧化物酶和乳过氧化物酶氧化亚硝酸盐的动力学研究。

DOI:
10.18388/abp.1999_4114
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发表时间:
1999
影响因子:
1.7
通讯作者:
L. Gebicka
L. Gebicka
中科院分区:
生物学4区
文献类型:
--
作者:
L. Gebicka

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本文研究了亚硝酸盐(NO2-)与辣根过氧化物酶和乳过氧化物酶的反应。顺序混合停流测量给出了亚硝酸盐与辣根过氧化物酶和乳过氧化物酶的化合物II(氧代铁血红素中间体)在pH 7.0下反应的速率常数,分别为13.3 +/- 0.07 mol(-1)dm 3 s(-1)和3.5 +/- 0.05 x 10(4)mol(-1)dm 3 s(-1)。在中性pH下,亚硝酸盐影响乳过氧化物酶与典型底物如2,2 '-连氮基-双[乙基-苯并噻唑啉-(6)-磺酸](ABTS)、愈创木酚或硫氰酸盐(SCN-)的活性测量。ABTS和愈创木酚氧化速率随亚硝酸盐浓度增加而线性增加,直至2.5-5 mmol dm(-3)。另一方面,在亚硝酸盐的存在下,双电子SCN-氧化被抑制。因此,亚硝酸盐竞争与研究的底物乳过氧化物酶。中间体,最有可能是二氧化氮(*NO2),更迅速地与ABTS或愈创木酚反应比乳过氧化物酶化合物II。然而,它没有有效地将SCN-氧化成OSCN-。NO2-对ABTS法和愈创木酚法测定辣根过氧化物酶活性无影响。
The reaction of nitrite (NO2-) with horseradish peroxidase and lactoperoxidase was studied. Sequential mixing stopped-flow measurements gave the following values for the rate constants of the reaction of nitrite with compounds II (oxoferryl heme intermediates) of horseradish peroxidase and lactoperoxidase at pH 7.0, 13.3 +/- 0.07 mol(-1) dm3 s(-1) and 3.5 +/- 0.05 x 10(4) mol(-1) dm3 s(-1), respectively. Nitrite, at neutral pH, influenced measurements of activity of lactoperoxidase with typical substrates like 2,2'-azino-bis[ethyl-benzothiazoline-(6)-sulphonic acid] (ABTS), guaiacol or thiocyanate (SCN-). The rate of ABTS and guaiacol oxidation increased linearly with nitrite concentration up to 2.5-5 mmol dm(-3). On the other hand, two-electron SCN- oxidation was inhibited in the presence of nitrite. Thus, nitrite competed with the investigated substrates of lactoperoxidase. The intermediate, most probably nitrogen dioxide (*NO2), reacted more rapidly with ABTS or guaiacol than did lactoperoxidase compound II. It did not, however, effectively oxidize SCN- to OSCN-. NO2- did not influence the activity measurements of horseradish peroxidase by ABTS or guaiacol method.
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影响因子: 7.4
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