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Purification and properties of 6-methylsalicylic acid synthase from Penicillium patulum.

机译：青霉青霉6-甲基水杨酸合酶的纯化及性质。

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摘要

6-Methylsalicylic acid synthase has been isolated in homogeneous form from Penicillium patulum grown in liquid culture from a spore inoculum. The enzyme is highly susceptible to proteolytic degradation in vivo and in vitro, but may be stabilized during purification by incorporating proteinase inhibitors in the buffers. The enzyme exists as a homotetramer of M(r) 750,000, with a subunit M(r) of 180,000. 6-Methylsalicyclic acid synthase also accepts acetoacetyl-CoA as an alternative starter molecule to acetyl-CoA. The enzyme also catalyses the formation of small amounts of triacetic acid lactone as an oligatory by-product of the reaction. In the absence of NADPH, triacetic acid lactone is the exclusive enzymic product, being formed at 10% of the rate of 6-methylsalicylic acid. The enzyme is inactivated by 1,3-dibromopropan-2-one, leading to the formation of cross-linked dimers similar to that observed with type I fatty acid synthases. Acetyl-CoA protects the enzyme against the inactivation and inhibits dimer formation. An adaptation of the purification method for 6-methylsalicylic acid synthase may be used for the isolation of fatty acid sythase from Penicillium patulum.
机译：6-甲基水杨酸合酶已经从孢子接种物的液体培养物中生长的青霉中分离出同质形式。该酶在体内和体外对蛋白水解降解高度敏感，但在纯化过程中可通过在缓冲液中掺入蛋白酶抑制剂使其稳定。该酶以M（r）750,000的同型四聚体形式存在，其亚基M（r）为180,000。 6-甲基水杨酸合酶也接受乙酰乙酰辅酶A作为乙酰辅酶A的替代起始分子。该酶还催化作为反应的寡聚副产物的少量三乙酸内酯的形成。在不存在NADPH的情况下，三乙酸内酯是唯一的酶产物，其形成率为6-甲基水杨酸的10％。该酶被1，3-二溴丙烷-2-酮灭活，导致形成类似于I型脂肪酸合酶的交联二聚体。乙酰辅酶A保护酶免于失活并抑制二聚体形成。对6-甲基水杨酸合酶的纯化方法的改进可用于从青霉菌中分离脂肪酸合酶。

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    Spencer, J B; Jordan, P M;

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  • 年度 1992
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