Matsumoto S, Ozawa R, Uchiumi K, Kurihara M
Insect Toxicology and Physiology, The Institute of Physical and Chemical Research, Saitama, Japan.
Biosci Biotechnol Biochem. 1996 Feb;60(2):369-73. doi: 10.1271/bbb.60.369.
Cell-free production of bombykol was done by incubating a pheromone gland homogenate in the presence of NADPH, ATP, and CoA. Addition of n-hexane to the reaction mixture stimulated bombykol production, resulting in production of 238 ng of bombykol from the homogenate equivalent to 2 pheromone glands after 23 h. Removal of either NADPH, ATP, or CoA resulted in no stimulation of bombykol production, suggesting that the final step of the bombykol biosynthetic pathway is done by acyl CoA synthetase and reductase, sequentially. Incubation first with ATP or high concentrations of ATP suppressed the production of bombykol. Since incubation with ATP also inhibited conversion of [1-14C]palmitoyl CoA into 1-hexadecanol, the inhibitory action of ATP seemed attributable to inactivation of the acyl CoA reductase by phosphorylation, as mediated by a protein kinase in the homogenate. Our results suggest that the activity of acyl CoA reductase in bombykol biosynthesis is regulated by phosphorylation/dephosphorylation, and that the activation occurs by dephosphorylation as mediated by phosphoprotein phosphatase.
通过在烟酰胺腺嘌呤二核苷酸磷酸（NADPH）、三磷酸腺苷（ATP）和辅酶A（CoA）存在的情况下孵育信息素腺匀浆来进行家蚕醇的无细胞生产。向反应混合物中添加正己烷刺激了家蚕醇的产生，在23小时后，从相当于2个信息素腺的匀浆中产生了238纳克家蚕醇。去除NADPH、ATP或CoA中的任何一种都不会刺激家蚕醇的产生，这表明家蚕醇生物合成途径的最后一步依次由酰基辅酶A合成酶和还原酶完成。首先用ATP或高浓度的ATP孵育会抑制家蚕醇的产生。由于用ATP孵育也会抑制[1-14C]棕榈酰辅酶A转化为1-十六醇，ATP的抑制作用似乎归因于匀浆中的一种蛋白激酶介导的磷酸化使酰基辅酶A还原酶失活。我们的结果表明，家蚕醇生物合成中酰基辅酶A还原酶的活性受磷酸化/去磷酸化调节，并且激活是由磷蛋白磷酸酶介导的去磷酸化发生的。
