Identifying a sigma class glutathione S-transferase 2 from the silkworm <i>Bombyx mori</i>
Identifying a sigma class glutathione S-transferase 2 from the silkworm <i>Bombyx mori</i>
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鉴定家蚕 Bombyx mori 中的 sigma 类谷胱甘肽 S-转移酶 2
DOI：
10.11416/jibs.86.1_001
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发表时间：
2017
影响因子：
--
通讯作者：
Kohji Yamamoto
中科院分区：
文献类型：
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作者：
Aiko Hirowatari;Sumiharu Nagaoka;Naotaka Yamada;Kohji Yamamoto
利用逆转录聚合酶链反应（RT-PCR）技术从家蚕（Bombyx mori）中克隆了一个编码sigma类谷胱甘肽S-转移酶（GST; bmGSTS 2）的cDNA，并进行了序列测定。推导的氨基酸序列与其它昆虫的sigma类GST序列同源性为68%~ 63%。bmGSTS 2 mRNA在各组织中分布广泛。该重组酶在大肠杆菌中以可溶性形式功能性过表达，纯化至同质并进行表征。bmGSTS 2的最适pH为7.0左右，在pH6.0 ~ 8.0条件下培养12 h后，bmGSTS 2的活性仍保持在75%以上。在低于40 ℃的温度下孵育30分钟不影响酶活性。bmGSTS 2能够催化谷胱甘肽与过氧化氢和4-羟基壬烯醛的反应。这些结果表明，bmGSTS 2可能在家蚕的抗氧化防御中发挥作用。
A cDNA that encodes a sigma class glutathione S-transferase (GST; bmGSTS2) from the silkworm (Bombyx mori) was cloned by reverse transcriptase polymerase chain reaction and sequenced. The deduced amino acid sequence revealed 68%-63% identity with the sigma class GSTs from other insects. bmGSTS2 mRNA was widely distributed in various tissues. The recombinant enzyme was functionally overexpressed as a soluble form in Escherichia coli, purified to homogeneity, and characterized. The optimum pH of bmGSTS2 was approximately pH 7.0, and bmGSTS2 retained> 75% of its original activity after incubation for 12 h at pH 6.0-8.0. Incubation for 30 min at temperatures below 40 C did not affect the enzymatic activity. bmGSTS2 was able to catalyze the reaction of glutathione with hydrogen peroxide, and 4-hydroxynonenal. These results indicate that bmGSTS2 may play a role in antioxidant defense in the silkworm.