Identification of key residues for activities of atypical glutathione S-transferase of Ceriporiopsis subvermispora, a selective degrader of lignin in woody biomass, by crystallography and functional mutagenesis
Identification of key residues for activities of atypical glutathione S-transferase of Ceriporiopsis subvermispora, a selective degrader of lignin in woody biomass, by crystallography and functional mutagenesis
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通过晶体学和功能诱变鉴定 Ceriporiopsis subvermispora（木质生物质中木质素的选择性降解剂）非典型谷胱甘肽 S-转移酶活性的关键残基
DOI：
10.1016/j.ijbiomac.2019.03.199
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发表时间：
2019
影响因子：
8.2
通讯作者：
Katahira Masato
中科院分区：
文献类型：
--
作者：
Osman Wan Hasnidah Wan;Mikami Bunzo;Saka Naoki;Kondo Keiko;Lin Meng-I;Nagata Takashi;Katahira Masato
Ceriporiopsis subvermispora（C.subvermispora）是木质素的选择性降解菌。谷胱甘肽S-转移酶（GST）是一种多功能酶，在细胞解毒和代谢中发挥重要作用。以1.95和2.19 μ m的分辨率分别解析了无GSH和结合GSH形式的C.subvermispora的GST CsGST 83044的晶体结构。GSH结合形式的结构显示CsGST 83044可以被归类为GST的一种代谢型。在GSH结合形式的CsGST 83044中，Asn 22、Asn 24和Tyr 46最靠近硫原子，并与巯基形成氢键。功能突变表明，它们对CsGST 83044的酶活性至关重要。第一次揭示了属于GSTFuA类的药物型GST的关键残基。先前的研究表明，CsGST 83044和另一种GST，CsGST 63524，不同的底物偏好; CsGST 83044喜欢较小的底物比CsGST 63524的酯酶活性。CsGST 83044的GSH结合口袋变小，这可以解释对较小底物的偏好。本研究结果为进一步利用木本植物资源开辟了一条道路。
Ceriporiopsis subvermispora(C.subvermispora) is a selective degrader of lignin in the woody biomass. Glutathione S-transferases (GSTs) are multifunctional enzymes that play important roles in cellular detoxification and metabolism. The crystal structures of a GST ofC.subvermispora, CsGST83044, in GSH-free and -bound forms were solved at 1.95 and 2.19 Å resolution, respectively. The structure of the GSH-bound form revealed that CsGST83044 can be categorized as an atypical-type of GST. In the GSH-bound form of CsGST83044, Asn22, Asn24, and Tyr46 are located closest to the sulfur atom and form hydrogen bonds with the thiol group. The functional mutagenesis indicated that they are critical for the enzymatic activities of CsGST83044. The critical residues of an atypical-type GST belonging to the GSTFuA class were revealed for the first time. A previous study indicated that CsGST83044 and another GST, CsGST63524, differ in substrate preference; CsGST83044 prefers smaller substrates than CsGST63524 for its esterase activity. The GSH-bound pocket of CsGST83044 turns out to be small, which may explain the preference for smaller substrates. Protein engineering of GSTs ofC.subvermisporain the light of the obtained insight may pave a path in the future for utilization of the woody biomass.