Microbial Biotechnology, Ruhr-Universität Bochum, Bochum, Germany.
X-Ray Structure Analysis of Proteins, Ruhr-Universität Bochum, Bochum, Germany.
Microbiol Spectr. 2021 Sep 3;9(1):e0047421. doi: 10.1128/Spectrum.00474-21. Epub 2021 Jul 28.
The glutathione -transferases carried on the plasmid for the styrene-specific degradation pathway in the Gordonia rubripertincta CWB2 were heterologously expressed in Escherichia coli. Both enzymes were purified via affinity chromatography and subjected to activity investigations. StyI and StyJ displayed activity toward the commonly used glutathione -transferase model substrate 1-chloro-2,4-dinitrobenzene (CDNB) with values of 0.0682 ± 0.0074 and 2.0281 ± 0.1301 mM and values of 0.0158 ± 0.0002 and 0.348 ± 0.008 U mg for StyI and StyJ, respectively. The conversion of the natural substrate styrene oxide to the intermediate (1-phenyl-2-hydroxyethyl)glutathione was detected for StyI with 48.3 ± 2.9 U mg. This elucidates one more step in the not yet fully resolved styrene-specific degradation pathway of Gordonia rubripertincta CWB2. A characterization of both purified enzymes adds more insight into the scarce research field of actinobacterial glutathione -transferases. Moreover, a sequence and phylogenetic analysis puts both enzymes into a physiological and evolutionary context. Styrene is a toxic compound that is used at a large scale by industry for plastic production. Bacterial degradation of styrene is a possibility for bioremediation and pollution prevention. Intermediates of styrene derivatives degraded in the styrene-specific pathways are precursors for valuable chemical compounds. The pathway in Gordonia rubripertincta CWB2 has proven to accept a broader substrate range than other bacterial styrene degraders. The enzymes characterized in this study, distinguish CWB2s pathway from other known styrene degradation routes and thus might be the main key for its ability to produce ibuprofen from the respective styrene derivative. A biotechnological utilization of this cascade could lead to efficient and sustainable production of drugs, flavors, and fragrances. Moreover, research on glutathione metabolism in is rare. Here, a characterization of two glutathione -transferases of actinobacterial origin is presented, and the utilization of glutathione in the metabolism of an is proven.
携带苯乙烯特异性降解途径质粒的戈登氏红球菌 CWB2 中的谷胱甘肽转移酶在大肠杆菌中异源表达。两种酶均通过亲和层析进行纯化，并进行了活性研究。StyI 和 StyJ 对常用的谷胱甘肽转移酶模型底物 1-氯-2,4-二硝基苯（CDNB）具有活性，其 值分别为 0.0682 ± 0.0074 和 2.0281 ± 0.1301 mM， 值分别为 0.0158 ± 0.0002 和 0.348 ± 0.008 U mg，StyI 和 StyJ 分别。检测到 StyI 将天然底物苯乙烯氧化物转化为中间体（1-苯基-2-羟乙基）谷胱甘肽，转化率为 48.3 ± 2.9 U mg。这阐明了戈登氏红球菌 CWB2 中尚未完全解析的苯乙烯特异性降解途径中的又一步骤。对两种纯化酶的特性分析为 actinobacterial 谷胱甘肽转移酶这一稀缺研究领域提供了更多的见解。此外，序列和系统发育分析将两种酶置于生理和进化背景下。苯乙烯是一种有毒化合物，工业上大量用于塑料生产。细菌降解苯乙烯是生物修复和污染预防的一种可能性。在苯乙烯特异性途径中降解的苯乙烯衍生物的中间体是有价值的化合物的前体。CWB2 中的途径已被证明比其他细菌苯乙烯降解菌具有更广泛的底物范围。本研究中表征的酶将 CWB2s 途径与其他已知的苯乙烯降解途径区分开来，因此可能是其从相应的苯乙烯衍生物生产布洛芬的能力的主要关键。该级联的生物技术利用可能导致药物、香料和香精的高效和可持续生产。此外，关于 中的谷胱甘肽代谢的研究很少。这里介绍了两种源自放线菌的谷胱甘肽转移酶的特性，并证明了谷胱甘肽在 的代谢中的利用。
