School of Chemical Engineering, University of Ulsan, Ulsan 44610, Republic of Korea.
Bioenergy and Biochemical Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 34141, Republic of Korea.
J Microbiol Biotechnol. 2017 Sep 28;27(9):1664-1669. doi: 10.4014/jmb.1611.11041.
Gamma-aminobutyric acid is a precursor of nylon-4, which is a promising heat-resistant biopolymer. GABA can be produced from the decarboxylation of glutamate by glutamate decarboxylase. In this study, a synthetic scaffold complex strategy was employed involving the Neurospora crassa glutamate decarboxylase (GadB) and GABA antiporter (GadC) to improve GABA production. To construct the complex, the SH3 domain was attached to the N. crassa GadB, and the SH3 ligand was attached to the N-terminus, middle, and C-terminus of GadC. In the C-terminus model, 5.8 g/l of GABA concentration was obtained from 10 g/l glutamate. When a competing pathway engineered strain was used, the final GABA concentration was further increased to 5.94 g/l, which corresponds to 97.5% of GABA yield. With the introduction of the scaffold complex, the GABA productivity increased by 2.9 folds during the initial culture period.
γ-氨基丁酸是尼龙-4 的前体，尼龙-4 是一种有前途的耐热生物聚合物。GABA 可以通过谷氨酸脱羧酶从谷氨酸脱羧产生。在这项研究中，采用了一种合成支架复合物策略，涉及蕈状芽孢杆菌谷氨酸脱羧酶（GadB）和 GABA 反向转运蛋白（GadC），以提高 GABA 的产量。为了构建复合物，将 SH3 结构域连接到 N. crassa GadB 上，将 SH3 配体连接到 GadC 的 N 端、中间和 C 端。在 C 端模型中，从 10 g/l 的谷氨酸中获得了 5.8 g/l 的 GABA 浓度。当使用工程化的竞争途径菌株时，最终的 GABA 浓度进一步增加到 5.94 g/l，对应于 GABA 产率的 97.5%。引入支架复合物后，在初始培养期间，GABA 的生产力提高了 2.9 倍。
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