Son Jina, Baritugo Kei-Anne, Sohn Yu Jung, Kang Kyoung Hee, Kim Hee Taek, Joo Jeong Chan, Park Si Jae
Department of Chemical Engineering and Materials Science, Graduate Program in System Health Science and Engineering, Ewha Womans University, Seoul 03760, Republic of Korea.
Center for Bio-based Chemistry, Division of Specialty and Bio-based Chemical Technology, Korea Research Institute of Chemical Technology, Daejeon 34602, Republic of Korea.
ACS Omega. 2022 Aug 4;7(33):29106-29115. doi: 10.1021/acsomega.2c02971. eCollection 2022 Aug 23.
γ-Aminobutyrate (GABA) is an important chemical by itself and can be further used for the production of monomer used for the synthesis of biodegradable polyamides. Until now, GABA production using harboring glutamate decarboxylases (GADs) has been limited due to the discrepancy between optimal pH for GAD activity (pH 4.0) and cell growth (pH 7.0). In this study, we developed recombinant strains expressing mutated GAD from (EcGADmut) and GADs from CICC20209 (LlGAD) and (LsGAD), all of which showed enhanced pH stability and adaptability at a pH of approximately 7.0. In shake flask cultivations, the GABA productions of H36EcGADmut, H36LsGAD, and H36LlGAD were examined at pH 5.0, 6.0, and 7.0, respectively. Finally, H36EcGADmut (40.3 and 39.3 g L), H36LlGAD (42.5 and 41.1 g L), and H36LsGAD (41.6 and 40.2 g L) produced improved GABA titers and yields in batch fermentation at pH 6.0 and pH 7.0, respectively, from 100 g L glucose. The recombinant strains developed in this study could be used for the establishment of sustainable direct fermentative GABA production from renewable resources under mild culture conditions, thus increasing the availability of various GADs.
γ-氨基丁酸(GABA)本身就是一种重要的化学物质,可进一步用于生产用于合成可生物降解聚酰胺的单体。到目前为止,由于谷氨酸脱羧酶(GADs)的最佳活性pH(pH 4.0)与细胞生长的pH(pH 7.0)之间存在差异,利用携带GADs生产GABA受到限制。在本研究中,我们构建了表达来自大肠杆菌(EcGADmut)的突变GAD以及来自嗜热栖热菌CICC20209(LlGAD)和嗜热栖热菌(LsGAD)的GAD的重组菌株,所有这些菌株在pH约为7.0时均表现出增强的pH稳定性和适应性。在摇瓶培养中,分别在pH 5.0、6.0和7.0下检测了H36EcGADmut、H36LsGAD和H36LlGAD的GABA产量。最后,H36EcGADmut(40.3和39.3 g/L)、H36LlGAD(42.5和41.1 g/L)和H36LsGAD(41.6和40.2 g/L)在pH 6.0和pH 7.0的分批发酵中,分别从100 g/L葡萄糖中产生了更高的GABA滴度和产量。本研究构建的重组菌株可用于在温和培养条件下从可再生资源建立可持续的直接发酵生产GABA,从而提高各种GADs的可用性。