Kim Changman, Kim Mi Yeon, Michie Iain, Jeon Byong-Hun, Premier Giuliano C, Park Sunghoon, Kim Jung Rae
School of Chemical and Biomolecular Engineering, Pusan National University, Busan, 609-735 Republic of Korea.
Sustainable Environment Research Centre (SERC), Faculty of Computing, Engineering and Science, University of South Wales, Pontypridd, Mid-Glamorgan CF37 1DL UK.
Biotechnol Biofuels. 2017 Aug 17;10:199. doi: 10.1186/s13068-017-0886-x. eCollection 2017.
3-Hydroxypropionic acid (3-HP) is an important platform chemical which can be produced biologically from glycerol. is an ideal biocatalyst for 3-HP because it can grow well on glycerol and naturally synthesize the essential coenzyme B. On the other hand, if higher yields and titers of 3-HP are to be achieved, the sustained regeneration of NAD under anaerobic conditions, where coenzyme B is synthesized sustainably, is required.
In this study, recombinant L17 overexpressing aldehyde dehydrogenase (AldH) was developed and cultured in a bioelectrochemical system (BES) with the application of an electrical potential to the anode using a chronoamperometric method (+0.5 V vs. Ag/AgCl). The BES operation resulted in 1.7-fold enhancement of 3-HP production compared to the control without the applied potential. The intracellular NADH/NAD ratio was significantly lower when the L17 cells were grown under an electric potential. The interaction between the electrode and overexpressed AldH was enhanced by electron shuttling mediated by HNQ (2-hydroxy-1,4-naphthoquinone).
Enhanced 3-HP production by the BES was achieved using recombinant L17. The quinone-based electron transference between the electrode and L17 was investigated by respiratory uncoupler experiments. This study provides a novel strategy to control the intracellular redox states to enhance the yield and titer of 3-HP production as well as other bioconversion processes.
3-羟基丙酸(3-HP)是一种重要的平台化学品,可由甘油生物合成。[具体菌株名称]是生产3-HP的理想生物催化剂,因为它能在甘油上良好生长并天然合成必需辅酶B。另一方面,若要实现更高的3-HP产量和滴度,则需要在可持续合成辅酶B的厌氧条件下持续再生NAD。
在本研究中,构建了过表达醛脱氢酶(AldH)的重组[具体菌株名称]L17,并使用计时电流法(相对于Ag/AgCl为+0.5 V)在生物电化学系统(BES)中对阳极施加电势来培养。与未施加电势的对照相比,BES操作使3-HP产量提高了1.7倍。当[具体菌株名称]L17细胞在电势下生长时,细胞内NADH/NAD比率显著降低。由HNQ(2-羟基-1,4-萘醌)介导的电子穿梭增强了电极与过表达的AldH之间的相互作用。
使用重组[具体菌株名称]L17通过BES实现了3-HP产量的提高。通过呼吸解偶联剂实验研究了电极与[具体菌株名称]L17之间基于醌的电子转移。本研究提供了一种控制细胞内氧化还原状态的新策略,以提高3-HP生产以及其他生物转化过程的产量和滴度。