Hengkai Meng1,2,3, Wei Zhang1,2, Huawei Zhu2,4, Fan Yang2,4, Yanping Zhang2, Jie Zhou5, Yin Li6. 1. Department of Cellular Biology, University of Science and Technology of China, Hefei, China. 2. CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, No. 1 West Beichen Road, Chaoyang District, Beijing, 100101, China. 3. State Key Laboratory of Transducer Technology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China. 4. University of Chinese Academy of Sciences, Beijing, China. 5. CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, No. 1 West Beichen Road, Chaoyang District, Beijing, 100101, China. jiezhouw@im.ac.cn. 6. CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, No. 1 West Beichen Road, Chaoyang District, Beijing, 100101, China. yli@im.ac.cn.
Abstract
BACKGROUND: An efficient supply of reducing equivalent is essential for chemicals production by engineered microbes. In phototrophic microbes, the NADPH generated from photosynthesis is the dominant form of reducing equivalent. However, most dehydrogenases prefer to utilize NADH as a cofactor. Thus, sufficient NADH supply is crucial to produce dehydrogenase-derived chemicals in cyanobacteria. Photosynthetic electron is the sole energy source and excess electrons are wasted in the light reactions of photosynthesis. RESULTS: Here we propose a novel strategy to direct the electrons to generate more ATP from light reactions to provide sufficient NADH for lactate production. To this end, we introduced an electron transport protein-encoding gene omcS into cyanobacterium Synechococcus elongatus UTEX 2973 and demonstrated that the introduced OmcS directs excess electrons from plastoquinone (PQ) to photosystem I (PSI) to stimulate cyclic electron transfer (CET). As a result, an approximately 30% increased intracellular ATP, 60% increased intracellular NADH concentrations and up to 60% increased biomass production with fourfold increased D-lactate production were achieved. Comparative transcriptome analysis showed upregulation of proteins involved in linear electron transfer (LET), CET, and downregulation of proteins involved in respiratory electron transfer (RET), giving hints to understand the increased levels of ATP and NADH. CONCLUSIONS: This strategy provides a novel orthologous way to improve photosynthesis via enhancing CET and supply sufficient NADH for the photosynthetic production of chemicals.
BACKGROUND: An efficient supply of reducing equivalent is essential for chemicals production by engineered microbes. In phototrophic microbes, the NADPH generated from photosynthesis is the dominant form of reducing equivalent. However, most dehydrogenases prefer to utilize NADH as a cofactor. Thus, sufficient NADH supply is crucial to produce dehydrogenase-derived chemicals in cyanobacteria. Photosynthetic electron is the sole energy source and excess electrons are wasted in the light reactions of photosynthesis. RESULTS: Here we propose a novel strategy to direct the electrons to generate more ATP from light reactions to provide sufficient NADH for lactate production. To this end, we introduced an electron transport protein-encoding gene omcS into cyanobacterium Synechococcus elongatus UTEX 2973 and demonstrated that the introduced OmcS directs excess electrons from plastoquinone (PQ) to photosystem I (PSI) to stimulate cyclic electron transfer (CET). As a result, an approximately 30% increased intracellular ATP, 60% increased intracellular NADH concentrations and up to 60% increased biomass production with fourfold increased D-lactate production were achieved. Comparative transcriptome analysis showed upregulation of proteins involved in linear electron transfer (LET), CET, and downregulation of proteins involved in respiratory electron transfer (RET), giving hints to understand the increased levels of ATP and NADH. CONCLUSIONS: This strategy provides a novel orthologous way to improve photosynthesis via enhancing CET and supply sufficient NADH for the photosynthetic production of chemicals.
Entities:
Keywords:
Cyanobacteria; Electron transfer; Electron transport protein; Lactate production; NADH availability; Photosynthetic electron
Authors: E K Khandogina; V A Ageĭkin; S V Zvereva; L F Marchenko; G R Mutovin; G P Snigireva; R V Lenskaia; V M Buiankin; V V Shakhtarin; A P Akif'ev Journal: Radiats Biol Radioecol Date: 1995 Sep-Oct
Authors: Peter J Dahl; Sophia M Yi; Yangqi Gu; Atanu Acharya; Catharine Shipps; Jens Neu; J Patrick O'Brien; Uriel N Morzan; Subhajyoti Chaudhuri; Matthew J Guberman-Pfeffer; Dennis Vu; Sibel Ebru Yalcin; Victor S Batista; Nikhil S Malvankar Journal: Sci Adv Date: 2022-05-11 Impact factor: 14.957
Authors: Jens Neu; Catharine C Shipps; Matthew J Guberman-Pfeffer; Cong Shen; Vishok Srikanth; Jacob A Spies; Nathan D Kirchhofer; Sibel Ebru Yalcin; Gary W Brudvig; Victor S Batista; Nikhil S Malvankar Journal: Nat Commun Date: 2022-09-07 Impact factor: 17.694