Archives

  • 2018-07
  • 2018-10
  • 2018-11
  • 2019-04
  • 2019-05
  • 2019-06
  • 2019-07
  • 2019-08
  • 2019-09
  • 2019-10
  • 2019-11
  • 2019-12
  • Itraconazole synthesis Industrial fermentation aims to produ

    2019-12-02

    Industrial fermentation aims to produce valuable products from cheap feedstocks by utilizing diverse microbial functions through aerobic or anaerobic fermentation [[8], [9], [10]]. During anaerobic fermentation, the reducing power is mostly directed to product synthesis rather than being oxidized, resulting in a higher yield of target products [11]. For example, organic acids such as lactic Itraconazole synthesis and succinic acid are efficiently produced through anaerobic fermentation [12,13]. However, the maximum yield of succinic acid production through the anaerobic fermentative pathway is restricted by NADH supply. A sufficient supply of reducing equivalents is essential for obtaining the maximum yield of target fermentation products. PDH converts pyruvate into acetyl-CoA with NADH being produced; if PDH is activated, the NADH availability can be improved, leading to increased production yield when NADH is required in the pathway [14]. It has been reported that activating PDH by increasing aceEF gene expression has improved ethanol production in E. coli [15]. Activating PDH has also satisfied the reducing equivalent requirement to increase anaerobic butanol production [16]. In addition, the theoretical maximum yield of 0.75 mol/mol for anaerobic 1,3-propanediol production from glycerol can be achieved after activating PDH [17]. A high-succinate-producing strain was screened by metabolic evolution. It turned out that PDH was greatly activated and that the sensitivity of PDH complex to NADH inhibition was attenuated due to three nucleotide mutations in lpd [17]. PDH activity increased under anaerobic conditions, which improve NADH supply for succinate production. Therefore, this could be a potential strategy to improve NADH supply under anaerobic conditions by regulating PDH activity through modifying its E3 component, LPD. Although there are few reports on LPD mutation, attempts to find more key sites that further increase PDH activity under anaerobic conditions are still highly desirable. Protein engineering for improving enzyme properties is playing increasingly important roles in chemical biotechnology, aiming to produce chemicals from biomass [18]. In this study, we first explored protein engineering of LPD by structural analysis to find more target sites and the IAA350/351/358VVV triple mutant was successfully verified to be the most effective at improving PDH activity. The mutated LPD attenuated sensitivity to NADH inhibition and also allowed the PDH complex to be more tolerant to NADH in anaerobic E. coli culture. The lpd mutation identified in this work may have wide applications in the production of reduced chemicals in E. coli, including succinate, ethanol, butanol, 1,3-propanediol and so on
    Materials and methods
    Results and discussion
    Conclusion In conclusion, the results presented here indicate that the IAA350/351/358VVV triple mutant of LPD resulted in a PDH complex that is less sensitive to inhibition by NADH. Notably, the altered LPD also maintain the high enzyme activity under the conditions of a higher NADH level. All these findings raise an interesting possibility, wherein parallel modification in lpd gene of other organisms can also alter PDH complexes that are less sensitive to NADH inhibition and still active under anaerobic conditions. Although some other mechanisms may result in PDH complex activity during anaerobic growth, the presence and functional activity of such an NADH-insensitive PDH complexes may have significant potential in biotechnological applications.
    Conflict of interest
    Acknowledgements This work was supported by the National Natural Science Foundation of China (31200078), the Shandong Province Science and Technology Project (No. ZR2017MC023 and 2017GSF217003).
    Gliomas and Aldehyde Dehydrogenase Aldehyde dehydrogenases (ALDHs) are key metabolic enzymes involved in regulation of glycolysis/gluconeogenesis pathways as well as in the detoxification of endogenous and exogenous aldehydes, regulating cell function and homeostasis. ALDHs play important roles in cell proliferation, differentiation, and survival and comprise a family of 19 members (Table 1), which are normally involved in the biosynthesis of retinoic acid (RA) and folate as well as in the formation of the neurotransmitter γ-aminobutyric acid (GABA; Box 1). Selective ALDH isoenzyme activity has been detected in cancer cells with increased aggressiveness, capacity for sustained proliferation, and tumor plasticity [1]. In addition, the ALDH isoenzymes are considered as functional biomarkers of CSCs, possessing an important mechanistic role in the metastatic activity of tumor cells as well as in modulating their response to therapy.