On the electrooxidation of glucose on gold: towards an electrochemical glucaric acid production as value-added chemical

The electrocatalytic oxidation of glucose to value-added chemicals, such as glucaric acid, has gathered increased interest in recent years. Glucose oxidation is a promising process which has the potential to contribute to establishing renewable resources as alternatives to fossil carbon sources. Herein, we present rotating disk electrode (RDE) studies on polycrystalline gold surfaces and subsequent Koutecký-Levich analysis as a benchmark to expand the understanding of reaction kinetics and competition between glucose, reaction intermediates and OH- at the catalyst surface. Combining electrochemical studies and Raman spectroscopy, it is shown that increasing glucose concentrations lead to a delayed oxidation of the gold catalyst surface, presumably by increased consumption rates of Au-hydroxide species.

Ginkgo Biloba, a famous living fossil tree and an ancient herbal traditional Chinese medicine

Background: Ginkgo (Ginkgo biloba) is a living fossil and a deciduous tree, which has extracts with antidiabetic, antioxidant anticancer, antihypertensive, immunestimulative, hepato-protective and antimicrobial activities, memory enhancement efficiency and beneficial effects against neurodegenerative disease. Objective: The goal of this manuscript is survey on medicinal values and natural benefits of Ginkgo. Methods: This review evaluated publication in MEDLINE/PubMEd database and Google Scholar. The keywords used to electronically search were Ginkgo biloba, living fossil, bioactive components and traditional Chinese medicine. Results: Its main active constituents extracts are flavones glycosides such kaempferol, quercetin and isorhamnetin, terpene lactones, alkyphenols, proanthocyanadins, rhamnose, glucose, D-glucaric acid, ginkgolic acid, organic acids such as hydroxykinurenic, kynurenic, protocatechic, shikimic and vanillic. Ginkgo kernels have been used as medicine or eaten as nut in traditional medicinal science. The most notable pharmaceutical application of Ginkgo are in cardiovascular disease, Alzheimer, impaired cerebral performance, vascular insufficiency, antidepressant-induced sexual dysfunction, premenstrual syndrome, liver fibrosis, vascular disease, tinnitus, macular degeneration, memory and vertigo. Conclusion: Development of modern drugs from Ginkgo by considering the importance of traditional medicinal Asian science with further researches should be emphasized.

Simultaneous Determination of 78 Compounds of Rhodiola rosea Extract by Supercritical CO2-Extraction and HPLC-ESI-MS/MS Spectrometry

The plant Rhodiola rosea L. of family Crassulaceae was extracted using the supercritical CO2-extraction method. Several experimental conditions were investigated in the pressure range of 200–500 bar, with the used volume of cosolvent ethanol in the amount of 1% in the liquid phase at a temperature in the range of 31–70°C. The most effective extraction conditions are pressure 350 bar and temperature 60°C. The extracts were analyzed by HPLC with MS/MS identification. 78 target analytes were isolated from Rhodiola rosea (Russia) using a series of column chromatography and mass spectrometry experiments. The results of the analysis showed a spectrum of the main active ingredients Rh. rosea: salidroside, rhodiolosides (B and C), rhodiosin, luteolin, catechin, quercetin, quercitrin, herbacetin, sacranoside A, vimalin, and others. In addition to the reported metabolites, 29 metabolites were newly annotated in Rh. rosea. There were flavonols: dihydroquercetin, acacetin, mearnsetin, and taxifolin-O-pentoside; flavones: apigenin-O-hexoside derivative, tricetin trimethyl ether 7-O-hexosyl-hexoside, tricin 7-O-glucoronyl-O-hexoside, tricin O-pentoside, and tricin-O-dihexoside; flavanones: eriodictyol-7-O-glucoside; flavan-3-ols: gallocatechin, hydroxycinnamic acid caffeoylmalic acid, and di-O-caffeoylquinic acid; coumarins: esculetin; esculin: fraxin; and lignans: hinokinin, pinoresinol, L-ascorbic acid, glucaric acid, palmitic acid, and linolenic acid. The results of supercritical CO2-extraction from roots and rhizomes of Rh. rosea, in particular, indicate that the extract contained all biologically active components of the plant, as well as inert mixtures of extracted compositions.

Consolidated bioprocessing of lignocellulose for production of glucaric acid by an artificial microbial consortium

Background The biomanufacturing of d-glucaric acid has attracted increasing interest because it is one of the top value-added chemicals produced from biomass. Saccharomyces cerevisiae is regarded as an excellent host for d-glucaric acid production. Results The opi1 gene was knocked out because of its negative regulation on myo-inositol synthesis, which is the limiting step of d-glucaric acid production by S. cerevisiae. We then constructed the biosynthesis pathway of d-glucaric acid in S. cerevisiae INVSc1 opi1Δ and obtained two engineered strains, LGA-1 and LGA-C, producing record-breaking titers of d-glucaric acid: 9.53 ± 0.46 g/L and 11.21 ± 0.63 g/L d-glucaric acid from 30 g/L glucose and 10.8 g/L myo-inositol in fed-batch fermentation mode, respectively. However, LGA-1 was preferable because of its genetic stability and its superior performance in practical applications. There have been no reports on d-glucaric acid production from lignocellulose. Therefore, the biorefinery processes, including separated hydrolysis and fermentation (SHF), simultaneous saccharification and fermentation (SSF) and consolidated bioprocessing (CBP) were investigated and compared. CBP using an artificial microbial consortium composed of Trichoderma reesei (T. reesei) Rut-C30 and S. cerevisiae LGA-1 was found to have relatively high d-glucaric acid titers and yields after 7 d of fermentation, 0.54 ± 0.12 g/L d-glucaric acid from 15 g/L Avicel and 0.45 ± 0.06 g/L d-glucaric acid from 15 g/L steam-exploded corn stover (SECS), respectively. In an attempt to design the microbial consortium for more efficient CBP, the team consisting of T. reesei Rut-C30 and S. cerevisiae LGA-1 was found to be the best, with excellent work distribution and collaboration. Conclusions Two engineered S. cerevisiae strains, LGA-1 and LGA-C, with high titers of d-glucaric acid were obtained. This indicated that S. cerevisiae INVSc1 is an excellent host for d-glucaric acid production. Lignocellulose is a preferable substrate over myo-inositol. SHF, SSF, and CBP were studied, and CBP using an artificial microbial consortium of T. reesei Rut-C30 and S. cerevisiae LGA-1 was found to be promising because of its relatively high titer and yield. T. reesei Rut-C30 and S. cerevisiae LGA-1were proven to be the best teammates for CBP. Further work should be done to improve the efficiency of this microbial consortium for d-glucaric acid production from lignocellulose.

Flux vs burden: biosensor optimization for dynamic pathway engineering

Recent progress in synthetic biology allows the construction of dynamic control circuits for metabolic engineering. This technology promises to overcome many challenges encountered in traditional pathway engineering, thanks to their ability to self-regulate gene expression in response to bioreactor perturbations. The central components in these control circuits are metabolite biosensors that read out pathway signals and actuate enzyme expression. However, the construction of metabolite biosensors is laborious and currently a major bottleneck for strain design. Here we present a general method for biosensor design based on multiobjective optimization. Our approach produces libraries of biosensors that optimally trade-off production flux against the genetic burden on the host. We explore properties of control architectures built in the literature, and identify their advantages and caveats in terms of performance and robustness to growth conditions or leaky promoters. We demonstrate the optimality of a control circuit for glucaric acid production in Escherichia coli, which has been shown to increase titer by 2.5-fold as compared to static designs. Our results lay the groundwork for the automated design of control circuits for pathway engineering, with applications in the food, energy and pharmaceutical sectors.