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Chain elongation of unprotected carbohydrates in water under mild conditions remains a challenge both in chemical and biochemical synthesis. The Knoevenagel addition or condensation enables transformations to bioactive scaffolds for pharmaceutical and agrochemical compounds. Unfortunately, the catalysts in use for these transformations often reduce the green metrics of the transformations. Here, we use in situ NMR visualizations to explore the prospective use of natural catalysts for the synthesis of triple- and quadruple-functionalized furan- or dihydrofuran-derivatives from glucose and malononitrile. The dihydrofuran derivatives are formed as kinetic, major intermediates in the pathway to furan derivatives when using naturally abundant MgO or bio-sourced chitosan and N-Methyl-d-glucamine (meglumine) as the catalysts in water. Both catalyst loading, solvent composition and pH can be adapted to populate dihydrofurans with four substituents by slowing down their further reactions. Higher temperatures and higher pH values favor the formation of triple-functionalized furans over quadruple-substituted dihydrofurans, which may be bicyclic or monocyclic. Compared to more traditional catalysts, nature-sourced options offer more sustainable options that emulate natural processes. Visualization with in situ NMR contributes to streamlining the development of cheap and environmentally benign procedures for carbohydrate chain elongation. [ABSTRACT FROM AUTHOR]

Many enzymes have latent activities that can be used in the conversion of non-natural reactants for novel organic conversions. A classic example is the conversion of benzaldehyde to a phenylacetyl carbinol, a precursor for ephedrine manufacture. It is often tacitly assumed that purified enzymes are more promising catalysts than whole cells, despite the lower cost and easier maintenance of the latter. Competing substrates inside the cell have been known to elicit currently hard-to-predict selectivities that are not easily measured inside the living cell. We employ NMR spectroscopic assays to rationally combine isomers for selective reactions in commercial S. cerevisiae. This approach uses internal competition between alternative pathways of aldehyde clearance in yeast, leading to altered selectivities compared to catalysis with the purified enzyme. In this manner, 4-fluorobenzyl alcohol and 2-fluorophenylacetyl carbinol can be formed with selectivities in the order of 90%. Modification of the cellular redox state can be used to tune product composition further. Hyperpolarized NMR shows that the cellular reaction and pathway usage are affected by the xenochemical. Overall, we find that the rational construction of ternary or more complex substrate mixtures can be used for in-cell NMR spectroscopy to optimize the upgrading of similar xenochemicals to dissimilar products with cheap whole-cell catalysts. [ABSTRACT FROM AUTHOR]