Alternative methods, including green synthetic approaches for the preparation of various types of nanoparticles are important to maintain sustainable development. Extracellular or intracellular extracts of fungi are perfect candidates for the synthesis of metal nanoparticles due to the scalability and cost efficiency of fungal growth even on industrial scale. There are several methods and techniques that use fungi-originated fractions for synthesis of gold nanoparticles. However, there is less knowledge about the drawbacks and limitations of these techniques. Additionally, identification of components that play key roles in the synthesis is challenging. Here we show and compare the results of three different approaches for the synthesis of gold nanoparticles using either the extracellular fraction, the autolysate of the fungi or the intracellular fraction of 29 thermophilic fungi. We observed the formation of nanoparticles with different sizes (ranging between 6 nm and 40 nm) and size distributions (with standard deviations ranging between 30% and 70%) depending on the fungi strain and experimental conditions. We found by using ultracentrifugal filtration technique that the size of reducing agents is less than 3 kDa and the size of molecules that can efficiently stabilize nanoparticles is greater than 3 kDa.
An improved sol-gel process involving the use of hollow silica microspheres as a supporting additive was applied for the co-immobilization of whole cells of Escherichia coli with Chromobacterium violaceum ω-transaminase activity and Lodderomyces elongisporus with ketoreductase activity. The co-immobilized cells with two different biocatalytic activities could perform a cascade of reactions to convert racemic 4-phenylbutan-2-amine or heptan-2-amine into a nearly equimolar mixture of the corresponding enantiomerically pure R amine and S alcohol even in continuous-flow mode. The novel co-immobilized whole-cell system proved to be an easy-to-store and durable biocatalyst.
Immobilization of transaminases creates promising biocatalysts for production of chiral amines in batch or continuous-flow mode reactions. E. coli cells containing overexpressed transaminases of various selectivities and hollow silica microspheres as supporting agent were immobilized by an improved sol-gel process to produce immobilized transaminase biocatalysts with suitable stability and mechanical properties for continuous-flow applications. The immobilized cell-based transaminase biocatalyst proved to be durable and easy-to-use in kinetic resolution of four racemic amines 1a–d. The batch and continuous-flow mode kinetic resolutions with transaminase biocatalyst of opposite stereopreference provided access to both enantiomers of the corresponding amines. By using the most suitable immobilized transaminase biocatalysts, this study describes the first transaminase-based approach for the production of both pure enantiomers of 1-(3,4-dimethoxyphenyl)ethan-1-amine 1d.
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