Biocatalytic Silylation: The Condensation of Phenols and Alcohols with Triethylsilanol

From MDPI via Jisc Publications RouterHistory: accepted 2021-07-20, pub-electronic 2021-07-22Publication status: PublishedFunder: Engineering and Physical Sciences Research Council; Grant(s): EP/S013539/1, EP/M506436/1Funder: Biotechnology and Biological Sciences Research Council; Grant(s): BB/L013649/1, BB/J014478/1, BB/M017702/1Funder: Tertiary Education Trust Fund; Grant(s): Graduate ScholarshipSilicatein-α (Silα), a hydrolytic enzyme derived from siliceous marine sponges, is one of the few enzymes in nature capable of catalysing the metathesis of silicon–oxygen bonds. It is therefore of interest as a possible biocatalyst for the synthesis of organosiloxanes. To further investigate the substrate scope of this enzyme, a series of condensation reactions with a variety of phenols and aliphatic alcohols were carried out. In general, it was observed that Silα demonstrated a preference for phenols, though the conversions were relatively modest in most cases. In the two pairs of chiral alcohols that were investigated, it was found that the enzyme displayed a preference for the silylation of the S-enantiomers. Additionally, the enzyme’s tolerance to a range of solvents was tested. Silα had the highest level of substrate conversion in the nonpolar solvents n-octane and toluene, although the inclusion of up to 20% of 1,4-dioxane was tolerated. These results suggest that Silα is a potential candidate for directed evolution toward future application as a robust and selective biocatalyst for organosiloxane chemistry

Biocatalytic Silylation: The Condensation of Phenols and Alcohols with Triethylsilanol

Silicatein-α (Silα), a hydrolytic enzyme derived from siliceous marine sponges, is one of the few enzymes in nature capable of catalysing the metathesis of silicon–oxygen bonds. It is therefore of interest as a possible biocatalyst for the synthesis of organosiloxanes. To further investigate the substrate scope of this enzyme, a series of condensation reactions with a variety of phenols and aliphatic alcohols were carried out. In general, it was observed that Silα demonstrated a preference for phenols, though the conversions were relatively modest in most cases. In the two pairs of chiral alcohols that were investigated, it was found that the enzyme displayed a preference for the silylation of the S-enantiomers. Additionally, the enzyme’s tolerance to a range of solvents was tested. Silα had the highest level of substrate conversion in the nonpolar solvents n-octane and toluene, although the inclusion of up to 20% of 1,4-dioxane was tolerated. These results suggest that Silα is a potential candidate for directed evolution toward future application as a robust and selective biocatalyst for organosiloxane chemistry

Improved Production and Biophysical Analysis of Recombinant Silicatein-α

Silicatein-α is a hydrolase found in siliceous sea sponges with a unique ability to condense and hydrolyse silicon–oxygen bonds. The enzyme is thus of interest from the perspective of its unusual enzymology, and for potential applications in the sustainable synthesis of siloxane-containing compounds. However, research into this enzyme has previously been hindered by the tendency of silicatein-α towards aggregation and insolubility. Herein, we report the development of an improved method for the production of a trigger factor-silicatein fusion protein by switching the previous hexahistidine tag for a Strep-II tag, resulting in 244-fold improvement in protein yield compared to previous methods. Light scattering and thermal denaturation analyses show that under the best storage conditions, although oligomerisation is never entirely abolished, these nanoscale aggregates of the Strep-tagged protein exhibit improved colloidal stability and solubility. Enzymatic assays show that the Strep-tagged protein retains catalytic competency, but exhibits lower activity compared to the His6-tagged protein. These results suggest that the hexahistidine tag is capable of non-specific catalysis through their imidazole side chains, highlighting the importance of careful consideration when selecting a purification tag. Overall, the Strep-tagged fusion protein reported here can be produced to a higher yield, exhibits greater stability, and allows the native catalytic properties of this protein to be assessed

Occurrence of Aflatoxin M1 in Milks of Five Animal Species in Iran: A Systematic Review and Meta-analysis

Consumption of milks contaminated with aflatoxin M1 (AFM1) may result in serious health problems in humans. In the present study, English and Persian electronic databases were comprehensively searched for publications from 2005 to 2018. Results indicated that pooled prevalence of AFM1 contamination in milks of buffalo, cow, sheep, goat, and camel were 86, 86, 42, 34, and 30%, respectively. Furthermore, average concentration of AFM1 were 78.73, 40.86, 26.71, 24.30, and 20.63 ng/L for milks in the same order. Therefore, continued monitoring of AFM1 contamination in milks and dairy foodstuffs deserves a serious governmental consideration