Anchor Peptide-based Immobilization Strategy Promote the Applications of Pickering Emulsion System in Natural Product Glycosylation.

Pickering emulsion systems are an advanced platform for efficient fine chemical production in biphasic enzymatic catalysis, while their applications are currently limited to a few commercial enzyme classes. Herein, we designed an anchor peptide-based immobilization strategy for Pickering emulsions to achieve efficient glycosylation of natural products by glycosyltransferases (GTs). Firstly, through enzyme mining, natural GTs were utilized to synthesize pharmaceutically important acacetin glucoside and galactoside.

Hook loop dynamics engineering transcended the barrier of activity-stability trade-off and boosted the thermostability of enzymes.

The improvement of enzyme thermostability often accompanies the decreased activity due to the loss of the key regions' flexibility. As a representative structure, unlocking the potential of loop dynamics will not only provide new ideas for stabilization strategies, but also help to deepen the understanding of the relationship between enzyme structural dynamics and function. In this study, a creative "hook loop dynamics engineering" (HLoD) strategy was successfully proposed for simultaneously improving the thermostability and maintaining activity of the model enzyme, Candida Antarctica lipase B.

The molecular basis and enzyme engineering strategies for improvement of coupling efficiency in cytochrome P450s.

Cytochrome P450s are heme-thiolate enzymes that have been broadly applied in pharmaceutical and biosynthesis because of their efficient oxidation at inert carbons. Extensive engineering campaigns are applied to P450s to explore new non-natural substrates and reactions; however, achieving high coupling efficiency is one of the main challenges. The undesirable uncoupling reactions result in the extra consumption of expensive cofactor NAD(P)H, and lead to the accumulation of reactive oxygen species and the inactivation of enzymes and organisms.

Modulating the Coupling Efficiency of P450 BM3 by Controlling Water Diffusion through Access Tunnel Engineering.

Cytochromes P450 have gained much interest for their broad substrate scope in the catalysis of oxidation reactions for pharmaceuticals, plastics, and hormones. However, achieving high coupling efficiency by the engineering of P450s is still a big challenge. The presence of extra water around the active site is deemed to be related to uncoupling.

Tunnel engineering for modulating the substrate preference in cytochrome P450HI.

An active site is normally located inside enzymes, hence substrates should go through a tunnel to access the active site. Tunnel engineering is a powerful strategy for refining the catalytic properties of enzymes. Here, P450HI (Q85H/V170I) derived from hydroxylase P450 from Bacillus subtilis was chosen as the study model, which is reported as a potential decarboxylase.

Enzymatic cascade biosynthesis reaction of musky macrolactones from fatty acids.

Musky macrolactones are an important group of compounds used in high-valued perfumery. An enzymatic cascade reaction including cytochrome P450 hydroxylase and lipase was explored to biosynthesize musky macrolactones. Firstly, fatty acids were hydroxylated by P450 hydroxylase to produce the corresponding ω-hydroxy fatty acids.

Chemoenzymatic Synthesis of Fragrance Compounds from Stearic Acid.

Fatty acids are versatile precursors for fuels, fine chemicals, polymers, perfumes, etc. The properties and applications of fatty acid derivatives depend on chain length and on functional groups and their positions. To tailor fatty acids for desired properties, an engineered P450 monooxygenase has been employed here for enhanced selective hydroxylation of fatty acids.