The invention relates to novel biocatalytic processes comprising the whole cell reduction of dehydrocholic acid (DHCA) compounds, novel 7-hydroxy steroid dehydrogenase mutants, the sequences coding for these enzyme mutants, methods for producing the enzyme mutants and use thereof in enzymatic conversions of cholic acid compounds, and in particular in the production of ursodesoxycholic acid (UDCA); also a subject of the invention are novel methods for the synthesis of UDCA using the enzyme mutants; and in particular a further improved method for producing UDCA using recombinant whole cell biocatalysts.

Methods of cross-linking polypeptide molecules are provided, where such methods include combining a cross-linking agent and polypeptide molecules in a solution under conditions suitable for a cross-linking reaction to occur. Also provided are preparations of cross-linked polypeptide molecules, where the polypeptide molecules are cross-linked by essentially unbranched cross-linking groups of at least 40 contiguous atoms. Further provided are test chemistry matrices and methods of making the same, where the matrices include a redox cofactor, an agent capable of eliciting a change in at least one measurable property of an indicator reagent in the presence of redox equivalents, an indicator reagent, and a preparation of cross-linked polypeptide molecules as described herein. Test elements and methods of using the same to diagnose diseases such as diabetes also are provided, where the test elements include a test chemistry matrix as described herein.

Methods are provided for stabilizing an enzyme by storing the enzyme in the presence of a stabilized coenzyme. In addition, enzymes are provided that are stabilized with a stabilized coenzyme, as well as the use thereof in test elements for detecting analytes. Other aspects include unique compositions, methods, techniques, systems and devices involving enzyme stabilization.

The invention relates to a glucose dehydrogenase showing an NADP/NAD activity ratio, namely the value obtained by dividing the enzyme activity value obtained by using NADP as a coenzyme by the enzyme activity value obtained by using NAD as a coenzyme, of not lower than 300, to a gene coding therefor, and to a transformant harboring that gene. The enzyme of the invention is very high in NADP specificity and can be suitably used, for example, for NADPH production, for coenzyme regeneration in enzymatic reduction reactions, and in biosensors for glucose concentration measurements.

The present invention relates to improved protein-based biosensors that are suitable for detection of one or more target molecules in a sample. The biosensors are fully-reversible with dynamic ranges suitable for analytic and diagnostic applications. The biosensors of the present invention may be used in synthetic biology, for example in constructing artificial cellular or extracellular signalling networks.

There is disclosed a highly stable biocatalytic multi-enzyme system on hydrophobic support for the efficient and continuous hydrogenation of carbon dioxide (CO.sub.2) to formate. The system immobilizes formate dehydrogenase (FDH), glucose dehydrogenase (GDH), and carbonic anhydrase (CA) enzymes on a hydrophobic surface-modified metal-organic framework (MOF), SA-HKUST-1. The hydrophobic surface modification with stearic acid enhances the enzyme stability and reusability, maintaining 95% activity after four cycles. The hydrophobicity of SA-HKUST-1 improves CO.sub.2 diffusion to the immobilized enzymes, significantly boosting the formate production. Enzyme specificity ensures selective reactions, with FDH facilitating CO.sub.2 bioconversion, CA accelerating CO.sub.2 hydration and GDH facilitating cofactor regeneration within the system. The system demonstrates superior performance, producing 255.8 mM formate per gram of MOF per hour. Operating under mild conditions with simple equipment, it reduces costs and eliminates harmful by-products. This invention offers an eco-friendly, sustainable approach for CO.sub.2 mitigation, with potential applications in industrial CO.sub.2 conversion processes.

Methods for stabilizing an enzyme by storing the enzyme in the presence of a stabilized coenzyme are disclosed. In addition, an enzyme stabilized with a stabilized coenzyme as well as the use thereof in test elements for detecting analytes are also disclosed. Other aspects include unique compositions, methods, techniques, systems and devices involving enzyme stabilization.

A method for preparing (S)-nicotine by reduction includes conducting a reduction process on an alkene compound as shown in Formula I and/or an iminium cation compound as shown in Formula II, thereby producing (S)-nicotine. The method is simple, safe, reliable, and yields both high purity and high quantities of (S)-nicotine production.

This invention presents a novel method for engineering glucose dehydrogenase (GDH) proteins by utilizing an atomistic grid-based computational method that analyzes and compares protein atomic compositions. The method involves constructing Localized spherical feature grids (LSFGs) centered around high-energy regions to store atomic properties, enabling comparison with a database of known protein grids. Two comparison techniques are applied: geometric alignment using rotation matrices and quaternions, and transformer-based similarity scoring. High-ranking matches guide functional and stability optimization through mutation design. Unlike conventional methods that require spatial alignment, this approach maps chemical properties directly onto the grid, enabling alignment-free comparisons based on chemical composition allowing comparison of specific protein regions even in the absence of structural similarity. This method provides alignment-free chemical profiling for structurally diverse proteins, facilitating advanced protein engineering and functional annotation.

The present disclosure encompasses a process for chiral resolution of racemic amines, particularly ethyl 2-(4-((tert-butoxycarbonyl)amino)phenyl)piperidine-3-carboxylate, into its desired isomer, ethyl (2R,3S)-2-(4-((tert-butoxycarbonyl)amino)phenyl)-piperidine-3-carboxylate, which is an intermediate useful in the synthesis of Avacopan.