Electric field orientations in solution and enzyme active site revealed by a two-directional vibrational probe

The catalytic power of an electric field depends on its magnitude and orientation with respect to the reactive chemical species. Understanding and designing new catalysts for electrostatic catalysis thus requires methods to measure the electric field orientation and magnitude at the molecular scale. We demonstrate that electric field orientations can be extracted using a two-directional vibrational probe by exploiting the vibrational Stark effect of both the C=O and C-D stretches of a deuterated aldehyde. Combining spectroscopy with molecular dynamics and electronic structure partitioning methods, we demonstrate that despite distinct polarities, solvents act similarly in their preference for electrostatically stabilizing large bond dipoles at the expense of destabilizing small ones. In contrast, we find that for an active site aldehyde inhibitor of liver alcohol dehydrogenase, the electric field orientation deviates markedly from that found in solvents, providing direct evidence for the fundamental difference between the electrostatic environments of solvents and a preorganized enzyme active site.

Cellulases: From Bioactivity to a Variety of Industrial Applications

Utilization of microbial enzymes has been widely reported for centuries, but the commercial use of enzymes has been recently adopted. Particularly, cellulases have been utilized in various commercial sectors including agriculture, brewing, laundry, pulp and paper and textile industry. Cellulases of microbial origin have shown their potential application in various commercial sectors including textile, pulp and paper, laundry, brewing, agriculture and biofuel. Cellulases have diversified applications in the food industry, food service, food supply and its preservation. Indeed, cellulases can tenderize fruits, clarify the fruit juices, reduce roughage in dough, hydrolyze the roasted coffee, extract tea polyphenols and essential oils from olives and can increase aroma and taste in food items. However, their role in food industries has by and large remained neglected. The use of immobilized cellulases has further expanded their application in fruit and vegetable processing as it potentiates the catalytic power and reduces the cost of process. Technological and scientific developments will further expand their potential usage in the food industry.

Catalytic Descriptors to Investigate Catalytic Power in the Reaction of Haloalkane Dehalogenase Enzyme with 1,2-Dichloroethane

Enzymes play a fundamental role in many biological processes. We present a theoretical approach to investigate the catalytic power of the haloalkane dehalogenase reaction with 1,2-dichloroethane. By removing the three main active-site residues one by one from haloalkane dehalogenase, we found two reactive descriptors: one descriptor is the distance difference between the breaking bond and the forming bond, and the other is the charge difference between the transition state and the reactant complex. Both descriptors scale linearly with the reactive barriers, with the three-residue case having the smallest barrier and the zero-residue case having the largest. The results demonstrate that, as the number of residues increases, the catalytic power increases. The predicted free energy barriers using the two descriptors of this reaction in water are 23.1 and 24.2 kcal/mol, both larger than the ones with any residues, indicating that the water solvent hinders the reactivity. Both predicted barrier heights agree well with the calculated one at 25.2 kcal/mol using a quantum mechanics and molecular dynamics approach, and also agree well with the experimental result at 26.0 kcal/mol. This study shows that reactive descriptors can also be used to describe and predict the catalytic performance for enzyme catalysis.

An electrostatic duel: subtle differences in catalytic performance of monoamine oxidase A and B isoenzymes elucidated at a residue level by quantum computations

The origin of the immense catalytic power of enzymes remains one of the biggest unresolved questions in biochemistry, with electrostatics being one of the main contenders. Herein we report results...

Review on Green Synthesized Nanocomposites and Their Biological Activities

In this paper, we reviewed the present status of synthesis of nano structured materials for technological development as bimetallic, trimetallic and various organic, inorganic polymers nanocomposites in the field of nanosciences. Nanocomposites play an important role in the field of sciences, engineering and industries due to their high catalytic power, high optical, electrical and mechanical properties, which can be very useful in the field of biosensors, nano medicines and many more as social welfare factor. There are many techniques used for the preparation of nanocomposites. Among them, green method is commonly used technique for the synthesis of nanomaterials which is cost-effective, eco-friendly and less hazardous materials for the environment. Here we attempt to present an elaborate work done in the field of nanocomposites dwelling upon their advantages, challenges and future prospects.

The physical origin of rate promoting vibrations in enzymes revealed by structural rigidity

Enzymes are the most efficient catalysts known to date. However, decades of research have failed to fully explain the catalytic power of enzymes, and most of the current attempts to uncloak the details of atomic motions at active sites remain incomplete. Here, a straightforward manner for understanding the interplay between the complex or irregular enzyme topology and dynamical effects at catalytic sites is introduced, by revealing how fast localized vibrations form spontaneously in the stiffest parts of the scaffold. While shedding light on a physical mechanism that allowed the selection of the picosecond (ps) timescale to increase the catalytic proficiency, this approach exposes the functional importance of localized motions as a by-product of the stability-function tradeoff in enzyme evolution. From this framework of analysis—directly accessible from available diffraction data—experimental strategies for engineering the catalytic rate in enzymatic proteins are proposed.

Rapid in vitro prototyping of O-methyltransferases for pathway applications in Escherichia coli

O-methyltransferases are ubiquitous enzymes involved in biosynthetic pathways for secondary metabolites such as bacterial antibiotics, human catecholamine neurotransmitters, and plant phenylpropanoids. While thousands of putative O-methyltransferases are found in sequence databases, few examples are functionally characterized. From a pathway engineering perspective, however, it is crucial to know the substrate and product ranges of the respective enzymes to fully exploit their catalytic power.In this study, we developed an in vitro prototyping workflow that allowed us to screen ~30 enzymes against five substrates in three days with high reproducibility. We combined in vitro transcription/translation of the genes of interest with a microliter-scale enzymatic assay in 96-well plates. The substrate conversion was indirectly measured by quantifying the consumption of the S-adenosyl-L-methionine co-factor by time-resolved fluorescence resonance energy transfer rather than time-consuming product analysis by chromatography. This workflow allowed us to rapidly prototype thus-far uncharacterized O-methyltransferases for future use as biocatalysts.