Thermal Sensitivity of the Enzymatic Activity of ß-Glucosidase: Simulations Lend Mechanistic Insights into Experimental Observations.

A crucial prerequisite for industrial applications of enzymes is the maintenance of specific activity across wide thermal ranges. ß-Glucosidase (EC 3.2.1.21) is an essential enzyme for converting cellulose in biomass to glucose. While the reaction mechanisms of ß-glucosidases from various thermal ranges (hyperthermophilic, thermophilic, and mesophilic) are similar, the factors underlying their thermal sensitivity remain obscure. The work presented here aims to unravel the molecular mechanisms underlying the thermal sensitivity of the enzymatic activity of the ß-glucosidase BglB from the bacterium Paenibacillus polymyxa. Experiments reveal a maximum enzymatic activity at 315 K, with a marked decrease in the activity below and above this temperature. Employing in silico simulations, we identified the crucial role of the active site tunnel residues in the thermal sensitivity. Specific tunnel residues were identified via energetic decomposition and protein-substrate hydrogen bond analyses. The experimentally observed trends in specific activity with temperature coincide with variations in overall binding free energy changes, showcasing a predominantly electrostatic effect that is consistent with enhanced catalytic pocket-substrate hydrogen bonding (HB) at Topt. The entropic advantage owing to the HB substate reorganization was found to facilitate better substrate binding at 315 K. This study elicits molecular-level insights into the associative mechanisms between thermally enabled fluctuations and enzymatic activity. Crucial differences emerge between molecular mechanisms involving the actual substrate (cellobiose) and a commonly employed chemical analogue. We posit that leveraging the role of fluctuations may reveal unexpected insights into enzyme behavior and offer novel paradigms for enzyme engineering.

Biocompatible nanoparticles for sustained topical delivery of anticancer phytoconstituent quercetin.

This study describes the release and retention of a herbal lipophilic drug in sustained and controlled manner in skin layers, given topically, intended for skin cancer. Quercetin -loaded nanoparticles were prepared by nanoprecipitation technique using ethylcellulose as polymer. Ethylcellulose was selected as it is biocompatible, but non-biodegradable and hence can act as a reservoir in skin furrows and ducts. It was observed that the Quercetin: Ethylcellulose: Tween 80 at different ratios affects particle sizes along with yield and entrapment efficiency. It was found that the size of nanoparticles could be varied by changing the speed of agitation and sonication. The nanoparticles were prepared in particle size range 228.77 +/- 2.0 nm and the zeta potential of the selected formulation were found to be -16.7 mV, which shows the stability of the preparation. The percent entrapment efficiency was found to be in the range from 51.96 to 53.93% and percent loading capacity in the range 34.19 to 5.12%. The amount of drug release from nanoparticles and of drug retained in skin was compared using ex vivo study which shows that the drug being lipophilic could be retained in the skin for longer duration thus reducing the dose and frequency of drug administration. Further the amount of drug reaching to other organs is also reduced since the systemic absorption of drug was low. Thus, Quercetin loaded nanoparticles were prepared for topical use.

Comparative measurement of hydration effects of herbal moisturizers.

Improvements of skin hydration properties by the use of polyherbal moisturizers are the recent advances in cosmetic preparations to avoid the harmful effects of chemical moisturizers. The main aim of the study was to establish selection preference of different available marketed herbal moisturizers on the basis of the efficiency of constituents for their hydration effects. The criteria for the selection of formulations were presence of herbal constituents, wheat germ oil and Aloe vera extract. Initially, physiochemical and psychometric studies were performed to visualize the compliance of moisturizers with the skin. The clinical study was carried out in six groups of six healthy human volunteers (aged 20-25 years) each applying moisturizers twice daily over a period of 3 weeks in their forearm. The skin properties measured were conductance, glow and appearance. The results indicated that all the moisturizers show moisturizing effect in a time-dependent pattern and the maximum increase in skin conductance was 168.125 and 165.24% for A2 and A1, respectively. Ranking of moisturizers based on conductance as well as physicochemical analysis is A2 > A1 > A4 > A3 > A5 > A6. It was found that the formulation A2 having wheat germ oil, Aloe vera extract and turmeric extract in combination showed best results due to their synergistic effect and wheat germ oil or Aloe extract, when present separately produced skin hydration to lesser extent.