Investigating Structural Property Relationships to Enable Repurposing of Pharmaceuticals as Zinc Ionophores

The importance of zinc in biology has gained greater recognition in recent years due to its essential contributions to the function of many endogenous enzymes. Disruption of zinc homeostasis may be useful in treating pathological conditions, such as Alzheimer’s, and for antiviral purposes. Despite the growth of knowledge and increased interest in zinc, little is known about the structure and function of zinc ionophores. In this study we analyse the Cambridge Structural Database and solution complexation studies found in the literature to identify key functional groups which may confer zinc ionophorism. Pharmaceuticals, nutraceuticals and amino acids with these functionalities were selected to enable us to explore the translatability of ionophoric activity from in vitro assays to cellular systems. We find that although certain species may complex to zinc in the solid and solution states, and may carry ions across simple membrane systems, this does not necessarily translate into ionophoric activity. We propose that the CSD can help refine key functionalities but that ionophoric activity must be confirmed in cellular systems.

Deciphering the effect of Natural Fermentation on Nutritional, Anti-nutritional compounds and Antioxidant Activity of Finger millet landraces

Fermentation is one of the most important bioprocessing techniques used to enhance nutritional components and a reduction in anti-nutritional compounds. In the present work, germinated and ungerminated grains of white and red –brown finger millet landraces were subjected to 8, 16 and 24 hours of fermentation to find out its response in terms of nutritional, anti-nutritional compounds and antioxidant activity. White and red-brown grains showed significant response to 16 and 24 hours process in terms of nutritional and anti-nutritional changes. Significant reduction in the tannin content (73%) was noted for germinated and un-germinated grains with the enhancement in nutritional compounds. Highest antioxidant activity was noted for germinated (91%) grains of white seed coat color grains. Process of natural fermentation and the microflora produces various endogenous enzymes which causes enhancement and reduction of nutritional and antinutritional compounds. Obtained results prove the importance of fermentation process and conservation of landraces for nutritional security.

Distinct Metalloproteinase Expression and Functions in Systemic Sclerosis and Fibrosis: What We Know and the Potential for Intervention

Systemic sclerosis (SSc) is a chronic debilitating idiopathic disorder, characterized by deposition of excessive extracellular matrix (ECM) proteins such as collagen which leads to fibrosis of the skin and other internal organs. During normal tissue repair and remodeling, the accumulation and turnover of ECM proteins are tightly regulated by the interaction of matrix metalloproteinases (MMPs) and endogenous tissue inhibitors of metalloproteinases (TIMPs). SSc is associated with dysregulation of the activity of these proteolytic and inhibitory proteins within the tissue microenvironment, tipping the balance toward fibrosis. The resultant ECM accumulation further perpetuates tissue stiffness and decreased function, contributing to poor clinical outcomes. Understanding the expression and function of these endogenous enzymes and inhibitors within specific tissues is therefore critical to the development of therapies for SSc. This brief review describes recent advances in our understanding of the functions and mechanisms of ECM remodeling by metalloproteinases and their inhibitors in the skin and lungs affected in SSc. It highlights recent progress on potential candidates for intervention and therapeutic approaches for treating SSc fibrosis.

The effects of camel chymosin and Withania coagulans extract on camel and bovine milk cheeses

Withania coagulans (W. coagulans) extract and camel chymosin have aspartic protease capable of coagulating milk for cheese production. This study investigated the quality of camel and bovine milk cheeses coagulated using Withania extracts, came chymosin, and their mixture in two experiments. In Experiment (1), a factorial design with four factors (W. coagulans, camel chymosin, incubation time, and incubation temperature) was performed. The effect of these factors on cheese’s yield and hardness were assessed. An enzyme concentration corresponding to a 36 µg/L of milk of W. coagulans, 50 IMCU/L of camel chymosin, holding time of 4 h, and incubation temperature of 60 °C provided the optimal textural hardness for both camel and bovine milk cheeses. Seven treatments were analyzed in experiment (2) were analyzed for physicochemical properties, yield, and sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGEitation). The results showed that pure Withania extract exhibited the lower coagulating effect resulting in cheeses with low yield, hardness, fat, protein, and total solids. The SDS-PAGE electropherograms of camel cheese showed several low molecular weight bands as compared to bovine cheese. This phenomenon is due to excessive proteolysis in camel cheese, which we believed is caused by the presence of endogenous enzymes.

Impact of combined α-galactosidase and xylanase enzymes on growth performance, nutrients digestibility, chyme viscosity, and enzymes activity of broilers fed corn-soybean diets

Two experiments were conducted to investigate the effects of a combined α-galactosidase and xylanase preparation on nutrients digestibility and growth performance in broiler chickens. Experiment 1 had 240 broilers allocated to 3 treatments with the dietary supplementation of 0, 300 and 500 g/t of the enzyme combination. Diet and amino acid (AA) digestibility were assessed. Experiment 2 was a 2 × 3 (enzyme × diet) factorial arrangement with 10 replicates of 12 male broilers per replicate. Diets were based on corn-Soybean meal (SBM) diet and had 3 nutritional levels (normal, 2% apparent metabolizable energy (AME) and crude protein (CP) reduction, and 4% AME and CP reduction). Each of these diets was fed with or without enzyme supplementation. Growth performance, chyme viscosity, nutrients digestibility, and endogenous enzymes activity were assessed. In experiment 1, enzyme supplementation improved the digestibility of Ca (P = 0.025) and ileal digestibility of total AA, Pro, Alu, Ile, Lys, His, Thr, Glu, Val, Leu, Tyr and Phe (P < 0.05), and also tended to increase the AME of diets (P < 0.10). In experiment 2, broilers fed the corn-SBM diet with 4% nutrient reduction had better growth performance (P < 0.05), jejunal digesta viscosity at 42 days (P < 0.01), and lower digestibility of gross energy (GE) (P < 0.05) when compared to those fed the normal nutrient diet. Enzyme inclusion increased digestibility of CP (P = 0.044), GE (P = 0.009), raffinose (P < 0.001) and stachyose (P < 0.001), improved average daily gain (P = 0.031), and reduced jejunal digesta viscosity at 42 days (P = 0.011). Besides, similar improvements trend in amylase, trypsin, sucrase, and maltase activity with enzyme inclusion were observed as with energy. These data support that the enzyme supplementation increased nutrients and ileal amino acid digestibility, improved performance and endogenous enzymes activity.

Cutin:cutin-acid endo-transacylase (CCT), a cuticle-remodelling enzyme activity in the plant epidermis

Cutin is a polyester matrix mainly composed of hydroxy-fatty acids that occurs in the cuticles of shoots and root-caps. The cuticle, of which cutin is a major component, protects the plant from biotic and abiotic stresses, and cutin has been postulated to constrain organ expansion. We propose that, to allow cutin restructuring, ester bonds in this net-like polymer can be transiently cleaved and then re-formed (transacylation). Here, using pea epicotyl epidermis as the main model, we first detected a cutin:cutin-fatty acid endo-transacylase (CCT) activity. In-situ assays used endogenous cutin as the donor substrate for endogenous enzymes; the exogenous acceptor substrate was a radiolabelled monomeric cutin-acid, 16-hydroxy-[3H]hexadecanoic acid (HHA). High-molecular-weight cutin became ester-bonded to intact [3H]HHA molecules, which thereby became unextractable except by ester-hydrolysing alkalis. In-situ CCT activity correlated with growth rate in Hylotelephium leaves and tomato fruits, suggesting a role in loosening the outer epidermal wall during organ growth. The only well-defined cutin transacylase in the apoplast, CUS1 (a tomato cutin synthase), when produced in transgenic tobacco, lacked CCT activity. This finding provides a reference for future CCT protein identification, which can adopt our sensitive enzyme assay to screen other CUS1-related enzymes.

FEATURES OF THE BIOCONVERSION OF SOY RAW MATERIALS INTO FERMENTED PRODUCTS

Comparative studies of biotechnological methods for obtaining fermented products based on the conversion of soy raw materials with a selected complex of endogenous enzymes and using a selected strain of Aspergillus oryzae - the producer of a complex of proteases and other hydrolases.

In situ Generation of Antibiotics using Bioorthogonal “Nanofactories”

Prodrug strategies use chemical modifications to improve the pharmacokinetic properties and therefore therapeutic effects of parent drugs. Traditional prodrug approaches use endogenous enzymes for activation. Bioorthogonal catalysis uses processes that endogenous enzymes cannot access, providing a complementary strategy for prodrug uncaging. Site-selective activation of prodrugs to drugs (uncaging) using synthetic catalysts is a promising strategy for localized drug activation. We discuss here recent studies that incorporate metal catalysts into polymers and nanoparticle scaffolds to provide biocompatible “enzyme-like” catalysts that can penetrate bacterial biofilms and activate prodrug antibiotics in situ, affording a new strategy to treat bacterial biofilm infections with the potential for reduced off-target effects.