Abstract
Garlic is a well-known example of natural self-defence system consisting of an inactive substrate (alliin) and enzyme (alliinase) which, when combined, produce highly antimicrobial allicin. Increase of alliinase stability and its activity are of paramount importance in various applications relying on its use for in-situ synthesis of allicin or its analogues, e.g., pulmonary drug delivery, treatment of superficial injuries, or urease inhibitors in fertilizers. Here, we discuss the effect of temperature, pH, buffers, salts, and additives, i.e. antioxidants, chelating agents, reducing agents and cosolvents, on the stability and the activity of alliinase extracted from garlic. The effects of the storage temperature and relative humidity on the stability of lyophilized alliinase was demonstrated. A combination of the short half-life, high reactivity and non-specificity to particular proteins are reasons most bacteria cannot deal with allicin's mode of action and develop effective defence mechanism, which could be the key to sustainable drug design addressing serious problems with escalating emergence of multidrug-resistant (MDR) bacterial strains.
Highlights
Rising awareness about sustainability and associated development of green chemistry lead to the growing popularity and demands for commercial products with minimal environmental impact, with present emphasis on the origin of material resources, bioaccumulation, biodegradability, and greenhouse gas and waste production [1]
The following chemicals were purchased from Sigma-Aldrich: (±)-L-alliin, Ethylenediaminetetraacetic acid (EDTA), Pyridoxal 50-phosphate hydrate (PLP), Dithiothreitol (DTT), N[Tris(hydroxymethyl)methyl]glycine (Tricine), Polyethylene glycol (PEG) 6000, Bovine serum albumin (BSA), Nutrient Broth (NB), Kanamycin sulfate, Fluorescein diacetate, Propidium iodide (PI), Polyethylenimine (PEI)
SDS-PAGE analysis revealed that alliinase obtained by PEG precipitation was of high purity without a significant presence of the accompanying proteins or degradation products
Summary
Rising awareness about sustainability and associated development of green chemistry lead to the growing popularity and demands for commercial products with minimal environmental impact, with present emphasis on the origin of material resources, bioaccumulation, biodegradability, and greenhouse gas and waste production [1]. Sustainable chemistry has become a key focus area for chemical companies [2], which motivates pharmaceutical, cosmetic and food industry to employ more extensively biocatalysis as an environmentally friendly and effective alternative to traditional synthetic preparation [3,4,5,6].
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