Beta-elimination of Hyaluronate by Red King Crab Hyaluronidase

Crustacean hyaluronidases are 1 poorly understood both in terms of their enzymatic properties and in terms of their structural features. In this work, we have shown that the hepatopancreas homogenate of the red king crab has a hyaluronidase activity that is an order of magnitude higher than its commercial counterpart. Zymography revealed the hyaluronidase activity of the protein roughly from40 to 50 kDa relative to the molecular marker used in electrophoresis. Analysis of the hepatopancreas transcriptome revealed a hyaluronidase sequence with an expected molecular weight of 42.5 kDa. It turned out that the reaction of cleavage of hyaluronate in the presence of a homogenate proceeds by the mechanism of b-elimination, which is well known for bacterial hyaluronidases. Thus, a new hyaluronidase of higher eukaryotes was found and described, which is not integrated into the modern classification of hyaluronidases.

Anti-Ageing Potential of S. euboea Heldr. Phenolics

In recent years, the use of Sideritis species as bioactive agents is increasing exponentially. The present study aimed to investigate the chemical constituents, as well as the anti-ageing potential of the cultivated Sideritis euboea Heldr. The chemical fingerprinting of the ethyl acetate residue of this plant was studied using 1D and 2D-NMR spectra. Isomeric compounds belonging to acylated flavone derivatives and phenylethanoid glycosides were detected in the early stage of the experimental process through 2D-NMR techniques. Overall, thirty-three known compounds were isolated and identified. Some of them are reported for the first time not only in S. euboea, but also in genus Sideritis L. The anti-ageing effect of the ethyl acetate residue and the isolated specialized products was assessed as anti-hyaluronidase activity. In silico docking simulation revealed the interactions of the isolated compounds with hyaluronidase. Furthermore, the in vitro study on the inhibition of hyaluronidase unveiled the potent inhibitory properties of ethyl acetate residue and apigenin 7-O-β-d-glucopyranoside. Though, the isomers of apigenin 7-O-p-coumaroyl-glucosides and also the 4′-methyl-hypolaetin 7-O-[6′′′-O-acetyl-β-d-allopyranosyl]-(1→2)-β-d-glucopyranoside exerted moderate hyaluronidase inhibition. This research represents the first study to report on the anti-hyaluronidase activity of Sideritis species, confirming its anti-inflammatory, cytotoxic and anti-ageing effects and its importance as an agent for cosmetic formulations as also anticancer potential.

Antihyaluronidase and Alkaline Phosphatase (ALP) Activities of Medicinal Plants to Combat Echis carinatus Venom-Induced Toxicities

Snakebite is one of the most neglected diseases of developing countries. Deaths due to snakebite envenoming are quite high in Pakistan, and many deaths are caused by Echis carinatus envenomation. Traditional use of medicinal plants against snakebites is a common practice in Pakistan due to countless benefits. The current study was performed with the objective to evaluate eighteen Pakistani medicinal plants inhibitory potential against hyaluronidase and alkaline phosphatase enzymes of Pakistani Echis carinatus venom. Hyaluronidase activity (0.2-1.6 mg/0.1 mL) and alkaline phosphatase activity (0.1-0.8 mg/0.1 mL) were measured in dose-dependent manner. Crude methanolic extracts of medicinal plants were used for in vitro investigation of their inhibitory activity against toxic enzymes. All active plants were fractioned using different solvents and were again analyzed for inhibitory activity of same enzymes. Results indicated all plants were able to neutralize hyaluronidase that Swertia chirayita (Roxb. ex Flem.) Karst., Terminalia arjuna Wight and Arn, Rubia cordifolia Thumb., and Matthiola incana (L.) R.Br. inhibited maximum hyaluronidase activity equivalent to standard reference ( p > 0.5 ). Pakistani medicinal plants are dense with natural neutralizing metabolites and other active phytochemicals which could inhibit hyaluronidase activity of Pakistani Echis carinatus venom. Further advanced studies at molecular level could lead us to an alternative for envenoming of Pakistani Echis carinatus venom.

Anti-snake venom and methanolic extract of Andrographis paniculata: a multipronged strategy to neutralize Naja naja venom acetylcholinesterase and hyaluronidase

The study investigates the ability of methanolic extract of Andrographis paniculata (MAP) to supplement polyvalent anti-snake venom (ASV) in inhibiting neurotoxic enzyme acetylcholinesterase (AChE) and ‘spreading factor’ hyaluronidase from Naja naja (N.N) venom. AChE and hyaluronidase activity were measured in 100 or 200 µg of crude venom, respectively, and designated as ‘control’. In Test Group I, enzyme assays were performed immediately after the addition of ASV/MAP/ASV + MAP to the venom. Inhibition of AChE by ASV (100–367 µg) was 12–17%, and of hyaluronidase (22–660 µg) was 33–41%. Under the same conditions, MAP (100–400 µg) inhibited AChE and hyaluronidase to the extent of 17–33% and 17–52%, respectively. When ASV (220 µg) and MAP (100–200 µg) were added together, AChE and hyaluronidase were inhibited to a greater extent from 39–63 to 36–44%, than when either of them was used alone. In Test Group 2, the venom was incubated with ASV/MAP/ASV + MAP for 10–30 min at 37 °C prior to the assay which enhanced AChE inhibition by 6%, 82% and 18% respectively, when compared to Test Group I. Though there was no change in inhibition of hyaluronidase in the presence of ASV, MAP could further increase the extent of inhibition by 27% and ASV + MAP upto 4%. In Test Group III, venom and substrate were incubated for 90 min and hyaluronidase activity was measured after the addition of inhibitors. Here, ASV + MAP caused increased inhibition by 69% compared to ASV alone. The study confirms the ability of phytochemicals in MAP to contribute to a multipronged strategy by supplementing, thereby augmenting the efficacy of ASV.

Assessment of Cosmeceutical Potentials of Selected Mushroom Fruitbody Extracts Through Evaluation of Antioxidant, Anti-Hyaluronidase and Anti-Tyrosinase Activity

Cosmeceutical formulations containing naturally derived active ingredients are currently preferred by consumers worldwide. Mushrooms are one of the potential sources for cosmeceutical ingredients but relevant research is still lacking. In this study, hot- and cold-water extractions were performed on four locally-cultivated mushrooms—Pleurotus ostreatus, Ganoderma lucidum, Auricularia polytricha and Schizophyllum commune—with the aim to assess the cosmeceutical potential of these mushroom fruitbody extracts. Total phenolics, polysaccharide and glucan content were determined. Antioxidant property of the mushroom extracts was assessed by determining the DPPH radical scavenging, ferric-reducing (FRAP) and superoxide anion (SOA) scavenging activity. Anti-hyaluronidase activity was used as an indicator for the anti-aging and anti-inflammatory property, while anti-tyrosinase activity was evaluated to assess the anti-pigmentation or whitening property of these extracts. Our results showed that total polysaccharide content of P. ostreatus extracts was the highest (235.8–253.6 mg GE/g extract), while extracts from G. lucidum contained the lowest glucan (10.12–10.67%). Cold-water extract from S. commune exhibited substantial tyrosinase inhibition activity (98.15%) and SOA scavenging activity (94.82%). The greatest hyaluronidase activity was exhibited by G. lucidum hot-water extract, with the value of 72.78%. The findings from the correlation analyses suggest that the cosmeceutical properties of these mushrooms can be attributed mainly to the combination of different types of compound such as polysaccharides and phenolics. Overall, cold-water extract of S. commune and hot-water extract of G. lucidum showed the best results and may be further investigated.