Enzymatic Protein Hydrolysates, and Ultrafiltered Peptide Fractions from Two Molluscs: Tympanotonus fuscatus var. radula (L.) and Pachymelania aurita (M.), with Angiotensin-I-Converting Enzyme Inhibitory and DPPH Radical Scavenging Activities.

CONTEXT: Multifunctional food protein-derived peptides attract a great deal of research interest due to their health-promoting benefits. Particularly, peptides that have both antihypertensive and antioxidant properties are desired, since both effects can be synergistic in prevention of cardiovascular diseases. AIM: The aim of this study was to investigate the angiotensin-I-converting enzyme (ACE) inhibitory and 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activities of two species of the Nigerian periwinkles: Pachymelania aurita and Tympanotonus fuscatus. METHODS: The ACE inhibitory and 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activities of simulated gastrointestinal digestion (SGID) hydrolysates and ultrafiltered (UF) fractions of T. fuscatus var. radula and P. aurita were determined. Human SGID of the protein extracts of T. fuscatus and P. aurita was carried out using pepsin, trypsin, and chymotrypsin, and the hydrolysates were fractionated into two by centrifugal ultrafiltration. The ACE inhibitory and DPPH radical scavenging activities of the crude hydrolysates and UF fractions were tested. The UF permeates were observed to have relatively higher activities and was subjected to gel filtration chromatography on Sephadex G-50. The chromatographic fractions showed absorbance at 215, 225, and 280 nm and were assayed for DPPH radical scavenging activity. RESULTS: The inhibitory effect of the fractions on ACE activity was reported as the minimum concentration of extract that caused 50% of the inhibition (IC50), where the IC50 values of P. aurita UF permeate and P. aurita UF retentate were 65.2 ± 6.4 and 301.9 ± 59.1 µg/ml, respectively, and that of T. fuscatus UF permeate (TFUFP) and T. fuscatus UF retentate were 54.93 ± 2.83 and 291.7 ± 8.6 µg/ml, respectively. CONCLUSION: This study suggests the potential health benefits of consuming T. fuscatus var. radula and P. aurita in health maintenance.

Antioxidant activity of the simulated gastro-intestinal digestion hydrolysate of two edible Nigerian marine molluscs: Tympanatonus fuscatus var radula (L.) and Pachymelania aurita (M.).

The multifunctional nature of antioxidant peptides makes them more attractive candidates as dietary ingredients in health maintenance. Therefore, food protein-derived antioxidant peptides are continuously investigated. This study investigated the in vitro antioxidant properties of hydrolysate and ultrafiltered peptide fractions of Pachymelania aurita and Tympanatonus fuscatus var radula-two commonly consumed marine molluscs known as periwinkles in southern Nigeria. Simulated gastrointestinal digestion (SGID) of soluble proteins of T. fuscatus and P. aurita was carried out using pepsin, trypsin and chymotrypsin, and the SGID hydrolysates were fractionated using a 3 kDa membrane filter. The hydrolysates and their fractions were investigated for anti-lipid peroxidation, hydroxyl radical scavenging activity (HRSA), ferric reducing antioxidant property (FRAP) and metal chelation activity, and they demonstrated clear antioxidant properties in all the assay models used. Low molecular weight fractions of the hydrolysates demonstrated more potent antioxidant activity than higher molecular weight fractions. This is profound in the metal chelation assay, where low molecular weight peptide fractions, T ≤ 3 kDa and P ≤ 3 kDa (IC50 values of 8.10 ± 0.011 and 5.56 ± 0.50 µg/ml respectively) had activity that is similar to that of EDTA (11.84 ± 0.89 µg/ml). Similar activity effects were observed in other assays where there was about 3-fold higher activity in low molecular weight fractions. These results demonstrate the presence of antioxidant peptide(s) in the protein hydrolysates of the periwinkles.

Evaluation of Analgesic Activities of Extracts of Two Marine Molluscs: Tympanotonus fuscatus var radula (Linnaeus) and Pachymelania aurita (Müller).

PURPOSE AND METHODS: In this study, the analgesic activity of the crude alcohol (acetone-methanol) and aqueous (in PBS, pH 7.2) extracts of the marine molluscs, Pachymelania aurita and Tympanotonus fuscatus, has been evaluated using the formalin test (for chronic antinociceptive) and the tail-flick (acute antinociceptive) pain models in male swiss albino mice. RESULTS: The results show that the extracts of P. aurita and T. fuscatus demonstrated high safety margins as single doses of up to 2000 mg/kg bwt proved to be well tolerated and non-lethal, although the alcohol extract of P. aurita caused necrosis in the liver and kidney when administered at a dose level of 2000 mg/kg bwt. In the formalin test, treatment with the aqueous extracts of P. aurita and T. fuscatus as well as the alcohol extract of T. fuscatus 30 min before the subcutaneous injection of 5% formalin to the paw of the mice resulted in a significant time- and dose-dependent reduction in total and phase 2a pain-related behavior and thus nociception. The extracts had no analgesic effect in tail-flick test up to the highest dose tested. CONCLUSION: Hence, the results from both models indicate that the site of their analgesic action is probably peripheral.

In vitro antiproliferative studies of extracts of the marine molluscs: Tympanatonus fuscatus Var radula (linnaeus) and Pachymelania aurita (muller).

This study aimed to investigate the antimitotic and antiproliferation activities of crude acetone-methanol and aqueous extracts of two marine molluscs commonly found in the Niger Delta region of Nigeria; T.fuscatus and P.aurita, against human cancerous cell lines (DU145, Hep-2, and HCC1395) cell lines in vitro. The antimitotic activity of the extracts was evaluated using Allium cepa root meristematic cells. Antiproliferative activity of the plant extracts against the cancerous cell lines was compared with normal cell line (VeroE6). Doxorubicin was used as a positive control. Gene expression studies using qPCR for the proapoptotic genes, CASP3, CASP8 and P53 were also carried out. The alcohol extract of T.fuscatus (TFAC) exhibited the most promising activity against all the cancer cell lines tested (DU145 IC50 = 96.48 ± 1.36 µg/ml, HCC 1395 IC50 = 61.44 ± 2.45 µg/ml, Hep2 IC50 = 0.52 ± 0.36 µg/ml) and also had the highest selectivity index of 4.94, 7.78 and 921.97 for DU145, HCC 1395 and Hep-2 cells respectively. Furthermore, TFAC was the only extract that significantly upregulated the expression of caspase 3, caspase 8 and P53. Thus, these findings suggest potential exploitation of TFAC as an anticancer agent.

Physicochemical properties and acute toxicity studies of a lectin from the saline extract of the fruiting bodies of the shiitake mushroom, Lentinula edodes (Berk).

A lectin (LEL) was isolated from the fresh fruiting bodies of the shiitake mushroom Lentinula edodes by a combination of gel filtration chromatography on Sephadex G-150 and affinity chromatography on an N-acetyl-Dgalactosamine-Sepharose 4B column. Its molecular mass, as determined by gel filtration, was estimated to be 71, 000 Daltons and its structure is homotetrameric with subunit molecular weight of approximately 18,000 Daltons. LEL agglutinated non-specifically red blood cells from the human ABO system as well as rabbit erythrocytes and in haemagglutination inhibition assays, exhibited sugar-binding specificity toward N-acetyl-D-galactosamine. EDTA had no inhibitory effect on its haemagglutinating activity, which was stable up to 70°C and was not affected by changes in pH. The lectin had no covalently-linked carbohydrate and amino acid composition analysis revealed that it contained 124 amino acid residues and was rich in tyrosine, proline, phenylalanine, arginine, glutamic acid and cysteine. LEL did not cause mortality neither was it observed to alter the morphology of key organs when administered intraperitoneally at concentrations up to 10,000 mg kg(-1) body weight of mice.