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Background: Oratosquilla woodmasoni is one of the marine squilla species, which is found in the entire Asia-Pacific region. This current study assesses the species as the main basis of both ACEi and antioxidant peptide. Objective: To isolate the ACEi peptide derived from O. woodmasoni and examine its ACE inhibition along with antioxidant potential. Materials and Methods: The squilla muscle protein was hydrolysed using alcalase and trypsin enzymes for 12 hours and tested for DH. The hydrolysates were examined for their ACEi activity and then the best hydrolysate was sequentially purified in various chromatographical methods. The purified peptide was studied for anti-oxidant and functional properties, followed by amino acid sequencing. The purified peptide was also evaluated for its toxicity by in vitro cell viability assay. Results: The DH% was found to be 47.13 ± 0.72% and 89.43 ± 2.06% for alcalase and trypsin, respectively. The alcalase 5th-hour hydrolysate was detected with potent activity (65.97 ± 0.56%) using ACEi assay and was primarily fractionated using ultrafiltration; the maximum inhibitory activity was found with 77.04 ± 0.52% in 3-10 kDa fraction. Subsequently, the fraction was purified using IEC and GFC, in which the AC1-A2 fraction had higher antihypertensive activity (70.85 ± 0.78%). The non-toxic fraction showed hexapeptide HVGGCG with molecular weight 529 Da with great potential of antioxidant activity along with functional property. Conclusion: This peptide could be developed as a potential ACE-inhibitory and antioxidant agent.

Biological waste from marine sources is discarded into various water bodies which leads to dramatic increase in the water pollution near coastal areas. This animal waste consists of bioactive compo...

Background: Green mussel Perna viridis is a bivalve mollusc which is native to the Indian coast and can be found in the Indo-Pacific as well as Asia-Pacific regions. This study evaluates the P. viridis foot (PVF) as a source of an anti-inflammatory peptide. Objective: To characterize and evaluate the possibility of pro-inflammatory cytokines, nitric oxide (NO) as well as cyclooxygenase (COX)-2 reduction in RAW264.7 cells and to analyze functional aspects of the derived peptide from PVF. Materials and Methods: The PVF was hydrolysed with different enzymes and the antiinflammatory activity of hour hydrolysates were evaluated using HRBC Membrane Stabilization (HMS) against hypotonicity induced haemolysis and Albumin Denaturation (AD) inhibition from induced heat assays. Later, the active hour hydrolysate was separated by ultrafiltration and purified using Size-Exclusion Chromatography (SEC). Further, the purified peptide’s sequence was identified using LC-MS/MS and functional properties were determined. Also, the peptide was observed for its anti-inflammatory effects in lipopolysaccharide (LPS)-stimulated RAW264.7 cells for pro-inflammatory cytokines, NO production and COX-2 activation. Results: Among the four enzymes 6th hour alcalase hydrolysate exhibited potent anti-inflammatory activity and was sequentially fractioned with molecular weight cut-offs; further active fraction (30- 10 kDa) was purified. The active peak-II was identified as EGLLGDVF (849.435 Da) and exhibited decent functional aspects. The peptide successfully reduced the production of pro-inflammatory cytokines, NO and COX-2 activation; and down-regulated the iNOS and COX-2 protein expression in LPS-stimulated RAW264.7 cells. Conclusion: Our study indicates that EGLLGDVF derived from PVF has potential antiinflammatory applications applicable in food and pharmaceutical industries.

To improve the quality of life of diabetic patients, oral delivery of insulin would be better than subcutaneous injection, and the encapsulation of insulin for its oral delivery is a promising alternative one. In this study, we prepared an oral insulin delivery system using thiolated chitosan nanoparticles (TCNPs) loaded with insulin (Ins) and tested under in vitro and in vivo systems. TCNPs prepared from CS and pentaerythritol tetrakis (3-mercaptopropionate) (PETMP) at 4:1 ratio showed 220 ± 4 nm, 2.3 ± 1 mV, and 119 ± 4 μmol g−1 in their size, charge and sulfhydryl content, respectively. There was a sustained release of insulin from the TCNPs at pH 5.3. TCNPs treatment did not alter cell viability in vitro and oral administration of TCNPs reached over the tip of the microvilli near the intestinal mucosa in vivo. There were increased and decreased the levels of insulin and glucose in the blood, respectively when Ins-TCNPs were orally administered in the diabetes induced rats. Thus, our results suggested that the insulin stays significantly for a prolonged period to make bio-distribution and bioavailability due to its interaction with the mucus of the intestine, thus offering a better oral insulin delivery system for diabetic patients.

The current objective was framed on isolation of two angiotensin-I converting enzyme inhibitory peptides from coastal trashes of squilla muscle (Harpiosquilla raphidea) prepared by enzymatic hydrolysis using alcalase, thermolysin and trypsin, which showed the highest degree of hydrolysate in 12th h hydrolysate 87.9 ± 1.6%, 63.1 ± 1.1% and 91.1 ± 2.7% respectively. Among these three hydrolysates, alcalase 5th h (64.8 ± 0.3) and thermolysin 6th h (68.4 ± 1.0) found to be maximum ACE-I inhibition activity. Initial separation was performed by ultra filtration (10–3 kDa) into three different fractions, alcalase (below 3 kDa) fraction observed maximum ACE-I inhibition activity with 75.2 ± 3.5% and thermolysin (3–10 kDa) with 79.5 ± 1.0% inhibition. Further, these two fractions have taken to ion exchange and gel filtration chromatography, the identified ACE-I inhibitory fractions (AC-A2 and TH-A2) also examined for its antioxidant properties. Further, these fractions were determined to be non-toxic against cell lines, functional activities by various assays and characterized into tri peptides MSN (Met-Ser-Asn) and MTH (Met-Thr-His) with a molecular weight of 350 Da and 388 Da respectively using LC–MS/MS. These low molecular weight peptides from natural sources may act as potential ingredient for nutraceutical and pharmaceutical industries.

The Meretrix meretrix is abundantly present in the Indian coastal areas which can be used as an important useful bioactive source for industrial applications. The M. meretrix visceral mass (MMV) was hydrolysed with four different enzymes and verified for anti-inflammatory activity with the help of HRBC membrane stabilization (HMS) and albumin denaturation (AD) assay. Among the hydrolysates, the tryptic 6th hour hydrolysate was selected for purification using ultrafiltration and size-exclusion chromatography (SEC). Further, the purified peptide was identified to have six amino acid sequence (HKGQCC, 675.582 Da). However, to confirm the anti-inflammatory effects of the purified peptide, it was investigated for nitric oxide synthase (iNOS), pro-inflammatory cytokines production as well as cyclooxygenase-2 (COX-2) activation in lipopolysaccharide (LPS)-stimulated RAW264.7 cells and also evaluated for its functional properties. The in-vitro gastrointestinal digestion was performed on the peptide which cleaved the peptide into two i.e. MMV1 (HK, 284.1 Da) and MMV2 (GQCC, 410.1 Da). The data suggested that the MMV2 peptide have maximum activity and was found to be stable at high temperatures. The MMV2 peptide demonstrated abrupt localization throughout the adult zebrafish body and successfully downregulated the mRNA levels of inflammation-related genes in LPS-induced adult zebrafish. This study indicates that the peptide MMV2 possesses anti-inflammatory activity by suppressing the induced inflammation and can be a strong competitor against non-steroidal anti-inflammatory drugs (NSAIDs).

Biological waste from marine sources is discarded into various water bodies which leads to dramatic increase in the water pollution near coastal areas. This animal waste consists of bioactive compounds such as fatty acids, amino acids, and chitin which can be used in agricultural and pharmaceutical sectors. The aim of the current study was to extract chitosan (CS) from the discarded shells of

Antioxidant peptides protect biological macromolecules against radical damages. The use of these peptides was evaluated using free radicals scavenging assays [2,2-diphenyl-1 picrylhydrazyl (DPPH) and hydroxyl] with the help of UV-visible and electron spin resonance (ESR) spectroscopy methods. The Octopus aegina mantle protein were tested upon hydrolysis using gastrointestinal enzymes up to 12 h, where pepsin hydrolysate exhibited superior properties (DPPH: 44.39±0.67% and hydroxyl: 38.84±1.07%) compared with trypsin and α-chymotrypsin. Consequently, the antioxidant activity of the purified hydrolysate increased on a successive purification, and the peptide sequence was determined to be 368.9 Da with Gly-Glu-Tyr amino acids. Tripeptide exerted free radical scavenging efficiency in DNA damage, lipid peroxidation and cellular destruction (MCF7 cells) under stress condition. The results obtained with octopus antioxidant peptide suggested its role as an adjunct in food and pharmaceutical industries.

The study aimed to identify bioactive peptide from Meretrix meretrix Linnaeus foot (MMF) and examine its potential of suppressing inflammation. In brief, the anti-inflammatory activity was identified by erythrocyte membrane protection and protein denaturation assay from MMF peptic 9th-h hydrolysate and was separated with three molecular weight cut-off units. The obtained four fractions were testified for activity and the fraction (10–3 kDa) with maximum activity was purified using gel permeation chromatography. Finally, the peptide sequence was identified as Asn–Pro–Ala–Gln–Asp–Cys (647.559 Da) by liquid chromatography–tandem mass spectrometry (LC–MS/MS). The hexapeptide was characterised for functional properties at different pH range. The non-toxic hexapeptide was able to reduce the cyclooxygenase (COX)-2 activation, pro-inflammatory cytokines and nitric oxide (NO) production significantly in RAW264.7 macrophage cells. The current results propose that the hexapeptide derived from MMF protein can act as an effective anti-inflammatory against pro-inflammatory cytokines, COX-2 and NO. Moreover, it could be used as an effective alternative source for drugs in pharma and also as an ingredient in food industries.

The visceral mass of the gastropod, Harpa ventricosa was hydrolysed using trypsin, alcalase and pepsin for 12 h to produce protein hydrolysates. Subsequently, the active hydrolysate was observed in the 3rd hour of tryptic hydrolysate (29.17 ± 0.62 and 34.85 ± 0.55 %) using human red blood cell (HRBC) membrane stabilization and albumin denaturation (AD) assays. The active hydrolysate was fractionated by membrane filtration unit, where

The aim was to drive the ACE-I inhibitory peptide from the dumped waste of O. woodmasoni muscle protein hydrolysate after 12 h of hydrolysis using thermolysin and pepsin. The degree of hydrolysis (DH) was observed till 12th hr and the maximum ACE-I inhibition activity was recorded in 2nd and 5th hr (73.51 ± 0.25% & 68.93 ± 0.23%) for thermolysin and pepsin respectively. Further, the higher active thermolysin hydrolysate was separated using ultrafiltration membrane unit and 10-3 kDa fraction showed chief ACE-I inhibition activity (IC50 value: 0.47 mg/mL). The active fraction was purified by consecutive chromatographic techniques using ion exchange chromatography (IEC) & gel filtration chromatography (GFC) using fast protein liquid chromatography (FPLC). Finally, the fraction TH1-A1 evaluated by ACE-I inhibition assay and showed 77.09 ± 2.18% inhibitions. The non-toxic peptide against MCF-7 cells was further characterized for functional properties at pH 2 to 10. The active purified fraction also exhibited antioxidant activity analyzed with various assays and found to have 5 amino acid sequence (Asn-Gly-Val-Ala-Ala) with molecular weight 431 Da through LC-MS/MS. In conclusion, the identified low molecular weight peptide from by catch O. woodmasoni waste has both ACE-I inhibition and antioxidant properties. Therefore, this peptide might be a better alternative nutraceutical and functional food after few more investigations.

Marine organisms provide an intriguing number of bioactive compounds. These bioactive compounds show immunomodulatory, antioxidant, anti-inflammatory and antimicrobial activities. Crabs rank third after shrimps and lobster as seafood delicacies and provide major health benefits when consumed due to presence of proteins, vitamins and unsaturated essential fatty acids. Due to their diverse biological activities, the peptides derived from marine sources may treat various diseases such as migraine, inflammatory bowel disease, rheumatoid arthritis etc. In this study the Crab Leg (CL) muscle was isolated and further hydrolysed using trypsin, alcalase and papain to produce enzymatic hydrolysates. The results obtained from degree of hydrolysis showed that the enzymes significantly cleave the proteins. Furthermore, the HPLC analysis for amino acid compositions of CL showed the sample contained both essential and non-essential amino acids where histidine (%) was highest. In vitro assays for anti-inflammatory activity like Albumin Denaturation (AD) and HRBC Membrane Stabilization (HMS) assay helped to determine the active hour hydrolysate showing the trypsin 3rd hr hydrolysate as having highest activity in CL (90.57 ± 0.95% and 49.94 ± 1.52%). This active hour hydrolysate was purified using gel filtration chromatography and the fractions were determined using AD and HMS assay which resolved fraction Ⅰ to be more active in CL. The low molecular weight peptide sequence was found to be L-G-L-G-A-A-V-L (713.456 Da) by using LC-MS/MS. Further, cell proliferation assay and ELISA were performed for the purified peptide to test the suppression of LPS-mediated induction of COX-2 in RAW264.7 macrophage cells.

In this current investigation, the antioxidant peptides were derived from marine Sepia brevimana mantle by trypsin, α-chymotrypsin and pepsin for 12 h hydrolysis. The active peptides were found in the hydrolysates of trypsin and α-chymotrypsin at the 7th hour and pepsin at the 8th hour. The highest activity was found in trypsin hydrolysate compared to others using various radical scavenging and metal ions transition assays, further the hydrophobic amino acids were observed in more quantity along with other amino acids. Active peptide was purified by consecutive chromatographic techniques and evaluated by DPPH (38.81 ± 1.07%) and reducing power assays (0.478 ± 0.03). The purified peptide exhibited significant inhibition for the linoleic acid auto-oxidation in the model system, protective effect on DNA damage due to hydroxyl radical induction and was non-toxic even at higher concentration. The mass and sequence of the peptide was found to be 679.5 Da and Ile/Leu-Asn-Ile/Leu-Cys-Cys-Asn, respectively. The current results suggested that cuttlefish peptide could be used as natural antioxidant in enhancing antioxidant properties of functional foods and in preventing oxidation reactions in food processing.

Antioxidant peptides were derived from the edible part of Indian squid, Loligo duvauceli, via the use of gastrointestinal enzymes for up to 12 hours. The active α-chymotrypsin hydrolysate exhibited the maximum activity compared to the trypsin and pepsin hydrolysates, as assessed by various radical scavenging and metal chelating assays. Moreover, essential and non-essential amino acids were present in the active hydrolysate. Furthermore, the hydrolysate was purified by ion exchange and gel filtration chromatography using fast protein liquid chromatography (FPLC). The purified peptide exhibited strong free radical scavenging, metal chelation and reducing power abilities that were confirmed with a UV-visible and electron spin resonance (ESR) spectrometer. The peptide also prevented DNA damage and inhibited lipid peroxidation due to its small size (682.5 Da) and content of the high redox potential amino acid sequence Trp-Cys-Thr-Ser-Val-Ser, which was confirmed by ESI-MS/MS. This peptide exhibited no cytotoxic effects on breast cancer cells (MCF7) and scavenged reactive oxygen species at the cellular level under H2O2-induced stress. These results suggest that the peptide derived from squid mantle protein acts as a potent antioxidant against oxidative stress and could be used as an efficient and safer adjunct in food processing.