Chang-Bum Ahn

Chitooligosaccharide and Its Derivatives: Preparation and Biological Applications

Chitin is a natural polysaccharide of major importance. This biopolymer is synthesized by an enormous number of living organisms; considering the amount of chitin produced annually in the world, it is the most abundant polymer after cellulose. The most important derivative of chitin is chitosan, obtained by partial deacetylation of chitin under alkaline conditions or by enzymatic hydrolysis. Chitin and chitosan are known to have important functional activities but poor solubility makes them difficult to use in food and biomedicinal applications. Chitooligosaccharides (COS) are the degraded products of chitosan or chitin prepared by enzymatic or chemical hydrolysis of chitosan. The greater solubility and low viscosity of COS have attracted the interest of many researchers to utilize COS and their derivatives for various biomedical applications. In light of the recent interest in the biomedical applications of chitin, chitosan, and their derivatives, this review focuses on the preparation and biological activities of chitin, chitosan, COS, and their derivatives.

Purification and antioxidant properties of octapeptide from salmon byproduct protein hydrolysate by gastrointestinal digestion

Pectoral fin protein from salmon processing byproduct was hydrolyzed using Alcalase, Flavourzyme, Neutrase, pepsin, Protamex, and trypsin, and the peptic hydrolysate showed the highest 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity. Antioxidant peptide was purified using consecutive chromatography. The purified antioxidant peptide was identified to be Phe-Leu-Asn-Glu-Phe-Leu-His-Val with molecular weight of 1018.48 Da by time of flight-mass spectrometry/mass spectrometry (TOF-MS) analysis. The IC50 values against DPPH and 2,2-azino-bis(3-ethylbenzthiazoline)-6-sulfonic acid (ABTS) cation radical scavenging activity were 486 and 152 μM, respectively, and the octapeptide showed strong ferric reducing power. In addition, the octapeptide showed significant (p<0.05) protection ability against hydroxyl radical-induced DNA damage and hydrogen peroxide-induced hepatic damage in Chang liver cells. Taken together, the pectoral fin protein hydrolysate and/or its active peptides may be useful ingredients in functional food.

Anti-asthmatic effect of marine red alga (Laurencia undulata) polyphenolic extracts in a murine model of asthma.

The aim of the present work is focused on protective effects of an edible red alga, Laurencia undulata ethanolic (EtOH) extracts (LU) containing a large amount of polyphenols against OVA-induced murine allergic airway reactions using in vivo histological and cytokine assay. Mice sensitized and challenged with ovalbumin (OVA) showed typical asthmatic reactions as follows: an increase in the number of eosinophil in bronchoalveolar lavage fluid; a marked influx of inflammatory cells into the lung around blood vessels and airways, and airway luminal narrowing; the development of airway hyperresponsiveness; the detection of TNF-alpha and Th2 cytokines, such as IL-4 and IL-5 in the bronchoalveolar lavage (BAL) fluid; and detection of allergen-specific IgE in the serum. The successive intraperitoneal administration of LU before the last airway OVA-challenge resulted in a significant inhibition of all asthmatic reactions. These results suggest that L. undulata polyphenolic extracts possess therapeutic potential for combating bronchial asthma associated with allergic diseases.

Purification and anti-inflammatory action of tripeptide from salmon pectoral fin byproduct protein hydrolysate.

In this study, the anti-inflammatory peptide from salmon pectoral fin byproduct protein hydrolysate by pepsin hydrolysis, was purified and identified using Sephadex G-25 gel permeation chromatography, high performance liquid chromatography and time-of-flight liquid chromatography/tandem mass spectrometry (TOF LC/MS/MS). The purified anti-inflammatory peptide was identified to be a tripeptide (PAY). Lipopolysaccharide treatment significantly (p<0.05) stimulated the production of nitric oxide (NO) and prostaglandin E2 (PGE2) in RAW264.7 cells. However, PAY treatment significantly (p<0.05) inhibited the production of NO by 63.80% and PGE2 by 45.33%. Western blotting analysis revealed that PAY significantly (p<0.05) suppressed the protein expression of inducible nitric oxide synthase and cyclooxygenase-2, which are responsible for the production of NO and PGE2. Additionally, PAY treatment also significantly (p<0.05) attenuated the production of pro-inflammatory cytokines, including tumour necrosis factor-α, interleukin-6 and -1β.

Chitosan–hydroxycinnamic acid conjugates: Preparation, antioxidant and antimicrobial activity

In this study, the antioxidant and antimicrobial activities of chitosan-caffeic acid, chitosan-ferulic acid, and chitosan-sinapic acid conjugates with different grafting ratios were investigated. The synthesized chitosan-hydroxycinnamic acid conjugates were verified by performing (1)H NMR and differential scanning calorimetry analysis. The antioxidant activities of the conjugates were increased compared to the unmodified chitosan, by 1.79-fold to 5.05-fold (2,2-diphenyl-1-picrylhydrazyl scavenging assay), 2.44-fold to 4.12-fold (hydrogen peroxide scavenging assay), 1.34-fold to 3.35-fold (ABTS(+) radical scavenging assay), and also exhibited an increased reducing power. The conjugates also showed excellent lipid peroxidation inhibition abilities in a linoleic acid emulsion system. The conjugates exhibited antimicrobial activity against 15 clinical isolates, two standard methicillin-resistant Staphylococcus aureus (MRSA) and three standard methicillin-susceptible S. aureus strains, as well as eight foodborne pathogens. Additionally, the conjugates showed no cytotoxic activity towards human Chang liver and mouse macrophage RAW264.7 cells.

Factors affecting anti-inflammatory effect of chitooligosaccharides in lipopolysaccharides-induced RAW264.7 macrophage cells.

In this study, factors affecting anti-inflammatory effect of chitooligosaccharides (COSs) in lipopolysaccharides (LPS)-induced RAW264.7 macrophage cells were investigated. The inhibition of NO secretion by COSs revealed that 90-COSs (90% N-deacetylation) significantly inhibited NO secretion than those of 50-COSs (50% N-deacetylation), and 90-HMWCOS (5000-10,000Da) in the 90-COSs showed the highest inhibition activity. Furthermore, 90-HMWCOSs also found to inhibit LPS-stimulated production of PGE(2), TNF-alpha and IL-6, as well as the expression of iNOS, COX-2, TNF-alpha, and IL-6. These results suggested that 90-HMWCOS may have anti-inflammatory effect via down-regulation of transcriptional and translational expression levels of TNF-alpha, IL-6 and iNOS and COX-2.

Anti-inflammatory action of high molecular weight Mytilus edulis hydrolysates fraction in LPS-induced RAW264.7 macrophage via NF-κB and MAPK pathways

Anti-inflammatory Mytilus edulis hydrolysates (MEHs) were prepared by peptic hydrolysis and MEH was further fractionated into three fractions based on molecular weight, namely >5kDa, 1-5kDa, and <1kDa. The >5kDa peptide fraction exerted the highest nitric oxide (NO) inhibitory activity and inhibited prostaglandin E2 (PGE2) secretion in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. Pretreatment with the >5kDa peptide fraction markedly inhibited LPS-stimulated inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) protein and gene expressions. Stimulation by LPS induced the production of pro-inflammatory cytokines, such as tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and -1β (IL-1β), whereas co-treatment with the >5kDa peptide fraction suppressed pro-inflammatory cytokine production. The >5kDa peptide fraction inhibited the translocation of NF-κB (nuclear factor-kappa B) through the prevention of IκBα (inhibitory factor kappa B alpha) phosphorylation and degradation and also inhibited the MAPK signaling pathway in LPS-stimulated RAW264.7 macrophages.

Antioxidant effects of fermented sea tangle (Laminaria japonica) by Lactobacillus brevis BJ20 in individuals with high level of γ-GT: A randomized, double-blind, and placebo-controlled clinical study

A randomized, double-blind, and placebo-controlled clinical study was performed to evaluate the antioxidant effects of fermented sea tangle (FST) on healthy volunteers with high levels of γ-glutamyltransferse (γ-GT). Forty-eight participants were divided into a placebo group and an FST group that received FST (1.5 g/day) for 4 weeks. Serum γ-GT, malondialdehyde (MDA), catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) activities were determined before and after the trial. Administering FST significantly decreased serum levels of γ-GT and MDA. Additionally, SOD and CAT activities were significantly augmented compared to those in the placebo group after 4 weeks, but no significant alteration was observed in GPx activity compared to that in the placebo group. Our findings indicate that FST enhanced the antioxidant defense system in a healthy population and may be useful as a functional food ingredient.

Protective effect of fermented sea tangle against ethanol and carbon tetrachloride-induced hepatic damage in Sprague-Dawley rats.

Sea tangle has long been used as Korean folk remedy to promote material health, and is one of the popular dietary supplement. This study was designed to evaluate the protective effect of fermented sea tangle (FST) against ethanol and carbon tetrachloride (CCl(4))-induced hepatotoxicity in rats. Sprague-Dawley rats were orally treated with FST (25, 250, 2500 mg/kg/day) with administration of ethanol (5 mL/kg) for 13 weeks and the single intraperitoneal (i.p.) dose of 50% CCl(4) (5 mL/kg/day, CCl(4) in olive oil) at 12 week, and repeated i.p. dose of 20% CCl(4) (2 mL/kg/day) for 1 week. Hepatotoxicity was evaluated by measuring the serum levels of glutamic pyruvate transaminase (GPT), gamma glutamyl transpeptidase (gamma-GT) and malondialdehyde (MDA) as well as the tissue levels of antioxidant enzyme such as superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx). Ethanol and CCl(4)-induced the rat liver damage, and significantly increased (p<0.05) the GPT, gamma-GT and MDA levels, and decreased the SOD, CAT and GPx levels. However, treatment with FST could decrease serum GPT, gamma-GT, and MDA levels significantly in plasma, and increase the activities of SOD, CAT, and GPx in liver tissues compared with ethanol and CCl(4)-treated group.

Protective effects against H2O2-induced damage by enzymatic hydrolysates of an edible brown seaweed, sea tangle (Laminaria japonica).

Enzymatic hydrolysates of Laminaria japonica were evaluated for antioxidative activities using hydroxyl radical scavenging activity and protective effects against H(2)O(2)-induced DNA and cell damage. In addition, activities of antioxidative enzymes, including catalase, glutathione peroxidase, and glutathione S-transferase, of the enzymatic hydrolysates from L. japonica were also estimated. L. japonica was first enzymatically hydrolyzed by seven carbohydrases (Dextrozyme, AMG, Promozyme, Maltogenase, Termamyl, Viscozyme, and Celluclast [all from Novo Co., Novozyme Nordisk, Bagsvaerd, Denmark]) and five proteinases (Flavourzyme, Neutrase, Protamex, Alcalase [all from Novo Co.], and pancreatic trypsin). The hydroxyl radical scavenging activities of Promozyme and pancreatic trypsin hydrolysates from L. japonica were the highest as compared to those of the other carbohydrases and proteinases, and their 50% inhibitory concentration values were 1.67 and 317.49 mug/mL, respectively. The pancreatic trypsin hydrolysates of L. japonica exerted a protective effect on H(2)O(2)-induced DNA damage. We also evaluated the protective effect on hydroxyl radical-induced oxidative damage in PC12 cells via propidium iodide staining using a flow cytometer. The AMG and pancreatic trypsin hydrolysates of L. japonica dose-dependently protected PC12 cells against cell death caused by hydroxyl radical-induced oxidative damage. Additionally, we analyzed the activity of antioxidative enzymes such as catalase, glutathione peroxidase, and the phase II biotransformation enzyme glutathione S-transferase in L. japonica-treated cells. The activity of all antioxidative enzymes was higher in L. japonica-treated cells compared with the nontreated cells. These results indicate that enzymatic hydrolysates of L. japonica possess antioxidative activity.