• YAKHAK HOEJI
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• v.38 no.5
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• pp.530-543
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• 1994

To study the feasibility of transmucosal delivery of leucine enkephalin (Leu-Enk) and $[D-ala^2]$-leucine enkephalinamide (YAGFL), their degradation extents and pathways in various rabbit mucosa extracts were investigated by high performance liquid chromatography. The degradation of Leu-Enk and YAGFL was observed to follow the first-order kinetics. The degradation half-lives of Leu-Enk in the nasal, rectal and vaginal mucosal extracts were 1.62, 0.37 and 1.12 hrs and those of YAGFL were 30.55, 9.70 and 6.82 hrs, respectively, indicating Leu-Enk was degraded in a more extensive and rapid manner than YAGFL. But the mucosal and serosal extracts of the same mucosa showed the similar degradation rates for both pentapeptides. The degradation was most rapid in the neutral pH and increasing concentrations of substrates retarded the degradation rates. The maior hydrolytic fragments of Leu-Enk were Des-Tyr-Leu-Enk and tyrosine, indicating the enzymatic hydrolysis by aminopeptidases. However, the data also suggested endopeptidases such as dipeptidyl carboxypeptidase and dipeptidyl aminopeptidase could play some role in the degradation of Leu-Enk. On the other hand, the hydrolytic fragments of YAGFL in all the mucosa extracts were mainly Tyr-D-Ala-Gly and Phe-Leu-Amide, demonstrating the hydrolytic breakdown by endopeptidases. The degradation pathways were further explored by concomitantly determining the formation of smaller metabolites of primary hydrolytic fragments of Leu-Enk and YAGFL in the mucosa extracts.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.9
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• pp.1139-1145
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• 2006

In this study, antioxidant activities of enzymatic and methanolic extracts from E. cava stem and leave were evaluated by measuring the scavenging activities on 1,1 diphenyl 2 picrylhydrazyl (DPPH), hydroxyl radical, hydrogen peroxide and the inhibitory effects on DNA damage induced by oxidative stress of cells. Enzymatic extracts were prepared by enzymatic hydrolysis of both stem and leave using food grade five different carbohydrases (Viscozyme, Celluclast, AMG, Termamyl, Ultraflo) and five proteases (Protamex, Kojizyme, Neutrase, Flavourzyme, Alcalase). The enzymatic extracts were lower than methanolic extracts in polyphenol contents, but higher in extraction yield by approximately 30%. The enzymatic extracts were superior to methanolic extracts in DPPH and H2O2 scavenging activities and DNA damage protective effect. There were no significant antioxidant activity difference between stem and leave, but the extracts of leave were relatively better than those of stem. In this study it is suggested that E. cava stem as well as its leave would be a good raw materials for antioxidants compound extraction and enzymatic hydrolysis would be a good strategy to prepare antioxidant extracts from seaweeds.

• Journal of Pharmaceutical Investigation
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• v.26 no.3
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• pp.175-185
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• 1996

To inhibit the enzymatic degradation of leucine enkephalin (Leu-Enk) and its synthetic analog. $[D-ala^2]$-leucine enkephalinamide (YAGFL), in the nasal, rectal and vaginal mucosal and serosal extracts of rabbits, effects of enzyme inhibitors such as amastatin (AM), puromycin (PM), thiorphan (TP), thimerosal (TM), EDTA, N-carboxymethyl-Phe-Leu (CPL), phenylethyl alcohol (PEA), phenylmercuric acetate (PMA), benzalkonium chloride (BC) and modified cyclodextrins, alone or in combination, were observed by assaying the pentapeptides staying intact during incubation. Mucosa extracts were prepared by exposing freshly-excised mucosal specimens mounted on Valia-Chien cells to isotonic phosphate buffer while stirring. The degradation of Leu-Enk and YAGFL followed the apparent first-order kinetics. The half-lives (mean) in the nasal, rectal and vaginal mucosal extracts were found to be 1.07, 0.33 and 1.14 hr for Leu-Enk, and 16.9, 6.2 and 6.8 hr for YAGFL, respectively. AM or PM, which is an aminopeptidase inhibitor, did not show a sufficient inhibition of Leu-Enk $(50\;{\mu}g/ml)$ degradation in all kinds of extracts. $Dimethyl-{\beta}-cyclodextrin\;(DM-{\beta}-CyD)$ decreased the degradation rate constants of Leu-Enk about 2 or 3 times, comparing with no additive. However, the use of mixed inhibitors of AM $(50\;{\mu}M)$/TM (0.25 mM)/EDTA (5 mM) resulted in a full stabilization of Leu-Enk by decreasing the degradation rate constants 67.3, 161.3 and 113.8 times far the nasal, rectal and vaginal mucosal extracts, respectively, comparing with no inhibitor. With mixed inhibitors, Leu-Enk remained intact more than 90% after 6 hr-incubation. In the stabilization of YAGFL, hM, TP or CPL alone showed little efffct, and some additives demonstrated a considerable inhibition of YAGFL degradation in the rank order of TM > BC > EDTA. However, the addition of mixed inhibitors such as TM (0.5 mM) and EDTA (5 mM) into the extracts protected YAGFL from the degradation by more than 85% even after 24 hr-incubation, suggesting almost complete inhibition of YAGFL degradation in the extract. On the other hand, $DM-{\beta}-CyD\;or\;hydroxypropyl-{\beta}-cyclodextrin$ (10%) were also found to retard enzymatic degradation rates of YAGFL markedly, and resulted in staying intact more than 80% of YAGFL in the nasal and vaginal mucosal extracts, and more than 60% in the rectal mucosal extract after 16 hr-incubation.

• Korean journal of food and cookery science
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• v.28 no.1
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• pp.51-55
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• 2012

This study was conducted to investigate the physicochemical properties of citrus peel extracts with different hot water extraction and enzymatic hydrolysis conditions. Enzymatic hydrolysis was also employed using Viscozyme L and results were compared with that of optimized hot water extract. Hot water extraction was performed under different parameters; the sample to solvent ratio(1:20, 1:15, 1:10), extraction time(2, 4 hrs), extraction temperature(85, $95^{\circ}C$) and enzymatic hydrolysis(0, 1%) and the subsequent extracts were used for determining their physicochemical properties, such as total yield, total phenolics, total flavonoids, and electron donating ability (EDA). With the increase in the sample to solvent ratio and extraction time, total yield, total phenolics, total flavonoids and EDA increased. But extraction temperature did not significantly affect the hot water extract. As hot water extract was hydrolyzed by the enzyme, total yield and active ingredients increased rapidly. In the result of total yield, total phenolics, total flavonoids and EDA, the activity of enzyme-treated extract was higher than those of enzyme-untreated extract. Based upon the overall hot water extraction efficiency, it was found that 20 times volume or 120 min at a time at $95^{\circ}C$ after enzyme treatment was optimal.

• KSBB Journal
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• v.28 no.6
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• pp.349-355
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• 2013

SC092 strain, producing a polysaccharide degrading enzyme, was isolated from the seawater. This strain was identified as Microbulbifer sp. using the comparative sequence analysis against known 16S rRNA sequence. A polysaccharide degrading enzyme from this strain was used to acquire the enzymatic extracts of Sargassum fulvellum. DPPH radical scavenging and SOD activity of the enzyme extracts of S. fulvellum were about 61.9% and 82.9% at 2 mg/mL, respectively. Nitrite scavenging activities was 52.5% at 2 mg/mL on pH 1.2. In addition, ${\alpha}$-glucosidase inhibitory activity was also increased in a dose-dependent manner and was about 52.7% at 2 mg/mL. To determine the influence of enzyme extracts of S. fulvellum on alcohol metabolism, the generating activity of reduced-nicotinamide adenine dinucleotide (NADH) by alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) were measured. ADH and ALDH activities were 118.0% and 177% at 2 mg/mL, respectively. ${\alpha}$-glucosidase inhibitory activity of enzyme extracts of S. fulvellum was remarkably increased in a dose-dependent manner and was about 52.7% at 2 mg/mL. These results indicate alcoholizing and ${\alpha}$-glucosidase inhibitory activities can be enhanced by the enzymatic extracts of S. fulvellum.

• Korean Journal of Medicinal Crop Science
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• v.20 no.5
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• pp.331-338
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• 2012

In the present work, mulberry fruit extracts by four extraction processes, namely wet pressing extraction (WPE), hot-water extraction (HWE), enzymatic hydrolysis (EH), and lactic-acid bacteria fermentation (LBF) by Lactobacillus plantarum TO-2100, were analyzed for nutrients and functional compounds. The sugar contents of extracts by WPE, HWE, EH, and LBF were 12.0, 10.9, 14.5, and 14.3 brix, respectively, and the extraction yields by EH and LBF were 1.65 and 1.50 times higher than those by WPE. Among the organic acids, tartaric acid and malic acid contents were the highest in the extracts by WPE. Acetic acid was best extracted by LBF, and citric acid was best extracted by EH. Lactic acid was detected only in LBF. The extracts by EH showed the highest contents of all vitamins with an exception that the extracts by LBF showed the highest contents of the folic acid, vitamin B12, and vitamin C. We also noted that vitamin B group was not detected in the extracts by LBF. The extracts by EH showed the highest contents of all the amino acids, whereas LBF showed the lowest. Polyphenol contents of extracts by EH and LBF were 3.05 and 2.51 times more than those by WPE respectively. Anthocyanin contents were 7.66, 7.14 times higher for EH and LBF compare to WPE. We manufactured mulberry fruit granular teas with different compositions and tested them for their sensory characteristics. We found that 15% mulberry fruit extracts by enzymatic hydrolysis and 85% dextrin composition gave the most satisfactory result.

• Proceedings of the KAIS Fall Conference
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• 2010.11b
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• pp.744-747
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• 2010

In this study, antioxidant activity of enzymatic, ethanolic and aqueous extract from Blueberry were evaluated by measuring the scavenging activity on 1,1-diphenyl-2-picrylhydrazyl (DPPH). Enzymatic extract were prepared by enzymatic hydrolysis of Blueberry using food grade five different carbohydrases (Viscozyme, celluclast, AMG, Termarmyl, Ultraflo) and five proteases (Protamex, Kojizyme, Neutrase, Flavourzyme, Alcalase). The ethanol extract were lower than enzymatic extracts in yield, but higher in ployphenolic contents. The 70% ethanolic extract of Blueberry exhibited better DPPH radical scavenging activity compared to those of other extracts. These results suggest that Blueberry would be a good raw materials for antioxidant.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.39 no.1
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• pp.19-24
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• 2013

To investigate the efficacy of enzymatic extract of Ecklonia cava and its polyphenol extract (AG-DK) as cosmetic ingredients, the anti-oxidative effect, anti-glycation effect, anti-melanogenic effect, and anti-inflammatory effect of the extracts were evaluated in vitro. The enzymatic extract of E. cava ($SC_{50}$ 42.9 ppm) and AG-DK ($SC_{50}$ 6.4 ppm) showed a strong DPPH free radical scavenging activity. The anti-glycation ability of the enzymatic extract of E. cava and AG-DK was tested using bovine serum albumin (BSA), which inhibited the formation of advanced glycation end-products (AGEs) in the BSA/glucose system. The enzymatic extract of E. cava ($IC_{50}$ 97.2 ppm) and AG-DK ($IC_{50}$ 7 ppm) had inhibitory effects on tyrosinase activity. Moreover, the enzymatic extract of E. cava and AG-DK had an anti-inflammatory effect through the inhibition of nitricoxide (NO) and prostaglandin E2 ($PGE_2$). These findings suggest that the enzymatic extract of E. cava and AG-DK can be applied to skin-care products as cosmetic ingredients.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.4
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• pp.494-499
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• 2010

In this study, Sargassum coreanum was enzymatically hydrolyzed to prepare water-soluble extracts by using five carbohydrates (Viscozyme, Celluclast, AMG, Termamyl and Ultraflo) and five proteases (Protamex, Kojizyme, Neutrase, Flavozyme and Alcalase) and their potential antioxidant activity were evaluated. The Celluclast and Neutrase extracts of Sargassum coreanum exhibited better DPPH radical scavenging activities (92.42% and 92.78%, respectively) and hydrogen peroxide ($H_2O_2$) scavenging activities (58.28% and 57.97%, respectively) compared to those of other enzymatic extracts. These results suggest that Sargassum coreanum would be a good raw materials for antioxidant and enzymatic hydrolysis would be a good strategy to prepare antioxidant extracts from seaweeds.

• BMB Reports
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• v.30 no.6
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• pp.438-442
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• 1997

${\gamma}-Glutamylamine$ cyclotransferase was purified to homogeneity from soybean (Glycine max) seeds. To our knowledge, it is the first purification of the enzyme from plant origins. The molecular weight of the enzyme estimated by Sephacryl S-300 gel filtration and SDS-PAGE was 27,000, indicating that the enzyme is a monomer. The optimal pH for activity was 8.6. The Km value for ${\gamma}-glutamyldansylcadaverine$ was 11 ${\mu}M$. The enzymatic activity was substantially inhibited by the addition of p-chloromercuribenzoate and partially inhibited by the $Cu^{2+}$ ion. However, neither other modification reagents nor other divalent metal ions affected the enzymatic activity. The comparison between the enzymatic activities of seed extracts treated with cycloheximide and control extracts, and the detection of the same single protein band by western blot analysis at the dormant stage without inhibition with distilled water indicate that ${\gamma}-Glutamylamine$ cyclotransferase is already present at the dormant stage and gradually activated during germination in soybean seeds.