• Korean Journal of Fisheries and Aquatic Sciences
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• v.25 no.3
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• pp.229-235
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• 1992

Fish protein hydrolysates(FPH) prepared from defatted mackerel meal by proteases such as complex enzymes, bromelain, alcalase, $\alpha-chymotrypsin,$ trypsin, papain and pepsin were tested for inhibitory activity against angiotensin-I converting enzyme(ACE). Among proteases tested, the hydrolysates obtained from the treatment of complex enzymes or bromelain showed relatively higher activity. ACE inhibitory activity of the hydrolysates increased until hydrolysis of 8 hrs, and was stable by heat treatment for 20min at $100^{\circ}C.$ From the profiles of fractionation of the hydrolysates with Bio-gel P-2, the most active fraction had about MW 1,450 and it's amino acid was abundant in Asp, Glu, Lys, Leu, Val and Ala. $IC_{50}\;(amounts\;of\;inhibitors\;needed\;for\;50\%\;inhibition)$ of the active fraction of the hydrolysates obtained from the treatment of complex enzyme and bromelain was 90 and $130 {\mu}g,$ respectively.

• Journal of agriculture & life science
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• v.43 no.6
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• pp.95-103
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• 2009

To develop natural intermediate flavoring substances, optimal hydrolysis conditions and taste compounds for two step enzyme hydrolysate(TSEH) using Grass puffer(Takifugu niphobles) were investigated. The optimal conditions for TSEH method were revealed in temperature at $55^{\circ}C$ 3 hour digestion with Alcalase (pH 7.5) at the 1st step and 2 hour at $45^{\circ}C$ digestion with Flavourzyme(pH 6.0~6.5) at the 2nd step. TSEH method was superior to hot-water extraction on the aspect of yield, nitrogen contents and organoleptic taste quality such as umami and control of bitter taste formation. In taste active-components in Glass puffer TSEH, total free amino acid content was 4,502 mg%, major free amino acids were Pro, Leu, Lys, Hypro, Tau, Arg, Phe, Ala, Glu and Val in ordor. As for nucleotides, IMP(372 mg%) was the principal component and also contents of TMAO, total creatinine, and betaine were 43, 278 and 41 mg% in Glass puffer TSEH, respectively. The major inorganic ions in TSEH were Na(949 mg%), K(222 mg%), Cl(1,180 mg%) and $PO_4$(1,081 mg%).

• Korean Journal of Fisheries and Aquatic Sciences
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• v.54 no.6
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• pp.849-860
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• 2021

This study investigated the preparation of seasoned anchovy sauce (SAS) and its functional characteristics by using aminopeptidase active fractions (AAFs) derived from squid Todarodes pacificus hepatopancreas as a bitter taste improver. As the base of the SAS, a hydrolysate (AAAH) prepared by continuously treating raw anchovies with Alcalase-AAF was used. The high-performance liquid chromatography profile of the AAAH suggested that the action of AAFs decreased the hydrophobicity of the N-terminal peptide related to bitterness in the protein hydrolysates. SAS was prepared by blending with the AAAH and other ingredients. The crude protein (2.5%), carbohydrates (18.4%), amino acid-nitrogen (1,325.1 mg/100 mL), and total free and released amino acids (FRAAs, 700.2 mg/100 mL) of SAS were higher than those of commercial anchovy sauce (CAS). Sensory evaluation revealed that SAS was superior to CAS in flavor, color, and taste. The main FRAAs of SAS were glycine (16.8%), alanine (13.2%), glutamic acid (7.8%), and leucine (7.3%). The amino acids that had a major influence on the taste according to the SAS taste values were glutamic acid, aspartic acid, alanine, and histidine. The angiotensin-converting enzyme inhibitory (2.21 mg/mL) and antioxidant activities (3.58 mg/mL) of SAS were superior to those of CAS.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.1
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• pp.97-104
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• 2002

Protein hydrolysate was prepared as a natural flavor stock from the sharp toothed eel (Muraenesox cinereus) mince using com-mercially available proteolytic enzymes, Alcalase, Neutrase, Protamex, and Flavourzyme. A 6 hr hydrolysis of mince, to which water of the equal weight of the mince was added, with $2\%$ (w/w, protein weight) Elavourzyme at $50^{\circ}$ yielded a hydrolysate of the highest acceptability. Removing the access lipid in liquified hydrolysate (not dehydrated) after enzyme hydrolysis, five times repetitive extraction using n-hexane (liquified hydrolysate : n-hexane=4 : 1, v/v) was effective, resulting in less than $1\%$ lipid content of the dehydrated-hydrolysate. The amino acid composition of the hydrolysate (prepared with Flavourzyme) was similar to that of the starting material. Hydrolysis led to an increase in concentration of not only total free amino acid but also free amino acid such as serine, glutamic acid, alanine, and methionine responsible for umami taste, especially up to about 40 times for methionine. Major free amino acids in amount were leucine, phenylalanine, valine, alanine, and isoleucine and they comprised about half of the total free amino acids, Moisture adsorption, fat adsorption, emulsifying capacity, and foaming capacity of the hydrolysate were 870.1 $\pm$ $7.9\%$, 352.0$\pm$ $5.3\%$, 50.3 $\pm$ $1.2\%$, and $87.5\pm$ $2.5\%$, respectively, and solubility was 83$\~$$84\%$ at acid pH range of 2$\~$4.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.3
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• pp.307-314
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• 2006

This study was conducted to screen in vitro angiotensin converting enzyme (ACE) inhibitory activities of methanol (MeOH) and aqueous extracts which were prepared by four different extractions-80% methanol extracts(ME) at $20^{\circ}C\;and\;70^{\circ}C$, respectively and aqueous extracts (AE) at both temperatures with the residue of the MEs-of ten marine green algae and nineteen brown algae collected along Jeju coast of Korea. Most marine brown algae extracts showed higher capacities than those of marine green algae in ACE inhibitory activity. Particularly, $70^{\circ}C$ MeOH extract (70ME) of Hizikia fusiforme showed the strongest inhibition activity (about 87%) among all the extracts. Also, 70 MEs of Enteromorpha linza, Ishige sinicola, Laminaria ochotensis, Petrospongium rugosum, Sagrassum horneri, Undaria pinnatifida and $20^{\circ}C$ MeOH extracts (20ME) of Myagropsis myagroides, Petrospongium rugosum, $20^{\circ}C$ aqueous extracts (20AE) of Codium contractum, Enteromorpha compressa, and $70^{\circ}C$ aqueous extracts (70AE) of Ecklonia cava, Petrospongium rugosum showed moderate ACE inhibitory activities more than 50% and the other extracts exhibited weak activities. On tile other hand, E. cava had the best ACE inhibitory activity among 70AEs. This indicates that 70AE of E. cava contains potential anti-ACE macromolecular. We tried to proteolytic digest 70AE of E. cava to induce production of anti-ACE peptides from E. cava 70AE. The enzymes used are five pretenses including Kojizyme, Flavourzyme, Neutrase, Alcalase, and Protamex, which are food grade-commercial enzymes from Novo Co. Flavourzyme-digest of E. cava 70AE showed the highest inhibitory activity about 90%. And the five different enzymatic digests of the E. cava 70AE ranged from 2.33 to 3.56 ${\mu}g/mL$, respectively in $IC_{50}$ values of anti-ACE activity.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.31 no.6
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• pp.791-798
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• 1998

To develop natural flavoring substances. optimal hydrolysis conditions for two stage enzyme hydrolysates (TSEH) using low-utilized shellfishes such as purplish clam and frozen oyster stored at $-20^{\circ}C$ for 60 days. The optimal conditions for TSEH method were revealed in temperature at $50^{\circ}C$ 3 hours digestion with alcalase (Aroase AP-10, $0.3%$ w/v, pH 8.0) at the 1st stage and $45^{\circ}C$ 2 hours digestion with neutrase (Pandidase NP-2, $0.3\%$ w/v, pH 6.0) at the 2nd stage. Among water extracts, autolytic extracts and 4 kinds of enzyme hydrolysates tests, TSEH method was superior to other methods on the aspect of yields, nitrogen contents, taste such as umami and control of off-flayer formation, and transparency of extracts. From the results of chemical experiments and sensory evaluation, we may conclude that TSEH from low-utilized marine products is more flavorable compared the conventional enzyme hydrolysates, it could be commercialized as the seasoning substances.

• Food Science of Animal Resources
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• v.32 no.5
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• pp.652-658
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• 2012

This study was carried out to identify the ACE (Angiotensin converting enzyme) inhibitory activity of casein hydrolysates for development of anti-hypertensive hydrolysates. Sodium caseinate was treated with six kinds of commercial proteases such as Flavourzyme, Protamex, Neutrase 1.5, Alcalase, Protease M, and Protease S for 8 h individually, and was then treated with the enzyme combination for 4 h at $45^{\circ}C$. The hydrolysate which had the highest ACE inhibitory effect was then hydrolysed successively with three digestive enzymes: pepsin, trypsin, and ${\alpha}$-chymotrypsin, at $37^{\circ}C$ for 4 h under conditions mimicking those of the gastrointestinal tract. UF (ultra filtration) treatment was applied to one of the secondary hydrolysates to determine ACE inhibitory activity. When sodium caseinate was hydrolysed by commercial proteases, the degree of hydrolysis (DH) showed 2.54 to 4.25% and after secondary hydrolysis, DH showed 4.30 to 5.22%. ACE inhibitory activity and $IC_{50}$ values decreased, and inhibition rates increased during hydrolysis. Protamex treatment showed the lowest $IC_{50}$ value ($516{\mu}g/mL$) and Flavourzyme hydrolysate showed the highest $IC_{50}$value ($866{\mu}g/mL$). As the first hydrolysate was treated with Flavourzyme, the ACE inhibitory activity increased. Neutrase hydrolysate had the highest activity with an $IC_{50}$ value ($282{\mu}g/mL$). When Neutrase plus Flavourzyme treatment was hydrolyzed by digestive enzymes, the $IC_{50}$ value ($597{\mu}g/mL$) was decreased statistically (p<0.05). As Neutrase plus Flavourzyme hydrolysate is treated by UF with MW cut-off 10,000, permeate showed $273{\mu}g/mL$ of $IC_{50}$ value, showed no difference, but retentate which has over MW 10,000 showed statistically different $IC_{50}$ value, $635{\mu}g/mL$ (p<0.05).

• Journal of Life Science
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• v.21 no.2
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• pp.309-315
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• 2011

Rice syrup meal (RSM) was enzymatically hydrolyzed using eight commercial proteases (Protamex, Neutrase, Flavourzyme, Alcalase, Protease M, Protease N, Protease A, Molsin F) for 4 hr at optimum pH and temperature. Proteolytic hydrolysates were examined in supernatant and precipitate using Lowry protein assay, semimicro Kjeldahl method and gravimetric method using weight difference before and after enzymatic hydrolysis. Although RSM contains a high amount of protein (71.2%), only a very small amount of protein was hydrolyzed. Two proteases (Protease M and Protease N) were found to be the most effective in the hydrolysis of RSM protein. In Lowry method, 57.5 and 59.0 mg protein/g RSM were hydrolyzed after Protease M and Protease N treatments, respectively. In gravimetric method, 80.0 and 85.4 mg protein/g RSM were hydrolyzed after Protease M and Protease N treatments. In Kjeldahl method, 67.43 and 70.43 mg protein/g RSM were hydrolyzed after Protamex and Protease N treatments, respectively. For synergistic effect, two or three effective commercial proteases (Protease M, Protease N and Protease A) were applied to RSM at one time. The highest hydrolysis of RSM protein was observed in both Lowry protein assay (80.3 mg protein/g RSM) and gravimetric methods (153.2 mg protein/g RSM) when three commercial proteases were applied at one time, suggesting the synergistic effect of those proteases.

• 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.