• Korean Journal of Food Science and Technology
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• v.20 no.5
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• pp.659-665
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• 1988

The ${\alpha}_{s1}$-and ${\beta}$-casein were purified by DEAE-cellulose chromatography and digested with alkaline protease from Bacillus subtilis. Bitter fractions from the hydrolyzates were isolated using n-butanol extraction, Sephadex G-25 gel chromatography, and high performance liquid chromatography. Peptide mixtures were separated by reverse-phase octadecyl silica column with linear gradient of 0-80% acetonitrile containing 0.1% trifluoroacetic acid. Major peaks were combined from replicate chromatographies and the bitterness of each peak was evaluated. The bitter-tasting peaks were rechromatograpied until isolated peaks were obtained. Three different bitter peptides(BP-I, BP-II, BP-III) were obtained from the ${\alpha}_{s1}$-casein hydrolyzate. BP-I was eluted at 34% acetonitrile and BP-II, 35%, BP-III, 26%, respectively. Two bitter peptides(BP-IV, BP-V) were isolated from the ${\beta}-casein$ hydrolyzate: BP-IV was eluted at 40% acetonitrile and BP-V, 42%. BP-V was the most hydrophobic peptide in the five bitter peptides. However, BP-I and BP-II tasted more bitter than BP-IV and BP-V.

• Korean Journal of Food Science and Technology
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• v.17 no.4
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• pp.276-282
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• 1985

To characterize bitter peptides in fermented protein foods, peptides were extracted with 2:1 (v/v) chloroform-methand from various samples and separated into fractions I, II, and III by Sephadex G-25 gel chromatography. Amino acid compositions of Mozzarella cheese, soybean paste, and each fraction from the two samples were analyzed to calculate the average hydrophobicity. All the solvent extracts of the food samples had strong bitter taste, although the original samples did not taste bitter. The yield of solvent extraction ranged from 0.08 to 62.50% of total nigrogen of food samples. The average hydrophobicity calculated from the amino acid composition of Mozzarella cheese was 1376 cal/mole, solvent extract 1,623 cal/mole, gel chromatography traction I, 1,797 cal/mole, fraction II, 2,454 cal/mole, and fraction III, 1,559 cal/mole. In the case of soybean paste, the average hydrophobicity of original sample, solvent extract, gel chromatography fraction I, II, and III wre 1,229, 1,654, 1,900,998 cal/mole, respectively. The important amino acids in bitter peptides were leucine, 2016, phenylalanine, proline, and voline.

• Korean journal of food and cookery science
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• v.10 no.1
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• pp.45-50
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• 1994

Conventional Doenjang, three kinds of Improved Doenjang(A; koji: soybean: salt=53: 100: 33, B; koji: soybean: salt=100: 100: 40, C; koji: soybean: salt= 200: 100: 40) prepared with different ratio of koji and salt were made to study the changes in the general contents, characteristics bitter peptides, correlations between bitterness and overall eating quality. 1. Total nitrogen contents increased a little, and amino nitrogen contents in all samples increase markedly. Especially, Amino nitrogen contents of conventrional Doenjang increased more than others. Reducing sugars of doenjang prepared with Asp. oryzae were higher than conventional Doenjang and increased throughout the aging period and Doenjang prepared with Asp. oryzae were more acidic. 2. To characterize bitter peptides in fermented Doenjang, peptides were extracted with 2: 1(v/v) chloroform-methanol and separated by Gel chromato-graphy with Sephadex and TLC. After Gel chromatography and TLC, each fraction examined presence of bitterness and evaluated intensity of bitterness. Amino acid composition of the fractions showing bitter tastes were as follows. Conv. peak 1-1 Trp-(Asp, Arg, Thr, Ser, Glu, Pro)-Phe Imp. A peak 1 Trp-(Glu, Val, Arg, lie)-Phe Imp. B peak 1 Trp-(lie, Pro, Asp, Lys, Val, Glu)-Trp. Imp. C peak 1-2 Trp-(Try, Thr, Glu, Pro, Gly)-Phe 3. Sensory evaluation revealed that correlation coefficient between bitterness and overall eating guality was not high.

• Fisheries and Aquatic Sciences
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• v.15 no.1
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• pp.15-19
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• 2012

A new cyclic peptide (six-membered amino acid), gamakamide-E (L-Leu-L-Met (SO)-L-Me-Phe-L-Leu-D-Lys-L-Phe), was isolated as a strongly bitter tasting compound from cultured oysters, Crassostrea gigas. The molecular formula of $C_{43}H_{61}N_7O_8S$ was deduced from high resolution fast atom bombardment mass spectrometry (HR FAB-MS) ($[M+H]^+$ m/z 836.4356 ${\Delta}$= -2.4 mmu). Its unique structure including a hydantoin structure was firstly elucidated by nuclear magnetic resonance (NMR) analysis. Stereochemistries of constituent amino acids were determined by chiral high performanced liquid chromatography analysis of natural and synthesized peptides.

• Fisheries and Aquatic Sciences
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• v.18 no.2
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• pp.131-135
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• 2015

Two new bitter-tasting cyclic peptides comprising six amino acids, namely gamakamide C and D, were isolated from cultured oysters Crassostrea gigas. Dimethylaminoazobenzene sulfonyl-amino acid analysis detected Val and Leu in gamakamide C and Ile and Leu in gamakamide D. The molecular formula of gamakamide C was determined as $C_{43}H_{60}N_{7}O_8S$ by high-resolution fast atom bombardment mass spectroscopy (HR FAB-MS) ($[M+H]^+m/z822.4200{\Delta}-2.4mmu$), and that of gamakamide D was determined as $C_{43}H_{62}N_7O_8S$ by HR FAB-MS ($[M+H]^+m/z836.4379{\Delta}-2.0mmu$). Comparison of amino acid analyses and fragment ions by MS/MS among gamakamide C, D, and E (known), the structures of gamakamide C and D were confirmed $as-{\small{L}}-Val-{\small{L}}-Met(SO)-{\small{L}}-NMe-Phe-{\small{L}}-Leu-{\small{D}}-Lys-{\small{L}}-Phe-$ and $-{\small{L}}-Ile-{\small{L}}-Met(SO)-{\small{L}}-NMe-Phe-{\small{L}}-Leu-{\small{D}}-Lys-{\small{L}}-Phe-$, respectively.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.47 no.2
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• pp.135-143
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• 2014

Exopeptidase active fractions from the hepatopancreas of the Argentina shortfin squid Illex argentinus, were obtained with acetone (AC 30-40%), ammonium sulfate (AS 60-70% saturation), anion exchange chromatography (AE-II, 0.2 M NaCl) and gel filtration chromatography (GF-I, 30-50 kDa) fractionation methods. A bitter peptide solution that has a bitterness equivalent to that of 2% glycylphenylalanine and prepared by tryptic hydrolysis of milk casein, was treated with the exopeptidase active fractions. The GF-I fraction was the best based on aminopeptidase activity (35.3 U/mg), percentage of recovery (30.7%) and a sensory evaluation (1.7). The amount of released amino acids increased as incubation time increased, and the bitterness of the enzyme reaction mixtures decreased. Incubation with the GF-I fraction for 24 h resulted in the hydrolysis of several peptides as revealed by the reverse-phase high performance liguid chromatography profile, with three peaks (3, 5 and 6) decreasing in area (%) and three peaks (1, 2 and 4) increasing in area (%). Therefore, the GF-I fraction appeared to be ideally suited to reduce bitterness in protein hydrolysates by catalyzing the hydrolysis of bitter peptides.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.28 no.1
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• pp.98-104
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• 1995

Five bitter taste compounds (OY-22, OY-23, OY-24, OY-25 and OY-26) were firstly isolated from cultured oyster (Crassostrea gigas) at Gamak Bay, Sourthern coast of Korea, between November, 1989 and January, 1990, and smoked-canned oyster, which were produced by the same oysters. They were presumed as cyclic peptides composed with 6 or 7 amino acids, including sulfur on the basis of NMR and MS spectra. Val and Leu in OY-24, leu and lie in OY-25 and tow leucines in OY-26 were detected from those each compounds, seperately, by the amino acid analysis. Another amino acids were thought as non-constitutional amino acids. They showed non-toxic to mice $(100{\mu}g/20g\;mice i.p.)$ and non actibacterial artivities to Asp. niger and B. subtilis $(10{\mu}g/disk)$. The chemical structures and other biological activities of them are now in studying.

• Food Science of Animal Resources
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• v.34 no.4
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• pp.543-551
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• 2014

The aim of this study was to evaluate the effect of proteolytic (Serratia liquefaciens, match %: 99.39) or lipolytic (Acinetobacter genomospecies 10, match %: 99.90) psychrotrophic bacteria (bacterial counts, analysis of free fatty acids (FFA) and analysis of free amino acids) on the microbial and chemical properties (yogurt composition), and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of yogurt during storage. Yogurts were prepared with raw milk preinoculated with each psychrotrophic bacteria. The total solid, fat, and protein content were not affected by preinoculation, but the pH of yogurt preinoculated with psychrotrophic bacteria was higher than in control. There was a dramatic increase in short chain free fatty acids among FFA in yogurt with Acinetobacter genomospecies 10. For 14 d of cold storage condition, SCFFA was 25.3 mg/kg to 34.4 mg/kg (1.36 times increased), MCFFA was 20.4 mg/kg to 25.7 mg/kg (1.26 times increased), and LCFFA was 240.2 mg/kg to 322.8 mg/kg (1.34 times increased). Serratia liquefaciens (match %: 99.39) in yogurt caused a greater accumulation of free amino acids (FAA), especially bitter peptides such as leucine, valine, arginine, and tyrosine, but SDS-PAGE showed that the inoculation of Serratia liquefaciens did not affect the degree of casein degradation during storage. Taken together, the excessive peptides and FFA in yogurt generated from psychrotrophic bacteria could develop off-flavors that degrade the quality of commercial yogurt products.

• Journal of Microbiology and Biotechnology
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• v.9 no.4
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• pp.509-513
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• 1999

The molecular size reduction and the formation of bitterness during a tryptic hydrolysis of soybean 11S glycinin were determined by using quantitative analysis and organoleptic evaluation. The 11S glycinin of 90% purity was prepared by cryoprecipitation and Con A Sepharose 4B affinity chromatography, and hydrolyzed with trypsin in a pH-stat reactor for 4 h. Bitterness was formed within 1 h of hydrolysis, and then slowly increased up to $3.5\times10^{-5}$ M quinine-HCl equivalent. The extent of hydrolysis (DH) was 7% at 1 h and increased up to 12% by the end of the reaction. The -amino nitrogen content increased from an initial 0.7 mM to 7 mM at the end of the period. The SDS-PAGE analysis showed that the acidic subunit of 11S glycinin was mostly hydrolyzed. The GP-HPLC analysis indicated that the bitterness was mainly contributed by the peptide fractions of molecular weights of 360-2,100 Da.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.4
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• pp.488-494
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• 1999

Hydrolytic peptides of salted and fermented anchovy sauce, and anchovy actomyosin for the development of a rapid fermentation method with conventional tastes and flavors were studied. The optimal temperatures of crude enzymes isolated from anchor, liver and viscera of squid were 55, 40$\~$45 and $45\~60^{\circ}C$, respectively. Crude enzyme isolated from anchovy was more effective on hydrolysis of anchovy actomyosin than that from squid liver and viscera. But the crude enzyme from squid liver was less effective on NaCl than that from anchovy. Three peptides occurred in anchovy actomyosin hydrolyzed with crude enzymes from anchovy and squid liver for 30 min. Their molecular weight were determined by Superdex 200 gel chromatography as 10,800, 5,800 and 2,600 dalton. When anchovy sauce was hydrolyzed with crude enzymes of anchovy, squid liver and viscera, and Protamex during 70 days, ranges of their low molecular weight of hydrolyzed peptides were 300$\~$1,000dalton detected by Sephadex G-50 and their major amino acid compositions were glutamic acid, glycine and alanine, which was related with conventional tastes. Those amino acid compositions were similar to those of anchovy sauce hydrolyzed with squid liver, In the case of Protamex treatment, hydrolyzed peptides had high levels of isoleucine and leucine, being associated with the bitter, but a low level of glutamic acid.