• Fisheries and Aquatic Sciences
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• 제14권3호
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• pp.174-178
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• 2011

This study aimed to determine the degree of hydrolysis and angiotensin-I-converting enzyme (ACE)-inhibitory activity of Giant Jellyfish Nemopilema nomurai (jellyfish) hydrolysates. The degree of hydrolysis using six proteolytic enzymes (Alcalase, Flavozyme, Neutrase, papain, Protamex, and trypsin) ranged from 13.1-36.8% and the inhibitory activities from 20.46-79.58%. Using papain hydrolysate, we newly isolated and characterized ACE-inhibitory peptides with a molecular weight of 3,000-5,000 Da that originated from jellyfish collagen. The purified peptide (FII-b) was predicted to be produced from an alpha-2 fragment of the type IV collagen of jellyfish. The N-terminal sequence of FII-b was Asp-Pro-Gly-Leu-Glu-Gly-Ala-His-Gly- and showed 87% identity to the collagen type IV alpha-2 fragment of Rattus norvegicus and a predicted protein from Nematostella vectensis, indicating that the ACE-inhibitory peptide originated from the collagen hydrolysate and had an $IC_{50}$ value of 3.8 ${\mu}g$/mL. The primary structure of the fragment is now being studied; this peptide represents an interesting new type of ACE inhibitor and will provide knowledge of the potential applications of jellyfish components as therapies for hypertension.

• Fisheries and Aquatic Sciences
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• 제15권4호
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• pp.265-273
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• 2012

The effects of processing and frozen storage conditions on the quality of anchovy Engraulis japonica fried surimi gels were investigated. Protein content decreased after surimi gel processing from 19.6% (raw meat) to 12.1% (kamaboko) due to the added ingredients and change in water content. Lipid content decreased from 2.8% (raw meat) to 1.3% in minced and 0.5% in surimi, but fried kamaboko showed a 6.9 % lipid level. Thiobarbituric acid values and thiobarbituric acid reactive substances levels were highest in kamaboko samples, 89.5 and 1.9 mg/g solid, and increased gradually with storage time to 101.8 and 4.6 mg/g solid, respectively. In vitro protein digestibility increased from 79.2% in raw anchovy to 88.5% in kamaboko samples. Levels of trypsin inhibitor decreased gradually with processing and during storage time from 2.43 in raw anchovy to 0.31 mg/g solid in the kamaboko sample after 60 days of frozen storage. No noticeable changes in total essential amino acid was observed during processing conditions. Computed protein efficiency ratio for kamaboko was highest (2.59) compared with whole anchovy (1.96), minced (1.94) and surimi (2.50). Fresh fried anchovy kamaboko showed similar values of hardness, springiness, gumminess and chewiness to commercial surimi gel, but a higher values were seen for fracturability and adhesiveness, and lower values for cohesiveness and resilience. The frozen and thawed anchovy kamaboko showed higher values for all of these rheological parameters compared with fresh and commercial kamaboko. Anchovy kamaboko showed the lowest lightness (62.9) and redness (0.16) and similar yellowness (11.9) compared with commercial kamaboko. Frozen storage and vacuum packaging were effective maintaining the shelf life of anchovy kamaboko within 30 days, but were not effective after 45 days due to fat oxidation.

• 한국수산과학회지
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• 제47권3호
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• pp.211-219
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• 2014

To confirm the food quality and storage stability of commercial Korean kamaboko, we experimented with the composition and textural properties using various surimis and kamaboko products. We also investigated changes in protein digestibility and lipid oxidation of vacuum packed products under chilled storage at $4{\pm}1^{\circ}C$. Among the fish meatbased surimi, vegetable mixed surimi had the lowest protein content (23.73 %), as compared to other surimi (51.9-73.6%). Siginificant (P<0.05) differences in protein, lipid content and degree of fat oxidation were noted between the fried kamaboko products of three companies. Adhesiveness, springiness, cohesiveness, gumminess, chewiness and resilience were similar in all samples, but there were notable differences in hardness and fractuability between samples. In vitro protein digestibility and trypsin indigestible substrate (TIS) were not inversely proportional in fried kamaboko products. The protein digestibility (80.30%) of steamed vegetable mixed fried kamaboko was lower than that of other fried samples (84.9-86.2%). Computed protein efficiency ratio (C-PER) of companies A and C's fried kamaboko was 2.6 but company B's was 1.9. There was no noticeable change in thiobarbituric acid reactive substances (TBARs) or protein digestibility for any of the vacuum packed fried kamaboko during 30 days of chilled storage.

• Fisheries and Aquatic Sciences
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• 제6권4호
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• pp.165-171
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• 2003

To investigate the effect of cooking methods on protein quality of domestic fresh monkfish meat (FMM) and imported frozen monkfish meat (IMM), in vitro protein qualities were determined by amino acid anlysis, trypsin indigestible substrate (TIS) formation, and protein digestibility using the four-enzyme method. Crude protein contents of the boiled FMM and IMM were $90\%$ of the dry base, which were higher than fresh FMM $(82\%)$ and IMM $(84\%)$. Profiles of total amino acid in FMM and IMM were not changed by cooking methods. Total free amino acid contents decreased to $ 29.0-33.6\%$ for boiled $(l00^{\circ}C,\;10 min)\;and\;24\%$ for steamed $(100^{\circ}C,\;10\;min)$ samples. In vitro protein digestibilities of boiled and steamed FMM incnased $86.6-86.8\%$, compared to raw IMM $(82.9\%)$, boiled and steamed IMM $85.1-85.5\%$ and raw IMM $(83.6\%)$. TIS of FMM (23.6 mg/g solid) and IMM (15.9 mg/g solid) showed no significant (p<0.05) difference in cooking methods. The C-PERs (computed protein efficiency ratio) of boiled FMM (2.63) and IMM (2.50) were significantly higher (<0.05) than raw (1.97) and steamed FMM(1.97) and IMM(1.94). These results demonstrate that boiling of FMM and IMM improves protein digestibility and C-PER when compared to steamed FMM and IMM. Therefore, boiling could be an excellent means to maintain high-protein quality of monkfish meat. Also, the cooking method may be applicable to the preparation of monkfish stew without any loss of free amino acids.

• Asian Pacific Journal of Cancer Prevention
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• 제15권21호
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• pp.9495-9498
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• 2014

Background: The difference in prognosis of adult and childhood acute lymphoblastic leukemia (ALL) can be attributed largely to variation in cytogenetic abnormalities with age groups. Cytogenetic analysis in acute leukemia is now routinely used to assist patient management, particularly in terms of diagnosis, disease monitoring, prognosis and risk stratification. Knowing about cytogenetic profile at the time of diagnosis is important in order to take critical decisions in management of the patients. Aim and Objectives: To determine the frequency of cytogenetic abnormalities in Pakistani adult patients with ALL in order to have insights regarding behavior of the disease. Materials and Methods: A retrospective analysis of all the cases of ALL (${\geq}15$years old) diagnosed at Aga Khan University from January 2006 to June 2014 was performed. Phenotype (B/T lineage) was confirmed in all cases by flow cytometry. Cytogenetic analysis was made for all cases using the trypsin-Giemsa banding technique. Karyotypes were interpreted using the International System for Human Cytogenetic Nomenclature (ISCN) criteria. Results: A total of 166 patients were diagnosed as ALL during the study period, of which 151 samples successfully yielded metaphase chromosomes. The male to female ratio was 3.4:1. The majority (n=120, 72.3%) had a B-cell phenotype. A normal karyotype was present in 51% (n=77) of the cases whereas 49% (n=74) had an abnormal karyotype. Of the abnormal cases, 10% showed Philadelphia chromosome; t(9;22)(q34;q11.2). Other poor prognostic cytogenetic subgroups were t(4;11)(q21;q23), hypodiploidy (35-45 chromosomes) and complex karyotype. Hyperdiploidy (47-57 chromosomes) occurred in 6.6%; all of whom were younger than 30 years. Conclusions: This study showed a relatively low prevalence of Philadelphia chromosome in Pakistani adults with ALL with an increase in frequency with age (p=0.003). The cumulative prevalence of Philadelphianegative poor cytogenetic aberrations in different age groups was not significant (p=0.6).

• KSBB Journal
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• 제13권1호
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• pp.83-89
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• 1998

Cellulose로 만들어진 다공성 미립담체를 이용하여 HeLa cell을 working volume 100mL의 spinner flask에서 배양하였으며, 세포가 완전히 자란 미립담제를 계대배양하기 위하여 담체간 세포 전이 배양 방법을 시도하였다. 부유세포의 농도는 다공성 미립담체-HeLa 시스템의 경우에 담체간 세포 전이 배양에 영향을 미치는 중요한 인자로 작용하였으며, 낮은 칼슘농도의 배지인 RPMI-1640과 빠른 교반 속도를 이용하여 활성을 유지한 많은 세포가 떨어지도록 유도하였으며, 담체간 세포 전이 배양을 효과적으로 3회 이상 실시할 수 있었다. 이렇게 배양한 세포에 재 조합 Vaccinia virus를 감염하여 그 수율을 비교한 결과 T-flask에서 떼어낸 세포로 접종한 미립담체 배양과 거의 비슷한 재조합 단백질($\beta$-galactosidase) 수율을 나타내었다. Trypsin 처리 방법에 의한 미립담체 계대 배양도 경우에 따라서는 유용한 미립담체 계대 배양 방법이 될 수 있지만 실제 생산 규모에의 적용에는 공정이 복잡해지고 정확한 제어가 필요하다는 등의 문제가 있다. 따라서, 추가적인 비용이나 공정이 필요 없는 간편한 방법인 담체간 세포 전이 배양은 동물세포 배양을 이용한 유용 단백질 및 바이러스 생산 공정의 규모 증대에 매우 유용한 수단이다.

• Fisheries and Aquatic Sciences
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• 제19권7호
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• pp.29.1-29.6
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• 2016

In the present study, we investigated to the antioxidant and angiotensin I-converting enzyme (ACE) inhibitory activities of the northern shrimp (Pandalus borealis) by-products (PBB) hydrolysates prepared by enzymatic hydrolysis. The antioxidant and ACE inhibitory activities of five enzymatic hydrolysates (alcalase, protamex, flavourzyme, papain, and trypsin) of PBB were evaluated by the 2, 2'-azino-bis [3-ethylbenzothiazoline-6-sulfonic acid] ($ABTS^+$) radical scavenging and superoxide dismutase (SOD)-like activities, reducing power and Li's method for ACE inhibitory activity. Of these PBB hydrolysates, the protamex hydrolysate exhibited the most potent ACE inhibitory activity with $IC_{50}$ value of $0.08{\pm}0.00mg/mL$. The PBB protamex hydrolysate was fractionated by two ultrafiltration membranes with 3 and 10 kDa (below 3 kDa, between 3 and 10 kDa, and above 10 kDa). These three fractions were evaluated for the total amino acids composition, antioxidant, and ACE inhibitory activities. Among these fractions, the < 3 kDa and 3-10 kDa fractions showed more potent $ABTS^+$ radical scavenging activity than that of > 10 kDa fraction, while the > 10 kDa fraction exhibited the significant reducing power than others. In addition, 3-10 kDa and > 10 kDa fractions showed the significant ACE inhibitory activity. These results suggested that the high molecular weight enzymatic hydrolysate derived from PBB could be used for control oxidative stress and prevent hypertension.

• BMB Reports
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• 제44권7호
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• pp.458-461
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• 2011

Enteropeptidase is a serine protease secreted by the pancreas and converts inactive trypsinogen to active trypsin. Enteropeptidase cleaves the C-terminal end of the substrate recognition sequence Asp-Asp-Asp-Asp-Lys ($D_4K$). The assay for enteropeptidase has utilized $GD_4K$-conjugated 2-naphthylamine ($GD_4K$-NA) as a fluorogenic probe over the last 30 years. However, no other $D_4K$-conjugated fluorogenic substrates of enteropeptidase have been reported. Furthermore, naphthalene is known as carcinogenic to humans. In this study, we used shift in the emission spectrum of $GD_4K$-conjugated 7-amino-4-methylcoumarin ($GD_4K$-AMC) as a fluorogenic method to measure enteropeptidase activity. The kinetic analysis revealed that enteropeptidase has a $K_M$ of 0.025 mM and a $k_{cat}$ of 65 $sec^{-1}$ for $GD_4K$-AMC, whereas it has a $K_M$ of 0.5 to 0.6 mM and a $k_{cat}$ of 25 $sec^{-1}$ for $GD_4K$-NA. The optimum pH of $GD_4K$-AMC hydrolysis was pH 8.0. Our data indicate that $GD_4K$-AMC is more suitable as a substrate for enteropeptidase than $GD_4K$-NA.

• Biomolecules & Therapeutics
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• 제3권2호
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• pp.165-170
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• 1995

The biochemical properties of the fibrinolytic protease of 50,800 Da isolated from the venom of Kgdistrodon blomhoffi brevicaudus were characterized. The enzyme hydrolyzed the carboxyl side of arginine in the synthetic chromogenic peptides, N-Benzoyl-Phe-Val-Arg-pNA and N-p-Tosyl-Gly-Pro-Arg-pNA, and the enzyme activity was inhibited by phenylmethylsulfonylfluoride indicating that the enzyme belongs to the serine protease family. The pretense showed maximum activity at pH 7.5 and inhibited by ZnCl$_2$, CuSO$_4$, but not by soybean trypsin inhibitor, pepstatin A, 2-mercaptoethanol and EDTA. The fm value determined with N-p-Tosyl-Gly-Pro-Arg-pNA was 0.2 mM. The thrombolytic activity of the purified enzyme was evaluated by platelet aggregation test in rabbits. While the platelet count ratio in blood of the rabbits injected with thrombin alone declined from 1.0 to 0.6 within 7 min and maintained around 0.6 for 24 hours thereafter, the ratio rapidly recovered from around 0.6 to 0.8 in 1 hr, to 1.0 in 24 hrs when the rabbits were sequentially treated with thrombin and the purified enzyme. The result showed that the serine protease from A. blomhoffi brevicoudus of 50,800 Da had a thrombolytic activity in vivo and the enzyme might be developed as a therapuetic agent for the treatment of thrombic disease.

• 생명과학회지
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• 제12권5호
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• pp.555-560
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• 2002

연접후치밀질(PSD)의 분자조성은 아직 대체로 알려져 있지 않다. 본 연구에서는 다양한 크기의 연접후치밀질 단백질들을 효율적으로 SDS-겔에서부터 추출하는 방법을 제시하고, 이를 이용하여 연접후치밀질의 1% n-octyl glucoside에 용해되지 않는 분핵에 존재하는 31kDa크기의 단백직(PSD31로 명명)을 SDS-겔로부터 분리하고, trypsin 처리하여 생성되는 펩티드의 아미노산 서열을 결정하였다. PSD31의 아미노산 서열은 adenine nucleotide translocator 1(ANT1)과 매우 상동성이 높았다. 연접후치밀질에 ANT1이 존재함은 신경조직의 세포외 공간에 adenosine nucleotide를 유리할 수 있는 기구가 연접에 존재함을 시사한다.