• International Journal of Oral Biology
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• 제43권2호
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• pp.61-68
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• 2018

Codium fragile (Suringar) Hariot is an edible green seaweed that belong to the Codiaceae family and has been used in Oriental medicine for the treatment of enterobiasis, dropsy, and dysuria. Methanol extract of codium fragile has anti-oxidant, anti-inflammatory and anti-cancer properties, although the anti-cancer effect on oral cancer has not yet been reported. In this study, we investigated the anti-cancer activity and the mechanism of cell death by methanol extracts of Codium fragile (MeCF) on human FaDu hypopharyngeal squamous carcinoma cells. Our data showed that MeCF inhibits cell viability in a dose-dependent manner, and markedly induced apoptosis, as determined by the MTT assay, Live/Dead assay, and DAPI stain. In addition, MeCF induced the proteolytic cleavage of procaspase -3, -7, -9 and poly(ADP-ribose) polymerase(PARP), and upregulated or downregulated the expression of mitochondrial-apoptosis factor, Bax(pro-apoptotic factor), and Bcl-2(anti-apoptotic factor). Futhermore, MeCF induced a cell cycle arrest at the G1/S phase through suppressing the expression of the cell cycle cascade proteins, p21, CDK4, CyclinD1, and phospho-Rb. Taken together, these results indicated that MeCF inhibits cell growth, and this inhibition is mediated by caspase- and mitochondrial-dependent apoptotic pathways through cell cycle arrest at the G1/S phase in human FaDu hypopharyngeal squamous carcinoma cells. Therefore, methanol extracts of Codium fragile can be provided as a novel chemotherapeutic drug due to its growth inhibition effects and induction of apoptosis in human oral cancer cells.

• Fisheries and Aquatic Sciences
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• 제20권7호
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• pp.13.1-13.7
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• 2017

In order to evaluate the maximal storable period of the raw crab for a non-thermal muscle separation, the quality changes of the leg meat of red snow crab (Chionoecetes japonicus) during freeze storage were investigated. Fresh red snow crabs were stored at $-20^{\circ}C$ for 7 weeks, and the leg muscle was separated by a no heating separation (NHS) method every week. During the storage, considerable loss of the leg muscle did not occur and microbiological risk was very low. In contrast, discoloration appeared at 2-week storage on around carapace and the leg muscle turned yellow at storage 3-week. In physiochemical parameters, protein and free amino acids gradually decreased with storage time, expected that proteolytic enzymes still activated at $-20^{\circ}C$. At 4-week storage, the sensory acceptance dropped down below point 4 as low as inedible and notable inflection points in pH and acidity were observed. The volatile base nitrogen was low, though a little increase was recorded. These results suggested that the maximal storable period at $-20^{\circ}C$ of the raw material was within 2 weeks and it was depended on external factor such as the discoloration. The present study might be referred as basic data for approaches to solve quality loss occurred in non-thermal muscle separation.

• 한국식품과학회지
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• 제18권4호
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• pp.270-277
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• 1986

Ammonium sulfate 염분획과 CM-cellulose chromatography로 Fig latex에서 ficin을 분리하여 다음과 같은 결과를 얻었다. 분리한 crude ficin을 전기영동한 결과 main band 와 3개의 minor band로 나타났으며 main peak의 정제에서 얻은 ficin은 단일 band로 나타났다. 분리한 ficin의 pH 및 열에 대한 영향을 PH7.0, $60^{\circ}C$에서 최적 작용을 나타내었다. 분리한 ficin의 pH에 대한 안정성은 pH $2{\sim}8$로 비교적 넓은 범위로 나타났고 열에 대한 안정성은 $0{\sim}55^{\circ}C$의 범위로 안정성이 나타났다. 분리한 ficin의 분자량은 19500으로 나타났다. 산성 아미노산 21.8%. 염기성 아미노산 3.5%, 중성아미노산74.7%로 나타났으며 cystine과 tryptophan을 제외한 아미노산 잔기수는 174로 나타났다.

• 한국식품과학회지
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• 제20권3호
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• pp.433-440
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• 1988

천연 멸치추출액의 수율과 관능적 품질의 향상을 위하여 마른 중멸치를 0.3N NaOH알칼리 용액으로 마쇄하고 $60^{\circ}C$에서 1시간, 3시 간, 5시간 처리한 뒤 중화시켜 알칼리성 및 중성단백질 분해효소로 4시간 동안 가수분해 시켰다. 알칼리 처리한 시료를 효소반응 시간별로 멸치 마쇄용액을 취하여 원심 분리시킨 상등액에 함유된 가용성 고형분, 단백질, 무기질 그리고 맛과 냄새에 대한 관능적 품질을 평가하였다. 그 결과 고형분의 수율은 0.3N NaOH로 5시간 처리한 시료에 alkaline protease로 1시간 반응시켰을때, 최고 76.4%가 회수되어 물로만 추출한 대조구보다 약 4배의 수율이 향상되었고. 조단백질의 수율은 최고 65.3%로 약 3.2배 였으며, 회분의 수율은 30-47%의 회분 수율범위를 보여 12.9%였던 대조구에 비하여 현저하게 증가하였다. 또한 맛이나 냄새와 같은 관능적 품질에서도 그 강도가 2-3배의 향상효과가 있었으며 효소를 처리한 시료의 기호도가 현저히 향상됨이 밝혀졌다. 이상의 결과에서 고형분, 단백질 및 회분수율과 관능적 품질을 고려할때 알칼리처리 3시간후 단백질분해 효소로 1-3시간 처리함이 멸치 추출액의 수율과 관능적 품질에 가장 좋은 것으로 평가되었다.

• 동의생리병리학회지
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• 제19권3호
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• pp.802-810
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• 2005

The tetrahydroisoquinolines included potent cytotoxic agents that showed antitumor activity,antimicrobial activity, and other biological properties. We studied the effect of CDST, 1-Chloromethyl-6,7-dimethoxy-3,4-dihydro-1H-isoquinoline-2-sulfonic acid amide, a newly synthesized anti-cancer agent. The cytotoxic activity of CDST in HL-60 cells was increased in a dose-dependent manner. CDST, tetrahydroisoquinolines derivative, was cytotoxic to HL-60 cells, with IC50 of $80{\mu}g/ml$. Treatment of CDST to HL-60 cells showed the fragmentation of DNA in a dose- and time dependent manner, suggesting that thesecells underwent apoptosis. Treatment of HL-60 cells with CDST was induced in a dose- and time-dependent activation of caspase-3, caspase-8 and proteolytic cleavage of poly(ADP-ribose) polymerase. In caspase activity assay, caspase-3 and -8 was activated after 12 h and 6 h posttreatment, respectively. CDST also caused the release of cytochrome c from mitochondria into the cytosol. CDST-induced cytochrome c release was mediated by caspase-8-dependent cleavage of Bid and Bax translocation. These results suggest that caspase-8 induced Bid cleavage and Bax translocation, caused mitochondrial cytochrome c release, and induce caspase-3 activationduring CDST-induced apoptosis in HL-60 cells.

• 한국식품과학회지
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• 제20권4호
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• pp.488-492
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• 1988

멸치추출액의 제조과정중 멸치를 0.3N NaOH용액으로 마쇄하여 1,3,5시간 동안 알칼리 처리한 뒤 알칼리와 중성 단백질 분해효소로 4시간 가수분해하면서 얻어진 멸치마쇄액의 물성을 측정하였다. 멸치추출액의 물리적 품질로서 점도, 색, 총산도. 및 상등액의 양을 측정하여 비교하였다. 그 결과 효소처리한 멸치 마쇄액의 상등액량을 전체무게에 대하여 89%로서 70%인 비교구보다 약 20%증가되는 결과를 보여 주었으며, 상대점도는 효소처리 시간이 경과함에 따라 약간 감소하는 경향이었다. 총산도는 반응초기에 현저히 증가하였다가 그 후 완만하여 졌으며 Hunter값으로 표시된 색의 변화는 ‘L’값이 점점 감소하여 어두워졌으며 이러한 현상은 알칼리처리를 많이한 시료일수록 더욱 현저하였다. 한편 ‘a’값의 변화는 효소를 반응시키는 동안 일정한 변화경향은 없었으나 엷은 붉은색 범위를 보여 주었고 'b'값은 효소처리에 의하여 감소하는 경향이 뚜렷하였다.

• 농업과학연구
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• 제40권3호
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• pp.221-226
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• 2013

Degrees of hydrolysis by alkaline protease produced from Serratia marcescens S3-R1 is 3.95-6.30% of whey proteins during 5, 15, 30, 60, 90, 120,180, 240 min incubation at $40^{\circ}C$. Proteolytic pattern of the whey proteins showed that various low molecular weight peptides were generated during the incubation periods. The biological function of in Raw 264.7 cells treated with whey protein hydrolytic peptides, anti-inflammatory effect showed exhibit in the expression of pro-inflammatory cytokines such as TNF-${\alpha}$, IL-6, COX-2 and iNOS by PCR analysis. COX-2 and iNOS gene expression inhibited in Raw 264.7 cells on whey protein hydrolysates below 3,000 dalton. The protease from Serratia marcescens S3-R1 showed a potential in production of low molecular weight whey protein hydrolysates which could be used for industrial application.

• 한국축산식품학회지
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• 제34권3호
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• pp.362-371
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• 2014

This study utilized commercially available proteolytic enzymes to prepare egg-white protein hydrolysates (EPHs) with different degrees of hydrolysis. The antioxidant effect and functionalities of the resultant products were then investigated. Treatment with Neutrase yielded the most ${\alpha}$-amino groups (6.52 mg/mL). Alcalase, Flavourzyme, Protamex, and Ficin showed similar degrees of ${\alpha}$-amino group liberation (3.19-3.62 mg/mL). Neutrase treatment also resulted in the highest degree of hydrolysis (23.4%). Alcalase and Ficin treatment resulted in similar degrees of hydrolysis. All hydrolysates, except for the Flavourzyme hydrolysate, had greater radical scavenging activity than the control. The Neutrase hydrolysate showed the highest 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity ($IC_{50}=3.6mg/mL$). Therefore, Neutrase was identified as the optimal enzyme for hydrolyzing egg-white protein to yield antioxidant peptides. During Neutrase hydrolysis, the reaction rate was rapid over the first 4 h, and then subsequently declined. The $IC_{50}$ value was lowest after the first hour (2.99 mg/mL). The emulsifying activity index (EAI) of EPH treated with Neutrase decreased, as the pH decreased. The EPH foaming capacity was maximal at pH 3.6, and decreased at an alkaline pH. Digestion resulted in significantly higher 1,1-diphenyl-2-picrylhydrazyl (DPPH) and ABTS radical scavenging activity. The active peptides released from egg-white protein showed antioxidative activities on ABTS and DHHP radical. Thus, this approach may be useful for the preparation of potent antioxidant products.

• Journal of Microbiology and Biotechnology
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• 제6권1호
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• pp.1-6
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• 1996

A bacterial strain NS70 producing an alkaline protease was isolated from soil samples taken near a hot spring and identified as Bacillus licheniformis by its morphological and physiological properties and cellular fatty acid analysis. The isolated alkaline protease was purified by ammonium sulfate fractionation, DEAE-, CM-, and Phenyl-Sepharose column chromatography. The molecular weight of the purified enzyme was estimated to be 32, 000 Da by sodium dodecylsulfate polyacrylamide gel electrophoresis. Its optimal pH and temperature for proteolytic activity against Hammarsten casein were 12 and $65^{\circ}C$, respectively. The enzyme was stable at alkaline pH range from 6.0 to 12.0, and fairly stable up to $65^{\circ}C$. The enzyme was inhibited by phenylmethylsulfonyl fluoride but not by EDTA and N-ethylmaleimide indicating that the enzyme is serine protease. Enzyme activity was markedly inhibited by $Hg^{2+}$ and $Cu^{2+}$. Autolytic phenomena were observed on purified protease NS70 but autolysis was reduced by the addtion of $Ca^{2+}$ ion or bovine serum albumin.

• Journal of Microbiology and Biotechnology
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• 제28권6호
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• pp.968-975
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• 2018

In the course of screening for microbes with nematicidal activity, we found that Enterobacter asburiae HK169 displayed promising nematicidal activity against the root-knot nematode Meloidogyne incognita, along with plant growth-promoting properties. Soil drenching of a culture of HK169 reduced gall formation by 66% while also increasing root and shoot weights by 251% and 160%, respectively, compared with an untreated control. The cell-free culture filtrate of the HK169 culture killed all juveniles of M. incognita within 48 h. In addition, the nematicidal activity of the culture filtrate was dramatically reduced by a protease inhibitor, suggesting that proteolytic enzymes contribute to the nematicidal activity of HK169. In order to obtain genomic information about the HK169 isolate related to its nematicidal and plant growth-promoting activities, we sequenced and analyzed the whole genome of the HK169 isolate, and the resulting information provided evidence that the HK169 isolate has nematicidal and plant growth-promoting activities. Taken together, these observations enable the future application of E. asburiae HK169 as a biocontrol agent for nematode control and promote our understanding of the beneficial interactions between E. asburiae HK169 and plants.