• 한국발생생물학회지:발생과생식
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• 제3권1호
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• pp.29-38
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• 1999

자궁내막증은 흔한 부인과적 질병이며 여성 불임의 한 원인이 되나 그 발생 원인에 대하여서는 아직 논란의 여지가 많다. 최근 월경혈의 역류가 한 원인이며 자궁내막증 환자가 정상여성에서 보다 역류되는 월경혈의 양이 많거나 침습성이 강한 것이 자궁내막증의 발생 원인이 될 수 있다는 이론들이 소개되었다. 종양이나 자궁내막 조직의 침습이나 전이에는 세포막외 기질 및 기저막의 파괴가 일어나야 하는데 이 과정에 plasminogen activators (PAs)나 matrix metalloproteinase (MMPs)같은 proteolytic enzyme이 관여한다. 이에 자궁 내막증환자와 대조군의 자궁내막에서 PA나 MMP를 억제하는 plasminogen activator inhibitor-1 (PAI-1)나 tissue inhibitor of metalloproteinase (TIMP-3)의 mRNA 발현의 차이를 quantitative competitive RT PCR로 연구하였으며, 그 결과 자궁내막증 환자의 황체기 자궁내막에서는 정상 대조군 환자에서 보다 PAI-1과 TIMP-3 mRNA발현이 낮음을 알 수 있었다. 따라서 자궁내막증 환자의 자궁내막에서는 PA와 MMP의 활성도가 증가할 수 있으며 이 증가된 proteolytic activity로 인하여 역류된 자궁내 막 조직의 복강내 침습이 보다 쉽게 일어날 가능성이 있다.

• 한국식품영양과학회지
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• 제21권4호
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• pp.398-406
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• 1992

어류가공시 부산물로 얻어지는 어피를 효율적으로 천연조미료로 이용하기 위한 기초 자료를 얻기 위해 가자미피로부터 알칼리 전처리법 (B-type)과 효소 전처리법 (E-type)으로 젤라틴을 추출하여 trypsin으로 가수분해할 때 가수분해조건과 그 가수분해물의 물성, 분자량 및 아미노산 조성을 비교 검토하였다. 젤라틴 가수분해물의 제조를 위한 가수분해 조건은 B-type과 E-type 젤라틴 모두 반응온도 55$^{\circ}C$, pH 9.0 및 효소농도 E/S (w/w) : 0.001이었고, 반응시간은 B-type은 4시간, E-type은 1시간이었다. 최적 가수분해 조건하에서 B-type및 E-type젤라틴의 가수분해도는 각각 63%와 82%였다. 가수분해물의 등전점은 B-type과 E-type이 각각 pH 5.30와 PH 5.27이었고, 전기전도도는 각각 352.8$\mu\textrm{m}$ho/cm와 268.8$\mu\textrm{m}$ho/cm로 시판 젤라틴의 193.2$\mu\textrm{m}$ho/cm보다 높았다 점도는 농도와 온도에 따라서는 다소 변화가 있었으나 pH의 변화에는 크게 영향을 받지 않았다. 분자량은 B-type이 6.5~25kDa으로 분포되어 있고 15kDa이 주종을 이루고 있었으나 E-type의 경우는 분자량이 6~20kDa이며 12.4kDa이 주종을 이루고 있었다. 아미노산 조성은 B-type과 E-type 간에 거의 차이가 없었으며 단맛을 내는 glycine, alanine, proline, hydroxyproline, serine 등이 전체아미노 산의 57%를 차지하였으며 쓴맛을 내는 valine, leu-cine, phenylalanine, tyrosine, methionine, arginine, histidine등의 함량은 전체아미노산에 대해 18%에 불과하였다.

• Journal of Microbiology and Biotechnology
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• 제24권6호
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• pp.795-802
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• 2014

The aim of this study was to determine the effects of temperature and supplementation with skim milk powder (SMP) on the microbial and proteolytic properties during the storage of cottage cheese. Cottage cheese was manufactured using skim milk with 2% SMP and without SMP as the control, and then stored at $5^{\circ}C$ or $12^{\circ}C$ during 28 days. The chemical composition of the cottage cheese and the survival of the cheese microbiota containing starter lactic acid bacteria (SLAB) and non-starter culture lactic acid bacteria (NSLAB) were evaluated. In addition, changes in the concentration of lactose and lactic acid were analyzed, and proteolysis was evaluated through the measurement of acid soluble nitrogen (ASN) and non-protein nitrogen (NPN), as well as electrophoresis profile analysis. The counts of SLAB and NSLAB increased through the addition of SMP and with a higher storage temperature ($12^{\circ}C$), which coincided with the results of the lactose decrease and lactic acid production. Collaborating with these microbial changes, of the end of storage for 28 days, the level of ASN in samples at $12^{\circ}C$ was higher than those at $5^{\circ}C$. The NPN content was also progressively increased in all samples stored at $12^{\circ}C$. Taken together, the rate of SLAB and NSLAB proliferation during storage at $12^{\circ}C$ was higher than at $5^{\circ}C$, and consequently it led to increased proteolysis in the cottage cheese during storage. However, it was relatively less affected by SMP fortification. These findings indicated that the storage temperature is the important factor for the quality of commercial cottage cheese.

• 한국미생물·생명공학회지
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• 제49권1호
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• pp.65-74
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• 2021

Serine proteases are the most versatile proteolytic enzymes with tremendous applications in various industrial processes. This study was designed to investigate the biochemical properties, critical residues, and the catalytic potential of alkaline serine protease using in-silico approaches. The primary sequence was analyzed using ProtParam, SignalP, and Phyre2 tools to investigate biochemical properties, signal peptide, and secondary structure, respectively. The three-dimensional structure of the enzyme was modeled using the MODELLER program present in Discovery Studio followed by Molecular Dynamics simulation using GROMACS 5.0.7 package with CHARMM36m force field. The proteolytic potential was measured by performing docking with casein- and keratin-enriched residues, while the effect of the inhibitor was studied using phenylmethylsulfonyl fluoride, (PMSF) applying GOLDv5.2.2. Molecular weight, instability index, aliphatic index, and isoelectric point for serine protease were 39.53 kDa, 27.79, 82.20 and 8.91, respectively. The best model was selected based on the lowest MOLPDF score (1382.82) and DOPE score (-29984.07). The analysis using ProSA-web revealed a Z-score of -9.7, whereas 88.86% of the residues occupied the most favored region in the Ramachandran plot. Ser327, Asp138, Asn261, and Thr326 were found as critical residues involved in ligand binding and execution of biocatalysis. Our findings suggest that bioengineering of these critical residues may enhance the catalytic potential of this enzyme.

• 한국식품조리과학회지
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• 제16권4호
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• pp.367-371
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• 2000

육류연화를 위해 조리실무현장에서 자주 이용하는 과일인 배, 키위, 무화과, 파인애플, 파파야에 함유된 단백질 분해효소을 이용한 육류연화를 비교한 결과는 다음과 같다. 1, 양념장:물이 혼합비율이 2:1, pH 5.0∼5.5 부근의 산성일 때 최상의 연육 효과가 나타났다. 2. Rheometer를 이용하여 전단력 측정을 한 결과, pineapple>papaya> fig> kiwifruit > pear의 순으로 전단력 감소가 나타났으며, 이때 저작 5회째까지의 전단력 감소를 이용한 과일즙간의 연도에 미치는 영향을 비교한 결과, p<0.001 수준에서 파일애플, 파파야, 무화과는배나 키위에 비해 유의적으로 연도에 더 큰 영향을 주는 것을 관찰할 수 있었다. 3. 과일즙 첨가를 하여 조리한 육류에 대해 관능검사를 한 결과, p<0.01에서 연도와 전체적인 기호도에 있어 파인애플, 파파야 처리군과 배 처리군간에 유의적인 차이를 나타냈으며, p<0.05에서는 배와 파파야가 맛에 있어 유의적인 차이를 보였다. 4. 과일즙에 따른 관능과 기계적인 연도간의 상관성을 보면, 과일즙에 따른 기계적 연도와의상관성은 r=-.877로 가장 큰 상관성을 보였고(p<0.01), 연도가 높아짐에 따라 기호도도 상승하는 상관성(r=.532, p<0.01)이 있었다. 또한 과일즙에 따라 관능적 연도는 r=.495의 상관성을 보였고, 기계적 연도와 관능적 연도간에는 r=.490으로 상관성이 있음을 알 수 있었다. 한편 p<0.05 수준에서 맛과 다즙성 사이에 r=.208로서 상관도는 낮으나 유의적인 차이를 나타냈다. 즉 단백질 분해효소를 갖고 있는 과일즙 단백질 연화에 미치는 효과는 매우 크며 이들을 이용한 레시피 표준화는 앞으로 필요한 연구과제라고 사료된다.

• 동아시아식생활학회지
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• 제4권3호
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• pp.87-96
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• 1994

This study was carried out to investigate the effect of hydrolysis by proteolytic enzymes on the functional properties of sesame protein concentrate. Sesame protein concentrate was hydrolyzed with papain, pepsin and trypsin to obtain 10% and 20% degree of hydrolysis. The nirogen solubility in water was increased with increasing the degree of hydrolysis. Bulk density was increased by enzymatic hydrolysis but water absorption capacity was increased only in the case of pepsin-hydrolyzed SPC. Higher fat absorption capacity was found in SPC with 10% DH than SPC with 20% DH. Emulsifying activity was also increased by enzymatic hydrolysis except SPC with 10% DH by papain.

• BMB Reports
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• 제30권2호
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• pp.125-131
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• 1997

This work reports studies of the catalytic and structural properties of pyridoxal kinase (ATP: pyridoxal 5' -phosphotransferase, EC. 2.7.1.35), Pyridoxal kinase catalyzes the phosphorylation of vitamin $B_6$ (pyridoxal, pyridoxamine, pyridoxine) using ATP-Zn as a phosphoryl donor. The enzyme purified from brain tissues is made up of two identical subunits of 40 kDa each. Native enzyme was inhibited by a substrate analogue, pyridoxal-oxime. Limited chymotrypsin digestion of pyridoxal kinase yields two fragments of 24 and 16 kDa with concomitant loss of catalytic activity. These fragments were isolated by DEAE ion exchange chromatography and used for binding studies with fluorescent ATP and pyridoxal analogues. The spectroscopic properties of both fluorescent pyridoxal analogue and Anthraniloyl ATP (Ant-ATP) bound to the 24 kDa fragment are indistinguishable from those of both pyridoxal analogue and Ant-ATP bound to the native pyridoxal kinase, respectively. The small 16 kDa fragment, generated by proteolytic cleavage of the kinase, does not bind any of the substrate analogues. Binding characteristics of Ant-ATP were extensively studied by measuring the changes in fluorescence spectra at various conditions. From the results presented herein, it is postulated that the structural domain associated with catalytic activity comprises approximately one-half of the molecular mass of pyridoxal kinase (24 kDa). whereas the remaining portion (16 kDa) of the enzyme contains a regulatory binding domain.

• Journal of Microbiology and Biotechnology
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• 제26권4호
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• pp.666-674
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• 2016

The bacterial strains were screened as potential starters for fermenting low-salt doenjang (a Korean traditional fermented soybean paste) using Korean doenjang based on proteolytic and antipathogenic activities under 6.5-7.5% NaCl conditions. Phylogenetic analysis based on 16S rRNA gene sequences showed that they all belonged to the genus Bacillus. Proteolytic and antipathogenic activities against Escherichia coli, Bacillus cereus, Staphylococcus aureus, Listeria monocytogenes, and Aspergillus flavus, as well as fibrinolytic, amylase, and cellulase activities of the 10 strains were quantitatively evaluated. Of these, strains D2-2, JJ-D34, and D12-5 were selected, based on their activities. The functional, phenotypic, and safety-related characteristics of these three strains were additionally investigated and strains D2-2 and D12-5, which lacked antibiotic resistance, were finally selected. Strains D2-2 and D12-5 produced poly-γ-glutamic acid and showed various enzyme activities, including α-glucosidase and β-glucosidase. Growth properties of strains D2-2 and D12-5 included wide temperature and pH ranges, growth in up to 16% NaCl, and weak anaerobic growth, suggesting that they facilitate low-salt doenjang fermentation. Strains D2-2 and D12-5 were not hemolytic, carried no toxin genes, and did not produce biogenic amines. These results suggest that strains D2-2 and D12-5 can serve as appropriate starter cultures for fermenting low-salt doenjang with high quality and safety.

• 한국식품영양과학회지
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• 제30권4호
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• pp.594-599
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• 2001

혈압상승을 주도하는 효소인 angiotensin converting enzyme(ACE)의 활성을 저해하는 펩타이드를 생산하는 protease 분비 균을 간장으로부터 분리하여 동정하였다. 여러대두 발효식품으로부터 단백질 분해능이 있는 균주를 분리하고, 분리된 균주들 중에서 ACE 저해활성이 가장 높은 균주인 SS103 균주를 선정하였다. 선정된 SS103 균주는 운동성이 있는 gram 양성 간균으로 내생포자를 형성하는 호기성균이었으며, pH 5~9 사이에서 생육하고 생육온도는 2$0^{\circ}C$~$50^{\circ}C$로 내열성이 약한 중온균의 특성을 보였으며, glucose, sucrose, mannitol 및 sorbitol 등의 당 이용 특성을 보였다. 그리고 세포의 지방산 조성은 $C_{15:0}$ anteiso, $C_{17:0}$ cis 형 및 $C_{17:0}$ iso 형이 주된 지방산으로 각각 33.9%, 18.8% 및 16.5%의 분포를 나타내었다. 이상과 같은 분석 겨과를 토대로 SS103 균주는 Bacillus subtilis로 동정되었다. Bacillus subtilis SS103 균주의 최적 배양 조건은 $37^{\circ}C$, 초기배양액 pH 8.0에서 배양시간 48시간으로 나타났으며, 이때 통기량이 높은 경우 효소활성도 높은 특성을 보였다.을 보였다.

• 약학회지
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• 제37권2호
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• pp.129-135
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• 1993

Serratia sp. Y-4 was isolated from soil. Many characteristics of the strain and optimum cultivation condition for protease production were investigated.,The protease from Serratia sp. Y-4 was purified and studied for the properties of the enzyme. The isolated strain was identified to the genus Serratia. The strain was cultivated in 1%-casein, 0.5%-Na$_{3}$PO$_{4}$.7H$_{2}$O, 0.1%-NaCl, 0.05%-KCI, 0.02%-MgSO$_{4}$.7H$_{2}$O, 0.02%-CaCl$_{2}$.2H$_{2}$O, 0.02%-ZnSO$_{4}$.7H$_{2}$O, 0.02%-MnCl$_{2}$.4H$_{2}$O and 0.5%-soy bean oil at pH 7.0 for 35 hrs. The enzyme was purified about 5.89 fold from the culture broth with 31.1% recovery and 19,613 u/mg through ultrafiltration, ammonium sulfate, DEAE-sephacel and Superose-12 chromatography. The purified enzyme was identified as one band by isoelectric focusing, SDS- and native-PAGE. It has maxium activity at $37^{\circ}C$ and pH 9.0. Molecular weight of it is approx. 50 kD and pl is about 6.70. Its Km value for casein was 20 mg/ml. 5 mM-EDTA, 5mM-SDS, Ag$^{+1}$, Cu$^{+2}$, Hg$^{+2}$ and Pb$^{+2}$ inhibited the enzyme.