• 한국수산과학회지
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• 제23권5호
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• pp.361-372
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• 1990

Processing conditions of whole sardine into modified fish sauce were investigated. Thawed and chopped sardine was homogenized and hydrolyzed using commercial proteolytic enzymes such as complex enzyme-2000($2.18{\cdot}10^4U/g solid$) and alcalase($1.94{\cdot}10^4\;U/g solid$) in a cylindrical vessel with 4 baffles and 6-bladed impeller. Optimal pH, enzyme concentration and temperature for the hydrolysis with complex enzyme-2000 were 7.0, $7\%$ (W/W) and $52^{\circ}C$, and-those with alcalase were 8.0, $6\%$ (W/W) and $60^{\circ}C$. In both cases, the reasonable amount of water for homogenization, agitation speed and hydrolyzing time were $100\%$ (W/W), 100 rpm and 210 minutes. Thermal treatment of the filtered hydrolysate at $90^{\circ}C$ for 2 hours with $6\%$ of invert sugar was adequated to inactivation of the enzymes and pasteurization of the hydrolysate. Flavor, taste and color of the hydrolysate were improved during the heating process in which the browning products might participate. The content of free amino nitrogen in the fish sauce seasoned with $15\%$ of table salt was ca. $1,640 mg\%$. Yield of the fish sauce based on the contents of proteinous and free amino nitrogen in the raw whole sardine was ca. $86\%$, and ca. $96\%$ of these compounds of the fish sauce was in the form of free amino nitrogen. The pH, salinity and histamine content of the fish sauce were $6.1\~6.3,\;14.2\~14.3\%$ and less than $10\;mg\%$.

• 농업과학연구
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• 제38권1호
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• pp.51-63
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• 2011

BoLA (bovine leukocyte antigens) have been known as gene complex related with bovine diseases and immunological traits. This study was conducted to find out the characteristics of BoLA-DRB3 gene related to mastitis and BL(bovine leukocyte) from 280 cattle [193 animals of Holstein cattle and 87 animals of Hanwoo]. As a result, five PCR-RFLP types (b, d, e, f and g) using HaeIII restriction enzyme, three BstYI restriction patterns (b, d and e) and eight RsaI restriction types(b, d, f, I, j, n, o and w) were identified. Moreover, we identified new d' type ($197{\rightarrow}175$/22), having one more cutting site by BstYI enzyme than d type allele and n' type ($180{\rightarrow}169$/11) having one more cutting site by RsaI enzyme than n allele was additionally identified. Next, we identified 53 SNPs in BoLA-DRB3 exon2 from 280 cattle. SNP frequency and heterozygosity of Holstein and Hanwoo were investigated in all the SNP genotype. These results might be based on research for identifying marker associated with bovine diseases.

• 한국미생물·생명공학회지
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• 제31권2호
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• pp.124-128
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• 2003

대전광역시 근교의 야산과 들판 등지에서 썩은 나뭇잎, 짚, 흙을 채취하여 각각을 배양한 다음 Congo red test에 의해 cellulase 활성을 보이는 균주를 선별하였다. Genomic DNA를 분리한 후 PCR을 수행하여 DNA sequence를 Gene Bank를 통해 분석한 결과 A. tubingensis로 밝혀졌다. 이것을 배양하여 상등액을 crude enzyme으로 사용하여 온도와 pH를 달리하면서 효소의 활성정도를 측정하였다. 대조균주로 A. oryzae KCTC 6291를 이용하였고, 본 연구를 통하여 분리한 균주인 A. tubingensis가 생산하는 cellulase는 A. oryzae의 cellulase에 비하여 각각 다른 온도와 pH에서 높은 안정성을 보여주었다. A. tubingensis는 각각의 온도에서 활성의 정도가 비슷했으며, 45$^{\circ}C$, 55$^{\circ}C$에서 높은 활성을 나타내고 있지만, 고르게 활성이 나타났다. 또한 pH 12.0에서 가장 높은 활성을 보여 주었고, pH 2.0, 3.0, 4.0에서는 양쪽 모두 거의 활성이 없었으며, 중성, 염기성에 대해서 활성에는 큰 변화가 없었다. 따라서, 분리 동정한 A. tubingensis는 온도와 pH에서 고르게 활성을 나타내므로 생균제로 활용할 수 있는 범위가 클 것으로 여겨진다.

• 대한화학회지
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• 제24권4호
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• pp.315-321
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• 1980

G. melanogenus로부터 분리한 폴리올 탈수소 효소는 이미 알려진 바의 다른 폴리올 탈수소 효소와는 달리 조효소로서 2,6-dichlorophenolindophenol (DPIP)와 같은 인위적 전자 수용체를 필수로 요구하고 있음으로 이 특수 효소의 반응메카니즘을 반응속도론적 연구를 통하여 규명코저 시도하였으며, 폴리올 산화반응에 대한 초기속도 측정실험과 효소반응 산물인 케토산에 의한 저해 실험을 통하여 이 반응은 Ping-Pong Bi-Bi형의 반응메카니즘으로 진행됨을 확인하였다. 따라서 두 기질 즉 포리올로서 D-mannitol 및 전자수용체로 DPIP가 효소에 의하여 반응이 진행될 경우 D-mannitol이 우선 효소와 작용하며 첫 반응산물로서 해당하는 케토산인 D-fructose가 생성될 것으로 기대되며 이 반응이 전체 반응속도를 조절하는 과정일 것이라고 추측하였다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권5호
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• pp.382-390
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• 2006

Various microorganisms were isolated from the surface waters and sediments of eutrophic lakes and reservoirs in Korea to enable an investigation of bacteria having algal lytic activities against Anabaena flos-aquae when water blooming occurs and to study enzyme profiles of algal lytic bacteria. Two bacterial strains, AFK-07 and AFK-13, were cultured, characterized and identified as Acinetobacter johnsonii and Sinorhizobium sp., respectively. The A. johnsonii AFK-07 exhibited a high level of degradatory activities against A. flos-aquae, and produced alginase, caseinase, lipase, fucodian hydrolase, and laminarinase. Moreover, many kinds of glycosidase, such as ${\beta}-galactosidase,\;{\beta}-glucosidase,\;{\beta}-glucosaminidase,\;and\; {\beta}-xylosidase$, which hydrolyzed ${\beta}-O-glycosidic$ bonds, were found in cell-free extracts of A. johnsonii AFK-07. Other glycosidases such as ${\alpha}-galactosidase,\;{\alpha}-N-Ac-galactosidase,\;{\alpha}-mannosidase,\; and\;{\alpha}-L-fucosidase$, which cleave ${\alpha}-O-glycosidic$ bonds, were not identified in AFK-07. In the Sinorhizobium sp. AFK-13, the enzymes alginase, amylase, proteinase (caseinase and gelatinase), carboxymethyl-cellulase (CMCase), laminarinase, and lipase were notable. No glycosidase was produced in the AFK-13 strain. Therefore, the enzyme system of A. johnsonii AFK-07 had a more complex mechanism in place to degrade the cyanobacteria cell walls than did the enzyme system of Sinorhizobium sp. AFK-13. The polysaccharides or the peptidoglycans of A. flos-aquae may be hydrolyzed and metabolized to a range of easily utilized monosaccharides or other low molecular weight organic substances by strain AFK-07 of. A. johnsonii, while the products of polysaccharide degradation or peptidoglycans were more likely to be utilized by Sinorhizobium sp. AFK-13. These bacterial interactions may offer an alternative effective approach to controlling the water choking effects of summer blooms affecting our lakes and reservoirs.

• Interdisciplinary Bio Central
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• 제5권2호
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• pp.3.1-3.7
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• 2013

Recently, the productivity of drug discovery has gradually decreased as the limitations of single-target-based drugs for various and complex diseases become exposed. To overcome these limitations, drug combinations have been proposed, and great efforts have been made to predict efficacious drug combinations by statistical methods using drug databases. However, previous methods which did not take into account biological networks are insufficient for elaborate predictions. Also, increased evidences to support the fact that drug effects are closely related to metabolic enzymes suggested the possibility for a new approach to the study drug combinations. Therefore, in this paper we suggest a novel approach for analyzing drug combinations using a metabolic network in a systematic manner. The influence of a drug on the metabolic network is described using the distance between the drug target and an enzyme. Target-enzyme distances are converted into influence scores, and from these scores we calculated the correlations between drugs. The result shows that the influence score derived from the targetenzyme distance reflects the mechanism of drug action onto the metabolic network properly. In an analysis of the correlation score distribution, efficacious drug combinations tended to have low correlation scores, and this tendency corresponded to the known properties of the drug combinations. These facts suggest that our approach is useful for prediction drug combinations with an advanced understanding of drug mechanisms.

• Journal of Chest Surgery
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• 제22권6호
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• pp.901-913
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• 1989

This study was investigated under the postulation that activation of intracellular calcium- calmodulin complex during ischemia-reperfusion leads to myocardial injury. The protective effects of calcium channel blocker, diltiazem and calmodulin inhibitors, trifluoperazine, flunarizine and calmidazolium from ischemic injury in rat hearts were observed by using Langendorff apparatus when the antagonists were infused for 3 min in the beginning of ischemia. Thereby, an increase in resting tension developed during 30-min ischemia was analyzed with regard to [1] the degree of cardiac functional recovery following 60-min reperfusion, [2] changes in biochemical variables evoked during 30-min ischemia. The results obtained were as follows: l. In the ischemic group, the resting tension was increased by 4.1*0.2 g at 30-min ischemia. However, the increase in resting tension was markedly reduced not only by pretreatment with diltiazem [3.3 p M] but also with calmodulin inhibitors, trifluoperazine [3.3 p M], flunarizine [0.5 p M] and calmidazolium [0.5 p M], respectively. 2. Recovery of myocardial contractility, +dF /dt and coronary flow were much reduced when evoked by reperfusion in the ischemic group. These variables were significantly improved either by pretreatment with diltiazem or with calmodulin inhibitors. 3. The resting tension increment evoked during ischemia was significantly inversely correlated with the degree of cardiac function recovered during reperfusion. 4. Following 30-min ischemia, the production of malondialdehyde and release of lysosomal enzyme were much increased in association with a decrease in creatine kinase activity. 5. The increases in malondialdehyde production and release of free lysosomal enzyme were suppressed by pretreatment with calmodulin inhibitors as well as diltiazem. Likewise, the decrease of creatine kinase activities was prevented by these calcium antagonists. With these results, it is indicated that a increase in resting tension observed during ischemia has an inverse relationship to the cardiac function recovered following reperfusion, and further, the later may be significantly dependent on the degree of biochemical alterations occurred during ischemia such as decrease in creatine kinase activity, increased production of malondialdehyde and increased release of free lysosomal enzyme. Thus it is concluded that calmodulin plays a pivotal role in the process of ischemic injury.

• 공업화학
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• 제23권4호
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• pp.394-398
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• 2012

아스트라갈린(AS)은 폴리페놀의 일종인 캠페롤(KR)에 글루코스가 하나 결합해있는 배당체로서, 천연에는 미량만이 존재하고 있다. 최근 tea seed extract (TSE)에 포함되어 있는 camelliaside A (CamA) 및 camelliaside B (CamB)로부터 복합효소인 Mash에 의한 부분가수분해에 의해 AS를 합성할 수 있다는 것이 알려졌다. 본 논문에서는, Mash에 의한 TSE의 선택적 가수분해에 의해 AS를 제조하는 공정에서 반응 온도, 효소량 및 반응물의 농도 등의 변수에 따른 반응성을 검토하였다. 반응 온도, 효소량 등이 증가할수록 AS의 생성 반응은 빨랐으나, 일단 생성된 AS가 급격하게 KR로 변환되는 문제점이 나타났다. 결론적으로, 반응 속도 및 AS의 선택성 측면을 고려하면, $50^{\circ}C$에서 TSE 대비 2배의 Mash를 사용하여 15%의 기질 농도에서 반응을 수행하는 것이 최적인 것으로 나타났다.

• 대한화학회지
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• 제34권3호
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• pp.232-238
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• 1990

Zn의 생화학적 역할을 연구하기 위하여, 간단한 모델시스템을 설정하였다. 즉, Zn가 리간드인 $OH_2나 NH_3$와 배위결합을 이루거나 carboxypeptidase A (CPA)의 기질에 해당하는 펩티드의 O=C-와 배위결합을 할 때, 그 기하학적 구조 및 net atomic charge의 변화를 조사하였다. Double Zeta basis set를 사용한 ab initio HF-SCF 계산에 의하면, Zn는 이들 리간드의 O-H, N-H, O=C-를 매우 polar하게 만든다. 특히, 펩티드의 탄소는 친전자성이 매우 증가하여, nucleophile의 공격을 용이하게 받을 수 있을 것으로 예측되었다. 또한, CPA에서 Zn 주위에 있는 리간드의 분자배위 상태를 조사하기 위하여, CPA+glycyltyrosine complex에 관한 molecular mechanics방법을 적용하였다. 이 결과를 X-ray 결과와 비교하였을 때, Zn는 4배위 결합 이외에 물분자도 관여할 것으로 예측된다.

• Journal of Microbiology and Biotechnology
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• 제11권5호
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• pp.763-771
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• 2001

Biphenyl dioxygenase (BPDO), which catalyzes the first step in the bacterial degradation of biphenyl and polychlorinated biphenyls, was characterized in Pseudomonas sp. B4. The bphA locus containing the four structural genes encoding BPDO were cloned and sequenced. A regulatory gene as well as a putative regulatory sequence were identified upstream of this locus. A transposase-like gene was found within a 1-kb region further upstream, thereby suggesting that the bphA locus may be carried on a transposable element. The three components of the BPDO enzyme have been separately overexpressed and purified from E. coli. The ferredoxin and terminal dioxygenase components showed biochemical properties comparable to those of two previously characterized BPDOs, whereas the ferredoxin reductase exhibited an unusually high lability. The substrate selectivity of BPDO was examined in vivo using resting cell assays performed with mixtures of selected polychlorinated biphenyls. The results indicated that para-substituted congeners were the preferred substrates. In vitro studies were carried out on a BPDO complex where the reductase from strain B4 we replaced by the more stable isoform from Comamonas testosteroni B-356. The BPDO enzyme had a specific activity of $0.26{\pm}0.02 {\mu}mol {min^-1}{mg^-1}\;of\;ISP_{BPH}$ with biphenyl as the substrate. The 2,3-, 4,4'-, and 2,4,4'-chlorobiphenyls were converted to single dihydrodiols, while 2,4'-dichlorobiphenyl gave rise to two dihydrodiols. The current data also indicated that 2,4,4'-trichlorobiphenyl was a better substrate than the 4,4'-dichlorinated congener.