• 한국식품영양학회지
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• 제28권6호
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• pp.1026-1032
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• 2015

본 연구는 ${\beta}$-galactosidase와 효모 발효에 의해 고순도 갈락토올리도당(HP-GOS)의 제조를 위한 효율적인 방법을 찾고자 수행하였다. 6종의 상업용 ${\beta}$-galactosidase를 이용해 효소반응 전후의 구성당을 각각 비교한 결과, Lactozym 3000L을 이용하여 제조한 GOS에서 효소반응을 통해 생성된 total GOS의 양이 가장 높은 것으로 나타났다. 생성된 GOS의 농도는 초기 lactose로부터 51%의 전환율을 나타냈다. 또한 GOS의 수율은 효모(Saccharomyces cerevisiae)의 발효에 의해 더욱 높아졌는데, 효모를 8% 첨가하여 36시간 발효 후 생성된 GOS의 농도는 71%까지 도달했음을 확인하였다. HPLC를 이용한 구성당 분석 결과, S. cerevisiae에 의한 발효를 통해 제조한 HP-GOS에는 발효 전에 존재했던 glucose의 함량이 급감되었을 뿐 아니라, 4'/6'-galactosyllactose와 total GOS의 양은 유의적으로 증가되었다. HP-GOS는 상업용 GOS보다 Lactobacillus 속(L. acidophilus and L. casei) 및 Bifidobacterium 속(B. longum and B. bifidum)의 성장을 촉진시키는 것으로 확인되었다. 이러한 결과를 종합해 볼 때, 효소 처리 및 효모 발효를 통해 고순도의 GOS가 제조되었으며, 제조된 HP-GOS는 상업용 GOS에 비해 장내 유용균으로 알려진 Bifidobacterium 속과 Lactobacillus 속의 생육을 증가시킴으로써 인간의 장내 건강에도 유익한 영향을 미칠 것으로 사료된다.

• 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 $10\;mg\%$.

• Journal of Periodontal and Implant Science
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• 제38권sup2호
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• pp.317-324
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• 2008

Purpose: Aggregatibacter actinomycetemcomitans was associated with localized aggressive periodontitis, endocarditis, meningitis, and osteomyelitis. The cytolethal distending toxin (CDT) of A. actinomycetemcomitans was considered as a key factor of these diseases is composed of five open reading frames (ORFs). Among of them, An enzymatic subunit of the CDT, CdtB has been known to be internalized into the host cell in order to induce its genotoxic effect. However, CdtB can not be localized in host cytoplasm without the help of a heterodimeric complex consisting of CdtA and CdtC. So, some studies suggested that CdtC functions as a ligand to interact with GM3 ganglioside of host cell surface. The precise role of the CdtC protein in the mechanism of action of the holotoxin is unknown at the present time. The aim of this study was to generate recombinant CdtC proteins expression from A. actinomycetemcomitans, through gene cloning and protein used to investigate the function of Cdt C protein in the bacterial pathogenesis. Materials and Methods: The genomic DNA of A. actinomycetemcomitans Y4 (ATCC29522) was isolated using the genomic DNA extraction kit and used as template to yield cdtC genes by PCR. The amplifed cdtC genes were cloned into T-vector and cloned cdt C gene was then subcloned to pET28a expression vector. The pET28a-cdtC plasmid expressed in BL21 (DE3) Escherichia coli system. Diverse conditons were tested to opitimize the expression and purification of functional CdtC protein in E. coli. Results: In this study we reconstructed CdtC subunit of A. actinomycetemcomitans Y4 and comfirmed the recombinant CdtC expression by SDS-PAGE and Western Blotting. The expression level of the recombinant CdtC was about 2% of total bacterial proteins. Conclusion: The lab condition of procedure for the purification of functionally active recombinant CdtC protein is established. The active recombinant CdtC protein will serve to examine the role of CdtC proteins in the host recognition and enzyme activity of CDT and investigate the pathological process of A. actinomycetemcomitans in periodontal disease.

• 고려인삼학회:학술대회논문집
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• 고려인삼학회 1984년도 학술대회지
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• pp.257-275
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• 1984

Enzymatic browning is considered desirable in tea and tobacco processing but undesirable in many fruits processing at the present time. It is necessary to understand the nature of the enzyme, phenoloxidase, in order to control browning reactions, and extend its effects to formation of browning products as antioxidants in ginseng. Ginseng exhibits antioxidant activity when incorporated with turkey dark meat patties. The activity in red ginseng showed about two times stronger than white ginseng. One of the phenolic antioxidants from fresh, white and reprocessed white ginseng was identified as phenol 2.6 Bis(1.1 dimethyl ethyl) 4-methyl among several unknown compounds by GC/mass spectrometer. In red ginseng, no phenol 2.6 Bis (1.1 dimethyl ethyl) 4-methyl was detected, the compound may be polymerized by phenoloxidase and form some higher molecular compounds which may possess high antioxidant activity. Phenoloxidase isozymes in fresh Korean ginseng (panax ginseng C.A. Meyer) were extracted with phosphate buffer at pH 7.3. The isozymes were purified through ammonium sulfate fractionation, dialysis and chromatography on a DEAE-cellulose column. Two groups of phenoloxidase were shown to be present, one in the floating agglomerated group and the other in the precipitate. group from the 0.85 saturation ammonium sulfate. The DEAE-cellulose column chromatography, the phenoloxidase isozyme present in the precipitate appears as the first peak (I), and that in the agglomerate in the second peak (II). Isozyme I showed higher activity with catechin and catechal, and isozyme II showed higher activity with p-cresol. The isozyme showed two optimum pH activity one at pH 4.5 and the other at 8.5 with catechin as substrate. Korean ginseng phenoloxidase has high heat stability. When heated at $75^{\circ}C$ for 2 hours, its activity remained $90\%\;and\;80\%$ on phenoloxidase I and II respectively. Phenoloxidase I was most active on (+) catechin followed by p-cresal, catechol and epicatechin. Phenoloxidase II was most active on p-cresal followed by (+) catechin, catechol, p-coumanic acid and epicatechin. Sodium bisulfite, sodium cyanide, ascorbic acid glutachion in the oxidized form, sodium diethyl dithiocarbomate and ethylendiamine tetra acetate (EDTA) acted as inhibitors. Red ginseng color development was initiated by phenoloxidase and finished by a followed sun drying process. The antiaging activity of ginseng may be initiated by the antioxidant in the ginseng.

• Journal of Ginseng Research
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• 제5권1호
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• pp.8-23
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• 1981

Color is one of the most important quality factors of red ginseng (Hong-sam) which is processed from fresh ginseng (Panax ginseng C. A. Meyer). Therefore, a study of characteristics of browning mixtures of aqueous fresh ginseng extracts, factors which accelerate the browning of the aqueous extracts, and the antioxidant activity of the browning mixtures may contribute to the improvement of the color and other quality of red ginseng and other ginseng products such as ginseng extracts. In the present study, factors which affect the Maillard-type browning reaction of aqueous extracts of fresh ginseng roots were investigated firstly by adding various concentrations (0.001-0.5M) of arginine or glucose solutions, by varying the browning reaction temperatures and durations. Secondly, some characteristics such as brown color intensity, amounts of water-soluble and ether-soluble extracts, amounts of non-dialyzable materials, pH, viscosity, and reactivity with 2,2'- diphenyl -1 - picrylhydrazyl and antioxidant activity of the browning mixtures of the aqueous fresh ginseng extracts with small amounts of 0.1 M arginine, 0.1 M glucose, and distilled water at various browning temperatures and reaction time were studied. The results of the present study are as follows. 1. Color intensity (absorbance at 470 nm) of the browning mixtures was increased by adding various concentrations of arginine solution to the fresh ginseng extract, but the addition of the same amount of glucose solution did not increase the color intensity. 2 The amounts of water- or ether-soluble extracts, amounts of non-dialyzable materials were slightly greater in case of the browning mixtures of the fresh ginseng extract with 0.1M arginine solution than in case of the browning mixtures of the fresh ginseng extract with the same amount of 0.1 M glucose solution. In the process of the browning reaction, the pH of the browning mixtures of the fresh ginseng extract with 0.1 M arginine solution decreased slightly, while that of the browning mixtures with 0. 1 M glucose solution was almost constant. 3. The color intensity (absorbance at 470 nm) of the browning mixtures of the fresh ginseng extract with 0.1 M arginine or 0.1 M glucose solutions did not correlate well with the reducing power or the antioxidant power of the browning mixtures. The antioxidant activity of 90% ethanol extracts from the earlier stages of the browning mixtures of the fresh ginseng extract with the arginine solution was almost comparable to that of the 90% ethanol extracts from the later stages of the corresponding browning mixtures. The browning mixtures of only the fresh ginseng extract or of the fresh ginseng extract with the glucose solution showed considerable antioxidant activity, although both showed less brown color intensity than the fresh ginseng extract with he arginine solution.

• 디지털융복합연구
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• 제14권1호
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• pp.253-262
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• 2016

연산오계는 오래전부터 건강기능 증진 및 치료 효능이 높은 것으로 알려져 왔다. 최근 건강 기능식품 소재로 기능성 펩타이드 효능이 알려짐에 따라, 연산오계 내장 부산물로 부터 올리고 펩타이드 최적 생산 공정 및 생성물 특성에 대하여 연구를 수행하였다. 최적 효소가수 분해 공정 표면반응 분석을 이용하여 수행하였다. 최적 공정 조건을 확립하기 위해서 온도 (40, 50, $60^{\circ}C$), pH (pH 6.0, 7.0, 8.0), 효소 (1, 2, 3%) 범위에서 수행을 하였다. 생성물에 대한 가수분해도, 아미노산, 분자량 분포를 분석하였다. 효소 가수분해 최적 온도는 $58^{\circ}C$, pH 7.5, 효소의 농도는 3% 이었다. 최적 조건에서 2 시간 효소 가수분해를 한 결과 75-80% 이었다. 구성 아미노산의 총 함량은 386.15 mg/100 g 이었고 유리 아미노산 총량은 155.26 mg/100 g. 분자량를 Maldi-TOF 으로 분석을 한 결과 90% 이상이 300-1,000 Da 분포를 보여주었다.

• Journal of Ginseng Research
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• 제40권3호
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• pp.260-268
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• 2016

Background: During the aging process, skin shows visible changes, characterized by a loss of elasticity and the appearance of wrinkles due to reduced collagen production and decreased elasticity of elastin fibers. Panax ginseng Meyer has been used as a traditional medicine for various diseases due to its wide range of biological activities including skin protective effects. Ginsenosides are the main components responsible for the biological activities of ginseng. However, the protective activities of an enzymatic preparation of red ginseng against human skin aging have not been investigated. Methods: The efficacy of an enzyme-treated powder complex of red ginseng (BG11001) in preventing human skin aging was evaluated by oral administration to 78 randomized individuals. All patients were requested to take three daily capsules containing either 750 mg of BG11001 or a placebo vehicle for 24 wk; at the end of the testing period, skin roughness, elasticity, and skin water content were measured. Results: BG11001 significantly reduced the average roughness of eye wrinkles and the Global Photo Damage Score compared with the placebo, although there were no significant differences in arithmetic roughness average between the groups. In addition, gross elasticity and net elasticity values increased, and transepidermal water loss level decreased, indicating improved skin elasticity and moisture content. Conclusion: In conclusion, enzyme-treated red ginseng extract significantly improved eye wrinkle roughness, skin elasticity, and moisture content. Moreover, enzyme-treated red ginseng extract would be useful substance as a bio-health skin care product.

• 한국미생물·생명공학회지
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• 제38권4호
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• pp.428-433
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• 2010

Agar로부터 agarose 제조 시 유기용매를 이용해서 황을 제거하는 방법이 일반적으로 많이 사용되어진다. 하지만 agar의 황을 가수 분해하는 효소를 사용할 경우 agarose 제조 시공정과정을 획기적으로 간소화할 수 있다. 따라서 arylsulfatase로 agaropectin에서 황 제거를 통해 agarose로 바꾸는 공정은 간단하고 높은 수율을 얻을 수 있기 때문에 agarose 생산에 효율적으로 적용할 수 있다. 본 연구에서는 arylsulfatase를 세포표면에 발현하는 효모생촉매를 이용하여 제조한 agarose의 황 함량과 gel 강도를 측정하였다. 처리한 효모(효)소의 농도가 증가할수록 증가된 탈황 반응에 의해 황함량이 줄어들었고, 특히 35 unit/mL의 효소 농도로 처리하였을 때 황 함량은 0.2%까지 감소하는 것을 확인할 수 있었다. 황 함량을 가장 낮출 수 있는 최적 조건은 0.6% agar(Junsei) 용액에 효모 표층 arylsulfatase 35 unit/mL로 처리하고 $40^{\circ}C$에서 3시간 반응시켰을 때 였다. 또한 1.0% DNA 전기 영동용 agarose의 gel 강도는 효모 표층 arylsulfatase 처리로 제조된 agarose의 경우 $559.8{\pm}0.12$로 상업적 agarose의 gel 강도($880.6{\pm}0.15\;g/cm^2$와) 보다는 낮았다. 따라서 효모 S. cerevisiae의 세포 표면에서 발현된 재조합 arylsulfatase 효소를 이용하여 agar로부터 전기영동용 agarose의 생산 공정에 적용 가능함을 알 수 있었다.

• 한국수산과학회지
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• 제27권1호
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• pp.7-12
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• 1994

건조온도(25, $45^{\circ}C$)를 달리한 건조새우를 수분활성을 달리하여 저장하였을 때의 갈변반응 및 shelf-life는 다음과 같다. 1. 저장중 갈변반응은 온도, 수분활성이 높을수록 촉진되었다. 2. Arrhenius식으로부터 구한 활성화 에너지는 $25^{\circ}C$에서 건조한 새우의 경우 $13.57{\sim}14.33kcal/mol$의 범위였으며, $45^{\circ}C$의 경우 $13.12{\sim}13.61kca1/mol$로서 $45^{\circ}C$의 경우가 $25^{\circ}C$보다 다소 낮은 값을 보였다. 3. Shelf-life는 온도와 수분 활성이 증가함에 따라 급격히 단축되었는데 $25^{\circ}C$에서 건조한 새우의 경우 수분 활성 0.65 저장 온도 $55^{\circ}C$에서 4일, 수분활성 0.33 저장 온도 $35^{\circ}C$에서 139일이었고, $45^{\circ}C$의 경우 수분 활성 0.65 저장 온도 $55^{\circ}C$에서 1일, 수분활성 0.33 저장 온도 $35^{\circ}C$에서 76일로서 $25^{\circ}C$에서 건조한 것의 shelf-life가 더 길게 나타났다. 4. 변온 조건에서의 저장 실험결과와 이론적으로 예측한 값사이의 유효 온도 차이는 예측치보다 실측치가 높게 나타났다.

• Interdisciplinary Bio Central
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• 제4권3호
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• pp.10.1-10.12
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• 2012

Introduction: We investigated frameshift suppressor tRNAs previously reported to use five-base anticodon-codon interactions in order to provide a collection of frameshift suppressor tRNAs to the synthetic biology community and to develop modular frameshift suppressor logic devices for use in synthetic biology applications. Results and Discussion: We adapted eleven previously described frameshift suppressor tRNAs to the BioBrick cloning format, and built three genetic logic circuits to detect frameshift suppression. The three circuits employed three different mechanisms: direct frameshift suppression of reporter gene mutations, frameshift suppression leading to positive feedback via quorum sensing, and enzymatic amplification of frameshift suppression signals. In the course of testing frameshift suppressor logic, we uncovered unexpected behavior in the frameshift suppressor tRNAs. The results led us to posit a four-base binding hypothesis for the frameshift suppressor tRNA interactions with mRNA as an alternative to the published five-base binding model. Conclusion and Prospects: The published five-base anticodon/codon rule explained only 17 of the 58 frameshift suppression experiments we conducted. Our deduced four-base binding rule successfully explained 56 out of our 58 frameshift suppression results. In the process of applying biological knowledge about frameshift suppressor tRNAs to the engineering application of frameshift suppressor logic, we discovered new biological knowledge. This knowledge leads to a redesign of the original engineering application and encourages new ones. Our study reinforces the concept that synthetic biology is often a winding path from science to engineering and back again; scientific investigations spark engineering applications, the implementation of which suggests new scientific investigations.