• Advances in environmental research
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• 제11권1호
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• pp.1-16
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• 2022

Amongst 27 isolates from deteriorated leather samples, Arthrobacter creatinolyticus KP015744 zzx28 was found to be an efficient collagenase producer. Collagenase production of 13.33 µmoles/min was shown at an optimum temperature at 37℃ after 72h and at pH 7.5 by using 2 ml/dL inoculum in 10 mg/ml collagen peptide type I as a substrate. In presence of Hg²⁺, EDTA and 𝛽-mercaptoethanol the collagenase production by the isolate was strongly inhibited however Fe²⁺, Ca²⁺and DMSO enhanced production of the enzyme. Specific activity was found to be 19.46×10³ U/mg and molecular weight 66 kD by SDS PAGE. Isolate also has potential to hydrolyze keratin which is another important protein found in leather. Experimental results propose that collagenase can be effectively used as a tool for collagen and keratin rich solid waste treatment.

• Journal of Plant Biology
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• 제39권3호
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• pp.173-177
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• 1996

A novel calcium binding protein (CaBP) was purified to electrophoretic homogeneity from Dunaliella salina. In the course of purification experiment, this CaBP was identified as a monomer and its molecular weight was about 21 kDand isoelectric point (pI) value was about 4.1 using isoelectrofocusing. This CaBP was able to bind Ca2+ even in the pressence of an excess MgCl2 and KCI both in solution. In the SDS-PAGE, the Ca2+-bound form was slower than the Ca2+-free form in the nondenaturing PAGE. This means that the CaBP undergoes conformational change in the Ca2+-bound condition. Furthermore, UV absorption spectrum and fluorescence intensity of this CaBP was investigated. UV absorption peak was appeared at about 258 nm and decreased somewhat in Ca2+-bound condition. In the measurement of fluorescence, maximum intensity was appeared at 303 nm and decreased in Ca2+-bound state, similarly as UV absorption spectrum. These show distinct changes upon Ca2+-binding, which indicate of structural and/or dynamic changes largely reminiscent of other members of the EF-hand Ca2+-binding protein family.

• Journal of Microbiology and Biotechnology
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• 제11권2호
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• pp.173-178
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• 2001

A monoclonal antibody (mAb) to the glucoprotein D (gD) of Herpes simplex virus type 2 (HSV-2) was successfully generated by hybridoma technology and characterized. The mAb, SKS2v, recognized a gD antigen with an apparent molecular mass of 60kDa in a Western blot analysis. The isotype was determined by a sandwich ELISA to be IgG2a. HSV-2 exhibited major antigens of 36, 43, 46, 47, 60, 69, 81, 96, 109, 112, 159, and 227 kDa among 25 protein profiles in SDS-PAGE, and among these antigens, those of 60, 112, 125, and 227 kDa were immunodominant in a Western blot analysis using antisera, thereby indicating that they play a role in inducing neutralizing antibodies in HSV-2 infection. When reacted with Vero cells infected with HSV-1 and HSV-2 SKSv2 showed a reactivity to the surface of the infected cells, and a gD antigen of 60 kDa appeared to be expressed in both types of HSV.

• Journal of Applied Biological Chemistry
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• 제43권3호
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• pp.141-146
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• 2000

Glycosidase activities were tested from the grape berries, Vitis labruscana B. Takasumi. Among various glycosidases, $\alpha$-D-galactosidase was found to be the most active in the flesh and other glycosidases were considerably active in the order of the following: $\alpha$-D-mannosidase>$\alpha$-D-glucosidase>$\beta$-D-glucosidase>$\beta$-D-galactosidase. In the seeds, $\alpha$-D-glucosidase activity was the highest and other glycosidases such as $\alpha$-D-galactosidase, $\beta$-D-glucosidase, and $\beta$-D-galactosidase were still significantly active. The $\alpha$-D-galactosidase in the grape flesh was purified over 83-folds through salting-out with $(NH_4)_2SO_4$ and a series of chromatographies employing Sephadex G-50, Octyl-Sepharose, Q-Sepha- rose, and Biogel P-100. The enzyme was a monomer of 45 kDs as determined through SDS-PAGE and Sephacryl S-200 chromatography. The purified enzyme showed a preference of $\alpha$-D-galactose to $\beta$-D-galactose as a substrate about 5.4 times. Sulfhydryl specific reagents such as N-ethylmaleimide and iodoacetamide significantly inhibited the enzyme activity to the extents of 48 and 52% of its initial activity, respectively. The optimumpH range of $\alpha$-D-galactosidase was around 6.5-7.0. The enzyme activity increased by 46% in the presence of 1mM $Fe^{2+}$.

• 식품저장과 가공산업
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• 제6권2호
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• pp.11-13
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• 2007

This study relates to low and medium molecular weight isoflavone-${\beta}$-D-glucan produced by submerged liquid culture of Agaricus blazei, a method of producing the isoflavone-B-D-glucan using autolysis enzyme of Agaricus blazei mycelia, and use of the isoflavone-B-D-glucan for anti-cancer and immunoenhancing effect. In acordance with one aspect of the present study, it deals with a method of producing isoflavone-${\beta}$-D-glucan, which comprises the followings; 1) culturing and separating mushroom mycelia, 2) producing low-medium molecular weight isoflavone-${\beta}$-D-glucan from high molecular weight one. The cytotoxicity on human cnacer cell line (Caco-2, MCF-7), the expression of Cyclin D, Bcl-2, Bax protein, p21 protein, p53 protein in MCF-7 cells assessed by SDS-PAGE and immunoblotting, and other immuno related factor such as TNF-a and IL-1B activities were examined. Structural identification of isoflavone-${\beta}$-D-glucan which shoed cytotoxicity against cancer cell and immunoenhancing effects was carried by separation with DEAE-cellulose column chromatography, TLC, HPLC, IR, NMR, Clinical test for the cancer patients (n=119) for 6 month was carried out, and immunoenhancing factors(NK cell number, ratio of T4/T8) were checked. We concluded the identified isoflavone-${\beta}$-D-glucan has immuno enhancing effects and could be useful for cancer chemoprevention.

• 한국동물학회지
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• 제35권2호
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• pp.211-218
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• 1992

한국산 노랑초파리 군에 속하는 8종의 유전적 유연관계를 밝히고자 생식적 격리 그리고 수용성 단백질을 전기영동법으로 분석하였다. 생리적 격리 실험에서 교배전격리 실험결과를 Watanane와 Kawanishi model에 근거하여 보면 D. aurario complex 3종 중 D. triauror물가 원시종이며 남 auraria는 팍생종으로 나타났다. 교배후 격리 실험에서 나. melonogaster와 D. simuions의 교배시, D. melonogoster를 수컷으로 하였을 때는 불임의 수컷만이, 또 D. melonogaster를 암컷으로 곯였을 때는 불임의 암컷만이 출현하였다. 그리고 남 ouraria complex 3종간의 교배에서는 수정 능력이 있는 수컷과 암컷이 출현하였는데, 이는 아직 남 auraria Complex가 semispecies단계에 있음을 나타내는 것이라 할 수 있다. SDS-PAGE로 분석한 노랑초파리 군 8종의 band pattern을 densitometer로 scanning한 결과 남 susu상가 가장 특이하였으며, TDE에 의한 유전적 거리(Aquadro and Avise's)는 남 auror지와 봉 triauror지사이가 0.155로 가장 낮았고, D. melonogaster와 고. mfa 사이가 0.422로 가장 높았다. 본 연구의 결과를 UPGMA법 뜨로 분석하면, 한국산 노랑초파리 군 8종은 4개의 아군으로 나뉘어지며 이들은 2개의 다른 큰무리로 구분되었는데, D. suzu가기 아군, D. lutescens의 아군, D. melanogoster와 봉 simulons의 아군이 속한 큰무리와, D. mfa,0. ouroria, D. biauroria 그리고 남 triourario가 속한 다른 큰무리로 나눌 수 있다.

• 한국식품저장유통학회:학술대회논문집
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• 한국식품저장유통학회 2007년도 학술발표회
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• pp.27-31
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• 2007

This study relates to low and medium molecular weight isoflavone-${\beta}$-D-glucan produced by submerged liquid culture of Agaricus blazei, a method of producing the isoflavone-B-D-glucan using autolysis enzyme of Agaricus blazei mycelia, and use of the isoflavone-B-D-glucan for anti-cancer and immunoenhancing effect. In acordance with one aspect of the present study, it deals with a method of producing isoflavone-${\beta}$-D-glucan, which comprises the followings; 1) culturing and separating mushroom mycelia, 2) producing low-medium molecular weight isoflavone-${\beta}$-D-glucan from high molecular weight one. The cytotoxicity on human cnacer cell line (Caco-2, MCF-7), the expression of Cyclin D, Bcl-2, Bax protein, p21 protein, p53 protein in MCF-7 cells assessed by SDS-PAGE and immunoblotting, and other immuno related factors such as TNF-${\alpha}$ and IL-1B activities were examined. Structural identification of isoflavone-${\beta}$-D-glucan which showed cytotoxicity against cancer cell and immunoenhancing effects was carried by separation with DEAE-cellulose column chromatography, TLC, HPLC, IR, NMR. Clinical test for the cancer patients (n=119) for 6 month was carried out, and immunoenhancing factors (NK. cell number, ratio of T4/T8) were checked. We concluded the identified isoflavone-${\beta}$-D-glucan has immuno enhancing effects and could be useful for cancer chemoprevention.

• 한국식품영양과학회지
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• 제24권4호
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• pp.636-641
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• 1995

Proteolytic activities were compared using three species involving in squid jeotkal fermentation and showing positive reaction upon casein test : Pseudomonas D2, Flavovacterium odoratum and Acinetobacter calcoaceticus. Pseudomonas D2 produced highest activity of protease at 72h when incubated in our own modified medium(polypeptone, 0.5% ; tryptone, 0.5% ; NaCl, 3% ; pH, 7.5). Thus, this specie was selected for the further study. The growth pattern was coincided with the production of protease. Thus purification of protease was proceeded by ethanol precipitation, sephadex G-100 gel filtration, and DEAE sepharose ion exchange chromatography. The purified protease showed highest activity at pH 7.0 and 5$0^{\circ}C$. The enzyme was very stable over the wide ragnes of the temperature ; even with one hour heat treatment at 7$0^{\circ}C$, the enzyme showed substantial amount of the activity toward casein. In addition, the enzyme was stable over the wide range of pH. Molecular weight of the protease was determined to be 17.4 kD by SDS-PAGE.

• Applied Biological Chemistry
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• 제36권3호
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• pp.178-183
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• 1993

D-xylose의 통성 발효성 효모, Candida sp. BT001로부터 D-xylose를 xylitol로의 전환을 촉매하는 효소, xylose reductase(alditol: $MADP^+$ 1-oxidoreductase, EC 1.1.1.21)를 salt fractionation, ion exchange, gel filtration과 affinity chromatography를 거쳐 정제하여 그 성질을 조사하였다. 정제된 xylose reductase는 보효소 NADPH 및 NADH에 모두 특이성을 나타내었으며, 또한 각각의 보효소에 대해 활성을 지니는 효소는 따로 분리되지 않았다. Specific activity는 NADPH에 대해 11.78 U/mg, MADH에 대해 6.01 U/mg이었으며, NADH/NADPH의 활성비는 0.51이었다. 정제된 xylose reductase의 분자량은 SDS-PAGE상에서 31,000, gel filtration상에서 61,000으로 2개의 subunit로 구성된 효소로 추정하였다. 정제된 xylose reductase의 D-xylose와 NADPH 및 NADH에 대한 Km값은 각각 $94.2{\times}10^{-3}M,\;0.011{\times}10^{-3}M$ 및 $0.032{\times}10^{-3}M$ 이었다. Aldose들에 대한 xylose reductase의 활성은 L-arabinose, D-xylose순으로 높았다. 최적 효소 반응의 pH 및 반응 온도는 각각 6.2와 $45^{\circ}C$이었으며, 이 효소는 $30^{\circ}C$에서 20분간 안정하였다.

• 한국식품저장유통학회지
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• 제15권6호
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• pp.903-908
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• 2008

본 연구는 두유와 전두유의 효소적 가수분해에 따른 단백질의 변화를 조사하였다. 총 유리아미노산 함량을 조사한 결과 두유(SM)과 전두유(WSM)에 비해서 저분자 두유(LSM)와 저분자 전두유(LWSM)에서 높게 나타났다. 필수아미노산 함량은 SM과 LSM에서 비슷하였으나 LWSM은 WSM보다 높게 나타났다. SDS-PAGE 전기영동 패턴 분석결과 SM과 WSM에서는 $33{\sim}72\;kDa$의 고분자가 존재하였으나 LSM와 LWSM에서는 17 kDa 이하의 저분자 단백질만 나타났다. 또한 이차원 전기영동한 결과 SM과 LSM에서의 고분자 단백질 spot이 WSM와 LWSM에서는 다양한 크기의 저분자 단백질 spot으로 변환되어 효소가수분해에 의해 고분자 단백질이 저분자화 되는 것으로 나타났으며, 향후 단백질 spot의 분리 및 특성에 관한 연구가 필요하다.