Bifidobacterium angulatum으로부터 α-galactosidase의 정제 및 특성

강대중,민해기,강국희 成均館大學校 科學技術硏究所 1993 論文集 Vol.44 No.1

α-galactosidase는 (α-D-galactoside galactohydrolase: EC 3.2.1.22)는 다당류나 올리고당인 melibiose, raffinose, stachyose와 guar gum 등의 α-galactosidic 결합을 하고 있는 당을 분해하는 효소이다. 장내세균중 우익균이며 우세균주인 Bifidobacteria로부터 α-galactosidase의 효소학적 특성을 알아보기 위하여 배양여액으로부터 ammonium sulfate분획, 핵산의 제거, Sephadex G-200 gel filtration 및 DEAE-Sephadex A-50 ion exchange chromatography 등의 4단계 정제과정을 거쳐 정제한 결과 약 8배로 정제되어 단일 단백질로 분리하였다. 또한, 효소분해산물을 알아보기 위하여 TLC에 의하여 당분해 및 올리고당의 생성을 알아보았다. 정제효소의 활성 최적온도는 37℃, 최적 pH는 7.0이었다. 효소활성이 K^+, Zn^2+과 같은 금속이온과 8-Hydroxyquinoline, Sodium Bisulfite 등에 의해선 그다지 저해되지 않았으나, Hg^+, Cu^2+과 p-Chloromercuribenzoic acid에 의해선 저해되었다. 효소의 분자량이 301, 995, 합성기질인 PNPG에 대한 K_m은 0.069mM, V_max는 347.22μmol/min.mg protein이었다. To elucidate enzymatic properties of α-galactosidase (EC 3.2.1.22) from Bif.angulatum. α-galactosidase was purified to homogeneity by ammonium sulfate fractionation, protamine sulfate precipitation, sephadex G-200 gel filtration, and DEAE-sephadex A-50 ion exchange chromatography. The purified α-galactosidase was found tobbe homogeneous by SDSpolyacrylamide gel electrophoresis. The molecular weight of this enzyme, estimated by sepadex G-200 gel filtration, was about 301,995. The optimum conditions for the enzyme reaction was pH 6.5 to 7.0 at 37℃. The purified enzyme was stable at 45℃ or below and in buffer at pH 6 to 7.0. The activity was inhibited by mercury, copper ion, and p-chloromercuribenzoic acid. The kinetics of this enzyme, with p-nitrophenyl-α-D-galactoside as substrate, was determined : K_m was about 0.069mM and V_mas was 347.22 μmoles/min mg protein.

로스팅 서리태를 이용한 음료 제조 및 품질 평가

정수옥,김혜연,한정순,김민주,강미숙,김애정,Jeong, Soo-ok,Kim, Hae-yean,Han, Jung-soon,Kim, Min-ju,Kang, Mi-sook,Kim, Ae-Jung 한국식품영양학회 2016 韓國食品營養學會誌 Vol.29 No.4

This study was performed to develop and evaluate beverage prepared with optimally roasted seoritae to maximize the isoflavone content and antioxidant activities of the beverage. Isoflavone content was maximized at the roasting temperature of $110^{\circ}C$ for 20 min. Both DPPH radical scavenging activity and ABTS radical scavenging activity along with total polyphenol content were highest when seoritae was roasted at $110^{\circ}C$ for 20 min. Western blotting was used to determine the level of nuclear factor of activated T-cells 1(NFATc1) involved in controlling osteoclast differentiation. The results showed that NFATc1 had a concentration-dependent inhibitory effect when the RoS110 (roasted seoritae at $110^{\circ}C$ for 20 min) samples were processed at varying concentrations (10, 50, and $100{\mu}g/mL$). Tea samples were prepared from optimally roasted seoritae by varying brewing times (5~90 min) at $65^{\circ}C$, and tea brewed for 60 min had the highest preference with $65^{\circ}C$ as the preferred temperature for drinking.

Bifidobacterium longum KCTC 3215의 β-Galactosidase 유전자의 Cloning 및 대장균에의 발현

민해기,강국희,성문희 成均館大學校 科學技術硏究所 1992 論文集 Vol.43 No.2

The β-D-galactosidase(β-gal) gene from Bifidobacterium longum KCTC 3215 was cloned to isolate and characterize it for potential use as a selection marker in food-grade cloning vector. Chromosomal DNA from Bif. longum KCTC 3215 was cleaved with the restriction enzyme Sau3AI and ligated to pBR 322 for transformation into E. coli JM109. A β-galactosidase positive clone was detected by its blue color on a medium supplemented with 5-bromo-4-chloro-3-indolyl-β-D-galactoside. This transformant possessed a single plasmid, designated pSK 100, which contained, in addition to the vector DNA, a 5.2 Kilobase(Kb) Sau3A1 insertion fragment coding for β-gal activity. An extract from JM 109(pSK100) contained a β-gal protein with the same electrophoretic mobility as the β-gal from Bif. longum KCTC 3215. A restriction enzyme map of pSK 100 was consisted of PvuII, HincII, PstI,AccI, EcoRI, etc.

산전 초음파로 진단된 선천성 수신증 태아의 경험 1례

강민아,김도균,최석철,양회생,배철성,윤혜원,심재철,하달봉 동국대학교 경주대학 1997 東國論集 Vol.16 No.1

Prenatal ultarsound represents an accurate and reliable method for the detection of intrauterine anomalies, especially urinary tract anomalies. Hydronephrosis is the most common congenital condition detected by prenatal ultrasound. Prenatal ultrasonographic diagnosis allows a good condition and therapeutic assessment of the patients prior to serious impairment of renal function so that in most cases the urological surgeon can opt for a parenchymal conserving procedure. The authors have experienced a case of congenital ureteropelvic obstruction with hydronephrosis diagnosed at 32 weeks of pregnancy and report our experience with a brief review of literature.

윤상 다륜형 아급성 피부 홍반성 루푸스 1예

강민정,박혜진,조소연,이미애,황규광,함정희 대한피부과학회 1999 대한피부과학회지 Vol.37 No.6

We report an annular type of subacute cutaneous lupus erythematosus(SCLE) which is rare in Korea, A 23-year-old female had erythematous annular rashes with a mild itching sensation on the face, trunk and both extremities. She also had painless oral mucosal ulceration, diffuse hair loss and mild arthralgia. Serologic abnormalities of the patient were positive anti-Ro Ab, ANA and Anti-ds-DNA Ab. The histopathologic findings showed vacuolar degenerations in the basal cell layer, mononuclear cell infiltrate in the upper dermis, and dermal edema. Direct IF revealed granular deposition of IgG and IgA at the dermo-epidennal junction.

α-Galactosidase 에 의한 Bifidobacteria 균수 측정에 관한 연구

강국희,이시경,백운화,민해기 成均館大學校 科學技術硏究所 1992 論文集 Vol.43 No.2

This method using the synthesis substrate of 5-bromo-4-chloro-3 indolyl-galactoside (X-a -Gal) was examined for the differential enumeration of Bifidobacteria and lactic acid producing bacteria. Bifidobacteria possess a high level of a-galactosidase activity. Bifidobacterium longum KCTC 3215 exhibited the highest a- galactosidase specific activity (8.57 units/mg protein). Determination of a -galactosidase activity using the PNPG procedure showed that Lactobacillus, Streptococcus, Pediococcus, and Leuconostoc strain had lower a-galactosidase activity as compared to Bifidobacteria. The X-a- Gal based medium is useful to identify Bifidobacteria among lactic acid producing bacteria since the enzyme action of a-galactosidase hydrolysis X-a-Gal substrate and releases indol which impacts a blue color to Bifidobacterial colonies on agar plates. All strains of Bifidobacteria appeared blue colonies on MRS agar medium supplemented with 100 uM X- a - Gal while colonies of other lactic acid producing bacteria appeared white or light blue.

Streptococcus thermophilus SKD 1006의 β-Galactosidase 유전자 구조

민해기,강국희,성문희 成均館大學校 科學技術硏究所 1992 論文集 Vol.43 No.2

The results can be concluded that the pSK 001 contains a gene for β-gal from Str. thermophilus SKD 1006, and expressed in E. coli JM 109. A 4.2 Kb of Hpa II-Hpa II fragment(lac Z of SKD 1006) of pSK 001 was subcloned into the corresponding site of pUC 19, and named pSK 002. A 1.2 Kb of Bgl II -Bgl II fragment of pSK 002 was ligated into the corresponding site of pUC 19, and named pSK 003. The nucleotide sequences of two portions(222bp close to 5' , 143 by close to 3' end) of Bgl II -Bgl II fragment of pSK 003 were determined by Sanger, Chen and Seeburg methods. The nucleotide sequences are identical to those of the gene for β-gal of Str. thermophilus A 054.

철결핍성빈혈 진단과 RDW, MCV

민해연,강연주,조경진,남영미 THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCINE 1995 Journal of biomedical laboratory sciences Vol.1 No.1

자동혈구계산기에서 산출되는 RDW나 MCV가 철결핍성빈혈의 구분진단에 어떠한 도움이 되는지를 보기 위하여 어는 대학병원의 최근 5년 동안의 의무기록과 건강진단결과를 이용하여 227명의 빈혈환자와 143명의 건강인을 선정하고 그들의 혈액검사 결과를 비교분석해 보았다. 분석결과 빈혈환자로서는 철결핍성빈혈과 재생불량성빈혈 환자가 많았으며, 그 외에 만성질환과 관련된 빈혈환자도 많은 것으로 나타났다. 정상인 들에 비하여 빈혈환자에서 RDW는 높게, 그리고 MCV는 낮게 나타났는데 특히 철겹핍성빈혈에서는 다른 빈혈에 비하여 19.3±4.8로 현저하게 높게 나타난 반면, MCV는 62.9±13.7fl로 현저하게 낮게 나타났으며, 그 빈혈의 정도에 따라 그 변화가 크게 나타났다. 빈혈관련수들을 이용하여 D.F.=0.26-0.012MCV-0.130MCH+0.073MCHC+0.052RDW+0.003PLT와 같은 판별함수가 도출되었고, 이를 토대로 철결핍성빈혈과 다른 빈혈을 구분하는데 있어서는 MCV, MCH, MCHC, RDW등이 판별력이 있다는 것을 확인 할수 있었다. RDW and MCV are thought to be the highly sensitive blood cell parameters in the differentiation of iron deficiency anemias. Through the medical records of 227 anemic patients and the physical checking results of 143 healthy persons in a General Hospital during the recent five years, the authors evaluated various blood cell parameters including RDW and MCV. Iron deficiency anemia, aplastic anemia and other anemias associated with chronic discase were shown as the three major causes of anemias in Korea. In the patients of iron deficiency anemia MCV was very low(62.9±13.7fL), while RDW was very high(19.3±4.8) showing much lower MCV and much higher RDW in severe IDA compared with in mild IDA. To differentiate iron deficiency anemias form other anemias, a discriminant function was developed from some blood cell parameters like MCV, MCH, MCHC, RDW and platelets(D.F.=0.26 -0.012MCV -0.130MCH+ 0.073MCHC +0.052RDW +0.003PLT).

Str.thermophilus 510의 β-Galactosidase 유전자의 Cloning 및 대장균에의 발현

강국희,민해기,이호근,성문희 成均館大學校 科學技術硏究所 1992 論文集 Vol.42 No.2

Chromosomal DNA from Str. thermophilus 510 was purified and cleaved with the restriction enzyme PstI. Chromosomal DNA was ligated to pBR322 for transformation into E. coli JM109. A β-galactosidase positive clone was detected by its blue color on a medium supplemented with 5-bromo-4-chloro-3-indolyl-β-D-galactoside. This transformat possessed a single plasmid, designated pSK 001, which contained, in addition of the vector DNA, A 7.0Kb Pst I insertion fragment coding for β-gal activity. A restriction enzyme map of pSK 001 was consist of Hind Ⅲ, EcoR I, Bgl Ⅱ, Sal I, Pvu Ⅱ, etc. An extract from E. coli JM109 (pSK001) contained a β-gal protein with the same electrophoretic mobility as the β-gal from Str. thermophilus 510.

김치로부터 분리한 Pediococcus halophilus에 의한 α-Glucosidase의 생산

강국희,민해기,홍승표 成均館大學校 科學技術硏究所 1992 論文集 Vol.42 No.2

Six strains able to efficiently produce α-glucosidase were isolated from kimchi by enrichment culture in APT medium. Among them, Strain No.2 was found to have levels of α-glucosidase activity. The morphological, physiological and biochemical characteristics of the strain No.2 were studied according to the methods of Bergey's manual. Based on the results obtained in these experiments, Stain No.2 was identified to be a similar species to Pediococcus halophilus. The optima conditions for α-glucosidase production were cultured 0.5% starch, 0.4% yeast extract, 0.4% trypticase peptone, initial pH 7.5 at 37℃ and 24 hours cultivation.