• Title/Summary/Keyword: isoelectric focusing

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Properties and Utilization of Undigested Peptides in Anchovy Sauces 2. Effect of Fermentation Periods on Undigested Peptides of Anchovy Sauces (멸치 액젓 중에 존재하는 미분해 펩티드의 특성과 이용 2. 미분해 펩티드에 미치는 숙성기간의 영향)

  • CHO Young-Je;KIM Se-Hwan;IM Yeong-Sun;KIM In-Soo;KIM Dong-Su;CHOI Yeung Joon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.3
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    • pp.393-398
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    • 1998
  • The effect of fermentation period on the accumulation of 55,600 dalton polypeptide was analyzed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) as a quality parameter of anchovy sauces. Also, proximate compositions, total nitrogen contents, amount of specfic pepited and isoelectric point(pI0 were investigated during fermentation periods. Total nitrogen contents significantly increased until 18 months. Polypeptide of 55,600 and 46,900 dalton on SDS-PAGE and pI 5.2, 5.6 and 6.0 on isoelectric focusing were identified in all the samples. Especially, the amount of 55,600 dalton had no important change during fermentation periods, and it had a high correlation with dilution degree of anchovy sauces diluted with water. The results could be suggest that the amonts of 55,600 dalton polypeptide will be index for quality estimation of commerical anchovy sauces.

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Interspecific Relationships within the Fungal Genus Pleurotus by Isozyme Analysis (동위효소 분석에 의한 느타리속의 종간 유연관계)

  • Lee, Hee-Kyung;Yoo, Young-Bok;Cha, Dong-Yeul;Min, Kyung-Hee
    • The Korean Journal of Mycology
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    • v.26 no.2 s.85
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    • pp.163-172
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    • 1998
  • Thirty six strains of Pleurotus spp., from world-wide nations, were examined for interspecific isozyme variation. A comparison of isozymes in mycelial extracts of the fungal genus Pleurotus was made by polyacrylamide gel isoelectric focusing. A total of one hundred and sixty six bands was resolved from six isozymes. A cluster analysis was done based on the zymograms for esterase, glucosephosphate isomerase, leucine aminopeptidase, malate dehydrogenase, peroxidase, and phosphoglucomutase. From the isozyme analysis, esterase showed higher degree of variability, while it was observed less variability for the enzymes such as glucosephosphate isomerase, malate dehydrogenase, and phosphoglucomutase. The species P. ostreatus, whose taxon is controversial, was discriminated from P. pulmonarius, while P. florida was classified as a distinct taxon. The clustering of P. sapidus and P. spodoleucus strains appeared to be more difficult. It was found that some strains were included to another cluster based on electrophoretic banding patterns. These results show that this lack of congruence among data sets may help explain the taxonomic difficulty within the genus Pleurotus. A dendrogram of genetic similarities was presented, and applications of isozyme data to the systematics of these commercially important fungi was discussed.

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Characterization of Enzymatic Properties of Streptomyces albus Amylase Expressed in Recombinant Bacillus subtilis (재조합 Bacillus subtilis 에서 발현된 Streptomyces albus 유래 amylase 의 효소공학적 특성)

  • Park, Geun-Woo;Kim, Myoung-Dong;Ahn, Jang-Woo;Kim, Young-Bae;Seo, Jin-Ho
    • Korean Journal of Food Science and Technology
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    • v.30 no.6
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    • pp.1426-1431
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    • 1998
  • The research was undertaken to characterize enzymatic properties of Streptomyces albus amylase expressed in recombinant Bacillus subtilis. Molecular weight and pI of the purified enzyme were estimated to be 50 kD by SDS-PAGE and 4.3 by isoelectric focusing. The optimum temperature and optimum pH were $45^{\circ}C$ and 6.0, respectively. D-and Z-value were estimated to measure thermostability of the purified enzyme. The Z-value was estimated $17.7^{\circ}C$, which is lower than typical amylase. Maltotetraose was produced as a major component from soluble starch in the early state of reaction but gradually degraded to maltose. Thin layer chromatography was also performed to analyze the reaction products. The parameters involved in Michaelis-Menten enzyme kinetics were found to be the maximum velocity of 0.37 mM/min and the Michaelis constant of 0.13%, respectively.

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Purification and Some Characteristics of the Proteolytic Enzyme in Fruitbody of Neungee [Sarcodon aspratus (Berk.) S. Ito] (능이 [Sarcodon aspratus (Berk.) S. Ito]중 단백질(蛋白質) 가수분해(加水分解) 효소(酵素)의 정제(精製) 및 성질(性質)에 관하여)

  • Lee, Tae-Kyoo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.15 no.3
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    • pp.276-285
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    • 1986
  • This study was undertaken to investigate the characteristics of the proteolytic enzyme extracted from Neungee mushroom [Sarcodon aspratus (Berk.) S. Ito]. The enzyme was purified by using Tris-acryl CM-cellulose ion exchange, gel filtration on Ultrogel AcA 54, Hydroxy apatite column chromatography and preparative isoelectic focusing. The specific activity of the purified enzyme increased 8 times as compared with that of the crude enzyme. The enzyme was homogeneous on polyacrylamide gel electrophoresis (PAGE). The optimum pH was 10.1, indicating the enzyme to be alkaline protease and the optimum temperature was $57^{\circ}C$. The enzyme was stable at temperatures lower than $50^{\circ}C$and at pH values ranging from 4.0 to 10.8. However, the enzyme activity decreased by 26 and 65% at 60 and $65^{\circ}C$, respectively, when incubated for 30 minutes. The enzyme activity was activated by $Mn^{++}$ and inhibited by $Cu^{++}$ and $Hg^{++}$. The enzyme was consisted of monomer and its molecular weight estimated to be about 30,100 when determined by sodium dodecyl sulfate PAGE. Isoelectric point of the enzyme was determined to be 9.80.

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Heterogeneity of Lactate Dehyrogenase Isozymes in tissues of Lampetra japonica (칠성장어(Lampetra japonica) 조직내 젖산수소이탈효소 동위효소들의 이질성)

  • 조성규;염정주
    • The Korean Journal of Zoology
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    • v.36 no.3
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    • pp.319-328
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    • 1993
  • All vertebrates other than lampreys exhibit multiple loci encoding lactate dehydrogenase (EC 1.1.1.27,LDH). From the result shown by cellulose acetate and starch gel electrophoresis, the lampreys were-reported to have only one isozyme. However in our results the LDH of skeletal muscle, heart and kidney in Lampetra japonica were separated into three isozymes and that of liver was separated into two isozymes by polyacrylamide gel electrophoresis. The LDH of skeletal muscle and heart were separated into four isozymes and that of liver was separated into two isozymes by polyacrylamide gel isoelectric focusing (PAGlEF). The LDH of skeletal muscle were separated into four isozymes through the chromatofocusing. The molecular weight of LDH isozymes in skeletal muscle was approximately estimated to be 140,000 by Sephadex G-200 gel filtration. The LDH isozymes of skeletal muscle, heart and liver were inhibited by pyruvate to the nearly similar degree. And the degree of inhibition by pyruvate showed the value between LDH A$_4$and LDH B$_4$isozyme. And the LDH isozymes in heart, liver and skeletal muscle were thermostable. The results mentioned above indicate that the LDH isozyme in lamprey (Lampetra japonica) has not one isozyme but isozymes. And it is also found out that the two structures of their subunits are similar each others.

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Morphological, Cytological and Molecular Evidence for Intersubgeneric F1 Hybrid between Glycine max x G. tomentella (콩 Glycine max와 G. tomentella의 종간교잡으로부터 얻은 Fl식물체 검증을 위한 형태적 · 세포학적 · 분자유전학적 연구)

  • Choi, In-Soo;Kim, Yong-Chul
    • Journal of Life Science
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    • v.18 no.4
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    • pp.454-460
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    • 2008
  • This study was carried out to demonstrate morphological, cytological and molecular evidence for intersubgeneric $F_1$ hybrid between Glycine tomentella and G. max cv. ‘Baemkong’. Morphological features of $F_1$ plant for pistil and stamen, flower color and growth habit showed intermediate type between G. tomentella and G. max cv. ‘Baemkong’. Chromosome number of $F_1$ plant was 2n=39, which explained the evidence of $F_1$ hybrid between G. tomentella (2n=38) and G. max cv. ‘Baemkong’ (2n=40). Polyacrylamide isoelectric focusing pattern for esterase and peroxidase also showed that the $F_1$ plant was true $F_1$ hybrid between G. tomentella and G. max cv. ‘Baemkong’. From RAPD analysis, we identified that 62 primers showing bands in $F_1$ hybrid had both bands from G. tomentella and G. max cv. ‘Baemkong’, which suggested that this was true $F_1$ hybrid. Based on our results from morphological, cytological and molecular analyses, we suggest that the $F_1$ plant was true intersubgeneric hybrid between G. tomentella and G. max cv. ‘Baemkong’. Our results still remain us further study to recover fertility of $F_1$ hybrids. The occurrence of maternal and/or paternal inheritance in $F_1$ hybrid from intersubgeneric cross between G. tomentella and G. max cv. ‘Baemkong’ need to be explained.

Extended-Spectrum $\beta$-Lactamase (ESBL) Typing of Klebsiella pneumoniae Isolated from Clinical Specimen in Pusan (부산시내 종합병원의 임상 검체에서 분리된 Extended -Spectrum $\beta$-Lactamase 생성 Klebsiella pneumoniae의 형별 분류)

  • 김윤태;이훈구
    • Korean Journal of Microbiology
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    • v.36 no.3
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    • pp.221-227
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    • 2000
  • This study was performed to investigate the biological characteristics of twenty isolates of extended spectnlm $\beta$-lactamase (ESBL) producing Klebsiellapnezm~onia collected kom the various clinical specimens of three hospitals in Pusan. Isoelectric focusing (IEF) and PCR were used to determine the types of $\beta$-lactamase gene in this study. Twenty isolates of ESBL producing Klebsiellnp~ieun~or~iae could be divided by PCR, such as TEM type (I1 strains), SHV type (8 strains); non TEM non SHV type (1 strain). In the isoelechic focusing test, the PI of TEM type was 5.2-6.0 and that of SHV type was 6.9-7.4. According to the pI value and PCR bands, twenty strains of ESBL Klebsiellapneumoniae were divided into 5 types: TEM type @I 5.2-6.0; 1080 bp on PCR band), TEM + SHV type (pI 5.2-6.0; andpI 7.0-7.4; 1080 bp and 599 bp on PCR band), SHV type (pl7.0-7.4; 599 bp on PCR band), non TEM non SHV type, and otber type (PCR result was SHV type but pI was not detected).

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Studies on the Transformation of Crop Plants. IV. Biochemical Characteristics of Embryogenic Callus in Rice (곡물류의 형질전환 유도에 관한 연구 (IV) 벼 배발생 세포의 생화학적 특징)

  • 정병균
    • Journal of Plant Biology
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    • v.36 no.4
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    • pp.377-382
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    • 1993
  • Rice (Oryza saliva L.) calli containing both embryogenic callus (EC) and non embryogenic callus (NEC) regions were initiated from the mature seed on MS medium supplemented with 2.0 mg/L 2,4-D, 0.5 mg/L kinetin. The calli were developed into two callus type which can be distinguished by visual examination depending on color and appearance. In order to illucidate the polypeptide composition between EC and NEC, the total protein extracted from two types of callus was analysed by electrophoresis. By one-dimesional anlaysis of SDS-PAGE and Isoelectric focusing, several protein bands showed quantitative and qualitative difference in each type of callus. The further analysis of the total protein with two-dimensional electrophoresis showed at least 20 EC specific protein and 10 NE specific protein. Also 3 specific protein spots showing micro heterogeneity of 90, 65, 50 kD were detected in EC, while a series of acidic heterologous protein spots were visualized in NEC.in NEC.

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Separation and Characterization of Two Forms of Acetolactate Synthase from Etiolated Pea Seedlings

  • Shin, Yong-Soo;Chong, Chom-Kyu;Choi, Jung-Do
    • BMB Reports
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    • v.32 no.4
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    • pp.393-398
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    • 1999
  • Acetolactate synthase (ALS) catalyzes the first reaction common to the biosynthesis of L-valine, L-leucine, and L-isoleucine. ALS is the target site of several classes of herbicides, including the sulfonylureas, the imidazolinones, and the triazolopyrimidines. Two forms of ALS (ALS I and ALS II) which have different affinity for Heparin have been separated from etiolated pea seedlings. The substrate saturation curves of both ALS I and ALS II were hyperbolic in contrast to previous reports. The two forms of ALS showed significant differences in their physical and kinetic properties. The values of $K_m$ for ALS I and ALS II were 9.0 mM and 4.8 mM, respectively. The pI values for ALS I and ALS II were determined to be 5.3 and 5.75 by isoelectric focusing, respectively. The native molecular weights for ALS I and ALS II obtained by nondenaturing gel electrophoresis and activity staining were 124 and 244 kDa, respectively. They also exhibited different sensitivity to feedback inhibition by end-product amino acids and inhibition by Cadre, an imidazolinone herbicide.

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Plasmid Profile and ${\beta}$-Lactamase Type of Multidrug-Resistant Salmonella typhi Isolated from Korea, 1997 (국내 분리 다제내성 장티프스균의 Plasmid 및 ${\beta}$-lactamase의 특성에 관한 연구)

  • Yoo, Jung-Sik;Shin, Young-Hack;Oh, Kyung-Soo;Lee, Jeom-Kyu;Kim, Ki-Sang
    • The Journal of the Korean Society for Microbiology
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    • v.34 no.5
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    • pp.445-452
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    • 1999
  • Eight strains of multidrug-resistant (MDR) Salmonella typhi were isolated from Kyonggi area during January-February, 1997. They were resistant to ampicillin, amoxicillin, carbenicillin, tetracycline, chloramphenicol, trimethoprim/sulfamethoxazole, trimethoprim. Eight strains had one plasmid respectively which size was approximately M.W 220 kb and showed same restriction pattern by endonuclease HindIII. The plasmid was similar to the plasmid in size that was related to multidrug resistant S. typhi isolated from southeast Asia. It were transferred by conjugation to recipient E. coli K-12 in frequency of $2.43{\times}10^4-1.73{\times}10^{-2}$ and transconjugant showed same drug-resistant pattern with donor cells. All of 8 strains produced ${\beta}$-lactamase that was assummed to TEM-l type by isoelectric focusing and PCR.

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