• Title/Summary/Keyword: 5-Amino-3H-1

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Cloning and Nucleotide Sequence Analysis of the Virulence Gene Cassette from Vibrio cholerae KNIH002 Isolated in Korea (국내 분리주인 Vibrio cholerae KNIH002로부터 독성 유전자 카세트의 클로닝 및 염기서열 분석)

  • 신희정;박용춘;김영창
    • Korean Journal of Microbiology
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    • v.35 no.3
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    • pp.205-210
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    • 1999
  • 16brio cholerne is an important pathogenic organism that causes dimhea in human beings. V ciaoleroe KNIH002 was isolated from patients suffering with dian.heal disease in Korea. From Southern hybridization using the amplified PCR product of 307 bp as a probe. which was obtained from PCR reaction using primer detecting cholera toxin gene, we have found that the c b gene located in 4.5-kb fragmenl double digested with Pstl and BgllI of the chromosome. Therefore, we made mini-libraries of the isolate using PstI and Bgm restriction endonuclease and pBluescript SKU(+) vector. As a result. we cloned 4.5-kb PstI-BglII fragment containing the c a gene encoding a cholera toxin from the constructed mini-libraries of V olzolerae KNlH002 by colony hybridization using the same probes. This recombinant plasmid was named pCTX75. E. coii XL1- Blue harboring pCTX75 showed the cytotoxicity on Chinese Hamster Ovary cells. From the sequencing of he cloned recombinant plasmid, we confinned that it has virulence gene cassette consisting of ace, zot, ctx.4 and cf"~B gene. The ace and zot genes were composed of 291 hp and 1.200 bp with ATG initiation codon and TGA lennination codon, respectively. Nucleotide sequence of the ace gene exhibited 100% identity with that of V cholera E7946 El Tor Ogawa strains. But, nucleolide and amino acid sequence comparison of the zot gene exhibited 99% and 98.8% identity with that of V cholerae 395 Classical Ogawa stram, respectively. Specially. the Ala-100, Ala-272 and Ala-281 sites of Zoi polypeptide presented in V choleme 395 Classical Ogawa strain are replaced by Val in V cholerae KNIH002.

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Biosorption of Cadmium by a Methanotrophs Exopolysaccharide (메탄산화세균의 EPS를 이용한 Cd의 생물흡착)

  • Lee, Hee-Ja;Kim, Kwang-Soo;Cho, Yang-Seok
    • Proceedings of the Korea Water Resources Association Conference
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    • 2006.05a
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    • pp.1415-1419
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    • 2006
  • 메탄을 탄소원 및 에너지원으로 이용하는 메탄산화균은 물질대사과정 중에 다량의 세포외 고분자물질인 Extracellular polymeric substances(EPS)를 생성하는데, EPS는 카르복실기와 같은 표면흡착 기능을 가지고 있어 생체흡착제로 사용이 가능하다. 따라서 본 연구에서는 메탄산화세균을 이용하여 중금속인 Cd의 흡착성능을 파악하여 활성슬러지의 흡착능과 비교하고, EPS 농도별, pH별 흡착량의 변화를 실험한 후 Freundlich 흡착모델식에 적용하여 흡착공정의 기본적인 설계인자를 도출하고자 하였다. 실험에 사용한 메탄산화세균은 매립지 복토층 상부 토양에서 분리하여 실험실에서 대량으로 배양하였으며, EPS 생성을 위해 메탄을 Head space의 20%를 주입하고 $30^{\circ}C$, 150rpm에서 질소원이 부족한 조건으로 48hr 동안 배양하였다. Cd의 흡착실험은 용액의 pH를 3에서 8까지 변화를 주면서 활성슬러지와 메탄산화세균의 시간별 흡착능을 측정하였다. 또한 중금속의 농도별 흡착능을 측정하여 흡착평형 상수를 파악하였으며, 중금속 흡착 전, 후 미생물의 SEM 촬영, FT-IR 분석, 전자현미분석(EPMA)을 통하여 무기성분 분석 및 표면관찰을 수행하였다. 실험결과 메탄산화세균에 의해 생성된 EPS 물질은 중금속에 대한 강한 결합능력이 있으며, Cd에 대한 최고 흡착능은 26mg Cd(Ⅱ)/g VSS의 값을 보였다. 이러한 미생물의 EPS의 흡착능은 pH와 칼슘이온의 영향을 많이 받았으며, 메탄산화세균의 FT-IR 분석결과 EPS에는 sulfate ester, pyruvate 등과 같은 작용기와 amino sugar, carboxyl 작용기들이 많이 존재하여 활성슬러지에 비해 중금속의 흡착능이 높은 것으로 사료되었다.X>${\mu}_{max,A}$는 최대암모니아 섭취률을 이용하여 구한 결과 $0.65d^{-1}$로 나타났다.EX>$60%{\sim}87%$가 수심 10m 이내에 분포하였고, 녹조강과 남조강이 우점하는 하절기에는 5m 이내에 주로 분포하였다. 취수탑 지점의 수심이 연중 $25{\sim}35m$를 유지하는 H호의 경우 간헐식 폭기장치를 가동하는 기간은 물론 그 외 기간에도 취수구의 심도를 표층 10m 이하로 유지 할 경우 전체 조류 유입량을 60% 이상 저감할 수 있을 것으로 조사되었다.심볼 및 색채 디자인 등의 작업이 수반되어야 하며, 이들을 고려한 인터넷용 GIS기본도를 신규 제작한다. 상습침수지구와 관련된 각종 GIS데이타와 각 기관이 보유하고 있는 공공정보 가운데 공간정보와 연계되어야 하는 자료를 인터넷 GIS를 이용하여 효율적으로 관리하기 위해서는 단계별 구축전략이 필요하다. 따라서 본 논문에서는 인터넷 GIS를 이용하여 상습침수구역관련 정보를 검색, 처리 및 분석할 수 있는 상습침수 구역 종합정보화 시스템을 구축토록 하였다.N, 항목에서 보 상류가 높게 나타났으나, 철거되지 않은 검전보나 안양대교보에 비해 그 차이가 크지 않은 것으로 나타났다.의 기상변화가 자발성 기흉 발생에 영향을 미친다고 추론할 수 있었다. 향후 본 연구에서 추론된 기상변화와 기흉 발생과의 인과관계를 확인하고 좀 더 구체화하기 위한 연구가 필요할 것이다.게 이루어질 수 있을 것으로 기대된다.는 초과수익률이 상승하지만, 이후로는 감소하므로, 반전거래전략을 활용하는 경우 주식투자기간은 24개월이하의 중단기가 적합함을 발견하였다. 이상의 행태적 측면과 투자성과측면의 실증결과를 통하여 한국주식시장에 있어서 시장수익률을 평균적으로 초과할 수 있는 거래전

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Growth Characteristics of Bifidobacteria and Quality Characteristics of Soy Yogurt Prepared with Different Proteolytic Enzymes and Starter Culture (단백분해효소와 배양방법의 종류에 따른 비피더스균의 생육특성 및 soy yogurt의 품질특성)

  • Lee, Jung-Eun;Lee, Sook-Young
    • Korean Journal of Food Science and Technology
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    • v.33 no.5
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    • pp.603-610
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    • 2001
  • The quality characteristics of soy yogurt prepared with different proteolytic enzymes and starter culture were evaluated. In order to facilitate the growth of lactic acid bacteria and subsequent production of lactic acid, soy protein isolate(SPI) was hydrolyzed using three kinds of proteases; one extracted from Aspergillus oryzae, bromelain and ${\alpha}-chymotrypsin$. The cultural systems employed thereafter for lactic fermentations were: 1) Bifidobacterium bifidum, 2) B. bifidum and Lactobacillus acidophilus, 3) B. bifidum and Lactobacillus bulgaricus. In soy yogurt, pH was more decreased by mixed culture method than single culture method with the accumulation of lactic acid. Viable cells of lactic acid bacteria in soy yogurts were measured $10^8$ CFU/g by the single culture method while $10^9$ CFU/g by the mixed culture method except ${\alpha}-chymotrypsin$ treatment. The amount of free amino acids in soy yogurts were substaintially increased by enzyme treatment. Viscosity was decreased by enzyme treatment, resulting in higher viscosity by ${\alpha}-chymotrypsin$ treatment. Water holding capacity was found to be higher in the single culture method in case of enzyme treatment. Among the various volatile flavor components isolated and identified after enzyme hydrolysis, acetaldehyde, ethanol, diacetyl, butyl alcohol contents tended to increase by lactic fermentation.

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Peptide Properties of Rapid Salted and Fermented Anchovy Sauce Using Various Pretenses 1. Hydrolysis of Anchovy Sauce and Actomyosin by Various Pretenses (단백질 분해효소를 이용하여 제조한 속성 멸치 액젓의 펩티드 특성 1. 단백질 분해효소에 의한 멸치 액젓 및 Actomyosin의 가수분해)

  • KIM In-Soo;CHOI Young-Joon;HEU Min-Soo;CHO Young-Je;IM Yeong-Sun;GU Yeun-Suk;YEO Saeng-Gyu;PARK Jae-Woon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.4
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    • pp.481-487
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    • 1999
  • The optimal conditions of enzymatic hydrolysis for preparation of rapid salted and fermented anchovy sauce (SFAS) using various pretenses such as trypsin, chymotrypsin, crude enzyme from squid liver and viscera, Alcalase, Neutrase and Protamex were studied. SFAS prepared with squid viscera had higher level of VBN (173.6 mg/100 g) when stored for 70 days than other samples, and peroxide values were almost equal among all samples during fermentation period. Total amino acids and nonprotein nitrogenous compounds remarkably increased as SFAS treated with Alcalase or Protamex which exhibited higher the hydrolysis rate of $57\%$ at 60 day than others. The optimal pHs of trypsin, chymotryosin, Alcalase, Neutrase and Protamex on anchovy actomyosin were 7.5, 6.5, 6.5, 7.0 and 5.0, respectively. Optimal temperatures of trypsin, chymotryosin, Alcalase and Neutrase were 55, 45, 60 and $55^{\circ}C$, respectively. Otherwise, Protamex activity increased as temperature increased from 20 to $70^{\circ}C$. Protamex had higher $K_m$ (3.545) and $V_{max}$ value (2.688) than others. Protamex affected less by NaCl had $52.5\%$ activity at the fermentation condition of $20^{\circ}C$ and $25\%$ NaCl. Protamex appeared to be very effective for the hydrolysis of crude actomyosin from ancnovy.

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Sphingomonas abietis sp. nov., an Endophytic Bacterium Isolated from Korean Fir

  • Lingmin Jiang;Hanna Choe;Yuxin Peng;Doeun Jeon;Donghyun Cho;Yue Jiang;Ju Huck Lee;Cha Young Kim;Jiyoung Lee
    • Journal of Microbiology and Biotechnology
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    • v.33 no.10
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    • pp.1292-1298
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    • 2023
  • PAMB 00755T, a bacterial strain, was isolated from Korean fir leaves. The strain exhibits yellow colonies and consists of Gram-negative, non-motile, short rods or ovoid-shaped cells. It displays optimal growth conditions at 20℃, 0% NaCl, and pH 6.0. Results of 16S rRNA gene-based phylogenetic analyses showed that strain PAMB 00755T was most closely related to Sphingomonas chungangi MAH-6T (97.7%) and Sphingomonas polyaromaticivorans B2-7T (97.4%), and ≤96.5% sequence similarity to other members of the genus Sphingomonas. The values of average nucleotide identity (79.9-81.3%), average amino acid identity (73.3-75.9%), and digital DNA-DNA hybridization (73.3-75.9%) were significantly lower than the threshold values for species boundaries; these overall genome-related indexes (OGRI) analyses indicated that the strain represents a novel species. Genomic analysis revealed that the strain has a 4.4-Mbp genome encoding 4,083 functional genes, while the DNA G+C content of the whole genome is 66.1%. The genome of strain PAMB 00755T showed a putative carotenoid biosynthetic cluster responsible for its antioxidant activity. The respiratory quinone was identified as ubiquinone 10 (Q-10), while the major fatty acids in the profile were identified as C18:1ω7c and/or C18:1ω6c (summed feature 8). The major polar lipids of strain PAMB 00755T were diphosphatidylglycerol, phosphatidylethanolamine, sphingoglycolipid, and phosphatidylcholine. Based on a comprehensive analysis of genomic, phenotypic, and chemotaxonomic characteristics, we proposed the name Sphingomonas abietis sp. nov. for this novel species, with PAMB 00755T as the type strain (= KCTC 92781T = GDMCC 1.3779T).

Enhancing the Enzymatic Activity of the Multifunctional β-Glycosyl Hydrolase (Cel44C-Man26AP558) from Paenibacillus polymyxa GS01 Using DNA Shuffling (DNA Shuffling을 이용한 Paenibacillus polymyxa GS01의 다기능 β-Glycosyl Hydrolase (Cel44C-Man26AP558) 효소 활성 증가)

  • Kang, Young-Min;Kang, Tae-Ho;Yun, Han-Dae;Cho, Kye-Man
    • Korean Journal of Microbiology
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    • v.48 no.2
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    • pp.73-78
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    • 2012
  • We previously reported that the truncated Cel44C-$Man26A_{P558}$ ${\beta}$-glycosyl hydrolase protein exhibits multifunctional activities, including cellulase, xylanase, and lichenase. DNA shuffling of the truncated Cel44C-$Man26A_{P558}$ enzyme was performed to enhance the enzymatic activity of the multifunctional ${\beta}$-glycosyl hydrolase. Two mutant enzymes, M2Cel44C-$Man26A_{P558}$ that carries one mutation (P438A) and M21Cel44C-$Man26A_{P558}$ that carries two mutations (A273T and P438A) were obtained. The enzymatic activity of the M21Cel44C-$Man26A_{P558}$ double mutant was lower than enzymatic activity of the single mutant (M2Cel44C-$Man26A_{P558}$). However, both mutants displayed the enhancements in their enzyme activities that were ${\approx}1.3$- to 2.2-fold higher than the original enzymatic activity in Cel44C-$Man26A_{P558}$. In particular, the mutant M2Cel44C-$Man26A_{P558}$ exhibited an approximate 1.5- to 2.2-fold increase in the cellulase, xylanase, and lichenase activities in comparison with the control (Cel44C-$Man26A_{P558}$). The optimum cellulase, linchenase, and xylanase activities of ${\beta}$-glycosyl hydrolase were observed at pH 7.0, pH 7.0 and pH 6.0, respectively. These results, therefore, suggest that the amino acid residue Ala438 plays important roles in the enhancement of the activity of multifunctional ${\beta}$-glycosyl hydrolase.

Immunogenicity of Synthetic Peptide Specific for Major Immunogenic Determinat of Hepatitis B Surface Antigen (B형간염(型肝炎) 표면항원(表面抗原)의 주면역원(主免疫原) 결정기(決定基)에 특이(特異)한 합성(合成) Peptide의 면역원성(免疫原性)에 관한 연구(硏究))

  • Shin, Kwang-soon;Han, Su-nam
    • Korean Journal of Veterinary Research
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    • v.25 no.1
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    • pp.7-17
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    • 1985
  • Many investigators have been pursuing various attempts so far to produce hepatitis B surface antigen(HBsAg) vaccines using the techniques such as isolation from plasma of chronic HBsAg carrier, recombinant DNA technique or preparation of synthetic peptides specific for immunogenic determinants. Hepatitis B virus can not grow on any cell lines by the tissue culture technique at the present time. The plasma of chronic HBsAg carrier is expensive and its source is limited. The HBsAg from the recombinant DNA technique gave still very low yield. Another approach, therefore, has been initiated to develop a synthetic hepatitis B virus vaccine. The possible use of several distinct synthetic vaccines in prophylaxis can be facilitated by availability of full synthetic immunogens. Peptides synthesized for potential application as antiviral vaccines have been mostly tested in the form of conjugates with carrier proteins, although the free synthetic peptide can be immunogenic. To understand basic knowledges on the antigenicity and immunogenicity of a synthetic peptide specific for major immunogenic determinant of HBsAg, a nonapeptide, $H_2N^{139}Cys-Thr-Lys-Pro-Thr-Asp-Gly-^{146}Asn-Aba$ COOH, which corresponds to HBsAg amino acid residues 139 to 147, was synthesized by the Merrifield's solid-phase method with a slight modification. The antigenicity and immunogenicity of this specific synthetic peptide were examined comparing with purified plasma-derived natural HBsAg. The results obtained are as follows; 1. The peptide synthesized showed the identical amino acid composition to the theoretical value. The degree of purification and molecular weight were acertained by methods of high performance liquid chromatography and mass spectrometry. 2. Using m-maleimidobenzoyl-N-hydroxysuccinimide ester as a conjugating agent, the synthetic peptide was conjugated to rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin. Their conjugation yields were 8.3, 9.5, 15.8, 13.5, and 11.2%, respectively. 3. The natural HBsAg was purified from plasma of chronic HBsAg carrier. By the electron microscopic observation of the purified natural HBsAg preparation, no Dane particles were observed and the preparation showed negative DNA polymerase activity. 4. Antigenicity of the synthetic peptide and the plasma-derived natural HBsAg was determined by competition radioimmunoassay using $^{125}I$-natural HBsAg. Their 50% inhibitions appeared as $90{\mu}g/ml$ and $0.12{\mu}g/ml$ for the synthetic peptide and the natural HBsAg, respectively. This indicates that the former was about 750-fold less antigenic than the latter. 5. Immunogenicity of the synthetic peptide was determined by administering the peptide-carrier conjugates into rabbits with and without Freund's complete adjuvant. Regardless the carrier proteins and adjuvant, positive immune responses to the synthetic peptide were observed. The higher antibody titers, however, were shown in the groups administered with Freund's complete adjuvant. 6. Immunizing dose 50% in mice of the various peptide-carrier conjugates was 5.47, 6.00, 65.16, 31.25 and $13.03{\mu}g/dose$ for rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin, respectively, while the natural HBsAg showed $0.65{\mu}g/dose$. 7. It was postulated that homologous proteins prefer to heterologous ones as the carriers.

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Process Optimization of Ginseng Berry Extract Fermentation by Lactobacillus sp. Strain KYH isolated from Fermented Kimchi and Product Analysis (발효 김치로부터 분리한 Lactobacillus sp. Strain KYH를 이용한 진생베리 추출물 최적 발효 공정 확립 및 생성물의 특성 분석)

  • Ha, Yoo-Jin;Yoo, Sun-Kyun;Kim, Mee Ree
    • Journal of the East Asian Society of Dietary Life
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    • v.26 no.1
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    • pp.88-98
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    • 2016
  • The pharmacological effects of ginseng berry have been known to improve psychological function, immune activities, cardiovascular conditions, and certain cancers. It is also known that fermentation improves the bioavailability of human beneficial natural materials. Accordingly, we investigated the optimal fermentation conditions of ginseng berry extract with strain isolated from conventional foods. We also analyzed the fermentation product and its antioxidant activity. The bacterium isolated from fermented kimchi was identified as Lactobacillus sp. strain KYH. To optimize the process, fermentation was performed in a 5 L fermenter containing 3 L of ginseng berry extract at 200 rpm for 72 hr. Under optimized conditions, batch and fed-batch fermentations were performed. After fermentation, organic acids, amino acids, sugars, ginsenosides, and antioxidant activity were evaluated. The optimum fermentation conditions were determined as pH 7.0 and a temperature of $30^{\circ}C$, respectively. After fermentation, the amounts and compositions of organic acids, amino acids, sugars, ginsenosides, and antioxidant activity were altered. In comparing the distribution of ginsenosides with that before fermentation, the ginsenoside Re was a major product. However, amounts of ginsenosides Rb1, Rc, and Rd were reduced, whereas amounts of ginsenosides Rh1 and Rh2 increased. Total phenol content increased to 43.8%, whereas flavonoid content decreased to 19.8%. The DPPH radical scavenging activity and total antioxidant activity increased to 27.2 and 19.4%, respectively.

Effect of Freezing and Thawing Methods on Duck Meat Characteristics (냉동과 해동 방법이 오리고기의 특성에 미치는 영향)

  • Heo, Kang-Nyeong;Kim, Ji-Hyuk;Kim, Sang-Ho;Kang, Bo-Seok;Kim, Chong-Dae;Cha, Jae-Beom;Hong, Eui-Chul
    • Korean Journal of Poultry Science
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    • v.43 no.4
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    • pp.197-206
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    • 2016
  • This work was carried out to investigate effects of the freezing/thawing method on duck meat kept in a freezer for a month. The meats used were breast muscle collected from Korean native ducks (KND) that were fed for 8 weeks (2.8 kg of live weight). Forty-five samples were used after being frozen in storage for one month and were then divided into 5 treatments (3 replications/treatment, 3 samples/replication). Five treatments (CON, FFFT, FFST, SFFT and SFST) were control groups (CON) and four were experimental groups, using $2{\times}2$ complex factors with two freezing methods (fast freezing, FF, $-50^{\circ}C$ in a deep freezer; slow freezing, SF, $-20^{\circ}C$ in a common freezer) and two thawing methods (fast thawing, FT, 5 h $12^{\circ}C$ with flow water; slow thawing, ST, 24 h $5^{\circ}C$ in a refrigerator). Lightness of KND meat in FF and FT groups was lower than that of control (P<0.05). Yellowness of KND meat of the ST group was higher than that of control (P<0.05). Cooking loss (CL) and water holding capacity (WHC) of KND meat in the control were lower than those of the freezing and thawing groups (P<0.01, P<0.05), but shear force (SF) of the control was higher than that of other groups (P<0.01). Moisture content of the ST group was higher than that of the FT group (P<0.05), and protein content of the FF group was higher than that of control (P<0.05). Stearic acid (C18:0) of the SF group was higher than that of the FF group (P<0.05). Arachidonic acid (C20:4n6) of control was higher than that of the SF and ST groups (P<0.01, P<0.05). Alanine, aspartic acid, glutamic acid, serine, and tyrosine content of the control were lower than that of the freezing and thawing groups (P<0.05). These results show that freezing and thawing methods affect meat color, shear force, cooking loss, and WHC-related water content.

Studies on electrophoretic pattern and amino acids of wild soybean protein[1] -Acrylamide gel electrophoretic pattern of seed protein- (야생대두단백질(野生大豆蛋白質)의 전기영동유형(電氣泳動類型)및 아미노산조성(酸組成)에 관(關)한 연구(硏究)[1] -종실단백질(種實蛋白質)의 Acrylamide gel 전기(電氣) 영동유형(泳動類型)-)

  • Park, H.;Lee, J.S.;Lee, C.Y.
    • Applied Biological Chemistry
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    • v.20 no.2
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    • pp.247-254
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    • 1977
  • Acrylamide gel electrophoretic pattern of 13 wild soybean (Glycine ussuriensis) was compared with that of G. gracilis and G. max var. Gwanggyo. Average Protein content (50%) of wild soybean was greater than that of C. gracilis (46%) and Gwanggyo (45%). Grain weight of wild soybean was one third of G. gracilis and one ninth of Gwanggyo. Electrophoresis of wild soybean protein showed total 16 different bands and three of which (Rm 0.09, 0.59 and 0.84) were specific and did not appeared in 86 var. of G. max which showed four specific bands (Rm 0.35, 0.45, 0.50 and 0.77) of total 17 bands. G. glacilis had all bands of Gwanggyo and two bands (Rm 0.53 and 0.59), one of which (Rm 0.59) was specific for wild soybean indicating that G. gracilis is middle type. Of 16 protein bands the third band (32%), the first band (28%) and the 5th band (13%) were main bands. Electrophoretic pattern could be sorted qualitatively into 4 groups, semiquantitatively into 6 groups and 2 or 4 groups depending on reference pattern by correlation or pattern similarity method. All sorting methods separated a wild soybean from Sogri mountain into a group and except that there were no similarity among methods but correlation methods seems more reasonable. Protein content was no relation with electrophoretic pattern but positively correlated with percent contribution of first band at 5% level suggesting that the first band may have a important role for protein synthesis.

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