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두유에서 Sacchasomyces uvarum 과 Lactobacillus acidophilus의 혼합배양
공인수,이정수,정용준,류인덕,오두환,유주현,Kong, In-Soo,Lee, Jung-Soo,Chung, Yong-Joon,Lew, In-Deok,Oh, Doo-Whan,Yu, Ju-Hyun 한국식품과학회 1987 한국식품과학회지 Vol.19 No.4
Sacchasomyces uvarum과 Lactobacillus acidophilus를 두유에서 단독 및 혼합배양하였을때 산생성에 미치는 조건은 검토하였다. 접종비율에 의한 산생성은 L. acidophilus와 S. uvarum의 접종비가 2 : 1일때 가장 높은 산도를 보였으며 배양시간은 16시간부터 높은 산도를 유지하였다. 배양온도는 $37^{\circ}C$가 가장 좋았으며 두유에 sucrose를 첨가하여 단독배양하면 산생성의 증가는 없었으나 혼합배양한 결과 산도가 증가하였다. Sucrose 2%와 skim milk 3%를 첨가하여 혼합배양하면 두유만을 사용하여 혼합배양한 것에 비하여 약4배의 산생성 증가를 보였다. S. uvarum의 두유 배양여액을 열처리하지 않고 두유에 첨가하여 L. acidophilus를 단독배양하면 단독배양만으로도 높은 산생성을 보여주었으나 열처리한 배양여액이 첨가된 두유에 L. acidophilus를 단독배양하면 산생성의 증가가 전혀 나타나지 않았다. Among the several lactic acid bacteria, Lactobacillus acidophilus showed the highest acid production when it was cultured mixed with Sacchasomyces uvarum in soymilk. The highest acid production was obtained in 16 hrs of cultivation when the inoculation ratio of L. acidophilus and S. uvarum was 2:1 and the temperature was $30{\sim}37^{\circ}C$. The acid production was greatly enhanced by the addition of 2.0% sucrose. However, skim milk was not stimulatory in mixed culture. During mixed culture in soymilk, acid production was affected by the enzymatic reaction of yeast.
공인수,박제현,한정현,최윤혁,이종희,진철호,이정기 한국미생물·생명공학회 2001 한국미생물·생명공학회지 Vol.29 No.2
Strong promoter로 밝혀진 alginate lyase 유전자의 promoter 부위에 대한 특성을 검토하기 위해 alginate lyase 유전자의 46개 N-terminal amino acid가 포함된 promoter 부분과, 같은 균으로부터 분리한 $\beta$-agarase의 유전자를 연결시켜 agarase의 activity를 평판배지상에서 보다 쉽게 확인하는 방법으로 promoter의 활성을 측정한 결과 alginate lyase 유전자 promoter에 의해서 $\beta$-agarase 유전자의 대량발현이 유도되고 있었으며 glucose의 존재하에서 $\beta$-agarase 유전자 발현이 일어나지 않는 catabolite repression 양상을 나타내고 있다. PCR로써 alginate lyase의 46개 N-terminal amino acid 부분이 순차적으로 제거된 plasmid를 제조하여 대량발현을 조사한 결과 46개의 아미노산이 제거된 후에도 $\beta$-agarase의 활성에는 변화가 없어 46개의 N-말단이 정상적인 상태에서 발현에는 영향을 미치고 있지 않음을 확인할 수 있었다. 또한 alginate lyase 유전자의 promoter region에 존재하는 가능한 2개의 promoter consensus sequence PI, PII를 subcloning한 결과 promoter PII만이 존재할 때도 대량발현이 유도되고 있음을 확인할 수 있었으며 동시에 glucose가 존재할 때 catabolite repression이 역시 나타나고 있어 이 부분이 발현 및 glucose에 의한 regulation에 매우 중요하게 작용하는 부분이라는 것을 확인할 수 있었다. Expression of f3 ~agarase Gene and Catabolite Repression in Escherichia coli by the Promoter of Alginate Lyase Gene Isolated from Marine Pseudomonas sp. Jin, Cheal~Ho, J~Hyeon Park, Jeong-Hyun Han, YoonM Hyeok Chae, Jong~Hee Lee, Jung-Kee Lee!, and In-800 Kong*. Faculty of Food Science and Biotechnology, Pukyong National UniversitYt Pusan 608-737, Korea, llnBioNet Co. 1690-3 Taejon 306-230, Korea - Promoter is a key factor for expression of the recombinant protein. There are many promoters for overexpression of protein in various organisms. The aly promoter of Pseudomonas sp. W7 isolated from marine environment was known to be a constitutive expression promoter of the alginate lyase gene, and it's promoter activity is repressed by glucose in Escherichia coli. To investigate the catabolite repression of the aly promoter ~md association between the promoter mutants, f3 agarase gene, which was also cloned from Pseudomonas sp. W7 was connected to the aly promoter with the sequence the coding 46 N-terminal amino acids ofthe alginate lyase gene. The constructed plasmid was introduced into E. coli and the agarase activity was measured. Fourty six amino acids of the alginate lyase gene was serially deleted using peR to the direction of 5' upstream region and subcloned. The agarase was overexpressed by the aly promoter and the production of agarase was repressed by the addition of glucose into culture media. Fourty six amino acids of alginate lyase did not affect the production of agarase at all. The deletion of a putative stem-loop structure in the aly promoter induced the decrease of f3 -agarase productivity.
서울특별시 강남구민의 관급수불소농도조정의식에 관한 조사연구
공인수,문혁수,백대일,김종배 대한구강보건학회 2003 大韓口腔保健學會誌 Vol.27 No.4
The purpose of this study was to obtain the information, for improvement of the support on water fluoridation in Korea. The author surveyed knowledge and awareness about water fluoridation of 3,378 people living in KangNam-Gu, Seoul. The people were selected by the technique of random sampling. Items of the questionnaire are as follows; the use of drinkable water, cookable water and the recognition of necessity to execute water fluoridation for the purpose of caries prevention, to prevent dental decay for dental health, to prevent dental decay for the reduction of health insurance expense and the recognition of the safety of water fluoridation and the possibility for the execution of water fluoridation. The obtained results are ; 1. There was some doubt about the safety of water fluoridation. As a result, only half of the people consider tap water as edible water and about 25% of the people consider tap water not suitable for drinking and cooking. 2. After reading an explanation about the concept, effect and the safety of water fluoridation the majority of the people approved the execution of it. 3. The people should be actively educated and promoted about the effectiveness and the significance of water fluoridation. Because they were not aware of water fluoridation for effective caries prevention as public oral health service even though they still tried to reduce the expenses of health insurance. 4. Despite of the necessity to support executing public health service such as water fluoridation, only 40% of the people recognized the fact that water fluoridation is safe. Therefore active education and promotion should be required for the safety of water fluoridation.
Chung, Yong Joon,Kong, In Soo,Kang, Yoon Suk,Yu, Ju Hyun 한국산업미생물학회 1990 한국미생물·생명공학회지 Vol.18 No.1
토양에서 분리한 호알카리성 Bacillus sp. YC-335가 생산하는 CGTase를 ethanol 침전법, DEAE-Toyopearl column chromatography, Sephadex G-100 column chromatography 방법 등에 의해 대량 정제하였다. 이 때 수율은 17.8%이었고 52.9배의 정제도를 가진 효소단백질을 얻을 수 있었다. 정제효소는 SDS-polyacrylamide gel 전기영동에 의해 분자량이 75,000 정도인 단일 peptide의 단백질임을 확인하였고 효소의 최적활성 pH는 6.0이었으며 pH 안정성은 pH 6-10까지 안정하였다. 최적활성온도는 50℃이었으며 열안정성은 50℃까지 안정하였고 이는 15mM CaCl_2에 의해 열안정성이 보호되었다. Alkalophilic Bacillus sp. YC-335 isolated from soil was capable of producing large amount of cyclodextrin glycosyltransferase(CGTase) in culture broth. This enzyme was successively purified 52.9 folds with 17.8 yield by ethanol precipitation, DEAE-Toyopearl column chromatography and Sephadex G-100 column chromatography. The purified enzyme have a molecular weight of approximately 75,000 estimated by SDS polyacrylamide gel electrophoresis. The optimum pH and temperature for the enzyme activity were 6.0 and 50℃, respectively. The enzyme was stable between pH 6 and 10, and up to 50℃. The thermostability of the enzyme was increased up to 60℃ by the addition of 15 mM CaCl_2.
Multi-copy Streptomyces 플라스미드, pJY501의 재조합 유도체의 특성
염도영,공인수,유주현 한국미생물 · 생명공학회 1990 한국미생물·생명공학회지 Vol.18 No.1
The restriction cleavage map of multi-copy recombinant plasmid, pJY502 (5.5 kb), carrying the thiostrepton resistance gene (tsr) was determined. Comparison of the restriction pattern with that of Streptomyces plasmids previously demonstrated that pJY502 was novel. The plasmid pJY502 had a broad host range in Streptomyces and contained single BgtII site for cloning purpose. Transformation frequency of pJY502 was $2.2 \times 10^5$ in S. lividans. E. coti-Streptomyces bifunctional plasmid, pJY504, was also constructed. Thiostrepton 내성 유전자(tsr)를 포함하는 multicopy 재조합 플라스미드 pJY502(5.5kb)의 제한효소 지도를 비교해본 결과 pJY502는 새로운 플라스미드로 확인되었다. pJY502는 Streptomyces에서 넓은 host range를 나타내었으며 cloning에 사용할 수 있는 단일 BglII 제한효소 인식부위를 갖고 있었다. pJY502의 형질전환 빈도는 S.lividans에서 $2.2 \times 10^5$이었다. 또한 E.coli-Streptomyces bifunctional 플라스미드 pJY504을 제조하였다.