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제조합 대장균에서 과발현된 Citrobacter freundii KCTC2006 유래의 β-Tyrosinase를 이용한 3,4-Dihydroxyphenyl-L-alanine의 생산
이승구,노현수,홍승표,이규종,왕지원,태동년,엄기남,방상구,김영준,성문희 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.1
재조합 대장균에서 대량발현 시킨 Citrobacter freundii KCTC 2006 유래이 효소 β-tyrosinase를 이용하여 pyrocatechol, sodium pyruvate, ammonium acetate로부터 3,4-dihydroxy phenyl-L-alanine을 생산하기 위한 연구를 수행하였다. 이 효소반응에 적합한 온도 및 pH 조건은 각각 18℃와 8.5로 결정되었고, 반응액 중의 ammonium acetate와 sodium pyruvate의 농도는 각각 300 mM, 50 mM 이상으로 조절하는 것이 적합하였다. Pyrocatechol의 경우는 20 mM에서 가장 높은 반응성을 나타냈으나, 기질을 반복적으로 첨가하며 장시간 동안 효소반응을 수행하는 경우에는 pyrocatechol의 고갈을 피하기 위하여, 20 mM에서 50 mM 사이로 조절하였다. 한편, 반응액 중에 ethanol을 10% 첨가한 경우에는 반응속도가약 20% 증가하였다. 이상과 같은 효소반응특성에 기초하여 조제한 기질용액에 β-tyrosinase를 1 unit/㎖ 농도로 가하고, pyrocatechol과 pyruvate가 고갈되지 않도록 간헐적으로 첨가하면서 효소반응을 수행한 결과, 24시간 만에 85.2%의 수율로 31.6g/ℓ의 3,4-dihydroxyphenyl-L-alanine를 생산할 수 있었다. By using the β-tyrosinase of Citrobacter freundii KCTC2006, which was cloned and overexpressed in Escherichia coli, 3,4-dihydroxy phenyl-L-alanine (L-DOPA) was synthesized efficiently from pyrocatechol, sodium pyruvate, and ammonium acetate. Optimal temperature and pH for the reaction were determined to be about 18℃ and 8.5, respectively. The effects of substrate concentrations were also examined at different concentrations of ammonium acetate, sodium pyruvate, and pyrocatechol. Ammonium acetate and sodium pyruvate increased the reaction rate until the concentrations reached to 300 mM and 50 mM, respectively. Although pyrocatechol showed the optimal concentration at 20 mM, it was controlled between 20 mM and 50 mM to avoid the depletion of substrate during the enzymatic synthesis. Based on above results, a reaction medium for the production of L-DOPA was prepared and incubated with 1 unit/㎖ of β-tyrosinase. Pyrocatechol and sodium pyruvate was added to the reaction solution intermittently to avoid the substrate depletion during the enzymatic reaction. After 24 hour of reaction, 31.6 g/ℓ L-DOPA was accumulated in the reaction solution as soluble and precipitated ones and the conversion yield was about 85.2%.
Wang Wei,Liang Xuanyu,Niu Qinghe,Wang Qizhi,Zhuo Jinyi,Su Xuebin,Zhou Genmao,Zhao Lixin,Yuan Wei,Chang Jiangfang,Zheng Yongxiang,Pan Jienan,Wang Zhenzhi,Ji Zhongmin 한국원자력학회 2023 Nuclear Engineering and Technology Vol.55 No.8
It is essential to evaluate the blasting-enhanced permeability (BEP) feasibility of a low-permeability sandstone-type uranium deposit. In this work, the mineral composition, reservoir physical properties and rock mechanical properties of samples from sandstone-type uranium deposits were first measured. Then, the reformability evaluation method was established by the analytic hierarchy process-entropy weight method (AHP-EWM) and the fuzzy mathematics method. Finally, evaluation results were verified by the split Hopkinson Pressure Bar (SHPB) experiment and permeability test. Results show that medium sandstone, argillaceous sandstone and siltstone exhibit excellent reformability, followed by coarse sandstone and fine sandstone, while the reformability of sandy mudstone is poor and is not able to accept BEP reservoir stimulation. The permeability improvement and the distribution of damage fractures before and after the SHPB experiment confirm the correctness of evaluation results. This research provides a reformability evaluation method for the BEP of the low-permeability sandstone-type uranium deposit, which contributes to the selection of the appropriate regional and stratigraphic horizon of the BEP and the enhanced ISL of the low-permeability sandstone-type uranium deposit.
Wang, Ji-Quan,Wang, Tao,Shi, Fan,Yang, Yun-Yi,Su, Jin,Chai, Yan-Lan,Liu, Zi Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.14
Background: We designed this randomized controlled trial (RCT) to assess whether lobaplatin-based concurrent chemotherapy might be superior to cisplatin-based concurrent chemotherapy for FIGO stage II and III cervical cancer in terms of efficacy and safety. Materials and Methods: This prospective, open-label RCT aims to enroll 180 patients with FIGO stage II and III cervical cancer, randomly allocated to one of the three treatment groups (cisplatin $15mg/m^2$, cisplatin $20mg/m^2$ and lobaplatin $35mg/m^2$), with 60 patients in each group. All patients will receive external beam irradiation (EBRT) and high-dose-rate intracavitary brachytherapy (HDR-ICBT). Patients in cisplatin $15mg/m^2$ and $20mg/m^2$ groups will be administered four cycles of $15mg/m^2$ or $20mg/m^2$ cisplatin intravenously once weekly from the second week to the fifth week during EBRT, while patients inthe lobaplatin $35mg/m^2$ group will be administered two cycles of $35mg/m^2$ lobaplatin intravenously in the second and fifth week respectively during pelvic EBRT. All participants will be followed up for at least 12 months. Complete remission rate and progression-free survival (PFS) will be the primary endpoints. Overall survival (OS), incidence of adverse events (AEs), and quality of life will be the secondary endpoints. Results: Between March 2013 and March 2014, a total of 61 patients with FIGO stage II and III cervical cancer were randomly assigned to cisplatin $15mg/m^2$ group (n=21), cisplatin $20mg/m^2$ group (n=21) and lobaplatin $35mg/m^2$ group (n=19). We conducted a preliminary analysis of the results. Similar rates of complete remission and grades 3-4 gastrointestinal reactions were observed for the three treatment groups (P=0.801 and 0.793, respectively). Grade 3-4 hematologic toxicity was more frequent in the lobaplatin group than the cisplatin group. Conclusions: This proposed study will be the first RCT to evaluate whether lobaplatin-based chemoraiotherapy will have beneficial effects, compared with cisplatin-based chemoradiotherapy, on complete remission rate, PFS, OS, AEs and quality of life for FIGO stage II and III cervical cancer.
Su-ji Wang,Bing Bai,Bo Gao,Jian Liu 한국응용곤충학회 2019 Journal of Asia-Pacific Entomology Vol.22 No.3
The soybean aphid, Aphis glycines Matsumura, is an important pest of soybean. During some years with occurrences of heavy aphid infestations in field, A. glycines can cause serious damage to soybeans. A. glycines has a heteroecious and holocyclic life cycle, with migration occurring between the primary and secondary hosts during each year. The primary hosts of A. glycines are Rhamnus spp., and the secondary hosts are soybean, Glycine max (L.) Merrill, and wild soybean species, Glycine soja Sieb & Zucc. In this study, A. glycines were fed on detached leaves and live plants of Trifolium repens L. and Metaplexis japonica (Thunb.) Makino, and their survival, development and reproduction were studied at five constant temperatures. These data were compared to those of controls fed on the known host plant G. max. When A. glycines were fed on detached leaves of T. repens and M. japonica, they showed vigorous development and reproduction. The novel and striking results were that: (1) A. glycines could not thrive on live plants of M. japonica, and only a few nymphs were deposited. (2) When A. glycines were fed on live plants of vegetative stage T. repens, adult longevity and fecundity were as long as 11.64 ± 1.17 days and as great as 12.92 ± 1.23 nymphs per adult, respectively. This work provides important evidence that M. japonica is not a host of A. glycines, but T. repens is probably an important host for this aphid.
Effects of various treatments for contaminated implants
( Ji Ho Jo ),( Su Gwan Kim ),( Ji Su Oh ),( Jae Seek You ),( Sung Chul Lim ),( Seong Yong Moon ),( Wang Shik Yang ),( Jin Sung Park ),( Dong Kook Seo ),( Jeong Eun Yang ),( Kyung Hwan Yu ),( Young Sil 조선대학교 치의학연구원 2015 Oral Biology Research (Oral Biol Res) Vol.39 No.2
Purpose: The purpose was to evaluate osseointegration of contaminated implants after sterilization. Materials and Methods: Resorbable blasting media surfaced implants (3.7×10 mm) were placed as a routine procedure in six mongrel adult dogs. In the control group, normal fixtures were placed. The implants were sterilized after contamination: ethylene oxide gas (group 1), normal saline (group 2), and without any treatment (group 3). Results: New bone formation rate (NBFR) of the control group was significantly lower than groups 1 and 3 at 4 weeks, and the control group was highest at 8 weeks. At 8 weeks, NBFR and bone to implant contact showed no significant difference among all groups, and osseointegration was also observed. Conclusion: Osseointegration can be observed in contaminated implants after sterilizing.
다중PCR증폭과 특이 DNA-chip을 활용한 꿀벌 주요 11종 병원체의 검출법 개발
왕지희(Ji-Hee Wang),이도부(Dobu Lee),구수진(Su-Jin Ku),백문철(Mun-Cheol Peak),민상현(Sang-Hyun Min),임수진(Su-jin Lim),이칠우(Chil-Woo Lee),윤병수(Byoung-Su Yoon) 한국양봉학회 2016 韓國養蜂學會誌 Vol.31 No.2
Against 11 major honeybee pathogens, new detection-method using multiplex PCR and specific DNA-chip were developed. 11 major pathogens were 2 species of bacteria, 3 of fungi and 6 of viruses, including Paenibacillus larvae, Melissococcus plutonius, Aspergillus flavus, Ascosphaera apis, Nosema ceranae, Black queen cell virus, Chronic bee paralysis virus, Deformed wing virus, Israeli acute paralysis virus, Sacbrood virus, and Korean sacbrood virus. Development of multiplex PCR and DNA-chip were well optimized just using DNA and cDNA isolated from a infected sample, such as larva or adult honeybee. Based on analysis of DNA chip, it could be easy recognized, which are dominant among 11 pathogens in test sample. It might be useful as confirmation-test for diagnosis of honeybee diseases.
꿀벌의 석고병, 백묵병 현장 진단을 위한 신속 Ascosphaera apis 및 Aspergillus flavus 검출법 개발
왕지희(Ji-hee Wang),민상현(Sang-hyun Min),임수진(Su-jin Lim),윤병수(Byoung-Su Yoon) 한국양봉학회 2016 韓國養蜂學會誌 Vol.31 No.1
Rapid detection methods against Ascosphaera apis and Aspergillus flavus were developed for Chalkbrood and Stonebrood in honeybee. Specific primer sets were selected which detect A. flavus or A. apis based on 18s ribosomal RNA independently and were evaluated using real-time PCR and Ultra-fast PCR (UF-PCR). As the result, Minimal detection time was reduced until 8 min 38 seconds for A. flavus and 12 min 57 seconds for A. apis under 30 cycles UF-PCR. The detections of these methods show that even 101 (100 atto gram) molecules of DNA template could be detected for Aspergillus flavus and that 103 (10 femto gram) molecules of DNA template could be detected for Ascosphaera apis. Specific UF-PCRs are not only useful to confirm pathogendetection with naked eye, but be easy to handle a mobile PCR instrument in field. To monitoring of Chalkbrood or Stonebrood in apiary field directly, applications of on-site pathogen detection would be expected.
Jang, Ji Yong,Min, Ji Hyun,Chae, Yun Hee,Baek, Jin Young,Wang, Su Bin,Park, Su Jin,Oh, Goo Taeg,Lee, Sang-Hak,Ho, Ye-Shih,Chang, Tong-Shin Mary Ann Liebert 2014 ANTIOXIDANTS AND REDOX SIGNALING Vol.20 No.16
<P>The collagen-stimulated generation of reactive oxygen species (ROS) regulates signal transduction in platelets, although the mechanism is unclear. The major targets of ROS include protein tyrosine phosphatases (PTPs). ROS-mediated oxidation of the active cysteine site in PTPs abrogates the PTP catalytic activity. The aim of this study was to elucidate whether collagen-induced ROS generation leads to PTP oxidation, which promotes platelet stimulation.</P>