• 제목/요약/키워드: AroK

검색결과 38건 처리시간 0.018초

Isolation and ars Detoxification of Arsenite-Oxidizing Bacteria from Abandoned Arsenic-Contaminated Mines

  • Chang, Jin-Soo;Yoon, In-Ho;Kim, Kyoung-Woong
    • Journal of Microbiology and Biotechnology
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    • 제17권5호
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    • pp.812-821
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    • 2007
  • The ecosystems of certain abandoned mines contain arsenic-resistant bacteria capable of performing detoxification when an ars gene is present in the bacterial genome. The ars gene has already been isolated from Pseudomonas putida and identified as a member of the membrane transport regulatory deoxyribonucleic acid family. The arsenite-oxidizing bacterial strains isolated in the present study were found to grow in the presence of 66.7 mM sodium arsenate($V;\;Na_2HAsO_4{\cdot}7H_2O$), yet experienced inhibited growth when the sodium arsenite($III;\;NaAsO_2$) concentration was higher than 26 mM. Batch experiment results showed that Pseudomonas putida strain OS-5 completely oxidized 1 mM of As(III) to As(V) within 35 h. An arsB gene encoding a membrane transport regulatory protein was observed in arsenite-oxidizing Pseudomonas putida strain OS-5, whereas arsB, arsH, and arrA were detected in strain OS-19, arsD and arsB were isolated from strain RW-18, and arsR, arsD, and arsB were found in E. coli strain OS-80. The leader gene of arsR, -arsD, was observed in a weak acid position. Thus, for bacteria exposed to weak acidity, the ars system may cause changes to the ecosystems of As-contaminated mines. Accordingly, the present results suggest that arsR, arsD, arsAB, arsA, arsB, arsC, arsH, arrA, arrB, aoxA, aoxB, aoxC, aoxD, aroA, and aroB may be useful for arsenite-oxidizing bacteria in abandoned arsenic-contaminated mines.

Application of Dynamic Regulation to Increase L-Phenylalanine Production in Escherichia coli

  • Wu, Jie;Liu, Yongfei;Zhao, Sheng;Sun, Jibin;Jin, Zhaoxia;Zhang, Dawei
    • Journal of Microbiology and Biotechnology
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    • 제29권6호
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    • pp.923-932
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    • 2019
  • Current strategies of strain improvement processes are mainly focused on enhancing the synthetic pathways of the products. However, excessive metabolic flux often creates metabolic imbalances, which lead to growth retardation and ultimately limit the yield of the product. To solve this problem, we applied a dynamic regulation strategy to produce $\text\tiny{L}$-phenylalanine ($\text\tiny{L}$-Phe) in Escherichia coli. First, we constructed a series of Phe-induced promoters that exhibited different strengths through modification of the promoter region of tyrP. Then, two engineered promoters were separately introduced into a Phe-producing strain xllp1 to dynamically control the expression level of one pathway enzyme AroK. Batch fermentation results of the strain xllp3 showed that the titer of Phe reached 61.3 g/l at 48 h, representing a titer of 1.36-fold of the strain xllp1 (45.0 g/l). Moreover, the $\text\tiny{L}$-Phe yields on glucose of xllp3 (0.22 g/g) were also greatly improved, with an increase of 1.22-fold in comparison with the xllp1 (0.18 g/g). In summary, we successfully improved the titer of Phe by using dynamic regulation of one key enzyme and this strategy can be applied for improving the performance of strains producing other aromatic amino acids and derived compounds.

Biosynthesis of Two Flavones, Apigenin and Genkwanin, in Escherichia coli

  • Lee, Hyejin;Kim, Bong Gyu;Kim, Mihyang;Ahn, Joong-Hoon
    • Journal of Microbiology and Biotechnology
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    • 제25권9호
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    • pp.1442-1448
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    • 2015
  • The flavonoid apigenin and its O-methyl derivative, genkwanin, have various biological activities and can be sourced from some vegetables and fruits. Microorganisms are an alternative for the synthesis of flavonoids. Here, to synthesize genkwanin from tyrosine, we first synthesized apigenin from p-coumaric acid using four genes (4CL, CHS, CHI, and FNS) in Escherichia coli. After optimization of different combinations of constructs, the yield of apigenin was increased from 13 mg/l to 30 mg/l. By introducing two additional genes (TAL and POMT7) into an apigenin-producing E. coli strain, we were able to synthesize 7-O-methyl apigenin (genkwanin) from tyrosine. In addition, the tyrosine content in E. coli was modulated by overexpressing aroG and tyrA. The engineered E. coli strain synthesized approximately 41 mg/l genkwanin.

MS 마커를 활용한 지역별 오계 유전자원의 다양성 및 유연관계 분석 (Genetic Diversity and Relationship of Ogye Population in Korea Using 25 Microsatellite Markers)

  • 노희종;김관우;이진욱;전다연;김승창;전익수;고응규;이준헌;김성희;백준종;오동엽;한재용;이승숙;조창연
    • 한국가금학회지
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    • 제45권3호
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    • pp.229-236
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    • 2018
  • 본 연구는 연산오계(천연기념물 제265호)와 이를 기원으로 하는 5개 지역별 오계 집단의 유전적 특성 및 차별성을 분석하기 위해 25개의 초위성체(MS) 마커를 이용하여 총 9개 집단 243수를 대상으로 유전자형을 분석하였다. 마커별 다형성 분석 결과, 총 153개의 대립유전자가 확인되었으며, $H_{\exp}$와 PIC의 경우 MCW0145에서 각각 0.640, 0.570으로 가장 높았고, $H_{obs}$는 MCW0252에서 0.607로 가장 높은 값을 나타내었다. 반면, LEI0166에서 $H_{\exp}$, $H_{obs}$, PIC가 각각 0.248, 0.204, 0.202로 가장 낮았다. 집단간 유전거리 분석 결과로는 9개 집단중 YSO 집단과 SUO 집단이 가장 가까운(0.073) 반면, LG 집단과 CBO 집단 사이에서 가장 먼(0.937) 것으로 확인되었다. 집단의 실제 구조를 확인하기 위한 집단별 균일도를 분석한 결과, 공시된 9개의 집단은 3개의 집단으로 구분했을 때 최적의 K값(7.96)을 얻을 수 있었으며, 5개의 오계 집단(YSO, ARO, CBO, CNO, SUO) 및 LG 집단과 CN RIR 집단은 각각 1, 2, 3번 군집에 분포하고 있는 것으로 나타났다. 한편, GBO 집단의 경우 1번과 3번 클러스터에 걸쳐서 분포하고 있는 것으로 보아 사육과정에서 타집단과의 교잡이 일어났을 것으로 추정된다. 이러한 결과를 통해 추후 오계 유전자원에 대한 국가 수준의 유전적 특성평가 및 관리의 기초 자료로 유용하게 활용될 것으로 기대된다.

Roundtable Discussion at the UICC World Cancer Congress: Looking Toward the Realization of Universal Health Coverage for Cancer in Asia

  • Akaza, Hideyuki;Kawahara, Norie;Nozaki, Shinjiro;Sonoda, Shigeto;Fukuda, Takashi;Cazap, Eduardo;Trimble, Edward L.;Roh, Jae Kyung;Hao, Xishan
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권1호
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    • pp.1-8
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    • 2015
  • The Japan National Committee for the Union for International Cancer Control (UICC) and UICC-Asia Regional Office (ARO) organized a Roundtable Discussion as part of the official program of the UICC World Cancer Congress 2014 in Melbourne, Australia. The theme for the Roundtable Discussion was "Looking Toward the Realization of Universal Health Care 'UHC' for Cancer in Asia" and it was held on December 5, 2014. The meeting was held based on the recognition that although each country may take a different path towards the realization of UHC, one point that is common to all is that cancer is projected to be the most difficult disease to address under the goals of UHC and that there is, therefore, an urgent and pressing need to come to a common understanding and awareness with regard to UHC concepts that are a priority component of a post-MDG development agenda. The presenters and participants addressed the issue of UHC for cancer in Asia from their various perspectives in academia and international organizations. Discussions covered the challenges to UHC in Asia, collaborative approaches by international organizations, the need for uniform and relevant data, ways to create an Asia Cancer Barometer that could be applied to all countries in Asia. The session concluded with the recognition that research on UHC in Asia should continue to be used as a tool for cancer cooperation in Asia and that the achievement of UHC would require research and input not only from the medical community, but from a broad sector of society in a multidisciplinary approach. Discussions on this issue will continue towards the Asia-Pacific Cancer Conference in Indonesia in August 2015.

Aspergillus nidulans의 광 조건하 유성분화에 관여하는 silA 유전자의 분리 및 기능분석 (Isolation and Functional Analysis of the silA Gene That Controls Sexual Development in Response to Light in Aspergillus nidulans)

  • 한상용;고진아;김종학;한규용;한갑훈;한동민
    • 한국균학회지
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    • 제36권2호
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    • pp.189-195
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    • 2008
  • Aspergillus nidulans는 빛이 없는 조건에서는 유성분화가 주로 일어나고 빛이 있는 조건에서는 유성분화가 억제되고 대신 무성분화가 유도된다. 빛에 의해서 유성분화가 억제되는 것은 빛에 반응하여 유성 또는 무성분화를 조절하는 유전자가 있다는 것을 시사한다. 따라서 빛에 의해서 조절되는 유전자를 연구하기 위하여 광 조건하에서 유성분화를 하는 silA98 돌연변이를 분리하였으며, 이를 보완하는 유전자를 분리 및 분석하고자 A. nidulans의 AMA-NotI genomic library로부터 silA98 돌연변이를 상보하는 유전자 silA를 분리하였다. silA 유전자의 예상 ORF는 2,388 bp의 염기로 구성되어지고 795개의 아미노산을 암호화하고 있었다. 이 유전자는 Saccharomyces cerevisiae의 ARO80 유전자와 상동성을 보이며 SilA 단백질의 N 말단에는 약 51.9%의 상동성을 가지는 ${Zn_2}{Cys_6}$ motif를 지니고 있었다. silA 유전자 결손돌연변이주는 광 존재 하에서뿐만 아니라 고농도의 sorbitol에서도 유성분화가 유도되었다. 이는 silA 유전자가 빛과 고삼투 조건에서 유성분화를 억제하는 조절과정에 관여하고 있음을 의미한다. silA 유전자를 niiA promoter로 과다 발현시켰을 때의 형질은 야생형과 큰 차이를 보이지 않았다.

대사공학에 의해 개발된 코리네박테리움 글루타미컴에 의한 4-히드록시벤질 알코올 생산 (Production of 4-Hydroxybenzyl Alcohol Using Metabolically Engineered Corynebacterium glutamicum)

  • 김부연;정혜빈;이지영;페러 레니;푸완토 헨리 슈쿠르;이진호
    • 한국미생물·생명공학회지
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    • 제48권4호
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    • pp.506-514
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    • 2020
  • 4-Hydroxybenzyl alcohol (4-HB alcohol)은 두통, 경련 행동, 현기증과 같은 신경계 질환에 유익한 효과를 나타내며 천마의 주요 생리활성 성분 중의 하나이다. 대사공학을 통해 4-hydroxybenzoate (4-HBA)를 생산하는 균주로부터 4-HB alcohol을 생산하는 재조합 Corynebacterium glutamicum을 개발하였다. 먼저 4-HBA를 생산하는 APS809로부터 염색체 내 NCgl2922 유전자에 Methanocaldococcus jannaschii 유래의 aroK 유전자를 삽입한 APS963을 개발하였다. 4-HBA의 카로복실 산을 4-hydroxybenzaldehyde (4-HB aldehyde)로의 환원을 촉매하는 Nocardia iowensis 유래의 car 유전자를 염색체에서 발현하는 균주를 개발하기 위해 NCgl1112 유전자 일부 단편에 car 유전자가 삽입된 GAS177를 개발하였다. 더 높은 농도의 4-HB alcohol을 생산하기 위해 4-HB alcohol을 aldehyde로 산화를 촉매하는데 관여하는 creG 유전자를 염색체상에서 제거된 GAS255를 개발하였다. 최종적으로 chorismate를 4-HBA로 전환하는 효소의 유전자 ubiCpr을 pcaHG에 삽입된 GAS355를 개발하였으며, 80 g/l 포도당을 함유한 삼각플라스크에서 발효하여 생산성을 평가한 결과, 2.3 g/l 4-HB alcohol이 생산되었으며 부산물로 0.32 g/l 4-HBA, 0.3 g/l 4-HB aldehyde가 축적되었다.

Synthesis of Chlorogenic Acid and p-Coumaroyl Shikimates from Glucose Using Engineered Escherichia coli

  • Cha, Mi Na;Kim, Hyeon Jeong;Kim, Bong Gyu;Ahn, Joong-Hoon
    • Journal of Microbiology and Biotechnology
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    • 제24권8호
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    • pp.1109-1117
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    • 2014
  • Chlorogenic acid and hydroxylcinnamoyl shikimates are major dietary phenolics as well as antioxidants, with recently discovered biological, activities including protection against chemotheraphy side effects and prevention of cardiovascular disease and cancer. Certain fruits and vegetables produce these compounds, although a microbial system can also be utilized for synthesis of chlorogenic acid and hydroxylcinnamoyl shikimates. In this study, we engineered Escherichia coli to produce chlorogenic acid and p-coumaroyl shikimates from glucose. For the synthesis of chlorogenic acid, two E. coli strains were used; one strain for the synthesis of caffeic acid from glucose and the other strain for the synthesis of chlorogenic acid from caffeic acid and quinic acid. The final yield of chlorogenic acid using this approach was approximately 78 mg/l. To synthesize p-coumaroyl shikimates, wild-type E. coli as well as several mutants were tested. Mutant E. coli carrying deletions in three genes (tyrR, pheA, and aroL) produced 236 mg/l of p-coumaroyl shikimates.

Fermentation and Metabolic Pathway Optimization to De Novo Synthesize (2S)-Naringenin in Escherichia coli

  • Zhou, Shenghu;Hao, Tingting;Zhou, Jingwen
    • Journal of Microbiology and Biotechnology
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    • 제30권10호
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    • pp.1574-1582
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    • 2020
  • Flavonoids have diverse biological functions in human health. All flavonoids contain a common 2-phenyl chromone structure (C6-C3-C6) as a scaffold. Hence, in using such a scaffold, plenty of high-value-added flavonoids can be synthesized by chemical or biological catalyzation approaches. (2S)-Naringenin is one of the most commonly used flavonoid scaffolds. However, biosynthesizing (2S)-naringenin has been restricted not only by low production but also by the expensive precursors and inducers that are used. Herein, we established an induction-free system to de novo biosynthesize (2S)-naringenin in Escherichia coli. The tyrosine synthesis pathway was enhanced by overexpressing feedback inhibition-resistant genes (aroGfbr and tyrAfbr) and knocking out a repressor gene (tyrR). After optimizing the fermentation medium and conditions, we found that glycerol, glucose, fatty acids, potassium acetate, temperature, and initial pH are important for producing (2S)-naringenin. Using the optimum fermentation medium and conditions, our best strain, Nar-17LM1, could produce 588 mg/l (2S)-naringenin from glucose in a 5-L bioreactor, the highest titer reported to date in E. coli.