• 제목/요약/키워드: afsR

검색결과 27건 처리시간 0.024초

Enhanced Clavulanic Acid Production in Streptomyces clavuligerus NRRL3585 by Overexpression of Regulatory Genes

  • Hung, Trinh Viet;Ishida, Kenji;Parajuli, Niranjan;Liou, Kwang-Kyoung;Lee, Hei-Chan;Sohng, Jae-Kyung
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • 제11권2호
    • /
    • pp.116-120
    • /
    • 2006
  • We constructed four recombinant plasm ids to enhance the production of clavulanic acid (CA) in Streptomyces clavuligerus NRRL3585: (1) pIBRHL1, which includes ccaR, a pathway-specific regulatory gene involved in cephamycin C and CA biosynthesis; (2) pIBRHL2, containing claR, again a regulatory gene, which controls the late steps of CA biosynthesis; (3) pGIBR containing afsR-p, a global regulatory gene from Streptomyces peucetius; and (4) pKS, which harbors all of the genes (ccaR/ claR/ afsR-p). The plasmids were expressed in S. clavuligerus NRRL3585 along with the $ermE^*$ promoter. All of them enhanced the production of CA; 2.5-fold overproduction for pIBRHL1, 1.5-fold for pIBRHL2, 1.6-fold for pGIBR, and 1.5-fold for pKS compared to the wild type.

Enhancement of Avermectin Production by Introduction of a Foreign Regulatory Gene in Streptomyces avermitilis

  • 황용순;이재영;김상수;김응수;최차용
    • 한국생물공학회:학술대회논문집
    • /
    • 한국생물공학회 2000년도 춘계학술발표대회
    • /
    • pp.195-198
    • /
    • 2000
  • Production of eight avermectin components was improved in Streptomyces avermitilis wild type strain (ATCC31267) and high producing mutant strain (ATCC31780) when transformed with a foreign regulatory gene, afsR2 of Streptomyces lividans. Wild type and the high producing strain of S. avermitilis transformed with multiple copies of afsR2 improved total avermectin productions by 2.3 fold and 1.5 fold, respectively. In both of wild type and the high producing transformants carrying afsR2, glycerol was proved to be the best carbon source for the stimulation of avermectin production.

  • PDF

Effects of Protein Kinase Inhibitors on In Vitro Protein Phosphorylation and on Secondary Metabolism and Morphogenesis in Streptomyces coelicolor A3(2)

  • Hong, Soon-Kwang;Sueharu, Horinouchi
    • Journal of Microbiology and Biotechnology
    • /
    • 제8권4호
    • /
    • pp.325-332
    • /
    • 1998
  • In vitro phosphorylation experiments with a cell extract of Streptomyces coelicolor A3(2) M130 in the presence of ${\gamma}-[^32P]$]ATP revealed the presence of multiple phosphorylated proteins, including the AfsR/AfsK kinases which control the biosynthesis of A-factor, actinorhodin, and undecylprodigiosin. Phosphorylation of AfsR by a cell extract as an AfsK source was significantly inhibited by Ser/Thr protein kinase inhibitors, staurosporine and K-252a, at concentrations giving 50% inhibition ($IC_50$) of $1{\mu}M\;and\;0.1{\mu}M$, respectively. Further in vitro experiments with the cell extracts showed that phosphorylation of multiple proteins was inhibited by various protein kinase inhibitors with different inhibitory profiles. Manganese and calcium ions in the reaction mixture also modulate phosphorylation of multiple proteins. Manganese at 10 mM greatly enhanced the phosphorylation and partially circumvented the inhibition caused by staurosporine and K-252a. A calcium-activated protein kinase(s) was little affected by these inhibitors. Herbimycin and radicicol, which are known as tyrosine kinase inhibitors, did not show any significant inhibition of AfsR phosphorylation. Consistent with the in vitro effect of the kinase inhibitors, they inhibited aerial mycelium formation and pigmented antibiotic production on solid media. On the contrary, when assayed in liquid culture, the amount of actinorhodin produced was increased by staurosporine and K-252a and greatly decreased by manganese. All of these data clearly show that the genus Streptomyces possesses several protein kinases of eukaryotic types which are involved in the regulatory network for morphogenesis and secondary metabolism.

  • PDF

Enhancement of Herboxidiene Production in Streptomyces chromofuscus ATCC 49982

  • Jha, Amit Kumar;Lamichhane, Janardan;Sohng, Jae Kyung
    • Journal of Microbiology and Biotechnology
    • /
    • 제24권1호
    • /
    • pp.52-58
    • /
    • 2014
  • Structurally, herboxidiene contains the tetrahydropyran acetic acid moiety and a side chain including a conjugated diene, and has been isolated from Streptomyces chromofuscus ATCC 49982. Its production was significantly elevated nearly 13.5-fold (0.74 g/l) in a medium supplemented with glycerol (medium No. 6A6), and was more efficacious (1.08 g/l; 19.8-fold) in fed-batch fermentation at 36 h in medium No. 6A6, from Streptomyces chromofuscus. For further enhancement, regulatory genes metK1-sp and afsR-sp from Streptomyces peucetius were overexpressed using an expression vector, pIBR25, and similarly ACCase from Streptomyces coelicolor and two genes, metK1-sp and afsR-sp, were also overexpressed using an integration vector, pSET152, under the control of the strong $ermE^*$ promoter in Streptomyces chromofuscus. Only the recombinant strains S. chromofuscus SIBR, S. chromofuscus GIBR, and S. chromofuscus AFS produced more herboxidiene than the parental strain in optimized medium No. 6A6 with an increment of 1.32-fold (0.976 g/l), 3.85-fold (2.849 g/l), and 1.7-fold(1.258 g/l) respectively.

Proteomics-Driven Identification of SCO4677-Dependent Proteins in Streptomyces lividans and Streptomyces coelicolor

  • Choi, Si-Sun;Kim, Seon-Hye;Kim, Eung-Soo
    • Journal of Microbiology and Biotechnology
    • /
    • 제20권3호
    • /
    • pp.480-484
    • /
    • 2010
  • AfsR2 is a global regulatory protein that stimulates antibiotic biosynthesis in both Streptomyces lividans and S. coelicolor. Previously, various afsR2-dependent genes including a putative abaA-like regulatory gene, SCO4677, were identified through comparative DNA microarray analysis. To further identify the putative SCO4677-dependent proteins, the comparative proteomics-driven approach was applied to the SCO4677-overexpressing strains of S. lividans and S. coelicolor along with the wild-type strains. The 2D gel electrophoresis gave approximately 277 protein spots for S. lividans and 207 protein spots for S. coelicolor, showing different protein expression patterns between the SCO4677-overexpressing strains and the wild-type strains. Further MALDI-TOF analysis revealed that only 18 proteins exhibited similar expression patterns in both S. lividans and S. coelicolor, suggesting that the SCO4677 could encode an abaA-like regulator that controls a few cross-species common proteins as well as many species-specific proteins in Streptomyces species.

Automotive Adaptive Front Lighting Requiring Only On/Off Modulation of Multi-array LEDs

  • Lee, Jun Ho;Byeon, Jina;Go, Dong Jin;Park, Jong Ryul
    • Current Optics and Photonics
    • /
    • 제1권3호
    • /
    • pp.207-213
    • /
    • 2017
  • The Adaptive Front-lighting System (AFS) is a part of the active safety system, providing optimized vision to the driver during night time and other poor-sight conditions of the road by automatic adaptation of lighting to environmental and traffic conditions. Basically, an AFS provides four different modes of the passing beam as designated in an United Nations Economic Commission for Europe regulation (ECE324-R123): neutral state or country light (Class C), urban light (Class V), highway light (Class E), and adverse weather light (Class W). In this paper, we first present an optics design for an AFS system capable of producing the Class C/V/E/W patterns requiring only on/off modulation of multi-array LEDs with no need for any additional mechanical components. The AFS optics consists of two separated modules, cutoff and spread; the cutoff module lights a narrow central area with high luminous intensity, satisfying the cutoff regulation, and the spread module forms a wide spread beam of low luminous intensity. Each module consists of two major parts; the first converts a discretely positioned LED array into a full-filled area emitting light source plane, and the second projects the light source plane to a 25 m away target plane. With the combination of these two optics modules, the four beam patterns are formed by simple on/off modulation of multi-array LEDs. Then we report the development of a prototype that was demonstrated to provide the four beam patterns.

방오페인트에 함유된 유기주석물질 분석방법 연구 (Study of an analytical method for determining organotin compounds in anti-fouling paints)

  • 김차수;이상은;윤주용;박일남;김만중;김인구;오현정
    • 해양환경안전학회:학술대회논문집
    • /
    • 해양환경안전학회 2007년도 추계학술발표회
    • /
    • pp.35-41
    • /
    • 2007
  • An International Convention on the control of harmful anti-fouling system on ships(AFS Convention) was adopted on 5 October 2001 at Diplomatic Conference in London, and is expected to be presently effectuated with ratification of more than 25-member nations possessing about 25% of total world tonnage. This convention regulates the operation of harmful anti-fouling system and especially prohibits the use of organotin compounds contained in anti-fouling paint. Organotin compounds have a tendency to be easily extracted by specific solvents and have high polarity and low volatility as specific characteristics. This drives us to attempt of going through the process named derivatization that is required in analysis using a gas chromatography(GC). This study was conducted to determine the proper pre-treatment method, ethylation in comparison with hydridization on the analysis of tributyltin in organotin compounds and to verify the application of the method through the experimental analysis practically used anti-fouling paint and painted layer sample of the served ship.

  • PDF