• 한국생물공학회:학술대회논문집
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• 2000.04a
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• pp.195-198
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• 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.

• Journal of Microbiology and Biotechnology
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• v.8 no.4
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• pp.325-332
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• 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.

• Proceedings of the Microbiological Society of Korea Conference
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• 1997.04a
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• pp.55.2-55.2
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• 1997

• Journal of Microbiology and Biotechnology
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• v.20 no.3
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• pp.480-484
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• 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.

• Journal of Microbiology and Biotechnology
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• v.24 no.1
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• pp.52-58
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• 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.

• Proceedings of the Microbiological Society of Korea Conference
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• 2004.05a
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• pp.85-86
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• 2004

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2002.10a
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• pp.101-101
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• 2002

• Journal of Food Hygiene and Safety
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• v.35 no.5
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• pp.438-446
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• 2020

Meju and nuruk (respectively soybean and malt) are traditional Korean fermentation starters that are vulnerable to contamination by harmful microorganisms such as aflatoxigenic fungi and their associated aflatoxins (AFs). In this study, Aspergillus spp. were isolated and identified from a total of 57 meju and 18 nuruk samples collected from Korean markets. Their potential aflatoxigenicity was investigated by examining the presence of three aflatoxin biosynthetic genes (aflO, aflP, and aflR) using multiplex polymerase chain reaction (mPCR) assays. Thereafter, aflatoxin production of isolates and the natural occurrence of AFs in meju and nuruk samples were analyzed by high-performance liquid chromatography (HPLC). A total of 177 Aspergillus isolates were identified and 130 isolates were obtained from meju samples. Of these, 25 isolates (19.2%) contained all three aflatoxin biosynthetic genes, and five (20%) of these isolates produced aflatoxins. Forty-seven of the Aspergillus isolates were obtained from nuruk samples, five of which (10.6%) expressed all three AF biosynthetic genes; however, none of these strains produced AFs. HPLC analysis showed that 88% (51/58) of the meju samples and 39% (7/18) of nuruk samples were not contaminated with AFs (below limit of detection). Among the isolates isolated from meju and nuruk, there were aflatoxigenic strains containing all three aflatoxin biosynthetic genes or producing aflatoxin in medium, but the frequency of aflatoxin contamination was low in the meju and nuruk samples.

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.420-422
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• 2005

TMC (Tautomycetin) is a liner polyketide immunosuppressive antifungal compound produced by Streptomyces spp. Inhibition of T cell proliferation with TMC was observed highly efficient at 100-fold lower than those needed to achieve maximal inhibition with cyclosporin A. To elucidate the biosynthetic pathway of TMC, a genomic DNA library was constructed using a E. coil-Streptomyces shuttle cosmid vector, pOJ446. The DNA libraries were screened by colony blot hybridization using several polyketide ${\beta}-ketosynthase$ (KS) probes amplified from TMC-producing Streptomyces genomic DNA using polymerase chain reaction (PCR), of which the degenerate primers were designed based on the highly conserved sequences present in KS domains of various type I polyketide synthase genes in Streptomyces species. This library construction and screening approach led to the isolation of several positive cosmid clones representing type I polyketide biosynthetic gene clusters. In addition, a Streptomyces regulatory gene called afsR2 (a global regulatory gene stimulating antibiotic production in both S. coelicolor and S. lividans) was successfully integrated into the TMC-producing Streptomyces chromosome via E. coil-Streptomyces heterologous conjugation mehtod. The more detailed results of production improvement and genetic characterization of TMC-producing Streptomyces spp. will be discussed.

• Proceedings of the Korean Society for Agricultural Machinery Conference
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• 2017.04a
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• pp.18-18
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• 2017

ISO11783은 농업 산업분야 통신 프로토콜의 국제 표준으로, 농용트랙터 및 작업기 ECU 간의 통신 프로토콜을 표준화한다. 이 표준은 서로 다른 제조사의 제품 간에 호환성을 갖게 하며, 정밀 농업에 대한 핵심 기반을 제공한다. 현재 해외에서는 이미 ISO11783 기반의 AFS(Advanced Farming System)를 통한 정밀농업이 상용화되어 농가에 보급되고 있다. 이에 비해, 국내에서 이러한 표준을 따르는 농기계들은 미비한 실정이며, 향후 농업의 정밀 농업화를 통한 고부가가치 창출 및 선진국의 무역 장벽에 대비와 해외 수출 판로 개척을 위해 ISO11783 표준에 대한 R&D가 필요로 한다. 이에 IsoAgLib를 분석하고 임베디드 보드에 Porting하여 ISO11783 기반 작업기 ECU를 구현하였고, 이를 기반으로 ISO11783 기반 작업기 ECU의 구현 방법을 발표한다. IsoAgLib의 시스템 아키텍처는 계층화 되어 있어, 타겟에 의존적인 계층만 수정하여, IAR 환경에서 Cortex M3 보드에 포팅을 완료하였다. 작업기 ECU들은 자신만의 인터페이스 화면을 갖으며, 이를 Object pool이라 한다. 이것을 Virtual Terminal(VT)에 업로드 하여, VT가 해당 작업기 ECU의 사용자 인터페이스 기능을 제공하도록 한다. 이에 작업기 ECU 구현 1 단계로, 'VT-Designer'를 통하여 Object pool를 설계한다. 2 단계, 'vt2iso'를 통해서 Object pool을 IsoAgLib 상에서 사용할 수 있도록 변환한다. 3 단계, 포팅된 IsoAgLib project에 변환된 파일을 포함 시킨다. 4 단계, 작업기 ECU의 주기적인 작업 및 각 메시지 수신시 수행할 작업을 코딩한다. 5 단계, 빌드 및 타겟 보드에 업로딩 한 후, New Holland 사의 $Intelliview^{TM}$ iv display (VT)과 연결하여 동작을 확인한다. 확인 결과로 VT에 디자인한 Object pool이 표시 되며 soft key 입력 시 작업기 ECU에서 LED가 변한다. 결론적으로, 연구 결과를 바탕으로 ISO11783 기반의 작업기 ECU의 디자인 및 구현이 가능하며, 이를 통해 향후 국내의 ISO11783 기반의 작업기 ECU의 개발에 도움을 줄 수 있다.