• 한국미생물·생명공학회지
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• 제25권3호
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• pp.253-257
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• 1997

The secE gene of Streptomyces lividans TK24 was cloned by the polymerase chain reaction method with synthetic oligonucleo- tide primers designed on the basis of the nucleotide sequences of Streptomyces coelicolor secE-nusG-rplK operon. The deduced amino acid sequences of the SecE were highly homologous to those of other known SecE protein, that is 36.8%, 30.4%, 80.0%, and 80.9%, similarity to E. coli, Bacillus subtilis, Streptomyces griseus, Streptomyces virginiae SecE, respectively and exactly same with Streptomyces coelicolor SecE. It means that in spite of evolutionary differences, the genes for protein translocation machinery are highly conserved in eubacteria. The gene organization of secE-nusG-rplK is also similar to that of E. coli, B. subtilis, and streptomycetes.

• Journal of Microbiology and Biotechnology
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• 제21권7호
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• pp.675-678
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• 2011

DNA methylation in Streptomyces griseus IFO 13350 was analyzed by high-performance liquid chromatographic analysis and bisulfite-based analysis to reveal two methylation sites, 5'-$GC^{5m}$ CGGC-3' and 5'-$GAG^{5m}$ CTC-3'. The methylation was reconstituted in Escherichia coli by simultaneous expression of S. griseus SGR4675 and S. achromogenes M.SacI. The E. coli cells produced plasmids that mimicked the methylation profile of S. griseus DNA, which was readily introduced into S. griseus. The results of this study raise the possibility of a promising approach to establish efficient transformation in several streptomycetes.

• 생명과학회지
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• 제7권3호
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• pp.186-191
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• 1997

수정된 선별법(modified selection method)을 사용하여 candiosis에 대한 새로운 생리활성물질(anticandiosis agent)을 생성하는 미생물을 분리하여Streptomyces sp. S627이라 명명하였으며, 이균의 발효액으로부터 methanol추출, diaion Hp-20, silica gel chromatography, 및 분취용 TLC 등의 방법을 사용하여 물리화학적 특성이 다른 S-637(A)와 S-627(B)을 분리하였다.

• 한국미생물·생명공학회지
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• 제39권2호
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• pp.104-110
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• 2011

방선균은 그램양성 토양 박테리아로서 항생제, 항암제, 항구충제, 면역억제제 등 유용한 2차 대사산물을 생산하는 유용 산업미생물이다. 비록 대부분의 방선균이 속해있는 스트렙토마이세스는 지난 수 십 년간 분자수준에서의 연구가 집중적으로 진행되어 왔으나, 최근에 분리된 잠재적 유용성을 갖는 스트렙토마이세스 이외의 희소방선균들은 유전자 조작시스템의 부재로 그 특성이 잘 규명되지 않고 있다. 본 연구에서는 독립적으로 분리된 180 여 방선균주들 중에서 희소방선균만을 선별하기 위하여 중합효소연쇄반응을 이용한 유전체 스크리닝 전략을 시도하였으며, 이 전략을 통하여 7종의 희소방선균을 성공적으로 분리하였다. 특히 여러 생리활성 테스트를 통하여, 항진균 및 항생제 활성을 띄는 잠재적 유용성이 높은 노카이디옵시스 균주 MMBL010을 선별하였다. 또한 전통적인 방선균 유전자 조작기법이 작동하지 않는 본 MMBL010 균주를 대장균 접합을 통한 유전자 전달 시스템도 최적화시킴으로써, 유전체 스크리닝을 통한 유용희소방선균의 선별 및 유전자 조작시스템 구축은 궁극적으로 희소방선균의 잠재적 유용성을 극대화시킬 수 있는 효율적인 전략으로 사료된다.

• Journal of Microbiology and Biotechnology
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• 제26권1호
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• pp.66-71
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• 2016

PikD is a widely known pathway-specific regulator for controlling pikromycin production in Streptomyces venezuelae ATCC 15439, which is a representative of the large ATP-binding regulator of the LuxR family (LAL) in Streptomyces sp. RapH and FkbN also belong to the LAL family of transcriptional regulators, which show greatest homology with the ATP-binding motif and helix-turn-helix DNA-binding motif of PikD. Overexpression of pikD and heterologous expression of rapH and fkbN led to enhanced production of pikromycin by approximately 1.8-, 1.6-, and 1.6-fold in S. venezuelae, respectively. Cross-complementation of rapH and fkbN in the pikD deletion mutant (ΔpikD) restored pikromycin and derived macrolactone production. Overall, these results show that heterologous expression of rapH and fkbN leads to the overproduction of pikromycin and its congeners from the pikromycin biosynthetic pathway in S. venezuelae, and they have the same functionality as the pathwayspecific transcriptional activator for the pikromycin biosynthetic pathway in the ΔpikD strain. These results also show extensive "cross-communication" between pathway-specific regulators of streptomycetes and suggest revision of the current paradigm for pathwayspecific versus global regulation of secondary metabolism in Streptomyces species.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XII)
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• pp.422-427
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• 2003

Streptomyces setonii (ATCC 39116) is a Gram-positive thermophilic soil actinomycetes capable of degrading single aromatic compounds including phenol and benzoate via ortho-cleavage pathway. we isolated approximately 6.3-kb S. setonii DNA fragment containing a thermophilic catechol 1,2-dioxygenase(C12O) gene. Here we further revealed that the 6.3-kb S. setonii DNA fragment was organized into two putative divergently-transcribed clusters with 6 complete and one incomplete open reading frames (ORFs). The first cluster with 3 ORFs showed significant homologies to previously known benA, benB, and benC, implying a part of benzoate catabolic operon. The second cluster revealed an ortho-cleavage catechol catabolic operon with three translationally-coupled ORFs (catR, catB, catA). Each of these individually-cloned ORFs was expressed in E. coli and identified as a distinct protein band with a theoretical molecular weight in SDS-PAGE. The expression of the cloned S. setonii catechol operon was induced in a heterologous S. lividans by specific single aromatic compounds including catechol, phenol, and 4-chlorophenol. The simitar induction pattern was also observed using a luciferase gene-fused reporter system, implying that S. setonii employs an inducer-specific regulatory mechanism for aromatic compound metabolism.

• Journal of Microbiology and Biotechnology
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• 제24권9호
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• pp.1226-1231
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• 2014

The rare actinomycete Pseudonocardia autotrophica was previously shown to produce a solubility-improved toxicity-reduced novel polyene compound named $\underline{N}ystatin$-like $\underline{P}seudonocardia$ $\underline{P}olyene$ (NPP). The low productivity of NPP in P. autotrophica implies that its biosynthetic pathway is tightly regulated. In this study, $wblA_{pau}$ was isolated and identified as a novel negative regulatory gene for NPP production in P. autotrophica, which showed approximately 49% amino acid identity with a global antibiotic down-regulatory gene, wblA, identified from various Streptomycetes species. Although no significant difference in NPP production was observed between P. autotrophica harboring empty vector and the S. coelicolor wblA under its native promoter, approximately 12% less NPP was produced in P. autotrophica expressing the wblA gene under the strong constitutive $ermE^*$ promoter. Furthermore, disruption of the $wblA_{pau}$ gene from P. autotrophica resulted in an approximately 80% increase in NPP productivity. These results strongly suggest that identification and inactivation of the global antibiotic down-regulatory gene wblA ortholog are a critical strategy for improving secondary metabolite overproduction in not only Streptomyces but also non-Streptomyces rare actinomycete species.

• Environmental Engineering Research
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• 제19권1호
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• pp.67-73
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• 2014

In India, disposal of vegetable market waste along with municipal solid waste in landfills or dumpsites is creating much nuisance in terms of odor nuisance, leachate production, and greenhouse gas emission into the atmosphere. Therefore, vegetable waste with high biodegradable and nutrient content is composted in a 550-L batch scale rotary drum composter to study the degradation process and its compost properties for its potential reuse as high quality compost. A total 150 kg of working volume was fixed for composting studies with two different ratios, trial A (6:3:1) of C/N 24 and trial B (8:1:1) of C/N 30, respectively. A maximum of $63.5^{\circ}C$ and $61.2^{\circ}C$ was observed in trials A and B; an average of $55^{\circ}C$ for more than 5 days, which helped in the degradation of organic matter and reduction of total and fecal coliform. The temperature dropped suddenly after the thermophilic stage in trial B, and leachate was observed due to insufficient amount of bulking agent. Mesophilic bacteria dominated during the initial stages of composting, and reduced considerably during the thermophilic stage. During the thermophilic stage, the rise in spore-forming organisms, including spore-forming bacteria, fungi, actinomycetes and streptomycetes, increased and these were predominant until the end of the composting process. By examination, it was observed that moisture and leachate production had adverse effects on the compost parameters with higher loss of micronutrients and heavy metals.

• 미생물학회지
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• 제43권4호
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• pp.321-324
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• 2007

Streptomyces 균주들의 진탕배양 시간에 따른 항원성 변화를 면역확산 침강법과 간접 ELISA법으로 측정하였다. 각 균주들에서 모두 색소 생산 시기에 나타난 새로운 침강선이 배양시간이 경과되면서 진해지는 양상을 보였다. S. coelicolor와 S. griseus의 항원성 변화는 상대적으로 약하게 나타난 반면 S. lavendulae와 S. viridochromogenes의 항원성 변화는 강하게 나타났다. 이 결과는 균주에 따라 정도의 차이는 있지만 Streptomyces 균체의 진탕배양에서의 생장이 균체의 외피 성분의 변화를 수반하며 이를 정량적으로 분식할 수 있음을 나타낸다.

• Journal of Microbiology
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• 제34권2호
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• pp.105-116
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• 1996

Of actinomycetes isolated from volcanic compost soils, 115 representative strains which showed distinctive morphologicla features were numerically classified, compared with reference strains of Streptomyces. One hundred and twenty unit characters were tested and the average probability of error was 4.27%. The cluster analysis resulted in two groups: group A included strains of actinomycetes except streptomycetes. Group A was divided into 2 major clusters (over 5 strains), 10-diaminopimelic acid. Group B was divided into 5 clusters, of which 4 clusters contained mesodiminopimelic acid and 1 cluster LL-diaminopimelic acid. The major clusters of group A showed higher abilities of substrate utilization and degradation, and higher resistance to inhibitors, whereas the minor and single member clusters of group A showed relatively higher antimicrobial activities. On the other hand, all clusters of group B showed relatively lower abilities of substrate utilization and degradation and lower resistance to inhibitors.