• 한국미생물·생명공학회지
• /
• 제22권4호
• /
• pp.333-339
• /
• 1994

In order to screen microorganisms producing phospholipase D (PLD) [EC 3.1.4.4], culture broths of about 900 strains of soil bacteria were subjected to examine for the PLD activity. When the hydrolytic activity of PLD (H-activity) in the supernatant was determined, 64 strains produced PLD more than 0.3 unit/ml and all of them were actinomycetes. Among 26 culture broths tested, 6 ones had transphosphatidylation activity (T-activity) of 30~68%. When the strains except one were cultivated on 3 different media at 30$\circ$C for 3 days under aerobic condition, strain # 1090 on medium B (yeast extract 1%, peptone 1%, glucose 1.5%, glycerol 1%, CaCO$_{3}$ 0.4%, and pH 7.2) produced PLD with much higher H- and T-activity, which were 8.3 units/ml and 76.3%, respectively. Subsequently, time course of PLD production of the strain # 1090 during cultivation with aeration of 1 v/v/m and agitation of 400 rpm at 30$\circ$C for 5 days on medium B in jar fermentor was investigated. H-activty of PLD reached almost maximum (about 9 units/ml) after 32 hours and maximal T-activity was found to be about 80%.

• 한국환경보건학회지
• /
• 제31권1호
• /
• pp.31-38
• /
• 2005

An anti-fungal material produced by actinomycetes was isolated from domestic soil. This actinomycetes was identified as Streptomyces albogriseus by 16S rDNA sequence. YEME (yeast extract 4 g, malt extract 10 g, glucose 4 g, D.W 1l, pH 7.00.2) medium was used for production of anti-fungal materials. S. albogriseus was cultured in a shaking incubator for 2 weeks at 150 rpm and $25^{\circ}C$. An anti-fungal material produced by S. albogriseus was identified at 340 nm by uv/vis- spectrometer and it showed powerful anti-fungal activity. This is the first report that secondary metabolite produced by S. albogriseus showed an activity against phytopathogenic fungi such as Collectrichum coccodes, Botrytis cinerea, Cladosporium cucumerinum, Didymella bryoniae.

• KSBB Journal
• /
• 제26권1호
• /
• pp.19-26
• /
• 2011

Of the 500 Actinomycetes isolates obtained from soil, one isolate grown on maltose as the sole carbon source produced compound BHK-P19, which inhibited the growth of multiple drug resistant P. aeruginosa 0245. Ultraviolet radiation mutagenesis curtailed production of BHK-P19. Mutation of the BHK-P19 producer using N-methyl-N'-nitro-N-nitroso-guanidine obviated the antibacterial activity to P. aeruginosa 0245, but not towards P. aeruginosa 0225. The mixing of BHK-P19 and BHK-S5 culture extracts inhibited P. aeruginosa 0254, 0225 and 1113. The combined application of BHK-P19 culture extract and Schizandra chinensis Baillon extract inhibited P. aeruginosa 0254, 0225, 0826, 1113, 1378, 1731 and 2492. Use of various concentrations of BHK-P19 culture extract and ampicillin markedly increased antibacterial activity against multi-drug resistant P. aeruginose 1113.

• The Plant Pathology Journal
• /
• 제22권2호
• /
• pp.107-114
• /
• 2006

The white cottony stem rot pathogen Sclerotinia scierotiorum was subjected to 70 different isolates of actinomycetes indigenous to Jordan as biological control agents. Forty of them demonstrated chitinase activity on crab shell chitin agay (CCA) media and they were segregated into three groups: 14 highly active, 12 moderately active, and 14 with low activity, with average clearing zones of (4.7-8.3), (3.7-4.3), and (2.3-3.3) mm surrounding colonies on CCA, respectively. Further, these isolates were able to inhibit radial mycelium growth of the pathogen and were categorized into three antagonistic groups: 13 strong, 13 moderate, and 14 weak antagonists, with antibiosis inhibition Bones of (32.0-45.7), (22.7-31.3), and (3.7-22.3) mm, respectively. High levels of chitinase activity of the isolates Ma3 (8.3 mm), Jul (7.7 mm), and Sa8 (7.7 mm) with their antagonistic activity against mycelium growth of 45.7, 44.3, and 40.7 mm were observed, respectively. These isolates exhibited fungicidal activity against sclevotia of S. sclerotiorum. On the other hand, isolates Na5, Aj3, and Aj2 that produced no chitinase showed fungistatic effect only.

• 한국유기농업학회지
• /
• 제19권4호
• /
• pp.589-602
• /
• 2011

In this study, experiments were conducted to determine the effect of different compost teas on plant growth reponses and yield of leaf lettuce. Compost tea is a liquid extract of compost obtained by mixing compost and water for a defined period of time. The pig manure and spent mushroom compost were made by steeping compost in water. Compost tea was aerated from 24 hours and molasses and kelp were added as supplements. The four types of compost were tested growth of lettuce. EC of animal manure compost tea was higher than that of spent mushroom compost tea. Mineral nutrients were significantly higher in animal manure compost tea compared with spent mushroom compost tea. Compost tea contains nutrient and a ranges of different organisms. The beneficial fungi and actinomycetes were prominent in a spent mushroom compost tea. Compost tea from animal manure had the higher numbers of total bacteria. The actinomycetes densities were high in spent mushroom compost tea. But actinomycetes were not founded in animal manure compost tea. The growth characteristics of lettuce in animal manure compost tea were higher than those of spent mushroom compost tea. And also SPAD value in leaf was high in plot treated with animal manure compost tea. The fresh yield of lettuce in animal compost tea was higher by 181% that of control plot. The effect of compost tea on growth of lettuce was largely attributable to mineral nutrient.

• Journal of Microbiology and Biotechnology
• /
• 제16권2호
• /
• pp.295-298
• /
• 2006

Actinomycetes are ubiquitous Gram-positive soil bacteria and a group of the most important industrial microorganisms for the biosynthesis of many valuable secondary metabolites as well as the source of various bioconversion enzymes. Cytochrome P450 hydroxylase (CYP), a hemebinding protein, is known to be involved in the modification of various natural compounds, including polyketides, fatty acids, steroids, and some aromatic compounds. Previously, six different novel CYP genes were isolated from a rare actinomycetes called Sebekia benihana, and they were completely sequenced, revealing significant amino acid similarities to previously known CYP genes involved in Streptomyces secondary metabolism. In the present study, these six CYP genes were functionally expressed in Streptomyces lividans, using an $ermE^{*}$ promoter-containing Streptomyces expression vector. Among six CYP genes, two S. benihana CYP genes (CYP503 and CYP504) showed strong hydroxylation activities toward 7-ethoxycoumarin. Furthermore, the recombinant S. lividans containing both the S. benihana CYP506-ferredoxin genes as well as the S. coelicolor feredoxin reductase gene also demonstrated cyclosporin A hydroxylation activity, suggesting potential application of actinomycetes CYPs for the biocatalysts of natural product bioconversion.

• 한국해양바이오학회지
• /
• 제14권2호
• /
• pp.143-154
• /
• 2022

We isolated marine actinomycetes, strain D-5 which produces anti-methicillin resistant Staphylococcus aureus (anti-MRSA) compound. Streptomyces sp. D-5 relatively grew well in the 20~25℃, pH 8.0, and NaCl 3.0%. The ethyl acetate extract of D-5 culture was separated by C₁₈ ODS open column and reverse phase HPLC to yield anti-MRSA compound. The molecular weight of this compound was determined to be 898 by a Liquid chromatograph-mass spectrometer (LC-MS). Compared with penicillin G, this compound showed significant anti-MRSA activity. It also exhibited an inhibition zone of 26 mm at a concentration of 64 ㎍/disk and an inhibition zone of 16 mm at a concentration of 16 ㎍/disk against the MRSA KCCM 40511. Furthermore, the co-treatment of HPLC peak 5 compound and vancomycin caused a more rapid decrease in MRSA cells than each compound alone. It showed 86.8% growth inhibition activity within 12 hours at a low concentration of 50 ㎍/mL during co-treatment, and 97.1% growth in-hibition activity within 48 hours against MRSA KCCM 40511. Taken together, our results suggest that Streptomyces sp. D-5 and its anti-MRSA compound could be employed as a potent agent in MRSA infection.

• 한국미생물·생명공학회지
• /
• 제32권1호
• /
• pp.78-83
• /
• 2004

국내 6개 지역에 걸쳐 채취한 토양 63점으로부터 1,000여 종의 방선균주를 분리하였다. 이들 균주의 배양 상징액 중 PLD의 생산이 양호한 균주를 탐색한 결과, 분해활성이 0.3U/$m\ell$이상인 131 균주를 일차로 선발하였다. 이중 전이 활성이 20% 이상인 균주는 23개였으며, 최종적으로 분해 및 전이활성이 가장 우수한 균주(방선균 KF923)를 선발하였다. 방선균 KF923 균주는 P 배지하에서 발효조 배양시 배양 48시간 후에 최고의 분해활성을 나타내었고(13U/$m\ell$) 전이활성은 95%로 나타났다. 방선균 KF923 균주가 생산한 PLD를 정제하여 비활성이 567U/mg의 PLD를 얻었다. 정제된 PLD는 분자량이 약 55kD으로 나타났고, 효소반응의 최적 pH는 6.0, 최적온도는 $60^{\circ}C$였으며, 효소의 안정성에 미지는 pH 및 온도의 영향은 pH 8.0부근 및 $40^{\circ}C$ 이하에서 가장 안정하였다. 또한, 특별한 금속이온의 요구성은 없는 것으로 나타났다. 방선균 KF923 균주가 생산한 PLD는 산업화에 충분히 활용 가능하며, 이를 위하여 효소의 대량생산 및 효소반응을 위한 공정개발 등의 연구가 필요하다고 생각한다.

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

• Applied Biological Chemistry
• /
• 제42권2호
• /
• pp.99-104
• /
• 1999

제주도내 25개 지점의 토양시료로부터 총 703 주의 방선균을 분리하였다. 네 종류의 배지 중 soil extract를 함유한 arginine glycerol salts agar가 다양한 방선균을 분리하는데 가장 적합하였으며, 목장초지 토양이 다른 지역의 토양에 비해서 방선균의 다양성과 밀도면에서 가장 좋은 분리원이었다 순수분리된 526개의 균주를 대상으로 Escherichia coli, Staphylococcus aureus 및 Pseudomonas solanacearum을 검정균으로 하여, 항균성 물질 생산여부를 검정한 결과, 생산균주의 발생빈도는 분리 및 배양에 사용한 배지의 종류에 따라서 $0{\sim}32.8%$로 많은 차이를 보였다. 세 종류의 검정균에 대하여 동시에 항균활성을 나타내는 10개 균주 중에서 pH 안정성과 열 안정성이 비교적 높은 물질을 생산하는 BL106Ba균주를 선택하여 등정한 결과, Streptomyces속에 속하며 S. albosporeus와 가장 유사한 것으로 추정되었다. B1106Ba의 배양액을 양이온 교환수지(AG MP-50)통과와 3차의 분자량 분획(Sephadex G-10)으로 정제하여 얻은 백색의 결정은 그람 음성균 및 양성균에 대해서는 강한 항균활성을 나타내었으나, 효모에 대해서는 약한 활성을 나타내었다. 이 결정 물질을 TLC와 HPLC로 분석한 결과 네 종류의 서로 다른 항균성 화합물이 혼합되어 있는 것으로 나타났다.