The beet armyworm, Spodoptera exigua, is one of the worldwide distributed agricultural pest insects and has been known to show high resistance to conventional chemical insecticides. Since it has been reported that secondary metabolites from actinomycetes show insecticidal activities against various insect pests, actinomycetes could be a potential source of insecticidal compounds. In this study, culture extracts of Streptomyces strains were tested for their insecticidal activity against Spodoptera exigua. Among them, culture extracts of IMBL-0003 strain which was identified as Streptomyces celluloflavus showed a high insecticidal activity (more than 90% mortality). These results suggested that secondary metabolites of this isolate could have potentials to be a efficient eco-friendly pesticide for controling Spodoptera exigua.

In order to investigate novel insect growth regulators (IGRs) from actinomycetes, 363 isolates of actinomycetes were tested for their IGR activities. Among them, Streptomyces sp. AN120537 showed the highest juvenile hormone antagonist (JHAN) activity and significant insecticidal activities against larvae of Aedes albopictus and Plutella xylostella. In addition, the dead larvae showed morphological deformities such as contraction of body segments or pigmentation of all body parts. Through liquid chromatography and bioassay-guided fractionation, 7 IGR compounds were identified from the crude hexane extract of AN120537. These results suggested that Streptomyces sp. AN120537 could be useful resources for development of eco-friendly insecticidal agents.

Streptomyces has been reported to produce various secondary metabolites which have the potential to become environmentally safe insecticides. In this study, 1,274 streptomyces culture filtrates were screened for their JHAN activity in order to identify novel insecticidal compounds. 34 isolates with high levels of JHAN activity were selected, and their insecticidal activities were tested against Plutella xylostella larvae. Among them, IMBL-263 which was revealed to be Streptomyces anulatus by 16s rRNA sequencing showed the highest insecticidal activity. Also, systemic activities of secondary metabolites extracted from the S. anulatus on plant, Brassica napus, were investigated. These results suggested that secondary metabolites from the S. anulatus might be useful for development of novel environmentally benign insecticides.

Beet armyworm, Spodoptera exigua, is known to be hard-to-kill pest by having high resistance to insecticides and its control is intensely dependent on fewer insecticides. In this study, the ethyl acetate extract of Streptomyces sp. was evaluated to find novel insecticidal agents against S. exigua. In order to determine the identity of insecticidal compounds, the crude ethyl acetate extract was fractionated based on the TLC profiling and bioassay-guided monitoring. These results indicated that the non-polar fraction have high level of insecticidal activity against second instars of S. exigua. These findings suggested that secondary metabolites produced by Streptomyces sp. could be considerable potential resources as novel insecticidal formulation candidates.

Streptomyces species have been studied to find potent pest control agents as an alternatives to chemical insecticides.Previously, one of the ethyl acetate extract of Streptomyces isolates cultured on unpolished rice medium showed highlevel of juvenile hormone antagonist and larvicidal activities against pest insects including Aedes albopictus, Plutella xylostellaand Laodelphax striatellus. It has been known that the biosynthesis of secondary metabolites of Streptomyces could beinfluenced by a variety environments such as nutritional composition and growth conditions. In this study, to optimizeculture conditions for stable production of insecticidal compounds from this isolate, binding assay and bioassay-guidedmonitoring were conducted using various culture conditions.

Alginate lyase from Streptomyces violaceoruber was purified by DEAE sephacel chromatography and SP sepharose chromatography. The specific activity of the purified enzyme was 14.6 units/mg protein, representing a 40.6-fold purification of the crude extract. The final preparation thus obtained showed a single band on Tricine-SDS polyacrylamide gel electrophoresis whose molecular weight was determined to be 23.3 kDa. The polyMG block of sodium alginate was hydrolyzed by the purified alginate lyase and then separated by activated carbon column chromatography and bio gel P-2 gel filtration. The main hydrolysates were composed of hetero type M/G-oligosaccharides with the degrees of polymerization (D.P.) being 6 and 8. To investigate the effects of hetero type M/Goligosaccharides from the sodium alginate on the growth of some intestinal bacteria, cells were cultivated individually on the modified-MRS medium containing D.P. 6 and 8 M/G-oligosaccharides. B. longumgrew 4.25-fold and 6.44-fold more effectively by the treatment of D.P. 6 and 8 M/G-oligosaccharides compared with those of standard MRS medium. In addition, B. bifidumgrew 3.3-fold and 5.4-fold more effectively by the treatment of D.P. 6 and 8 M/G-oligosaccharides. In conclusion, D.P. 8 was more effective than D.P. 6 hetero M/G-oligosaccharides as regards the growth of Bifidobacteriumspp. and Lactobacillus spp

Alginate lyase from Streptomyces violaceoruber was purified by DEAE sephacel chromatography and SP sepharose chromatography. The specific activity of the purified enzyme was 14.6 units/mg protein, representing a 40.6-fold purification of the crude extract. The final preparation thus obtained showed a single band on Tricine-SDS polyacrylamide gel electrophoresis whose molecular weight was determined to be 23.3 kDa. The polyMG block of sodium alginate was hydrolyzed by the purified alginate lyase and then separated by activated carbon column chromatography and bio gel P-2 gel filtration. The main hydrolysates were composed of hetero type M/G-oligosaccharides with the degrees of polymerization (D.P.) being 6 and 8. To investigate the effects of hetero type M/Goligosaccharides from the sodium alginate on the growth of some intestinal bacteria, cells were cultivated individually on the modified-MRS medium containing D.P. 6 and 8 M/G-oligosaccharides. B. longumgrew 4.25-fold and 6.44-fold more effectively by the treatment of D.P. 6 and 8 M/G-oligosaccharides compared with those of standard MRS medium. In addition, B. bifidumgrew 3.3-fold and 5.4-fold more effectively by the treatment of D.P. 6 and 8 M/G-oligosaccharides. In conclusion, D.P. 8 was more effective than D.P. 6 hetero M/G-oligosaccharides as regards the growth of Bifidobacteriumspp. and Lactobacillus spp. Key words: hetero M/G-oligosaccharides, Streptomyces violaceoruber

Streptomyces is the largest genus of Actinobacteria that forms fungus-like branched networks of hyphae. Streptomyces has been clinically important because they produce various secondary metabolites with antibacterial, antifungal, and nematocidal activities. In order to explore novel insecticidal compounds, extracts from 363 strains of Actinobacteria were screened for their juvenoid and anti-juvenoid activities using yeast-two hybrid system. Among them, extract of Streptomyces spp. showed high anti-juvenoid activity. This extract also showed high level of insecticidal activities against larvae of Aedes albopictus, Laodelphax striatellus, and Ostrinia furnacalis. These results suggested that the secondary metabolites of Streptomyces could be natural sources of novel insecticidal compounds.

소나무재선충에 대하여 살선충 활성을 가지는 친환경 물질을 탐색하기 위하여 생리활성물질 분리율이 높은 방선균을 이용하였다. 방선균은 충북 영동, 충남 태안도, 경북 문경과 상주의 산림에서 채집한 토양으로부터 분리, 배양하여 실험에 이용하였다. 분리한 방선균은 모두 32개이며 방선균 배양여액을 소나무재선충에 직접 노출시켜 48시간 후 소나무재선충의 치사 여부를 조사하였고 방선균이 소나무재선충의 증식에 미치는 영향을 알아보기 위하여 직접 노출 후 잿빛곰팡이병균(Botrytis cinerea)이 충분히 자란 PDA(potato dextrose agar)에 접종한 후 배양기에서 15일에서 30일 동안 보관 후 소나무재선충의 증식 밀도를 조사하였다. 방선균에 직접 노출시켜 살선충 활성을 알아본 결과 상주 갑장산 지역에서 분리한 5개 방선균 균주의 보정사충률이 100%를 나타내었으며 32개의 방선균 중 21개가 보정사충률이 90% 이상이었다. 증식에 미치는 영향을 알아본 결과 소나무재선충의 증식률이 AM210, SG16, YD116, YD315 균주에서 10% 미만으로 나타났다. 그 중 YD116 균주를 형태적, 분자적으로 동정한 결과 Streptomyces atratus YD116이었으며 S. atratus에서 분리된 물질들 중 hydrazidomycin과 유사한 hydrazine hydrate로 실험한 결과 1000ppm에서 97.8%의 보정사충률을 나타내었다.

A xylanolytic microorganism, strain DY-7, was isolated from the gut of the mole cricket, Gryllotalpa orientalis. The result of phylogenetic analysis based on its 16S rDNA sequence revealed that the isolate was a Gram-positive bacterium belonging to the genus Streptomyces. The cloned gene (1350-bp) encoding a GH family 10 β -1,4-xylanase (XylA) from Streptomyces sp. strain DY-7 was overexpressed in Escherichia coli BL21 and its gene products were characterized. The hydrolysis activities of rXylA and rXylAΔCBD II against xylosidic materials were maximum at pH 5.5 and 65oC. However, deletion of CBD II in the C-terminus region of XylA significantly increased the thermal stability of the enzyme at high temperatures above 50oC. The xylanolytic activity of rXylA was slightly enhanced in the presence of 1 mM Mn2+ and 5 mM sodium azide but it was completely inactivated by 1 mM Hg2+ and 5 mM N-bromosuccinimide. rXylA was capable of efficiently decomposing various xylosidic compounds, PNP-cellobioside, and PNP-xylopyranoside, whereas other hexose-based compounds were insensitive to the enzyme. The specific activities of rXylA toward oat spelts xylan and PNP-cellobioside were 649.8 U/mg and 328.1 U/mg, respectively. Enzymatic degradation of birchwood xylan and xylooligosaccharides (xylotriose to xylohexaose) resulted in the production of xylobiose (>75%) as the main hydrolysis product together with a small amount (4%<) of xylose as the final hydrolysis product.

Streptomyces padanus IA70-5 has been shown to be a promising biological control agent for the suppression of pepper anthracnose. In this study, we assessed the potential use of strain IA70-5 as a biological control agent for Phytophthora blight caused by Phytophthora capsici. Strain S. padanus IA70-5 was found to inhibit the mycelial growth and zoosporangium formation of P. capsici causing Phytophthora blight on pepper plants. In experiments with hot pepper fruit, IA70-5 suppressed the progression of Phytophthora rot by over 90% in pre-inoculated treatments with culture suspension. In experiments with 60-day-old pepper plants, IA70-5 suppressed Phytophthora blight by over 90%. These results demonstrated the potential for S. padanus IA70-5 to provide a practical biological agent for the control of Phytophthora blight in the field.

고추에 큰 피해를 일으키는 탄저병에 대한 강한 길항력이 있는 세균을 선발하기 위해 선행연구에서 식물뿌리 시료로부터 분리하여 보관중인 세균들을 대상으로 검정에 사용하였다. 총 457균주로부터 IA70-5균주를 최종 선발하였고, 16S rDNA 염기서열 분석을 통해 Streptomyces padanus로 동정하였다. S. padanus IA70-5는 색소를 분비하지 않고 운동성이 없으며 전형적인 Streptomyces속에 속하는 세균들처럼 나선형의 형태를 이루고 있었다. S. padanus IA70-5 균주는 in vitro에서 Colletotrichum acutaum의 균사생장, 포자발아, 그리고 부착기 형성을 효과적으로 억제하였다. 실내 고추 과실에 대하여 병원균 접종 전 IA70-5 배양액 처리 시 약 90%의 탄저병 억제효과를 나타내었다. 본 연구결과를 통하여 길항방선균 S. padanus IA70-5는 고추 탄저병을 억제하는 효과가 있음을 확인하였다.

방선균은 토양 속에 다양하게 존재하는 미생물의 일종으로 그람 양성 진정세균으로 이차 대사산물을 생산하는 시기와 포자 착생이 시작되는 세포분화의 시기가 밀접한 관련이 있다. S. griseus는 streptomycin을 비롯한 다양한 종류의 endopeptidase 및 exopeptidase들을 생산한다. 방선균에서의 protease 생산은 많은 경우에 이차대사산물이 형성되거나 형태분화가 유도되는 시기에 동시에 시작된다는 점에서 protease가 이차대사물질 생산 및 세포분화에 일정한 기능을 수행할 것이라는 점을 시사하고 있다. 본 연구에서는 S. griseus IFO 13350에서 클로닝한 SGPE protease가 각 strain에서 형태학적으로나 생리적으로 어떠한 gene dosage 효과를 미치는지 조사하는 것이었다. sprD 유전자가 S. lividans를 숙주로 사용한 시스템에서 대량발현이 성공적으로 되는 것을 확인한 후, 본 유전자를 클로닝한 S. griseus IFOI3350 균주와 이의 A-factor 결손주인 S. griseus HHI에 형질전환하였다. S. griseus HHI과 S. griseus IFO13350 에서는 protease activity가 벡터만 도입된 대조군과 sprD 유전자가 들어간 형질전환체에서 큰 차이를 보이지 않았다. 또한 S. griseus IFO13350 및 HHI 모두에서 생리학적·형태학적 분화의 차이를 발견하지 못하였다. Chymotrypsin 계열의 protease를 암호화하는 유전자만이 S.griseus에서 발현이 repression 된다는 사실을 본 연구 결과를 통하여 알게 되었다. 이를 바탕으로 sprD 유전자와 동일계열의 chymotrypsin 계열의 유전자들이 공통적으로 S. griseus 에서 repression되는 일반적인 기전이 있을 것으로 판단, chymotrypsin 계열 유전자들의 promoter 부분의 염기 상동성을 조사하였다. 번역개시부위 바로 상부 유전자부터 상동성을 조사한 결과 적어도 상당부분의 염기배열이 잘 보존되는 지역이 존재함을 알게 되었다. 향후 이들 발현 기구의 조절기구를 연구함으로서 protease의 기능을 밝히는데 좋은 단서를 제공할 것으로 판단된다.

항진균 활성 관련 단백질을 탐색하기 위해 미역류로부터 식물병원성 곰팡이의 생장을 저해하는 SAR01 균주를 분리하였고, FAME (fatty acid methyl ester) 분석 결과, Streptomyces sp.로 동정되었다. 방사선 조사(60Co)를 실시한 결과, Botrytis cinerea를 포함한 5종의 식물병원성 곰팡이에 대한 항진균 활성을 소실한 SAR535 균주 외 6종의 돌연변이 균주가 유도되었다. SAR01 야생형 균주와