• 한국미생물·생명공학회지
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• 제20권2호
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• pp.169-177
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• 1992

토양으로부터 alkaline protease 생성능이 강한 Streptomyces griseus HC-1141을 분리하였으며, 효소생산의 최적 배양조건은 0.5 casein, 0.05 ammonium chloride, 0.1 ferrous sulfate, 2.0의 lactose, pH 8.0에서 84시간 배양했을 때이다. 효소의 정제는 ammonium sulfate 침전, DEAE-cellulose ion exchange chromatography, Sephadex G-150 gel filtration, crystallization으로 하여 53.23배 정제할 수 있었으며 polyacrylamide gel 전기영동상 단일밴드를 나타내었다.

• The Plant Pathology Journal
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• 제36권2호
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• pp.185-191
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• 2020

Streptomyces griseus S4-7, a well-characterized keystone taxon among strawberry microbial communities, shows exceptional disease-preventing ability. The whole-genome sequence, functional genes, and bioactive secondary metabolites of the strain have been described in previous studies. However, proteomics studies of not only the S4-7 strain, but also the Streptomyces genus as a whole, remain limited to date. Therefore, in the present study, we created a proteomics reference map for S. griseus S4-7. Additionally, analysis of differentially expressed proteins was performed against a wblE2 mutant, which was deficient in spore chain development and did not express an antifungal activity-regulatory transcription factor. We believe that our data provide a foundation for further in-depth studies of functional keystone taxa of the phytobiome and elucidation of the mechanisms underlying plant-microbe interactions, especially those involving the Streptomyces genus.

• KSBB Journal
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• 제4권2호
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• pp.162-166
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• 1989

스트렘토마이신 생산균주들인 Streptomyces griseus와 S.galbus strain들은 tryptic soy (TS) broth에 배양하여 항생제 생산을 조사하였을 때 S. griseus ATCC 27001이 가장 좋은 균주로 나타났다. 또한 S. griseus ATCC 12475와 ATCC 23345에서도 생장과 항생물질의 생산이 뛰어난 것으로 나타났다. 문헌에 보고된 streptomycin 생산배지들을 조사하여 본 결과 glucose-soybwan meal-sodium chloride (GSS) broth와 K (Chucken) broth에서, 조사한 다른 배지들에 비해 더 많은 양의 스트렙토마이신이 생성되었다. 이들 배지에 든 성분 중에 콩속에 든 특정한 성분이 스트렙토마이신의 생성을 활성화 시켰다. Meat extract를 soybean meal과 함께 가하여 주면 항생제 생성이 더 증가되었다. distillers' solubles도 meat extract와 유사한 활성이 관찰되나 com steep liquor를 가하여 주면 항생제 생산이 감소되었다.

• Journal of Microbiology and Biotechnology
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• 제19권10호
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• pp.1191-1196
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• 2009

The production of Streptomyces griseus trypsin (SGT) by S. griseus IFO13350 transformed with the expression vector pWHM3-TR1R2, containing sprT encoding SGT and the two positive regulatory genes sgtR1 and sgtR2, was investigated in various media. Cultivation in Ferm-0 gave 1.4 times more trypsin activity than in C5/L medium. In addition, replacement of 2% glucose and 1% skim milk in Ferm-0 with 2% dextrin and 1% tryptone (designated Ferm-II) enhanced trypsin activity 4.1-fold. To simplify the purification process, the supernatant from the S. griseus transformant cultured in Ferm-II medium was fractionated with ammonium sulfate (25-55%), then subjected to Hitrap Benzamidine FF affinity column chromatography. The specific activity of SGT purified by one-step chromatography was 69,550 unit/mg protein and the overall purification yield was above 8%, indicating that this method is more effective than those previously reported. Purified SGT was most active at pH 8.0 and $50^{\circ}C$, and it maintained activity between pH 7.0 and 9.0 and at temperatures up to $70^{\circ}C$. These enzymatic properties are very similar to those of authentic eukaryotic trypsin purified from bovine pancreas.

• Journal of Microbiology
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• 제41권4호
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• pp.289-294
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• 2003

Gelatinase is a proteolytic enzyme that hydrolyzes gelatin. Gelatinolytic activity was detected from culture broths of Streptomyces griseus IFO13350 and HH1 by paper disc assays on 0.5% agar plates containing 1% gelatin. The concentrated extracellular protein from the S. griseus was analyzed by SDS polyacrylamide gel, and two proteins, with molecular weights of 30 and 28 kDa, respectively, were identified to have gelatinase activity by gelatin zymography. The protein with a molecular weight of 28 kDa was confirmed to be S. griseus trypsin (SGT). The effects of metal ions and metal chelators on the protease activity of the SGT were studied. Of the metal ions tested, only manganese was found to enhance the protease activity, 2.6 times, however, $Co^{2+},\;Cu^{2+},\;and\;Zn^{2+}$, and metal chelators, such as EDTA and EGTA, inhibited the SGT activity. When the protease activity of the SGT was measured at various pHs, in the presence of 5 mM $MnCl_2$, its highest activity was at pH 11.0, whereas only 60% of the maximum activity was observed between pHs 4.0 and pH 6.0, and almost 80% activity between pHs 7.0 to pH 10.0. The protease activity was measured at various temperatures in the presence of 5 mM $MnCl_2$. The SGT was found to be stable up to $60^{\circ}C$ for 30 min, while only 16% of the enzyme activity remained at $60^{\circ}C$, and at $80^{\circ}C$ almost all the activity was lost. The optimal temperature for the protease activity was $50^{\circ}C$.

• 생명과학회지
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• 제19권5호
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• pp.676-679
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• 2009

미생물들 사이에 존재하는 수평적 유전자 전달을 이용하여 Jurkat T cell에 대하여 새로운 항생물질을 생산하기 위해 토양박테리아 AY2000과 여러 종류의 항생물질 생산 균주인 Streptomyces griseus의 공배양을 수행하였다. MTT assay를 수행하여 세포 독성을 실험을 하였을 때 공배양 상등액은 각각 배양 하였을 때보다 높은 세포독성을 보였고 또한 48시간 배양 하였을 때 가장 높은 활성을 나타내는 것으로 나타났다. 더욱이 DAPI 염색을 하였을 때 Jurkat T cell 세포의 세포핵의 변화도 관찰 되었다. 이런 결과는 공배양 상등액에 새로운 항생물질이 생성되었음을 보여주었고 이런 방법으로 새로운 항생물질 생산에 이용되어 질수 있음을 의미한다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제6권1호
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• pp.18-24
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• 2001

The advantage of using Streptomyces griseus HUT 6037 in the production of chitinase or chitosanase is that the organism is capable of hydrolyzing amorphous or crystalline chitin and chitosan according to the type of the substrate used. We investigated the effects of the enzyme induction time and chitin sources, CM-chitosan and deacetylated chitosan (degree of deacetylation 75-99%), on production of chitosanase. We found that this strain accumulated chitosanase when cells were grown in the culture medium containing chitosanaceous substrates instead of chitinaceous substrates. The highest chitosanase activity was obtained at 4 dyas of cultivation with 99% deacetylated chitosan. The specific activities of chitinase and chitosanase were 0.91 and 1.33 U/mg protein at 3 and 5 days, respectively. From the study of the enzymatic digestibility of various degrees of deacetylated chitosan, it was found that (GlcN)$_3$, (GlcN)$_4$and (GlcN)(sub)5 were produced during the enzymatic hydrolysis reaction. The results of this study suggested that the sugar composition of (GlcN)$_3$was homogeneous and those of (GlcN)$_4$and (GlcN)(sub)5 were heterogeneous.

• Journal of Applied Biological Chemistry
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• 제43권3호
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• pp.136-140
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• 2000

A novel 6-deoxyhexose biosynthetic gene cluster different from the one for the biosynthesis of streptomycin was isolated from Streptomyces griseus using specifically designed PCR primers to compare the sequence of known dTDP-glucose synthase genes. We cloned a 5.8-kb DNA from Streptomyces griseus ATCC10137, which contained the 4-ketoreductase homologue (grsB), dTDP-glucose synthase (grsD), and dTDP-glucose 4, 6-dehydratase (grsE) genes. Escherichia coli cultures containing plasmid of the PCR product which encoded the grsE region under the controUed T7 promoter were able to catalyze the formation of dTDP-4-keto-6-deoxy-D-glucose from TDP-glucose. The enzyme showed high substrate specificity, being specific to only dTDP-glucose that is known to be incorporated into secondary metabolites such as antibiotics.

• 농약과학회지
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• 제16권4호
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• pp.383-386
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• 2012

비농경지의 식물 근권토양에서 식물체의 초기생육촉진 효과가 있는 방선균을 분리하였다. 분리된 미생물의 16S rDNA 염기서열을 분석한 결과 Streptomyces spp.로 동정되었다. Streptomyces griseus (MSS181), Streptomyces griseoaurantiacus (MSS269), Streptomyces microflavus (MSS275), Streptomyces herbaricolor (MSS276)의 배양액을 오이, 고추, 담배와 토마토의 생육초기단계에 관주 처리하여 식물체의 초장, 건조중량을 측정하였다. 방선균 처리에 의해 오이의 초장은 대조구에 비해 16-29% 증가하였으나 건조중량에는 통계적으로 유의한 차이가 없었다. 같은 방선균을 고추에 처리하였을 때 고추의 초장은 대조구에 비해 10-19%, 건조중량은 19-25% 증가하였다. 담배의 건조중량은 44-73% 증가하였고 토마토의 건조중량도 65-165% 증가하였다. 공시균주의 식물병원균에 대한 항균활성을 검정한 결과, MSS275의 Phytophthora capsici, Fusarium oxysporum, Rhizoctonia solani와 Sclerotinia sclerotiorum에 대한 강한 항균활성을 확인하였다.

• Journal of Microbiology
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• 제39권4호
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• pp.305-313
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• 2001

The spr D gene encoding Strptomyces griseus protease D(SGPD); a chymotrypsin-like proteae, was cloned from Strptomyces griseus IFO13350 and sequence. Most of the amino-acid sequence deduced from the nucleotide sequence is idential to that Strptomyces griseus IMRU3499 except that one amino acid has been deleted and Trp 369 has been substituted into Cys369 in the SGPD from S. griseus IFO13350 without affecting the protease activity. The spr D gene was overexpressed in Streptomyce liv-idans TK24 as a heterologous host. Various media with different compositions were also used to max-imize the productivity of SGPD inthe heterologous host. The SGPD productivity was best when the transformant S. lividans TK24 was cultivated in R2YE medium. The relative chymotrypsin activity of the culture broth measured with an artificial chromogenic substrate, N-scuccinyl-ala-ala-pro-phe-p-nitroanilide, was 16 units/ml. A high level of SGPD was also produced in YEME and SAAM medial but it was relatively lower that in R2YE medium and negligible amounts of SGPD were produced in GYE, GAE and Benedict media. The growth of S. lividans reacted the maximum level of cell mass at days 3 and 4 of the culture, but SGPD production started in the stationary phase of cell growth and kept increase in till the 10$^{th}$ day of culture in R2YE and YEME medium, but in GYE media the productivity reached maximum level at 8days of cultivation. The introduction of the spr D gene into S. lividans TK24 triggered biosyntheis of the pigmented antibiotic , actinorhodin, which implies some protease may paly a very improtant role in secondary-metabolite formation in sStreptomyces.