• Microbiology and Biotechnology Letters
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• v.20 no.2
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• pp.169-177
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• 1992

An alkaline protease producing microorganism was isolated from soil and identified as Streptomyces griseus HC-1141. The optimum culture condition of Streptomyces griseus HC- 1141 for the production of alkaline protease was as follows; 0.5% casein, 0.05% ammonium chloride, 0.1% ferrous sulfate. 2.0% lactose, pH 8.0 and 84 hrs. The enzyme was purified about 53 folds by ammonium sulfate treatment, DEAE-cellulose ion exchange chromatography and gel filtratioo on Sephadex G-150. The homogeneity of the purified enzyme was verified by polyacrylamide gel electrophoresis. The molecular weight was estimated to be 31,000 on sodium dodecyl sulfate polyacrylamide gel electrophoresis. This enzyme consists of glycine and glutamic acid as major amino acids. The N-terminal and C-terminal residues of the alkaline protease were leucine and histidine respectively.

• The Plant Pathology Journal
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• v.36 no.2
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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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• v.4 no.2
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• pp.162-166
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• 1989

When various strains of Streptomyces griseus and S. galbus were examined for the ability on the production of streptomycin in tryptic soy(TS) broth, S. griseus ATCC 27001 was found to be the best. S. griseus ATCC 12475 and ATCC 23345 showed also good growth and favorable production of streptomycin. Examination of various complex media reported in fermentation literatures for the industrial production of streptomycin indicated that glucose-soybean media-sodium chloride (GSS) broth and K (Chucken) broth gave higher yields of streptomycin than others studied by us. Examination of the ingredients of media producing streptomycin in high yield indicated that some components in soybean activated the production of streptomycin. Addition of meat extract enhanced the yield of streptomycin but it could be substituted with distillers solubles without much effect on the yield. Addition of corn steep liquor decreased the production of streptomycin.

• Journal of Microbiology and Biotechnology
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• v.19 no.10
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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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• v.41 no.4
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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 $50^{\circ}C$.

• Journal of Life Science
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• v.19 no.5
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• pp.676-679
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• 2009

In order to produce a new antibiotic material against Jurkat T cells using horizontal gene transfer among microbes, co-cultures between soil bacteria AY2000 and the multiple antibiotic producer S. griseus was carried out. It showed that the highest active substance against Jurkat T cells was produced at 48 hr of co-culture time with MTT assay. Moreover, a morphological change of nuclear of Jurkat T cells treated with co-cultured substance was observed in DAPI staining. This result suggests that a new material was produced with co-culture supernatant, and that co-culture between microboes can develop new antibiotic materials.

• Biotechnology and Bioprocess Engineering:BBE
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• v.6 no.1
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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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• v.43 no.3
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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.

• The Korean Journal of Pesticide Science
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• v.16 no.4
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• pp.383-386
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• 2012

Streptomyces spp. were isolated from rhizosphere in fallow lands. The Streptomyces spp. were identified as Streptomyces griseus (MSS181), Streptomyces griseoaurantiacus (MSS269), Streptomyces microflavus (MSS275), Streptomyces herbaricolor (MSS276) based on 16S rRNA gene sequences. Afterwards, cucumber, pepper, tobacco and tomato were drenched with the isolates at early growth stages and plant growth such as height and dry weight of plants was measured. By treatment of Streptomyce spp., plant height of cucumber was increased by 16-29% compared to the control, But there were no statistically significant differences in dry weight. When the same isolates were treated on chili-pepper, plant height and dry weight of chili-pepper were increased respectively by 10-19% and 19-25% compared to the control. The dry weight of tobacco and tomato were increased by 44-73% and 65-165%, respectively compared to the control. When antifungal activities of the isolates were tested against plant pathogenic fungi, Streptomyces microflavus (MSS275) effectively inhibited the mycelial growth of Phytophthora capsici, Fusarium oxysporum, Rhizoctonia solani and Sclerotinia sclerotiorum.

• Journal of Microbiology
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• v.39 no.4
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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.