• Title/Summary/Keyword: substrate mycelium

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Physiological importance of trypsin-like protease during morphological differentiation of streptomycetes

  • Kim, In-Seop;Kang, Sung-Gyun;Lee, Kye-Joon
    • Journal of Microbiology
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    • v.33 no.4
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    • pp.315-321
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    • 1995
  • The relationship between morphological differentiation and production of trypsin-like protease (TLP_ in streptomycetes was studied. All the Streptomyces spp.In this study produced TLP just before the onset of aerial mycelium formation. Addition of TLP inhibitor, TLCK, to the top surface of colonies inhibited aerial mycelium formation as well as TLP inhibitor, TLCK, to the top surface of colonies inhibited aerial mycelium formation as well as TLP activity. Addition of 2% glucose to the Bennett agar medium repressed both the aerial mycelium formation and TLP production in S. abuvaviensis, S. coelicolor A3(2), S exfoliatus, S. microflavus, S. roseus, s. lavendulae, and S. rochei. However the addition of glucose did not affect S. limosus, S. felleus, S. griseus, S. phaechromogenes, and S. rimosus. The glucose repression on aerial mycelium formation and production of TLP was relieved by the addition of glucose anti-metabolite (methyl .alpha.-glucopyranoside). Therefore, it was concluded that TLP production is coordinately regulated with morphological differentiation and TLP activity is essential for morphological differentiation in streptomycetes. The proposed role of TLP is that TLP participates in the degradation of substrate mycelium protein for providing nutrient for aerial mycelial growth.

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Nutritional Regulation of Morphological and Physiological Differentiation on Surface Culture of Streptomyces exfoliatus SMF13

  • KYE JOON LEE;KIM, IN SEOP
    • Journal of Microbiology and Biotechnology
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    • v.5 no.4
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    • pp.200-205
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    • 1995
  • Nutritional factors regulating the morphological differentiation and physiological differentiation of Streptomyces exfoliatus SMF13 on surface cultures were evaluated. S. exfoliatus SMF13 produced leupeptin and chymotrypsin-like protease (CTP) at the stage of substrate mycelium growth, and leupeptin-inactivating enzyme (LIE) and trypsin-like protease (TLP) at the stage of aerial mycelium growth. The activity of leupeptin and CTP was high in the region of active growing substrate mycelium, whereas the activity of LIE and TLP was high in the region of aerial mycelium or spores. The differentiations were induced in glucose-limited conditions or by the addition of glucose anti-metabolite (methyl $\alpha$-glucopyranoside), but repressed by high concentrations of glucose or casamino acids. Morphological differentiation (formation of aerial mycelia and spores) was closely related with physiological differentiation (formation of brown-pigment, LIE and TLP). The local distribution of leupeptin, CTP, LIE, and TLP in a developing colony showed that colony development correlated with the production and functions of the compounds: CTP is essential for providing a nitrogen source for mycelium growth: leupeptin regulates TLP activity: LIE inactivates leupeptin: TLP hydrolyzes nongrowing mycelium.

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The Effects of Temperature and Nutritional Conditions on Mycelium Growth of Two Oyster Mushrooms (Pleurotus ostreatus and Pleurotus cystidiosus)

  • Hoa, Ha Thi;Wang, Chun-Li
    • Mycobiology
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    • v.43 no.1
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    • pp.14-23
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    • 2015
  • The influences of temperature and nutritional conditions on the mycelium growth of oyster mushroom Pleurotus ostreatus (PO) and Pleurotus cystidiosus (PC) were investigated in laboratory experiment during the summer season of 2014. The results of the experiment indicated that potato dextrose agar (PDA) and yam dextrose agar (YDA) were the most suitable media for the mycelium growth of oyster mushroom PO while four media (PDA, YDA, sweet potato dextrose agar, and malt extract agar medium) were not significantly different in supporting mycelium growth of oyster mushroom PC. The optimal temperature for mycelium growth of both oyster mushroom species was obtained at $28^{\circ}C$. Mycelium growth of oyster mushroom PO was improved by carbon sources such as glucose, molasses, and at 1~5% sucrose concentration, mycelium colony diameter of mushroom PO was achieved the highest value. Whereas glucose, dextrose, and sucrose as carbon sources gave the good mycelium growth of oyster mushroom PC, and at 1~3% sucrose concentration, mycelium colony diameter of PC was achieved the maximum value. Ammonium chloride concentrations at 0.03~0.09% and 0.03~0.05% also gave the greatest values in mycelium colony diameter of mushroom PO and PC. Brown rice was found to be the most favourable for mycelium growth of two oyster mushroom species. In addition, sugarcane residue, acasia sawdust and corn cob were selected as favourable lignocellulosic substrate sources for mycelium growth of both oyster mushrooms.

The change of ascorbate oxidase isozyme pattern during mycelial development of streptomyces lavendulae (Streptomyces lavendulae의 균사체 발달에 따른 ascorbate oxidase 동위효소 양상 변화)

  • 이현무;김재헌
    • Korean Journal of Microbiology
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    • v.26 no.2
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    • pp.137-142
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    • 1988
  • pH decreased as the substrate mycelium developed, $\Delta$pH was 1.05-1.15, but increased after the aerial mycelium formation. The lactic acid content in culture solution showed no difference between 0.2% and 5% glucose, at which the aerial mycelium formation was repressed. The growth and development of mycelium was delayed by the lactate treatment. The activity of catalase was maximum in 24 hours after inoculation, and the wuperoxide dismutase activity showed a constant level during the developmental phases. The ascorbic acid accumulated after the aerial mycelium formation. The ascorbate oxidase isozyme of Rf 0.44 appeared, while the isozyme of Rf 0.36 desappeared during the development.

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Proteases and Protease Inhibitors Produced in Streptomycetes and Their Roles in Morphological Differentiation

  • KIM DAE WI;KANG SUNG GYUN;KIM IN SEOP;LEE BYONG KYU;RHO YONG TAIK;LEE KYE JOON
    • Journal of Microbiology and Biotechnology
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    • v.16 no.1
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    • pp.5-14
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    • 2006
  • Streptomycetes are Gram-positive microorganisms producing secondary metabolites through unique physiological differentiation [4]. The microbes show unusual morphological differentiation to form substrate mycelia, aerial mycelia, and arthrospores on solid medium [19]. Substrate mycelium growth is sustaining with sufficient nutrients in the culture medium. The concentration of a specific individual substrate in the culture environment is the most important extracellular factor allowing vegetative mycelia growth, where extracellular hydrolytic enzymes participate in the utilization of waterinsoluble substrates. However, with starvation of nutrients in the culture medium, the vegetative mycelia differentiate to aerial mycelia and spores. It has been considered that shiftdown of essential nutrients for mycelia growth is the most important factor triggering morphological and physiological differentiation in Streptomyces spp. Since proteineous macromolecule compounds are the major cellular components, these are faced to endogenously metabolize following a severe depletion of nitrogen source in culture nutrients (Fig. 1). Various proteases were identified of which production was specifically related with the phase of mycelium growth and also morphological differentiation. The involvement of proteases and protease inhibitor is reviewed as a factor explaining the mycelium differentiation in Streptomyces spp.

The characteristics of fungal-mycelium-based composite materials using spent mushroom substrates of Flammulina velutipes (팽이버섯 수확후배지를 이용한 곰팡이 균사체 기반 복합소재의 특성)

  • Gi-Hong An;Du-Ho Choi;Jae-Gu Han;Kang-Hyo Lee
    • Journal of Mushroom
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    • v.21 no.3
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    • pp.185-189
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    • 2023
  • This study was conducted to develop a renewable and sustainable bio-material to replace polystyrene (EPS) in fungal-mycelium-based composite using agricultural by-products. Four mushrooms (Ganoderma lucidum, Fomitella fraxinea, Phellinus linteus, and Schizophyllum commune) were cultured in an oak sawdust plus rice bran substrate to select the mushroom with the best growth. The mycelia of G. lucidum showed the best growth. To investigate the optimal mixing ratio with spent mushroom substrate (SM) and oak sawdust (OS), samples were prepared by mixing SM and OS at ratios of 50%:50%, 60%:40%, and 80%:20% (w/w). Each substrate was then inoculated with G. lucidum. G. lucidum showed the best mycelial growth of 140.0 mm in the substrate with SM and OS mixed at a 60%:40% ratio. It was also found that the substrate with SM and OS mixed at a 60%:40% ratio had the best handling properties. The compressive strength of mycelial materials inoculated with G. lucidum was in the range of 300-302 kgf mm-1, and the materials were four times stronger than polystyrene materials. These results indicate that substrates comprising spent mushroom substrate mixed with oak sawdust can be successfully upcycled to mycelium-based composite materials using G. lucidum. This represents a sustainable approach.

Characteristics of Trypsin-like Protease and Metalloprotease Associated with Mycelium Differentiation of Streptomyces albidoflavus SMF301

  • Kang, Sung-Gyun;Kim, In-Seop;Jeong, Byung-Cheol;Ryu, Jae-Gon;Rho, Yong-Taik;Lee, Kye-Joon
    • Journal of Microbiology
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    • v.33 no.4
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    • pp.307-314
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    • 1995
  • Trypsin like protease (TLP) and metalloprotease (MTP) were induced in associated with the mycelium differentiation in Streptomyces albidoflavus SMF301. TLP and MTP were purified and characterized from the culture. The molecular mass of TLP and MTP were estimated to be 32 kDa and 18 kDa, respectively. The molecular mass of TLP and MTP were estimated to be 32 kDa and 18 kDa, respectively. The optimum pH and temperature of TLP were 10 and 40.$^{\circ}C$ Those of MTP were 8 and 55 $^{\circ}C$ TLP was stable at alkaline pH (6-9) and unstable above 45.$^{\circ}C$and MTP was stable at alkaline pH and unstable above 80.$^{\circ}C$ Km and Vmax values with benzoyl-arginyl p-nitroanilide of TLP were 139 $\mu$M, and 10 nmole of nitroanilide released per min per$\mu\textrm{g}$ protein, respectively. Km, and Vmax values with a synthetic substrate, leucine p-nitroanilide, or MTP were 58.9 $\mu$M, 3.47 nmol of nitroanilide released per min per$\mu\textrm{g}$protein, respectively. TLP was inhibited competitively by leupeptin; the inhibition constant was 0.0031 $\mu$M. MTP was inhibited by EDTA, phenonthroline and bestatin.

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The Hydrolysis of Dimethyl-cis-1,3-dibenzyl-2-oxoimidazolidine-4,5-dicarboxylate by Immobilized Whole Cells of Chromobacterium chocolatum (고정화된 Chromobacterium chocolatum의 Whole Cell을 이용한 Dimethyl-cis- 1,3-dibenzyl-2-oxoimidazolidine-4,5-dicarboxylate의 가수분해)

  • Lee, Youn Jin;Shim, Sang Kyun;Ahn, Yong Hyun
    • Journal of the Korean Chemical Society
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    • v.41 no.9
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    • pp.483-487
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    • 1997
  • The whole cells of Chromobacterium chocolatum was immobilized in the matrix of polyacrylamide and then used for the hydrolysis of dimethyl-cis-1,3-dibenzyl-2-oxoimidazolidine-4,5-dicarboxylate. This hydrolysis yielded the optically active monoester ( > 96% ee) which is useful as an synthetic intermediate of (+)-biotin. We have studied the optimum condition of hydrolysis by using immobilized cells under variable concentration of substrate, reaction time and pH levels. The activity of lipase in immobilized cell was retained for longer than 4 weeks. The best conversion yield of product was obtained when 2 g of wet cell was immobilized and then reacted with 200 mg of substrate at pH 7.

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Breadcrumb as a New Substrate for Trametes versicolor and Schizophyllum commune Submerged Cultivation

  • Ivanova, Tetiana S.;Bisko, Nina A.;Krupodorova, Tetiana A.;Barshteyn, Victor Yu.
    • Microbiology and Biotechnology Letters
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    • v.42 no.1
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    • pp.67-72
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    • 2014
  • A new substrate, breadcrumbs, was investigated for biomass accumulation, the pH of the cultural broth, the formation of primary metabolites such as the proteins and endopolysaccharides of Schizophyllum commune 1768 and Trametes versicolor 353, as well as its bioconversion efficiency. The results showed that S. commune gives more mycelial mass ($23.96{\pm}0.8g/l$) and in a shorter period (4 days) than T. versicolor ($15.76{\pm}0.5g/l$ in 5 days). The pH values changed from the initial 6.1 to 3.6 in S. commune cultural broth and to 4.4 in T. versicolor cultural broth. Maximal endopolysaccharide content in the mycelia of S. commune and T. versicolor were 7.13% and 6.42%, correspondingly. Crude protein content in S. commune mycelium was 18.83 % on the 4th day of cultivation, and 20.03%, in the mycelium of T. versicolor, on the 6th day of cultivation. Kinetic parameters for the quantitative estimation of cultivation efficiency were calculated for biomass, endopolysaccharide, and crude protein concentrations.

Spore Production of Entomopathogenic Fungus, Beauveria bassiana 726, Using Molasses (당밀을 이용한 곤충병원성 곰팡이 Beauveria bassiana 726의 포자생산)

  • 김병혁;강성우;윤철식;성재모;홍석인;김승욱
    • KSBB Journal
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    • v.14 no.3
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    • pp.365-370
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    • 1999
  • To optimize the culture conditions for Beauveria bassiana 726, the effects of culture medium, pH, and temperature on mycelium and spore production were investigated. The optimum temperature and pH for the cultivation of B. bassiana 726 were 28 $^{\circ}C$ and 5.0, respectively. The optimized medium was composed of 1.0~2.0% total sugar from molasses, 0.5% corn steep liquor and 0.05% KH$_2$PO$_4$. In the cultivation of B. bassiana 726 with the optimum medium, the specific growth rate and substrate utilization were well-fitted with the proposed kinetic model in the shake flask and stirred tank reactor. When the fed-batch cultivation using carbon suorce, nitrogen source, and mineral salt as a feeding medium was compared with batch cultivation in stirred tank reactor, mycelium (12.7 g/L) and spore production (5.4$\times$$10^8/mL$) were enhanced up to 110% and 85%, respectively.

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