• Applied Biological Chemistry
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• v.37 no.4
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• pp.221-225
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• 1994

For producing single cell protein from the agricultural waste, heat treatment and antibiotics on the growth of Cellulomonas sp. KL-6, isolated in rotting leaf and the adjacent soil mixture, were examined. The organism was able to grow until 5 min. at $65^{\circ}C$, 1 min. at $75^{\circ}C$ and 1/4 min. at $85^{\circ}C$ in gradually rising temperatures. It can be Seen that preheating the suspension at $48^{\circ}C$ results in a marked decrease in heat resistance. On heating at temperature of $55^{\circ}C$ for 30 min., strain KL-6 was more resisted in the 0.1 M phosphate buffer when such substrates as casamino acid (1%), yeast extract (1%) or xylose (5%) were added to it whereas this organism was appeared weaker resistances in 0.1 M phosphate buffer when cysteine (0.03 M), sodium citrate (1%) or casein (1%) were in fused into it. Test strain was susceptible to penicillin-G $(1.563\;{\mu}g/ml)$ and ampicillin $(3.125\;{\mu}g/ml)$, but the organism was resisted to kanamycin $(>200\;{\mu}g/ml)$. The treatment of strain KL-6 with sodium dodecyl sulfate (SDS) resulted in the elimination of R-plasmid from the host strain and the elimination rate with SDS $(10{\sim}30\;{\mu}g/ml)$ was about $9.2{\sim}31.2%$, respectively.

• Microbiology and Biotechnology Letters
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• v.16 no.2
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• pp.119-125
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• 1988

$\beta$-Glucosidase of Cellulomonas sp. CS1-1 in cellular compartment was localized with cell-bound form while Avicelase and carboxymethylcellulase (CMCase) were appeared with extracellular enzyme. Cell growth on cellulose or CMC minimal broth was increased by glucose addition. $\beta$-Glucosidase production on cellobiose or CMC minimal broth was repressed by the addition of glucose. However, on CMC minimal broth, the enzyme production was specially stimulated by cellobiose addition. $\beta$-Glucosidase production was also induced by CMC, starcth and maltose compared with glycerol, arabinose, xylose and trehalose. From the above results, it was concluded that glucose effect on $\beta$-glucosidase biosynthesis showed catabolite repression, but enzyme production was induced by cellobiose, CMC, and starch, indicating that $\beta$-glucosidase is inducible enzyme. Yeast extract stimulated $\beta$-glucosidase production more than peptone and ammonium sulfate. $\beta$-Glucosidase activity was increased with 50mM MgCl$_2$in 10mM potassium phosphate buffer (pH 7.0). Optimum conditions for enzyme activities were pH 6.0 and 42$^{\circ}C$, Km value of $\beta$-glucosidase for p-nitrophenyl-$\beta$-D-glucosidase was 0.256mM and Ki for $\beta$-D(+)-glucose was 9.0mM.

• Korean Journal of Agricultural Science
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• v.24 no.2
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• pp.275-282
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• 1997

The prcx.iuction of extracellular 1,4-${\beta}$-glucanase by Cellulomonas sp. CS1-1 on microcrystalline cellulose, sigmacell was maximal after 5-day cultivation as 280 units/mL, which was three times higher than the level produced on carboxymethyl cellulose. A carboxymethyl cellulase containing the carbohydrate of 8.2% was purified from the culture filtrate by successive procedures of column chromatographies. Purification factor was calculated as 22-folds with the specific carboxymethyl cellulase activity of 31.9 units/mg. The molecular weight and isoelectric point of the purified enzyme were 54,000 and pI 5.4, respectively. The optimal pH and temperature were 6.0 and $45^{\circ}C$, and the enzyme was stable between pH 6.5 and 7.5 and below $50^{\circ}C$. The estimated Km and Vmax were 10 mg/mL and $6.25{\mu}mol/min$ for carboxymethyl cellulose and 30.3 mg/mL and $2.85{\mu}mol/min$ for sigmacell, respectively. The enzyme was partially inhibited by $Ag^+$, $Zn^{+{+}}$, $Fe^{+{+}}$ and EDTA, while completely inhibited by $Cd^{+{+}}$ and $Hg^{+{+}}$ at 1 mM concentration.

• Korean Journal of Microbiology
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• v.24 no.2
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• pp.154-160
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• 1986

In order to develope interspecific fusion of the genus Cellulomonas capable of assimilation cellulose, the optimun conditions for the protoplast formation was investigated to examine the susceptibility of cell wall, between different species of the same genus using scanning electron microscope. The variation in the susceptibilities of Cellulomonas sp. CS 1-1 and C. flavigena to lysozyme treatment were considerably remarkable, although they belong to the same genus. The rate of protoplast formation of CS1-1 was 99.9% being treated with lysozyme $(100{\mu}g/ml)$ for 30 minute and that of C. flavigena was about 80% being treated at the concentration of $600{\mu}g/ml$ of lysozyme for 6 hours. The susceptibility of cell wall to the lysozyme treatment on protoplast formation of the strain, CS1-1 seems not to be depend on the cultural periods of cells. On the contrary, that of C. flavigena was considerably depend on the periods. Cells of C. flavigena at mid exponential phase could be more efficiently converted to protoplast cells than those at late exponential phase be done. The rate of the protoplast formation was 95%, when cells of C. flavigena at mid exponential phase were treated with lysozyme $600{\mu}g/ml$ for 6 hours and observed by SEM. In the evalution of protoplast formation of the CS1-1 results of counting method in plate after osmotic shock treatment were similar to the results of the direct observation method by means of SEM. But in the case of C. flavigena the latter method was much more reliable than the former, because the differences between the number of spheroplasts and protoplasts were not able to figure out on conuting the number of protoplast after osmotic shock tretment.

• Microbiology and Biotechnology Letters
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• v.16 no.4
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• pp.303-309
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• 1988

In order to establish the process of interspecific protoplast fusion of the genus Cellulomonas capable of utilizing of cellulose, C. flavigena NCIB 12901 and Cellulomonas sp. CSI-1, the optimum conditions for the regeneration and fusion were examined. The condition of suitable osmotic stabilizer for the protoplast regeneration of C. flavigena was established by using 0.4M sorbitol. And then, by addition of 3% po]yvinyl pyrrolidone (PVP) to cell wall regeneration medium, regeneration frequency was increased 3 times higher than that without PVP addition. The optimum conditions for the interspecific protoplast fusion between auxotrophic and antibiotics resistant mutants were obtained with 40%(W/V) of PEG (polyethylene glycol) 6000 as the fusogenic agent and 25mM of CaCl$_2$on treating time for 15 min. The fusion frequency between mutants was from 2.0$\times$10$^{-4}$ to 4.0$\times$10$^{-4}$ under the optimum conditions. The fusants were confirmed to revert from protoplast to cells of rod type during regeneration process and the aggregation of protoplast by PEG was observed. Also the progress of fusion was observed by scanning electron microscopy, Many isolated fusants were shown to be complement clones of both parents which occured at a high frequency among the isolated clones.

• Korean Journal of Agricultural Science
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• v.4 no.2
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• pp.167-172
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• 1977

Colony morphology, growth responses on some simple carbon sources and taxonomic position were established for three stock cultures of National Collection of Industrial Bacteria, Scotland. It was confirmed that NCIB 8077 belonged to the Cellulomonas species and that NCIB 8633 and NCIB 8634 belonged to the Pseudomonas species. Taxonomy of other cellulolytic bacteria published on various journals was also discussed.

• Microbiology and Biotechnology Letters
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• v.24 no.5
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• pp.613-618
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• 1996

Microorganisms capable of producing ascorbic acid-2-phosphate (AsA2P) from ascorbic acid (AsA) and pyrophosphate (PPi) were screened from the culture collection of this laboratory. Among them, Cellulomonas sp. AP-7 showed the highest productivity of AsA2P. The optimal conditions for the production of AsA2P from AsA and PPi with cell-free extract as an enzyme source were investigated. The reaction mixture for the maximal production of AsA2P consisted of 21 g protein of cell-free extract per liter as the enzyme source, 250 mM AsA, 200 mM sodium pyrophosphate, 150 mM sodium acetate buffer (pH 4.5). By using this reaction mixture, 31.9 mM of AsA2P, which corresponded to a 12.76% yield based on AsA, was produced after incubation of 48 hr at 33$\circ$C.

• Journal of the Korea Organic Resources Recycling Association
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• v.19 no.4
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• pp.84-89
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• 2011

Cellulose-degrading bacteria were isolated and identified from cocopeat which has a good quality as a bulking agent in composting. Various bacteria from different sourecs of cocopeat were detected on CMC agar media, and these were found to be Burkholderi2a sp., Bacillu subtilis, Sphingomonas sp., Rhodotorula sp. & Pseudomonas sp. etc. Among these, four bacteria were further selected and analyzed for their biochemical characteristics and CMCase activities. CMCase activities of four bacteria, P. aeruginosa, P. stutzeri, B. subtilis, and P. luteola were found to be 83%, 40%, 8%, 6%, respectively, compared with that of the standard strain Cellulomonas sp.

• Applied Biological Chemistry
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• v.39 no.5
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• pp.343-348
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• 1996

Experiments were carried out to find possibility of economic production of SCP in mixed culture by Cellulomonas sp. KL-6 and E. coli LI-10. The best cell growth was obtained at the ratio of 1 : 1(v/v) in mixed culture. When these strains were mixed culture, cell growth was increased to about 63%, compared with those of single culture of strain KL-6. It was found that the majority of the population during growth in mixed culture consisted of strain KL-6. $CaCO_3$ added to the medium as the ratio of 0.1% was enhanced medium pH. Cell growth increased in that circumstances. These strains produced much amounts of cellobiose, but glucose was not detected in filter paper medium. When these organisms were cultured under the optimal medium for 4 days, cell mass was produced $1.0\;g/{\ell}$. The results showed the increase of cell mass up to 53% than those produced in CMC medium.

• Applied Biological Chemistry
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• v.38 no.6
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• pp.496-501
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• 1995

Experiments were carried out to find out the optimal condition of SCP produced by Cellulomonas sp. KL-6 and to evaluate nutritional value for the protein of this organism Intracellular- and extracellular proteins produced by this strain were estimated to be nearly maximum, $266\;{\mu}g/m{\ell}\;and\;37\;{\mu}g/m{\ell}$, in the medium containing 0.001% of thiamin after 5 days cultivation. When used rice straw as carbon source for the cell growth of this organism after crusing them by cutting mill, and treating them with 1.0% of NaOH and 10.0% of $NH_4OH\;at\;80^{\circ}C$ for 30 minutes and neutralizing continuatively them with 85% of $H_3PO_4$, SCP production rates were very increased to $1.63\;g/{\ell}$ (NaOH) and $1.47\;g/{\ell}$ (NH4OH), respectively than $0.5\;g/{\ell}$ produced in untreated rice straw. We compared their amino acids patterns with that of FAO provisional patterns. Amino acids content of strain KL-6 was excellents. However. when intended these cell mass to use in practical animal feeding test it would be advisable that destruction or lysis of cell wall should be done.