• Korean Journal of Food Science and Technology
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• v.29 no.5
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• pp.1021-1027
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• 1997

We examined some properties of yeast cell wall lytic enzyme produced by Dicyma sp. YCH-37. Several metal ions, reducing reagents, and chemical modifiers have little effects on the lytic activity, except guanidine-HCl. Yeast cells of early log phase were more susceptible to the enzyme than those of stationary phase, and heat-treated cells were more easily lysed than intact living ones. Yeast cells pretreated with organic solvents such as butanol and acetone were more susceptible to the enzyme than intact living ones. Yeast cells cultured in Yeast extract-Malt extract medium containing 0.5 M ammonium sulfate were easily lysed by the lytic enzyme, and yeast cells cultured without shaking were more easily lysed by the enzyme than those with shaking. When SDS, ${\beta}-mercaptoethanol$, Triton X-100, sodium sulfite, and KCl were added to enzyme reaction mixture each, lysis of yeast cells was more effective.

• Microbiology and Biotechnology Letters
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• v.21 no.3
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• pp.276-280
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• 1993

Cell lytic enzyme, 5'-phosphodiesterase, and AMP-deaminase were used to produce yeast extract as a natural seasoning from beer yeast cells. Prior to the addition of cell lytic enzyme, heat treatment was performed to increase the cell wall degradation` the optimum condition of the cell lytic enzyme was 50C at pH 7.0. The production yields by the enzymatic method and conventional autolysis method were 42% and 35%, respectively. The total quantity of 5'-nucleotides, GMP and IMP, produced by enzymatic method was increased by 45% than that by the conventional method. Futhermore, the operation time of enzymatic method was only 6.5 hrs, significantly reduced from 24 hrs of the conventional method.

• Korean Journal of Microbiology
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• v.21 no.4
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• pp.213-220
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• 1983

Conditions for isolation of protoplasts from conidiospores of Trichoderma koningii ATCC 26113 were tested. Maximum production of conidial protoplasts was obtained by preincubation of conidiospores on liquid minimal medium for 8 1/2 hrs. and by reaction with cell wall lytic enzyme for 3 hrs. Among effective cell wall lytic enzymes (Driselase, p-Glucuronidase, Novozyme and Zymolyase), Driselase was the most effective one on the production of conidial protoplasts. The production of conidial protoplasts was also enhanced by addition of 2-Deoxy-D-Glucose $(25{\mu}g/ml)$ into liquid minimal medium. Over 70% of the initial swollen conidia, preincubated in liquid minimal medium supplemented with 2-Deoxy-D-Glucose $(25{\mu}g/ml)$, were converted to protoplasts by incubation with 2% (w/v) commercial lytic enzyme Driselase at $28^{\circ}C$ for 3 hrs. The reversion frequency of the conidial protoplasts was about 30 times (25-50%) higher than that of mycelial protoplasts (0.6-1.3%).

• Journal of Microbiology and Biotechnology
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• v.5 no.4
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• pp.206-212
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• 1995

A strain BL-29, which produces a extracellular lytic enzyme on E. coli was isolated from the soil. The strain was identified as belonging to the genus Bacillus sp. The lytic enzyme was purified to homogeneity by ion exchange chromatography and gel filtration. Specific activity of the purified enzyme was 28, 850 U/mg protein and yield of the enzyme was 5$%$. The purified enzyme showed a single band on SDS-PAGE and its molecular weight was estimated to be 31, 000 by SDS-polyacrylamide gel electrophoresis and gel filtration column chromatography. The optimum temperature and pH were $55^{\circ}C$ and pH 10.0, respectively. The enzyme was stable at $45^{\circ}C$ but enzyme activity was reduced by up to 50$%$ when the temperature was raised to $55^{\circ}C$ for 15 min. Stable range of pH was from 5.0 to 11.0. but Enzyme activity was inhibited by lead-acetate, mercuric chloride, ethylene glycol-bis-[$\beta$-aminoethyl ether]-N, N, $N^1, $N^1$-tetraacetic acid (EGTA), and ethylenediamine tetraacetic acid (EDTA), but not affected considerably by treatment with other chemical reagents.

• Proceedings of the Zoological Society Korea Conference
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• 2000.10a
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• pp.139.2-140
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• 2000

No Abstract, See Full Text

• Korean Journal of Food Science and Technology
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• v.9 no.2
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• pp.97-105
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• 1977

1) In order to obtain a microbial strain having a strong yeast cell wall lytic activity, about 156 isolates capable of forming clear zones on baker's yeast-peptone-bouillon agar plate were obtained from soil, mud and water samples and a strain K-42 with the highest lytic activity was identified as Bacillus circulans. 2) Effect of carbon sources on the lytic enzyme production by the K-42 strain was in the decreasing order of maltose>glucan>xylose>control in 2-day culture and of lactose>galactose>glucan>control in 3-day culture. Effect of inorganic nitrogen sources was in the decreasing order of ammonium acetate>sodium nitrate>control in 2-day culture and of ammonium chloride>ammonium oxalate>control in 3-day culture, whereas organic nitrogen sources except milk casein showed an increase in 2-day culture and a decrease in 3-day culture. Synergistic effect of carbon sources and nitrogen sources was not observed. 3) The enzyme production by the K-42 strain was greatly affected by pH change of the culture medium, thus a high lytic activity could be maintained by keeping the pH range of $7{\sim}8$ and adding carbon or nitrogen sources. 4) Optimum conditions for the lytic activity of the K-42 strain were obtained at $pH\;7{\sim}8$ and $60^{\circ}C$ and the extent of hydrolysis toward heated yeast cell wall was 65%.

• Korean Journal of Food Science and Technology
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• v.35 no.6
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• pp.1143-1149
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• 2003

To elucidate a method of preventing dental caries, strains producing lytic enzymes were isolated and their characteristics were investigated. Among 5,00 alkalophilic strains isolated from soil, 22 strains showed lytic activity against Streptococcus mutans. Strain No. 4830, with the highest lytic activity, was selected for further study. Strain 4830 showed 94% sequence homology with the 16S rDNA sequence of Bacillus alcalophilus, but it was concluded to be different from Bacillus alcalophilus because of its biochemical characteristics. The strain was named Bacillus sp. 4830. The lytic enzyme from Bacillus sp. 4830 was purified by ethanol precipitation and CM-agarose column chromatography. The molecular weight of the lytic enzyme was determined to be 28 kDa by SDS-PAGE. The lytic enzyme was stable between pH 5.0 and pH 11 and up to $40^{\circ}C$. The optimal pH and temperature for the lytic activity was 9.0 and $50^{\circ}C$, respectively.

• The Korean Journal of Mycology
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• v.14 no.4
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• pp.273-280
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• 1986

The degree of autolysis and lytic enzyme production in the culture filtrate of Rhizopus oryzae was investigated. The formation of protoplast by using autolytic enzymes from Rh. oryzae was also attempted. Protoplasts were liberated from Rh. oryzae mycelium by lytic enzymes present in autolytic-phase culture filtrate. Maximum release of chitosanase and proteolytic enzyme into culture filtrate during autolysis was corresponded to maximum protoplast-liberating activity. High yields of protoplasts were obtained from 10 hr-age of Rh. oryzae mycelium with 0.5 M mannitol as osmotic stabilizer. The optimum temperature and pH for mycelium digestion were $25{\sim}30^{\circ}C$ and $6.0{\sim}6.5$ respectively. The mycelium of the 18 hours cultures were treated with autolytic enzyme in same volume of osmotic stabilizer at $30^{\circ}C$ for 5 hours and then it was confirmed by scanning electoron microscope that protoplast were produced beside the digesting cell wall of the fungi.

• Applied Microscopy
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• v.13 no.1
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• pp.49-61
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• 1983

Protoplast releasing mechanisms from Trichoderma koningii, fine structures of the released protoplsts, and their regeneration mode were studied by scanning and transmission electron microscopy. Two types of protoplast releasing mechanisms were observed. In one mechanism, cytoplasm emerged through a cell wall pore developed by cell lytic enzymes and formed a spherical protoplast. In the other mechanism, as the cell wall became progressively thinner, the inner cytoplasm partially rounded to form nonspherical bodies which became spherical protoplasts after being released into the enzyme solution. But, these two types of protoplast releasing mechanisms did not seem to be. mutually exclusive but could occur on the same mycelium simultaneously. And it appeared that cytoplasm which did not become a protoplast by the first mechanism could from a protoplast by the second mechanism. The preparations contained two types of protoplasts, released from different sites of the mycelia. Those released from younger mycelia had dense cytoplasm and small vesicles. Those released from the older mycelia had less dense cytoplasm and larger vacuoles. In the case of regeneration, before producing normal mycelia, most of the protoplasts assumed aberrant tube and yeast-like-forms. Normal mycelia were produced at the end of the yeastlike-forms and sometimes in the middle of the aberrant tube.

• Microbiology and Biotechnology Letters
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• v.10 no.4
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• pp.245-252
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• 1982

The enzymes lytic to red yeast cell wall, which were produced by Penicillium lilacinum ATCC 36010 and Bacillus pumilus No 41, hydrolyzed an extracellular mannan from Rhodotonla glutinis IFO 0695. mannan was arranged with $\beta$-1,3 and $\beta$-1,4 linkages alternatively. Using this mannan, substrate specificities of these enzymes were investigated. The one from Penicillium lilacinum was an unique mannanase which hydrolyzed $\beta$-1,3 mannoside bond and the other from B. pumilus was a new type of mannanase which cleaved $\beta$-1,4 mannoside bond with requirement of the existence of $\beta$-1,3 linkage on the reducing side. Both enzymes released two kinds of oligosaccharide from mannan, respectively. However, the enzyme from Pen lilacinum produced tetrasaccharide and disaccharide and one of them, tetrasaccharide, was hydrolyzed to disaccharide further. The one from B. pumilus released tetrasaccharide and hexasaccharide from mannan finally.