• Microbiology and Biotechnology Letters
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• v.14 no.6
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• pp.467-471
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• 1986

Yeast cell wall lytic enzyme was purified from Fusarium moniliforme by ammonium sulfate fractionation and gel column chromatography. The lytic activity was found to consist of three enzyme activities which were resolved on Sephadex G-100. The first peak on chromatogram exhibited proteolytic, lytic and laminarinase activities, and the second had both lytic and laminarinase activities, whereas the third peak was shown to contain lytic activity only. Three enzyme activities showed the synergistic effect and reducing agents accelerated the yeast roil wall lysis. This indicates that lytic, proteolytic and laminarinase activity acted cooperatively in the lysis of intact cells. Tannic acid precipitate of crude enzyme constituted of three enzyme activities had a high lytic activity on viable yeast cell and has proved useful in yeast protoplast formation.

• Microbiology and Biotechnology Letters
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• v.23 no.6
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• pp.671-677
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• 1995

An extracellular enzyme showing lytic activity on E. coli peptidoglycan had been isolated from Bacillus sp. BL-29. The lytic enzyme was purified to homogeneity by ion-exchange chromatography and gel filtration, with a recovery of 5%. The enzyme was monomeric and had an estimated molecular weight of 31,000 Da. The mode of action of the purified enzyme was also investigated. When the purified lytic enzyme was incubated with cell wall peptidoglycan, N-terminal amino groups were released without the release of reducing groups. The N-terminal amino acid released was identified as dinitrophenylalanine (DNP-alanine) by analysis of terminal amino acid by dinitrophenylation method. This result suggests that the lytic enzyme should be a kind of N-acetylmura-myl-L-alanine amidase.

• Applied Biological Chemistry
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• v.20 no.1
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• pp.123-129
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• 1977

A potent lytic strain was selected by an extensive screening test of microorganisms isolated from soils and sewages on the medium containing baker's yeast as a carbon source. This strain (M-10) was identified to a strain of Humicola sp. by the Genera of Fungi (Clements, 1964). The strain was cultured on the basal medium composed of 2% of baker's yeast, 0.3% of $K_2HPO_4$, 0.01% of $MgSO_4{\cdot}7H_2O$, 0.1% of yeast extract in a shaking incubator. Cultural conditions for lytic enzyme production has been studied, and the results obtained were as follows: 1. The Optimal conditions for lytic enzyme production were: initial pH 5.5 to 6.0, temperature $33^{\circ}C$ in shaking culture. 2. Among the various carbon sources, baker's yeast (4%) was the best for lytic enzyme production, increasing the level of activity eight, times higher than when grown on glucose (1%). 3. The most effective concentration of $K_2HPO_4\;and\;MgSO_4{\cdot}7H_2O$ in the basal medium for lytic enzyme production was 0.1% and 0.01% respectively. 4. When the strain was cultured under the optimal conditions, the production of lytic enzyme was maximized in 72 hours.

• Journal of Microbiology and Biotechnology
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• v.11 no.6
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• pp.957-963
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• 2001

Bacillus subtilis YL-1004 was isolated from soil for the development of agents to control dental caries. This strain produced an extracellular lytic enzyme that hydrolyzed the Streptococcus mutans cell wall. The lytic enzyme was purified to homogeneity by affinity chromatography and gel permeation chromatography to give a single band on SDS-PAGE and non-denaturing polyacrylamide gel electrophoresis. The molecular weight of the enzyme was deduced from SDS-PAGE and gel chromatography to be 38 kDa and the PI to be 4.3 from isoelectric focusing. Sirty $\%$ of its lytic activity remained after incubation at $50^{\circ}C$ for 30 min, and its optimal temperature was $37^{\circ}C$ . The enzyme showed its highest activity at pH 8.0 and was stable at pHs ranging from 4.0 to 9.0. Treatment with several modifiers showed that a cysteine residue was involved in the active site of the enzyme. This lytic enzyme from Bacillus subtilis YL-1004 exhibited specificity towards Streptococci and also showed autolytic activity on Bacillus subtilis YL-1004.

• Microbiology and Biotechnology Letters
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• v.24 no.3
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• pp.299-303
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• 1996

Isolation, identification, and culture conditions of a lytic enzyme producing microorganism against Lacto- bacillus plantarum were investigated. The selected strain was gram-positive, rod (0.7 $\times$ 2.7 $\mu$m in size), and non-motile. The strain did not have any flagella and spores. According to its cultural and physiological characteristics, the strain was identified as Bacillus sp. The optimal pH and temperature for the production of lytic enzyme were 8.0 and 30$\circ$C, respectively. The maximum enzyme activity showed 1.5 units/ml in the medium composed of 1% peptone and 0.1% NaCl.

• Microbiology and Biotechnology Letters
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• v.24 no.6
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• pp.678-686
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• 1996

To improve the preservation of Kimchi, we isolated Lactobacillus plantarum lytic enzyme-producing strain from soil, and the enzyme was purified and characterized. From the observation of cultural and morpho- logical characteristics, the isolated strain was identified as Metarrisium anisopliae (Metschn.) Sorok. The enzyme was purified to 75-folds with 40% yields through affinity adsorption and CM-Sephadex C-50 column chromatog- raphy. The optimum pH and temperature for lytic activity are 4.0 and 40$\circ$C, respectively, and the enzyme acitvity is stable between pH 3.0 and 9.0, and up to 50$\circ$C. The enzyme is a monomeric protein with molecular weight of 40,000 daltons by SDS-PAGE and gel filtration. The enzyme is endopeptidase which breaks the peptide linkage of Lactobacillus plantarum peptidoglycan. The lytic action spectra confirmed that Leuconostoc mesenteroides, a useful strain for the fermentation of Kimchi, is not lysed by the enzyme. The enzyme activity is inhibited by N-bromosuccinimide (NBS), which probably indicates the involvement of tryptophan residue in active site of the enzyme, and also inhibited by Ag$^{+}$. The amino acid composition analysis showed that the enzyme contains more acidic amino acids than basic ones, and composition of alanine, glycine, proline and tyrosines was very high.

• Microbiology and Biotechnology Letters
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• v.24 no.4
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• pp.445-451
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• 1996

The strain YCH-37, which produces yeast cell wall lytlc enzyme, was isolated from soil. From the microscopic observation, morphological and cultural characteristics, this strain was identified to fungus, Dicyma sp. So, we named this strain as Dicyma sp. YCH-37. The lytic enzyme effectively lysed Salmonella typhimurium among intact living bacteria and Torulopsis, Hansenula, Zygosaccharomyces among intact living yeast, as well as autoclaved yeast strains. The yeast cell wall lytic enzyme was succesively purified to 204 folds with 13% yields through yeast glucan affinity adsorption and DEAE-cellulose column chromatography. The enzyme was identified to monomeric protein with molecular weight of 25,000 daltons from the results of SDS-PAGE and gel filtration. The optimum pH and temperature for the yeast lytic activity were 8.0 and 50$\circ$C, respectively. The enzyme was stable up to 40$\circ$C, and between pH 4.0-pH 10.0.

• Microbiology and Biotechnology Letters
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• v.13 no.2
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• pp.137-144
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• 1985

Studies were conducted on the conditions for preparation of yeast protoplasts utilizing Hansenula anomala var. anomala FRI YO-32 as well as Saccharomyces cerevisiae KFCC 32356 and a lytic enzyme from the snail Helix pomatia. The cell wails of the strain FRI YO-32 and S cerevisiae were found to be resistant to activity of the snail lytic enzyme if they were not treated with thiol compounds. Dithiothreitol was found to be more effective than 2-mercaptoethanol, but the latter was considered to be practical. As factors influencing the formation of yeast protoplast, it was considered to be concentration and incubation time of 2-mercaptoethanol or the lytic enzyme, growth stages in yeast cultivation, initial number of yeast cells, and concentration of osmotic stabilizer (KCI). Optimum conditions for the preparation of yeast protoplasts were determined.

• Journal of Microbiology and Biotechnology
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• v.16 no.10
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• pp.1613-1621
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• 2006

Isolation and identification of algal lytic bacteria were carried out. Nine strains of algal lytic bacteria were isolated by the double-layer method using Anabaena flos-aquae as a sole nutrient. The isolate, AFK-13, showing the highest algal lytic activity was identified as Sinorhizobium kostiense based on the l6S rDNA sequence. The algal lytic experiments of the culture supernatants of AFK-13 demonstrated that the bacterial cell growth reached a maximum at 36-h culture, but the supernatant of 72-h culture exhibited the highest activity. Components among the extracellular products in the crude enzyme of the supernatant from S. kostiense AFK-13 culture were responsible for degradation of cell walls of Anabaena flos-aquae. Algal lytic assay tests of the culture supernatants suggest that the main substances for algal lytic activity could be proteinaceous. The activity of glucosidase was observed highly by polysaccharolytic analysis using the crude enzyme from S. kostiense AFK-13, whereas activities of galactosidase, mannosidase, rhamnosidase, and arabinosidase were also detected in low levels. The molecular weights (MW) of ${\alpha}-\;and\;{\beta}$-glucosidases were estimated to be approximately 50-100 kDa by the ultrafiltration method.

• Korean Journal of Microbiology
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• v.22 no.4
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• pp.215-222
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• 1984

A fungus producing cell wall lytic enzyme for Rhodotorula glutinis was isolated from local soil and identified partially as a species of Aspergillus fumigatus group. Thd cell wall lytic enzyme was an inducible exoenzyme and composed of at least lytic polysaccharidase and protease which act cooperatively in the lysis of intact cells. The lytic polysaccharidase was not able to hydrolyze ${\beta}-1,\;3\;and\;{\beta}-1$, 6-glucan which have the same types of bond as found in the cell wall of Ascomycetous yeasts. The lytic polysaccharidase alone was sufficient to hydrolyze the fractionated cell wall (alkali-insoluble residues) of R. glutinis, whereas it showed low activity against intact cells.