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KMID : 0379119940220020138
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Korean Journal of Mycology
1994 Volume.22 No. 2 p.138 ~ p.144
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Isolation of Protoplasts from Rhizopus nigricans
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Abstract
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Conditions for isolation of protoplasts from spores and mycelia of Rhizopus nigricans were studied. Larger amount of protoplasts was obtained from swollen spores in liquid medium contained with 5% of 2-deoxy-D-glucose for 4 hours than from mycelia. Enzyme mixture of Novozym 234(2%) and β-giucuronidase(5000 unit/㎖) wes most effective for the isolation of protoplasts from swollen spores and from mycelia. The solution of 0.6 M MgSO₄ or mannitol and pH 6.0 showed good results as the osmotic stabilizer and the optimal condition of pH of the enzyme solution for the isolation of protoplast from the swollen spores, respectively. At this condition, 8.0 × 106 cells/㎖ of protoplasts was obtained from swollen spores by digestion with lytic enzyme mixture for 2 hours.
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