Study on the protoplast preparation and regeneration of Gliocladium sp. producing antiinflammation compound 6,22-diene-5,8-epidioxy ergosta-3-hydroxy / 中国生化药物杂志

ABSTRACT

Purpose　The aim is to study the conditions of preparation and regeneration of Gliocladium sp. (F) protoplast. Methods　 Different enzyme systems, enzymolysis time, osmotic pressure stabilizers were studied to investigate their influence on the productivity and regenerating rate of Gliocladiumsp. F protoplasts. The HPLC method was used to determine EP (6,22-diene-5,8-epidioxy ergosta-3-hydroxy) content.Results　The higher productivity of protoplasts was obtained when mycelia of strain F growing for 60 hours was digested at 28℃ for 4 hours by solution containing 2% cellulase and 2% helicase dissolved in 0.5 mol/L mannitol and the medium containing 0.5mol/L mannitol as osmotic pressure stabilizer would be suitable for protoplast regeneration. According to the EP productivity detected by HPLC, high positive rate of the regenerated strains growing in the medium containing 0.5mol/L mannitol could be got. Conclusion　The results will promote the research of strain F mutantgenesis and will be helpful for obtaining the strain more effectively biosythesizing compound EP.