title: Isolation, purification and regeneration of protoplasts from Sporotrichum thermophile conidiospores.
abstract: Protoplasts of uniform size were prepared from mononucleated conidiospores of Sporotrichum thermophile. Conidia were preincubated in glucose yeast extract medium at 45 C for 4 h. The conidia were collected resuspended in buffer containing 0.6 M KCl (as stabilizer), and incubated with Novozyme SP249 and Cellulase CP at 37 C for 6 h. The protoplasts were separated from cell wall fragments and intact conidia by centrifugation over 50% sucrose. The purified protoplasts were regenerated in glucose yeast extract broth after 7 h of incubation at 45 C.
authors.affiliation: Department of Microbiology, CBS&H, G.B. Pant University of Agriculture and Technology, Pantnagar, India.
authors.forename: V K
authors.initials: VK
authors.lastname: Sharma
authors.forename: R
authors.initials: R
authors.lastname: Goel
authors.forename: B N
authors.initials: BN
authors.lastname: Johari
mesh_terms: D002473:Cell Wall / Q000378:metabolism; D002480:Cellulase; D002498:Centrifugation / Q000379:methods; D005947:Glucose / Q000378:metabolism; D006868:Hydrolysis; D007202:Indicators and Reagents; D008358:Mannose / Q000378:metabolism; D011523:Protoplasts / Q000378:metabolism* / Q000502:physiology; D013172:Spores, Fungal / Q000378:metabolism; D013175:Sporothrix / Q000235:genetics / Q000378:metabolism*; D013395:Sucrose / Q000378:metabolism; D013696:Temperature
chemical_list: D007202:Indicators and Reagents; D013395:Sucrose; D002480:Cellulase; D005947:Glucose; D008358:Mannose