6533b82bfe1ef96bd128d5fb

RESEARCH PRODUCT

Protoplasts fromPodospora anserina: Isolation, purification, and transformation

Joan SalomEduardo VicenteFederico UruburuSergi FerrerDaniel Ramón

subject

ChromatographybiologyRegeneration (biology)fungifood and beveragesGeneral Medicinebiochemical phenomena metabolism and nutritionProtoplastequipment and suppliesbiology.organism_classificationApplied Microbiology and BiotechnologyMicrobiologyPodospora anserinaMicrobiologylaw.inventionCell wallTransformation (genetics)lawbacteriaCentrifugationMyceliumFiltration

description

Protoplasts fromPodospora anserina mycelium were produced using the commercially available enzyme Novozym 234. Different parameters involved in protoplast isolation were analyzed in order to establish optimal conditions, and protoplast production was notably increased. For the purification of protoplasts, several techniques based on both centrifugation and filtration were assayed, with filtration yielding the best results. Regeneration of protoplasts was studied on different media and osmostic stabilizers, and about 80% regeneration was obtained. The good physiological condition of the protoplasts produced with this method is demonstrated by the lack of cell wall and high regeneration rate and transformation frequencies.

https://doi.org/10.1007/bf01567982