A novel directed evolution approach for co-evolution of β-glucosidase activity and organic acid tolerance

Journal of Biotechnology, Год журнала: 2025, Номер 401, С. 1 - 10

Опубликована: Фев. 19, 2025

Язык: Английский

Optimization of Protoplast Preparation and Establishment of PEG-Mediated Genetic Transformation Method in Cordyceps cicadae

Journal of Fungi, Год журнала: 2025, Номер 11(3), С. 219 - 219

Опубликована: Март 13, 2025

Cordyceps cicadae (C. cicadae) is an important edible medicinal fungus; however, owing to its wild growth and lack of genome annotation, construction a stable genetic transformation system in C. greatly limited, impeding the extensive exploitation industry. Here, we successfully established efficient plasmid method within protoplasts by PEG mediation using pCas9-EGFP as marker plasmid. In order overcome low efficiency acquire sufficient for transformation, influence enzyme species, enzymatic hydrolysis time, temperature, fungal age on protoplast preparation were analyzed sequentially, optimal conditions determined follows: 2-day-old mycelia with 1.5% lywallzyme at 34 °C 5 h. Our results indicate that no less than 5.1 × 107 CFU/mL could be acquired. Additionally, five osmotic pressure stabilizers including potassium chloride (KCl), sodium (NaCl), glucose, mannitol, sucrose employed enhance regeneration protoplasts, among which exhibited highest rate 10.43%. The was 37.3 CFU/µg DNA. On this basis, constructed, laying foundation further gene editing metabolic engineering cicadae.

Язык: Английский

CRISPR/Cas9: A cutting-edge tool for cellulase enhancement in fungi

Fungal Biology Reviews, Год журнала: 2024, Номер 50, С. 100394 - 100394

Опубликована: Окт. 2, 2024

Язык: Английский