Protoplast fusion and gene recombination in the uncommon actinomycete Planobispora rosea producing GE2270

作者：Beltrametti Fabrizio^*; Barucco Daniele; Rossi Roberta; Selva Enrico; Marinelli Flavia

来源：Journal of Antibiotics, 2007, 60(7): 447-454.

DOI：10.1038/ja.2007.57

• planobispora rosea
• protoplast
• genome shuffling
• GE2270
• antibiotic
• genetic improvement
• actinomycetes

An efficient method for protoplast generation for the uncommon actinomycete Planobispora rosea, the producer of the thiazolylpeptide antibiotic GE2270, was developed using a combination of hen egg white lysozyme and Streptomyces globisporus mutanolysin. This method converted more than 70% of vegetative mycelium to protoplasts, which were then regenerated with 50% efficiency in an optimized medium. When P rosea protoplasts were efficiently fused, recombination between different antibiotic (streptomycin and gentamicin) resistance markers originated sensitive strains (str(s)gen(s)) at frequencies as high as 18% and double resistant fusants (strr(r)gen(r)) at frequencies as high as 29%. Double resistant fusants showed GE2270 productivity intermediate between the productivity of the parental strains. Protoplast generation and fusion in P rosea makes whole genome shuffling feasible as an approach to be used alternately with classical random mutagenesis in industrial strain improvement programs.

• 出版日期2007-7