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Protoplast production and transformation of morphological mutants of the Quorn® myco-protein fungus, Fusarium graminearum A3/5, using the hygromycin B resistance plasmid pAN7-1

Published online by Cambridge University Press:  01 July 1997

MARILYN G. WIEBE ,
MICHAELA NOVÁKOVA ,
LAURIE MILLER ,
MARGARET L. BLAKEBROUGH ,
GEOFFREY D. ROBSON ,
PETER J. PUNT  and
ANTHONY P. J. TRINCI
MARILYN G. WIEBE
Affiliation:
School of Biological Sciences, 1.800 Stopford Building, University of Manchester, Manchester M13 9PT
MICHAELA NOVÁKOVA
Affiliation:
Institute of Microbiology, Czech Academy of Sciences, Videňská 1083, 142 20 Prague 4, Czech Republic
LAURIE MILLER
Affiliation:
Microbiology Section, Department of Molecular Biology & Biotechnology, University of Sheffield, Sheffield S10 2TN
MARGARET L. BLAKEBROUGH
Affiliation:
School of Biological Sciences, 1.800 Stopford Building, University of Manchester, Manchester M13 9PT
GEOFFREY D. ROBSON
Affiliation:
School of Biological Sciences, 1.800 Stopford Building, University of Manchester, Manchester M13 9PT
PETER J. PUNT
Affiliation:
Molecular Genetics & Gene Technology, TNO Medical Biological Laboratory, PO Box 45, 2280 AA Rijswijk, The Netherlands
ANTHONY P. J. TRINCI
Affiliation:
School of Biological Sciences, 1.800 Stopford Building, University of Manchester, Manchester M13 9PT
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Abstract

A protocol for the generation of high yields of viable protoplasts has been developed for several highly branched (colonial) strains of the Quorn® myco-protein fungus, Fusarium graminearum A3/5. Driselase was found to produce higher protoplast yields (ca 109 g−1 wet weight) than the other lytic enzymes tested (Glucanex, Novozyme, β-glucuronidase, Sigma lytic enzyme, or ICN lytic enzyme), although yields differed for the various strains. Protoplast regeneration frequencies of 25–50% were observed when glucose (1·0 M) or sucrose (1·0 M) was used as the osmotic stabilizer. A highly branched strain of F. graminearum CC1-5, which grows better in submerged culture than the more sparsely branched wild-type strain (A3/5) was transformed using the hygromycin B resistance plasmid pAN7-1.

Type
Research Article
Information
Mycological Research , Volume 101 , Issue 7 , July 1997 , pp. 871 - 877
Copyright
The British Mycological Society 1997

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