title: Heterologous protein expression in Hypocrea jecorina: a historical perspective and new developments.
abstract: Hypocrea jecorina, the sexual teleomorph of Trichoderma reesei, has long been favored as an industrial cellulase producer, first utilizing its native cellulase system and later augmented by the introduction of heterologous enzymatic activities or improved variants of native enzymes. Expression of heterologous proteins in H. jecorina was once considered difficult when the target was an improved variant of a native cellulase. Developments over the past nearly 30 years have produced strains, vectors, and selection mechanisms that have continued to simplify and streamline heterologous protein expression in this fungus. More recent developments in fungal molecular biology have pointed the way toward a fundamental transformation in the ease and efficiency of heterologous protein expression in this important industrial host. Here, 1) we provide a historical perspective on advances in H. jecorina molecular biology, 2) outline host strain engineering, transformation, selection, and expression strategies, 3) detail potential pitfalls when working with this organism, and 4) provide consolidated examples of successful cellulase expression outcomes from our laboratory.
authors.affiliation: National Bioenergy Center, National Renewable Energy Laboratory, 15013 Denver West Parkway, Golden, CO 80401, United States.
authors.forename: Arjun
authors.initials: A
authors.lastname: Singh
authors.affiliation: Biosciences Center, National Renewable Energy Laboratory, 15013 Denver West Parkway, Golden, CO 80401, United States.
authors.forename: Larry E
authors.initials: LE
authors.lastname: Taylor
authors.affiliation: Biosciences Center, National Renewable Energy Laboratory, 15013 Denver West Parkway, Golden, CO 80401, United States.
authors.forename: Todd A
authors.initials: TA
authors.lastname: Vander Wall
authors.affiliation: National Bioenergy Center, National Renewable Energy Laboratory, 15013 Denver West Parkway, Golden, CO 80401, United States.
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Linger
authors.affiliation: Biosciences Center, National Renewable Energy Laboratory, 15013 Denver West Parkway, Golden, CO 80401, United States.
authors.forename: Michael E
authors.initials: ME
authors.lastname: Himmel
authors.affiliation: Biosciences Center, National Renewable Energy Laboratory, 15013 Denver West Parkway, Golden, CO 80401, United States.
authors.forename: Kara
authors.initials: K
authors.lastname: Podkaminer
authors.affiliation: Biosciences Center, National Renewable Energy Laboratory, 15013 Denver West Parkway, Golden, CO 80401, United States.
authors.forename: William S
authors.initials: WS
authors.lastname: Adney
authors.affiliation: Biosciences Center, National Renewable Energy Laboratory, 15013 Denver West Parkway, Golden, CO 80401, United States. Electronic address: Steve.decker@nrel.gov.
authors.forename: Stephen R
authors.initials: SR
authors.lastname: Decker
mesh_terms: D002480:Cellulase / Q000235:genetics / Q000378:metabolism*; D025141:Drug Resistance, Fungal / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D005800:Genes, Fungal; D056426:Genetic Loci; D035901:Hypocrea / Q000235:genetics / Q000378:metabolism*; D007218:Industrial Microbiology; D010802:Phylogeny; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014242:Trichoderma / Q000235:genetics / Q000378:metabolism*
chemical_list: D005656:Fungal Proteins; D011994:Recombinant Proteins; D002480:Cellulase
keywords: Cel7A; Cellobiohydrolase; Cellulase expression systems; Fungal transformation; Hypocrea jecorina; QM6a; Rut-C30; Trichoderma reesei; cbh1; cbh1 deletion