Erschienen:
Springer Science and Business Media LLC, 2019
Erschienen in:Scientific Reports
Sprache:
Englisch
DOI:
10.1038/s41598-019-41573-8
ISSN:
2045-2322
Entstehung:
Anmerkungen:
Beschreibung:
<jats:title>Abstract</jats:title><jats:p><jats:italic>Trichoderma reesei</jats:italic> is an established protein production host with high natural capacity to secrete enzymes. The lack of efficient genome engineering approaches and absence of robust constitutive gene expression systems limits exploitation of this organism in some protein production applications. Here we report engineering of <jats:italic>T</jats:italic>. <jats:italic>reesei</jats:italic> for high-level production of highly enriched lipase B of <jats:italic>Candida antarctica</jats:italic> (calB) using glucose as a carbon source. Multiplexed CRISPR/Cas9 in combination with the use of our recently established synthetic expression system (SES) enabled accelerated construction of strains, which produced high amounts of highly pure calB. Using SES, calB production levels in cellulase-inducing medium were comparable to the levels obtained by using the commonly employed inducible <jats:italic>cbh1</jats:italic> promoter, where a wide spectrum of native enzymes were co-produced. Due to highly constitutive expression provided by the SES, it was possible to carry out the production in cellulase-repressing glucose medium leading to around 4 grams per liter of fully functional calB and simultaneous elimination of unwanted background enzymes.</jats:p>