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Andrey M. Chulkin, Valeriy Yu. Kislitsin, Ivan N. Zorov, Igor A. Shashkov, Aleksandra M. Rozhkova
Influence
of the tacA transcription factor knockout on the transcription and expression
of the cbhi gene in the Penicillium verruculosum strain
Abstract
Annotation. The tacA
gene, encoding the TacA repressor protein, was cloned by “walking the uncloned
DNA” method from the genomic DNA of the fungus Penicillium verruculosum
B1-221-151. Knockout of the tacA and niaD genes by the
CRISPR/CAS9 led to the production of a new host strain P. verruculosum
ΔniaDΔtacA, characterized by a higher rate of extracellular protein
biosynthesis. Analysis of the transcription and expression of the cbhI
gene in the original P. verruculosum B1-221-151 strain and in the P.
verruculosum ΔniaDΔtacA strain showed a sharp increase in the
level of cbhI gene transcription 2 h after the start of induction with
cellobiose, cellotriose, gentiobiose, and a mixture of di- and tri-saccharides
in comparison with the transcription of the cbhI gene in the original
strain. The specific activity of cellobiohydrolase I, the main enzyme of the
cellulolytic complex of the fungus P. verruculosum, by 96 h of
fermentation of the ΔtacA strain increased 3 times compared to the
original strain.
Key words: transcription, CRISPR/CAS9,
cellobiohydrolase I
Copyright (C) Chemistry Dept., Moscow State University, 2002
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