Keyword search (4,163 papers available)

"Cas9" Keyword-tagged Publications:

Title Authors PubMed ID
1 CRISPR/Cas9 mediated gene editing of transcription factor ACE1 for enhanced cellulase production in thermophilic fungus Rasamsonia emersonii Singh V; Raheja Y; Basotra N; Sharma G; Tsang A; Chadha BS; 37658430
CSFG
2 CRAPS: Chromosomal-Repair-Assisted Pathway Shuffling in Yeast Dykstra CB; Pyne ME; Martin VJJ; 37584634
BIOLOGY
3 Rapid, scalable, combinatorial genome engineering by marker-less enrichment and recombination of genetically engineered loci in yeast Abdullah M; Greco BM; Laurent JM; Garge RK; Boutz DR; Vandeloo M; Marcotte EM; Kachroo AH; 37323580
BIOLOGY
4 The MyLo CRISPR-Cas9 Toolkit: A Markerless Yeast Localization and Overexpression CRISPR-Cas9 Toolkit Bean BDM; Whiteway M; Martin VJJ; 35708612
BIOLOGY
5 The chimeric GaaR-XlnR transcription factor induces pectinolytic activities in the presence of D-xylose in Aspergillus niger Kun RS; Garrigues S; Di Falco M; Tsang A; de Vries RP; 34236481
CSFG
6 Identification of a Novel Biosynthetic Gene Cluster in Aspergillus niger Using Comparative Genomics Evdokias G; Semper C; Mora-Ochomogo M; Di Falco M; Nguyen TTM; Savchenko A; Tsang A; Benoit-Gelber I; 34064722
BIOLOGY
7 Seamless site-directed mutagenesis of the Saccharomyces cerevisiae genome using CRISPR-Cas9. Biot-Pelletier D, Martin VJ 27134651
BIOLOGY
8 W361R mutation in GaaR, the regulator of D-galacturonic acid-responsive genes, leads to constitutive production of pectinases in Aspergillus niger. Alazi E, Niu J, Otto SB, Arentshorst M, Pham TTM, Tsang A, Ram AFJ 30298571
CSFG

 

Title:The chimeric GaaR-XlnR transcription factor induces pectinolytic activities in the presence of D-xylose in Aspergillus niger
Authors:Kun RSGarrigues SDi Falco MTsang Ade Vries RP
Link:https://pubmed.ncbi.nlm.nih.gov/34236481/
DOI:10.1007/s00253-021-11428-2
Publication:Applied microbiology and biotechnology
Keywords:Aspergillus nigerCRISPR/Cas9Chimeric transcription factorD-xylosePectinases
PMID:34236481 Category: Date Added:2021-07-08
Dept Affiliation: CSFG
1 Fungal Physiology, Westerdijk Fungal Biodiversity Institute & Fungal Molecular Physiology, Utrecht University, Uppsalalaan 8, 3584, CT, Utrecht, The Netherlands.
2 Centre for Structural and Functional Genomics, Concordia University, 7141 Sherbrooke Street West, Montreal, Quebec, H4B 1R6, Canada.
3 Fungal Physiology, Westerdijk Fungal Biodiversity Institute & Fungal Molecular Physiology, Utrecht University, Uppsalalaan 8, 3584, CT, Utrecht, The Netherlands. r.devries@wi.knaw.nl.

Description:

Aspergillus niger is a filamentous fungus well known for its ability to produce a wide variety of pectinolytic enzymes, which have many applications in the industry. The transcriptional activator GaaR is induced by 2-keto-3-deoxy-L-galactonate, a compound derived from D-galacturonic acid, and plays a major role in the regulation of pectinolytic genes. The requirement for inducer molecules can be a limiting factor for the production of enzymes. Therefore, the generation of chimeric transcription factors able to activate the expression of pectinolytic genes by using underutilized agricultural residues would be highly valuable for industrial applications. In this study, we used the CRISPR/Cas9 system to generate three chimeric GaaR-XlnR transcription factors expressed by the xlnR promoter by swapping the N-terminal region of the xylanolytic regulator XlnR to that of the GaaR in A. niger. As a test case, we constructed a PpgaX-hph reporter strain to evaluate the alteration of transcription factor specificity in the chimeric mutants. Our results showed that the chimeric GaaR-XlnR transcription factor was induced in the presence of D-xylose. Additionally, we generated a constitutively active GaaR-XlnR V756F version of the most efficient chimeric transcription factor to better assess its activity. Proteomics analysis confirmed the production of several pectinolytic enzymes by ?gaaR mutants carrying the chimeric transcription factor. This correlates with the improved release of D-galacturonic acid from pectin by the GaaR-XlnR V756F mutant, as well as by the increased L-arabinose release from the pectin side chains by both chimeric mutants under inducing condition, which is required for efficient degradation of pectin. KEY POINTS: • Chimeric transcription factors were generated by on-site mutations using CRISPR/Cas9. • PpgaX-hph reporter strain allowed for the screening of functional GaaR-XlnR mutants. • Chimeric GaaR-XlnR induced pectinolytic activities in the presence of D-xylose.





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