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Absence of the highly expressed small carbohydrate-binding protein Cgt improves the[...]
Results
Analysis of cgt expression during growth on different carbon sources
Bioinformatics
Media and growth conditions of Actinoplanes sp. SE50/110
Preparation of spore solutions of Actinoplanes sp. SE50/110
Preparation of media
Shake flask cultivation
Miniaturized cultivation in the BioLector system of m2p-labs GmbH (Baesweiler, Germany)
Acarbose quantification from the supernatant by high-performance liquid chromatography
Starch-binding assay
SDS-polyacrylamide gel electrophoresis
Matrix-assisted laser desorption ionization-time of flight-mass spectrometry
Recombinant DNA work
Deletion of the gene cgt by CRISPR/Cas9 technique
Conjugal transfer to Actinoplanes sp. SE50/110 and plasmid curing
Screening experiments in the Biolog® OmniLog Phenotypic Microarray System
Sampling and RNA isolation
Reverse transcription quantitative PCR
Results
Distribution of single-domain CBM-20 proteins in the eubacterial world
Confirmation of the starch-binding function by an invitro assay
Analysis of cgt expression during growth on different carbon sources
The gene deletion mutant ∆cgt has no apparent growth phenotype on different carbon sources or under carbon-limited conditions
Cgt has no impact on osmolality- or pH-tolerance
The ∆cgt mutant displays an improved acarbose formation on maltose minimal medium
Deletion of cgt has no impact on the expression of acarbose biosynthesis genes
Discussion
References
Aufsatz in einer Zeitschrift
Absence of the highly expressed small carbohydrate-binding protein Cgt improves the acarbose formation in Actinoplanes sp. SE50/110
Entstehung
2020
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