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http://dx.doi.org/10.18419/opus-14679
Autor(en): | Guitart Font, Emma Sprenger, Georg A. |
Titel: | Opening a novel biosynthetic pathway to dihydroxyacetone and glycerol in Escherichia coli mutants through expression of a gene variant (fsaAA129S) for fructose 6-phosphate aldolase |
Erscheinungsdatum: | 2020 |
Dokumentart: | Zeitschriftenartikel |
Seiten: | 22 |
Erschienen in: | International journal of molecular sciences 21 (2020), No. 9625 |
URI: | http://nbn-resolving.de/urn:nbn:de:bsz:93-opus-ds-146982 http://elib.uni-stuttgart.de/handle/11682/14698 http://dx.doi.org/10.18419/opus-14679 |
ISSN: | 1422-0067 |
Zusammenfassung: | Phosphofructokinase (PFK) plays a pivotal role in glycolysis. By deletion of the genes pfkA, pfkB (encoding the two PFK isoenzymes), and zwf (glucose 6-phosphate dehydrogenase) in Escherichia coli K-12, a mutant strain (GL3) with a complete block in glucose catabolism was created. Introduction of plasmid-borne copies of the fsaA wild type gene (encoding E. coli fructose 6-phosphate aldolase, FSAA) did not allow a bypass by splitting fructose 6-phosphate (F6P) into dihydroxyacetone (DHA) and glyceraldehyde 3-phosphate (G3P). Although FSAA enzyme activity was detected, growth on glucose was not reestablished. A mutant allele encoding for FSAA with an amino acid exchange (Ala129Ser) which showed increased catalytic efficiency for F6P, allowed growth on glucose with a µ of about 0.12 h-1. A GL3 derivative with a chromosomally integrated copy of fsaAA129S (GL4) grew with 0.05 h-1 on glucose. A mutant strain from GL4 where dhaKLM genes were deleted (GL5) excreted DHA. By deletion of the gene glpK (glycerol kinase) and overexpression of gldA (of glycerol dehydrogenase), a strain (GL7) was created which showed glycerol formation (21.8 mM; yield approximately 70% of the theoretically maximal value) as main end product when grown on glucose. A new-to-nature pathway from glucose to glycerol was created. |
Enthalten in den Sammlungen: | 04 Fakultät Energie-, Verfahrens- und Biotechnik |
Dateien zu dieser Ressource:
Datei | Beschreibung | Größe | Format | |
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ijms-21-09625.pdf | 2,97 MB | Adobe PDF | Öffnen/Anzeigen |
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