Recombinant expression of an l-amino acid oxidase from the fungus Hebeloma cylindrosporum in Pichia pastoris including fermentation.

Heß M, Bloess S, Risse JM, Friehs K, Fischer von Mollard G (2020)
MicrobiologyOpen 9(10): e1112.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
 
Download
OA 136.30 KB
Abstract / Bemerkung
l-amino acid oxidases (LAAOs) are flavoenzymes that catalyze the oxidative deamination of l-amino acids to the corresponding alpha-keto acids, ammonia, and hydrogen peroxide. Here, we show the overexpression, purification, and the characterization of LAAO4 from the fungus Hebeloma cylindrosporum in the yeast Pichia pastoris with a 9His-tag and compare this with the recently characterized 6His-hcLAAO4 expressed in E. coli. The expression of the enzyme with an ER-signal sequence in P.pastoris resulted in a glycosylated, secreted protein. The enzymatic activity without activation was higher after expression in P.pastoris compared to E. coli. Due to treatment with acidic pH, a striking increase of activity could be detected for both expression systems resulting in similar specific activities after acid activation. Regarding the substrate spectrum, temperature stability, Km, and vmax values, hcLAAO4 showed very few differences when produced in these two expression systems. A higher yield of hcLAAO4 could be obtained by fermentation. © 2020 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.
Erscheinungsjahr
2020
Zeitschriftentitel
MicrobiologyOpen
Band
9
Ausgabe
10
Art.-Nr.
e1112
eISSN
2045-8827
Finanzierungs-Informationen
Open-Access-Publikationskosten wurden durch die Universität Bielefeld im Rahmen des DEAL-Vertrags gefördert.
Page URI
https://pub.uni-bielefeld.de/record/2945772

Zitieren

Heß M, Bloess S, Risse JM, Friehs K, Fischer von Mollard G. Recombinant expression of an l-amino acid oxidase from the fungus Hebeloma cylindrosporum in Pichia pastoris including fermentation. MicrobiologyOpen. 2020;9(10): e1112.
Heß, M., Bloess, S., Risse, J. M., Friehs, K., & Fischer von Mollard, G. (2020). Recombinant expression of an l-amino acid oxidase from the fungus Hebeloma cylindrosporum in Pichia pastoris including fermentation. MicrobiologyOpen, 9(10), e1112. https://doi.org/10.1002/mbo3.1112
Heß, Marc, Bloess, Svenja, Risse, Joe Max, Friehs, Karl, and Fischer von Mollard, Gabriele. 2020. “Recombinant expression of an l-amino acid oxidase from the fungus Hebeloma cylindrosporum in Pichia pastoris including fermentation.”. MicrobiologyOpen 9 (10): e1112.
Heß, M., Bloess, S., Risse, J. M., Friehs, K., and Fischer von Mollard, G. (2020). Recombinant expression of an l-amino acid oxidase from the fungus Hebeloma cylindrosporum in Pichia pastoris including fermentation. MicrobiologyOpen 9:e1112.
Heß, M., et al., 2020. Recombinant expression of an l-amino acid oxidase from the fungus Hebeloma cylindrosporum in Pichia pastoris including fermentation. MicrobiologyOpen, 9(10): e1112.
M. Heß, et al., “Recombinant expression of an l-amino acid oxidase from the fungus Hebeloma cylindrosporum in Pichia pastoris including fermentation.”, MicrobiologyOpen, vol. 9, 2020, : e1112.
Heß, M., Bloess, S., Risse, J.M., Friehs, K., Fischer von Mollard, G.: Recombinant expression of an l-amino acid oxidase from the fungus Hebeloma cylindrosporum in Pichia pastoris including fermentation. MicrobiologyOpen. 9, : e1112 (2020).
Heß, Marc, Bloess, Svenja, Risse, Joe Max, Friehs, Karl, and Fischer von Mollard, Gabriele. “Recombinant expression of an l-amino acid oxidase from the fungus Hebeloma cylindrosporum in Pichia pastoris including fermentation.”. MicrobiologyOpen 9.10 (2020): e1112.
Alle Dateien verfügbar unter der/den folgenden Lizenz(en):
Creative Commons Namensnennung 4.0 International Public License (CC-BY 4.0):
Volltext(e)
Access Level
OA Open Access
Zuletzt Hochgeladen
2020-10-19T09:59:09Z
MD5 Prüfsumme
1899b5116f9f0ccd5f83709fd78e5920


Export

Markieren/ Markierung löschen
Markierte Publikationen

Open Data PUB

Web of Science

Dieser Datensatz im Web of Science®
Quellen

PMID: 32852125
PubMed | Europe PMC

Suchen in

Google Scholar