Extracellular production of the recombinant bacterial transglutaminase in Pichia pastoris.
Bioreactor
GAP promoter
Microbial transglutaminase
Pichia pastoris
Recombinant protein production
Journal
Protein expression and purification
ISSN: 1096-0279
Titre abrégé: Protein Expr Purif
Pays: United States
ID NLM: 9101496
Informations de publication
Date de publication:
07 2019
07 2019
Historique:
received:
13
10
2018
revised:
31
12
2018
accepted:
08
03
2019
pubmed:
16
3
2019
medline:
28
4
2020
entrez:
16
3
2019
Statut:
ppublish
Résumé
Microbial pro-transglutaminase (pro-MTGase) from Streptomyces mobaraensis was expressed in Pichia pastoris (Komagataella phaffii) under the control of constitutive GAP promoter. The single copy of the gene containing clone was grown in shake flasks to determine the optimum conditions for the production of recombinant pro-MTGase. Three temperature (20 °C, 25 °C, 28 °C) and four pH (5, 6, 7, 7.5) values were evaluated at the shake flask level for the extracellular production of pro-MTGase. The highest enzyme activity was obtained with low temperature (20 °C) and high pH (7.5). The maximum yield was 9120 U/L. For the large-scale extracellular production of pro-MTGase, the clone was cultivated in 5 L bioreactor. The fermentation process was carried out at 20 °C, pH 7 and 20% dissolved oxygen for 79 h. The enzyme activity was calculated as 37640 U/L for large-scale production. These results indicate that P. pastoris expression system is very suitable for recombinant MTGase production under the control of the GAP promoter.
Identifiants
pubmed: 30872133
pii: S1046-5928(18)30528-X
doi: 10.1016/j.pep.2019.03.003
pii:
doi:
Substances chimiques
Bacterial Proteins
0
Recombinant Proteins
0
Transglutaminases
EC 2.3.2.13
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
83-90Informations de copyright
Copyright © 2019 Elsevier Inc. All rights reserved.