Heterologous expression of Neurospora crassa cbh1 gene in Pichia pastoris resulted in production of a neutral cellobiohydrolase I.
Neurospora crassa
Pichia pastoris
cellobiohydrolase I
cellulase
glycosylation
Journal
Biotechnology progress
ISSN: 1520-6033
Titre abrégé: Biotechnol Prog
Pays: United States
ID NLM: 8506292
Informations de publication
Date de publication:
05 2019
05 2019
Historique:
received:
28
07
2018
revised:
14
01
2019
accepted:
20
02
2019
pubmed:
1
3
2019
medline:
16
4
2020
entrez:
1
3
2019
Statut:
ppublish
Résumé
The high production cost of cellulase is one of the limitations that hinder the commercialization of lignocellulose-based biorefineries. As one of the important cellulases, Neurospora crassa cellulase is not so intensively investigated as T. reesei cellulase. In this study, the cbh1gene (NCU07340) cloned from N. crassa was expressed in Pichia pastoris under the control of alcohol oxidase 1 (AOX1) promotor. Six transformants with the highest resistance to G418 were selected by two rounds of transformant screening, among which the most robust producer of the recombinant cellobiohydrolase I (CBHI) has an Avicelase activity of 0.22 U/mL. After fermentation optimization, it was improved to 0.30 U/mL. Interestingly, the optimal temperature and pH of the recombinant CBHI were 60°C and 7.2, respectively, and it was quite stable within the wide ranges of temperature and pH. This work is a good example for the future improvement and optimization of N. crassa cellulase.
Substances chimiques
Fungal Proteins
0
Recombinant Proteins
0
Cellulase
EC 3.2.1.4
Cellulose 1,4-beta-Cellobiosidase
EC 3.2.1.91
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e2795Informations de copyright
© 2019 American Institute of Chemical Engineers.