Influence of codon optimization, promoter, and strain selection on the heterologous production of a β-fructofuranosidase from Aspergillus fijiensis ATCC 20611 in Pichia pastoris.

Autor: Coetzee G; Department of Process Engineering, Stellenbosch University, Private Bag X1, Matieland, 7602, South Africa. coetzeeg@sun.ac.za., Smith JJ; Department of Process Engineering, Stellenbosch University, Private Bag X1, Matieland, 7602, South Africa., Görgens JF; Department of Process Engineering, Stellenbosch University, Private Bag X1, Matieland, 7602, South Africa.
Jazyk: angličtina
Zdroj: Folia microbiologica [Folia Microbiol (Praha)] 2022 Apr; Vol. 67 (2), pp. 339-350. Date of Electronic Publication: 2022 Feb 08.
DOI: 10.1007/s12223-022-00947-8
Abstrakt: Fructooligosaccharides (FOS) are compounds possessing various health properties and are added to functional foods as prebiotics. The commercial production of FOS is done through the enzymatic transfructolysation of sucrose by β-fructofuranosidases which is found in various organisms of which Aureobasidium pullulans and Aspergillus niger are the most well known. This study overexpressed two differently codon-optimized variations of the Aspergillus fijiensis β-fructofuranosidase-encoding gene (fopA) under the transcriptional control of either the alcohol oxidase (AOX1) or glyceraldehyde-3-phosphate dehydrogenase (GAP) promoters. When cultivated in shake flasks, the two codon-optimized variants displayed similar volumetric enzyme activities when expressed under control of the same promoter with the GAP strains producing 11.7 U/ml and 12.7 U/ml, respectively, and the AOX1 strains 95.8 U/ml and 98.6 U/ml, respectively. However, the highest production levels were achieved for both codon-optimized genes when expressed under control of the AOX1 promoter. The AOX1 promoter was superior to the GAP promoter in bioreactor cultivations for both codon-optimized genes with 13,702 U/ml and 2718 U/ml for the AOX1 promoter for ATUM and GeneArt ® , respectively, and 6057 U/ml and 1790 U/ml for the GAP promoter for ATUM and GeneArt ® , respectively. The ATUM-optimized gene produced higher enzyme activities when compared to the one from GeneArt ® , under the control of both promoters.
(© 2022. Institute of Microbiology, Academy of Sciences of the Czech Republic, v.v.i.)
Databáze: MEDLINE
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