Recombinant Schizophyllum commune α-glucuronidase expression and production in the methylotrophic yeast Pichia pastoris
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Typ
Examensarbete för masterexamen
Master Thesis
Master Thesis
Program
Biotechnology (MPBIO), MSc
Publicerad
2014
Författare
Noormohammadian, Hormoz
Modellbyggare
Tidskriftstitel
ISSN
Volymtitel
Utgivare
Sammanfattning
The α-glucuronidase of Schizophyllum commune was expressed heterologously in Pichia pastoris. The α- glucuronidase of the wood degrading fungus S. commune belongs to glycoside hydrolase (GH) family 115 and its extracellular activity release D-glucuronic and 4-O-methyl-D-glucuronic acid residues from polymeric xylan backbone and aldouronic acids. The recombinant P. pastoris strain was grown in repressive BMGY medium that supported biomass generation and induced in BMMY medium for extracellular secretion of the recombinant α-glucuronidase. The activity of the secreted enzyme was assayed using α-glucuronidase measurement kit on a mixture of tri:tetra:penta aldouronic acid substrates. The glicosidic hydrolysis activity of the enzyme was assayed at pH 6.0 an and reached 1.08 nkat mL-1. Also, the enzyme was stable for 18 h at The fed-batch fermentation of the recombinant of P. pastoris resulted in production of 0.723 mg of the crude recombinant enzyme per liter of the fermentation culture. The engineered P. pastoris expression system produced in this project can be used for development and optimization of a cost effective cell line for production of high yiel of correctly fol e an functional α-glucuronidase.
Beskrivning
Ämne/nyckelord
Energi , Livsvetenskaper , Annan kemi , Biologiska vetenskaper , Industriell bioteknik , Biomaterialvetenskap , Energy , Life Science , Other Chemistry Topics , Biological Sciences , Industrial Biotechnology , Biomaterials Science