Expression of trehalose synthase gene from Pseudomonas putida P06 in Pichia pastoris
Journal Title: Pakistan Journal of Pharmaceutical Sciences - Year 2014, Vol 27, Issue 3
Abstract
The trehalose synthase (TreS) gene from Pseudomonas putida P06 was successfully ligated with pPICZaA expression vector by the EcoRI and XbaI and was transformed into Pichia pastoris GS115 by electrotransformation. The trehalose synthase gene was fused to the genome of Pichia pastoris GS115 and was controlled by AOX1 promoter. The TreS protein was successfully expressed in intracellularly. SDS-PAGE results illustrated that a specificity protein band was observed at about 76 kDa. The cell lysates could convert 60% maltose into trehalose at 50°C and pH 7.5 in 10% maltose substrate for 24 h. The Pichia pastorisas exogenous gene expression host is safer to produce endotoxin free TreS than E.coli.
Authors and Affiliations
Wang Tengfei , Hongjuan Liu , Kun Dai , Ruiming Wang , Shiru Jia
Keywords
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