Abstract: We report a novel fusion tag for expressing
recombinant proteins in E. coli. The fusion tag is the C-terminus part
of the human GMCSF gene comprising 45 amino acids, which aid in
over expression of otherwise non expressible genes. Expression of
hIFN a2b with this fusion tag also escapes the requirement of rare
codons for expression. This is also a first report of a small fusion tag
of human origin having affinity to heparin sepharose column
facilitating the purification of fusion protein.
Abstract: A. niger XP isolated from Vietnam produces very low amount of acidic phytase with optimal pH at 2.5 and 5.5. The phytase production of this strain was successfully improved through gene cloning and expression. A 1.4 - kb DNA fragment containing the coding region of the phyA gene was amplified by PCR and inserted into the expression vector pPICZαA with a signal peptide α- factor, under the control of AOX1 promoter. The recombined plasmid was transformed into the host strain P. pastoris KM71H and X33 by electroporation. Both host strains could efficiently express and secret phytase. The multicopy strains were screened for over expression of phytase. All the selected multicopy strains of P. pastoris X33 were examined for phytase activity, the maximum phytase yield of 1329 IU/ml was obtained after 4 days of incubation in medium BMM. The recombinant protein with MW of 97.4 KW showed to be the only one protein secreted in the culture broth. Multicopy transformant P. pastoris X33 supposed to be potential candidate for producing the commercial preparation of phytase.