Isolation, characterization and evaluation of the Pichia pastoris sorbitol dehydrogenase promoter for expression of heterologous proteins.
Sorbitol is used as a non-repressive carbon source to develop fermentation process for Mut(s) recombinant clones obtained using the AOX1 promoter in Pichia pastoris. Sorbitol dehydrogenase is an enzyme in the carbohydrate metabolism that catalyzes reduction of D-fructose into D-sorbitol in the presence of NADH. The small stretch of 211bps upstream region of sorbitol dehydrogenase coding gene has all the promoter elements like CAAT box, GC box, etc. It is able to promote protein production under repressive as well as non-repressive carbon sources. In this study, the strength of the sorbitol dehydrogenase promoter was evaluated by expression of two heterologous proteins: human serum albumin and erythrina trypsin inhibitor. Sorbitol dehydrogenase promoter allowed constitutive expression of recombinant proteins in all carbon sources that were tested to grow P. pastoris and showed activity similar to GAP promoter. The sorbitol dehydrogenase promoter was active in all the growth phases of the P. pastoris.
Biocon Research Limited, SEZ Unit, Plot Nos. 2 & 3, Phase IV, B.I.A., Bommasandra-Jigani Link Road, Bengaluru 560 099, India.
Sankar
S
Periyasamy
Nagaraj
N
Govindappa
Suma
S
Sreenivas
Kedarnath
K
Sastry
D001483:Base Sequence; D002244:Carbon / Q000378:metabolism; D003001:Cloning, Molecular; D004895:Erythrina / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D007064:L-Iditol 2-Dehydrogenase / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D010843:Pichia / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics; D012709:Serum Albumin / Q000235:genetics
D010940:Plant Proteins; D011994:Recombinant Proteins; D012709:Serum Albumin; C053884:trypsin inhibitor, Erythrina caffra; D002244:Carbon; D007064:L-Iditol 2-Dehydrogenase
Carbon sources; Constitutive expression; Pichia pastoris BICC 9450; Promoter elements; Sorbitol dehydrogenase promoter