US EPA

1227360 

Journal Article 

High-level expression and production of human lactoferrin in Pichia pastoris 

Jiang, T; Chen, L; Jia, S; Ma, Y 

2008 

Yes 

Dairy Science & Technology
ISSN: 1958-5586 

88 

2 (Apr 2008) 

173-181 

Lactoferrin (LF) is a multifunctional iron-binding glycoprotein which is found in high concentrations in milk. Recombinant human LF (rhLF) may provide health benefits. In the present study, we report an optimization of the production system of recombinant human lactoferrin that has enabled the production of recombinant protein at levels up to 1200 mg divided by L super(-1). The hLF was expressed in the methylotrophic yeast Pichia pastoris using the pPIC9K vector. The expression level of recombinant hLF (rhLF) was improved significantly by mixed methanol/glycerol feeding at the ratio of 4:1 during the induction phase of high cell-density fermentation. A yield of approximately 1200 mg times L super(-1) was obtained in fed-batch fermentation with this system, which was much higher than the reported values for other systems (1 mg times L super(-1) to 115 mg times L super(-1)). The rhLF was purified via ion-exchange chromatography using SP Sepharose Fast Flow and has a similar molecular mass of 80 kg times mol super(-1) to native hLF. The level of glycosylation of the recombinant protein is similar to that of the native protein. 

Substance P; Feeding; Glycosylation; lactoferrin; Methanol; Batch culture; Glycerol; Pichia pastoris; Cell density; Glycoproteins; Milk; Fermentation