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Pichia pastoris produced large quantities of a wheat 9 kDa lipid transfer protein as a recombinant secreted protein. A single step procedure applying the yeast culture directly to expanded-bed Streamline SP-XL was used to purify it 4 times. From 700 ml of culture, 476 mg of highly purified LTP were obtained in one day. This represents a 6-fold higher yield in comparison to the five days purification scheme it replaces.