Banana lectin (BanLec) was isolated from slightly overripe bananas (PCI 6-7) by homogenation in NaCl solution, followed by extraction in the presence of glucose, ammonium sulfate precipitation, and affinity chromatography. Yields were approximately 10-fold greater that those of previously published methods using acidic extraction from very overripe fruit (Peel Color Index [PCI] 7+). By dilution of added isotopically labeled recombinant lectin, the content of total exchangeable BalLec was shown to be constant or to slightly decrease with increasing stage of ripeness, even though extractable BanLec increased, followed by rapid decrease in overripened fruit. In the course of this study we observed that recombinant BanLec expressed in Escherichia coli, although chemically and functionally identical to native BanLec, differed slightly in its apparent molecular size on gel filtration, probably due to differences in its native folding.