Gc-ms and hplc have been employed to identify 5-hydroxy-L-norvaline (L-2-amino-5-hydroxypentanoic acid) in the culture fluids of pyridoxine-starved cultures of two pyridoxine auxotrophs of Escherichia coli B. The production of 5-hydroxynorvaline is shown to be secondary to pyridoxine starvation of the cultures, and the quantities of 5-hydroxynorvaline are similar to those of the common protein amino acids present in these cultures. This is the first time that 5-hydroxynorvaline has been identified as a product of bacterial metabolism.