A novel biological method was developed for the production of L-3,4-dihydroxyphenylalanine (L-DOPA) from L-tyrosine by p-hydroxyphenylacetate 3-hydroxylase of Escherichia coli strain W (ATCC 11105). About 48 mM (or 1% w/v) L-DOPA was obtained by a fed batch operation in 50 h when a recombinant strain constitutively producing the enzyme was used.