PROBLEM TO BE SOLVED: To provide an enzyme electrode which can generate electric energy efficiently to the maximum from a fuel such as glucose, and to provide a manufacturing method, and a biofuel cell.SOLUTION: A glucose multistep oxidative enzyme electrode where a coenzyme pyrroloquinoline quinone type glucose dehydrogenation enzyme and a coenzyme pyrroloquinoline quinone type gluconic acid dehydrogenation enzyme are fixed, as an electrode catalyst, to an electrode material, and two step glucose oxidative by coenzyme PQQ type glucose dehydrogenation enzyme and coenzyme PQQ type gluconic acid dehydrogenation enzyme are made to conjugate to the electrode reaction is provided. Consequently, the number of electrons that can be taken out from a fuel such as coke is doubled from two to four per molecule compared with a conventional enzyme electrode using only the glucose dehydrogenation enzyme as the electrode catalyst, and the output of enzyme electrode is enhanced.
