1.1.99.35 D275E substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 6%, D-mannose 11%, D-allose 44%, 3-O-methyl-D-glucose 45%, D-galactose 12%, D-xylose 7%, D-lactose 52%, D-maltose 44%, respectively 639216 1.1.99.35 D276E substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 13%, D-mannose 19%, D-allose 54%, 3-O-methyl-D-glucose 57%, D-galactose 26%, D-xylose 20%, D-lactose 75%, D-maltose 66%, respectively 639216 1.1.99.35 D340F/Y418F increase in temperature stability 655127 1.1.99.35 D340F/Y418I increase in temperature stability 655127 1.1.99.35 D448N substrate specificity: D-glucose 100% activity, 3-O-methyl-D-glucose 72%, D-allose 39%, D-galactose 14%, D-maltose 36%, D-lactose 48%, respectively 654398 1.1.99.35 D456N substrate specificity: D-glucose 100% activity, 3-O-methyl-D-glucose 78%, D-allose 43%, D-galactose 16%, D-maltose 41%, D-lactose 59%, respectively 654398 1.1.99.35 DD457N substrate specificity: D-glucose 100% activity, 3-O-methyl-D-glucose 80%, D-allose 46%, D-galactose 11%, D-maltose 32%, D-lactose 43%, respectively 654398 1.1.99.35 E277A substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 11%, D-mannose 33%, D-allose 142%, 3-O-methyl-D-glucose 91%, D-galactose 46%, D-xylose 23%, D-lactose 83%, D-maltose 39%, respectively 639216 1.1.99.35 E277D substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 9%, D-mannose 19%, D-allose 68%, 3-O-methyl-D-glucose 58%, D-galactose 21%, D-xylose 15%, D-lactose 87%, D-maltose 73%, respectively 639216 1.1.99.35 E277G substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 15%, D-mannose 38%, D-allose 189%, 3-O-methyl-D-glucose 113%, D-galactose 59%, D-xylose 28%, D-lactose 100%, D-maltose 52%, respectively 639216 1.1.99.35 E277H substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 3%, D-mannose 11%, D-allose 67%, 3-O-methyl-D-glucose 45%, D-galactose 12%, D-xylose 9%, D-lactose 57%, D-maltose 39%, respectively 639216 1.1.99.35 E277K substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 7%, D-mannose 18%, D-allose 116%, 3-O-methyl-D-glucose 84%, D-galactose 28%, D-xylose 18%, D-lactose 79%, D-maltose 74%, respectively 639216 1.1.99.35 E277N substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 8%, D-mannose 29%, D-allose 148%, 3-O-methyl-D-glucose 91%, D-galactose 43%, D-xylose 18%, D-lactose 86%, D-maltose 59%, respectively 639216 1.1.99.35 E277Q substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 10%, D-mannose 31%, D-allose 134%, 3-O-methyl-D-glucose 80%, D-galactose 35%, D-xylose 25%, D-lactose 68%, D-maltose 54%, respectively 639216 1.1.99.35 E277V substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 8%, D-mannose 29%, D-allose 150%, 3-O-methyl-D-glucose 101%, D-galactose 25%, D-xylose 19%, D-lactose 114%, D-maltose 65%, respectively 639216 1.1.99.35 I278F substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 4%, D-mannose 14%, D-allose 54%, 3-O-methyl-D-glucose 47%, D-galactose 15%, D-xylose 12%, D-lactose 64%, D-maltose 49%, respectively 639216 1.1.99.35 additional information constructed of heterodimeric PQQGDH-B composed of native wild-type and inactive mutant H168Q subunits. The heterodimeric wild-type/H168Q shows slightly decreased GDH activity and almost identical substrate specificity profile to the wild-type enzyme. The Hill coefficient of the heterodimer is calculated as 1.13, indicating positive cooperativity 655929 1.1.99.35 additional information expression in Pichia pastoris using the Saccharomyces cerevisiae alpha-factor signal sequence for secretion. The productivity of secreted PQQGDH-B achieves 218 kU/liter, i.e. 43 mg/liter. The secreted PQQGDH-B in Pichia pastoris is glycosylated but shows similar enzymatic properties as compared with those of recombinant PQQGDH-B produced in Escherichia coli 655363 1.1.99.35 N279H substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 2%, D-mannose 6%, D-allose 49%, 3-O-methyl-D-glucose 50%, D-galactose 13%, D-xylose 8%, D-lactose 64%, D-maltose 61%, respectively 639216 1.1.99.35 N428C the catalytic efficiency is increased by 47% compared to the wild type enzyme, the mutant shows increased affinity for pyrroloquinoline quinone and is twice more active toward D-glucose and more selective toward maltose than the wild type 724172 1.1.99.35 N428C the mutant shows almost twice the maximum catalytic activity compared to the wild type 723943 1.1.99.35 N452T substrate specificity: D-glucose 100% activity, 3-O-methyl-D-glucose 59%, D-allose 33%, D-galactose 5%, D-maltose 30%, D-lactose 31%, respectively 654398 1.1.99.35 N462H substrate specificity: D-glucose 100% activity, 3-O-methyl-D-glucose 53%, D-allose 32%, D-galactose 2%, D-maltose 25%, D-lactose 31%, respectively 654398 1.1.99.35 S231C substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 3%, D-mannose 8%, D-allose 46%, 3-O-methyl-D-glucose 76%, D-galactose 14%, D-xylose 8%, D-lactose 69%, D-maltose 69%, respectively 639218 1.1.99.35 S231D substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 2%, D-mannose 9%, D-allose 38%, 3-O-methyl-D-glucose 71%, D-galactose 12%, D-xylose 8%, D-lactose 54%, D-maltose 38%, respectively 639218 1.1.99.35 S231H substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 2%, D-mannose 12%, D-allose 57%, 3-O-methyl-D-glucose 86%, D-galactose 17%, D-xylose 7%, D-lactose 56%, D-maltose 38%, respectively 639218 1.1.99.35 S231K increase in thermal stability. Substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 5%, D-mannose 10%, D-allose 43%, 3-O-methyl-D-glucose 82%, D-galactose 15%, D-xylose 5%, D-lactose 59%, D-maltose 70%, respectively 639218 1.1.99.35 S231L substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 6%, D-mannose 13%, D-allose 62%, 3-O-methyl-D-glucose 105%, D-galactose 20%, D-xylose 12%, D-lactose 73%, D-maltose 76%, respectively 639218 1.1.99.35 S231M substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 5%, D-mannose 9%, D-allose 43%, 3-O-methyl-D-glucose 80%, D-galactose 10%, D-xylose 8%, D-lactose 56%, D-maltose 41%, respectively 639218 1.1.99.35 S231N substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 5%, D-mannose 12%, D-allose 61%, 3-O-methyl-D-glucose 109%, D-galactose 18%, D-xylose 15%, D-lactose 66%, D-maltose 51%, respectively 639218 1.1.99.35 T416V/T417V increase in temperature stability 655127