1.4.1.16 A69R mutation to the correspondung residue of Symbiobacteium thermophilum DAPDH. Mutation improves the catalytic efficiencies toward 2-keto acids and does not affect the catalytic efficiency toward meso-DAP -, 762982 1.4.1.16 F146W site-directed mutagenesis, the mutant shows altered activity levels with meso-2,6-diaminoheptanedioate and pyruvate as substrates 724026 1.4.1.16 F146W/M152Q site-directed mutagenesis, the mutant shows altered activity levels with meso-2,6-diaminoheptanedioate and pyruvate as substrates 724026 1.4.1.16 H227C site-directed saturation mutagenesis, the mutant shows 15.1fold increased activity with phenylpyruvate compared to the wild-type enzyme 724034 1.4.1.16 H227I mutant accepts substrates D-2-phenylglycine and D-homophenylalanine 762879 1.4.1.16 H227V mutant shows high activity toward phenylpyruvic acid and 2-oxo-4-phenylbutyric acid 762879 1.4.1.16 H227V site-directed saturation mutagenesis, the mutant shows 35.1fold increased activity with phenylpyruvate compared to the wild-type enzyme 724034 1.4.1.16 K159R mutation decreases the kcat/KM value with meso-DAP, the catalytic efficiency toward pyruvic acid increases by 24% 763313 1.4.1.16 M152A the mutant shows increased activity towards meso-2,6-diaminoheptanedioate and reduced activity towards pyruvate compared to the wild type enzyme 742276 1.4.1.16 M152D inactive 742276 1.4.1.16 M152E inactive 742276 1.4.1.16 M152H the mutant shows increased activity towards meso-2,6-diaminoheptanedioate and no activity towards pyruvate compared to the wild type enzyme 742276 1.4.1.16 M152K the mutant shows increased activity towards meso-2,6-diaminoheptanedioate and reduced activity towards pyruvate compared to the wild type enzyme 742276 1.4.1.16 M152L the mutant shows increased activity towards meso-2,6-diaminoheptanedioate and pyruvate compared to the wild type enzyme 742276 1.4.1.16 M152N the mutant shows increased activity towards meso-2,6-diaminoheptanedioate and reduced activity towards pyruvate compared to the wild type enzyme 742276 1.4.1.16 M152Q site-directed mutagenesis, the mutant shows altered activity levels with meso-2,6-diaminoheptanedioate and pyruvate as substrates 724026 1.4.1.16 M152S the mutant shows increased activity towards meso-2,6-diaminoheptanedioate and reduced activity towards pyruvate compared to the wild type enzyme 742276 1.4.1.16 M152W inactive 742276 1.4.1.16 additional information in order to enlarge the substrate binding pocket of the meso-diaminopimelate dehydrogenase from Symbiobacterium thermophilum to accommodate larger 2-keto acids, e.g. phenylalanine, four amino acid residues, Phe146, Thr171, Arg181, and His227, are targeted for site saturation mutagenesis 724034 1.4.1.16 additional information systems-wide metabolic pathway engineering in Corynebacterium glutamicum for bio-based production of diaminopentane. Superior strains with desirable properties such as (i) the release from unwanted feedback regulation at the level of aspartokinase and pyruvate carboxylase by introducing the point mutations lysC311 and pycA458, (ii) an optimized supply of the key precursor oxaloacetate by amplifying the anaplerotic enzyme, pyruvate carboxylase, and deleting phosphoenolpyruvate carboxykinase which otherwise removes oxaloacetate, (iii) enhanced biosynthetic flux via combined amplification of aspartokinase, dihydrodipicolinate reductase, diaminopimelate dehydrogenase and diaminopimelate decarboxylase, and (iv) attenuated flux into the threonine pathway competing with production by the leaky mutation hom59 in the homoserine dehydrogenasegene 712938 1.4.1.16 P69G mutation results in decrease of kcat/KM with meso-DAP and pyruvic acid 763313 1.4.1.16 Q154L/D158G/T173I/R199M/H249N construction of a thermostable, NADP+-dependent D-amino acid dehydrogenase (DAADH) from the meso-diaminopimelate dehydrogenase of strain A1 by introducing five point mutations into amino acid residues located in the active site. In the presence of NADP+, the mutant enzyme catalyzes the oxidative deamination of several D-amino acids, including D-cyclohexylalanine, D-isoleucine, and D-2-aminooctanoate, but not of meso-diaminopimelate. The corresponding 2-oxo acids are aminated in the presence of NADPH and ammonia in the reverse reaction, mutant substrate specificity, overview. The mutant enzyme is also more thermostable than its parental meso-diaminopimelate dehydrogenase -, 724643 1.4.1.16 Q154L/T173I/R199M/P248S/H249N/N276S the mutant shows strongly reduced deamination activity with meso-2,6-diaminoheptanedioate and increased animation activity with pyruvate, phenylpyruvate, and 4-methyl-2-oxopentanoic acid compared to the wild type enzyme -, 742282 1.4.1.16 R181F site-directed saturation mutagenesis, the mutant shows 6.4fold increased activity with phenylpyruvate compared to the wild-type enzyme 724034 1.4.1.16 R181F/H227V site-directed saturation mutagenesis, the mutant shows 19.3fold increased activity with phenylpyruvate compared to the wild-type enzyme 724034 1.4.1.16 R35S the mutant shows reduced catalytic efficiency with meso-2,6-diaminoheptanedioate and NADP+ compared to the wild type enzyme -, 741705 1.4.1.16 R35S/R36V the mutations significantly lower the specific activity toward NADP+ compared to the wild type enzyme. The mutant enzyme favors NAD+ over NADP+ 742303 1.4.1.16 R71A mutation destroys the cation-pi interaction between residues R71 and Y205 762982 1.4.1.16 R71A the mutant shows about wild type catalytic efficiency with meso-2,6-diaminoheptanedioate and increased catalytic efficiency with NADP+ compared to the wild type enzyme -, 741705 1.4.1.16 R71E for reductive amination, the mutant shows a 1.5fold increase in Km and 0.24fold decrease in kcat. For oxidative deamination, a 50% decrease in the kcat/Km value is observed 762982 1.4.1.16 R71I R71I shows 0.37fold decrease in kcat/Km value for pyruvic acid 762982 1.4.1.16 R71K the kcat/Km values toward pyruvic acid and meso-DAP increase by 1.4fold and 1.8fold, respectively 762982 1.4.1.16 R71Q the kcat/Km values toward pyruvic acid and meso-DAP both decrease by 30% 762982 1.4.1.16 R71S for pyruvic acid, R71S shows few changes in kcat/Km values 762982 1.4.1.16 S90T mutation results in decrease of kcat/KM with meso-DAP and pyruvic acid 763313 1.4.1.16 T171H mutant accepts substrates D-2-phenylglycine and D-homophenylalanine 762879 1.4.1.16 T171P site-directed saturation mutagenesis, the mutant shows 2.2fold increased activity with phenylpyruvate compared to the wild-type enzyme 724034 1.4.1.16 T171S site-directed saturation mutagenesis, the mutant shows 2.8fold increased activity with phenylpyruvate compared to the wild-type enzyme 724034 1.4.1.16 T171S/H227V site-directed saturation mutagenesis, the mutant shows 10.6fold increased activity with phenylpyruvate compared to the wild-type enzyme 724034 1.4.1.16 T171S/R181F site-directed saturation mutagenesis, the mutant shows 4.0fold increased activity with phenylpyruvate compared to the wild-type enzyme 724034 1.4.1.16 T171S/R181F/H227V site-directed saturation mutagenesis, the mutant shows 14.6fold increased activity with phenylpyruvate compared to the wild-type enzyme 724034 1.4.1.16 T70S catalytic ability of T70S does not change much 763313 1.4.1.16 V14L mutation results in decrease of kcat/KM with meso-DAP and pyruvic acid 763313 1.4.1.16 V156D mutation results in decrease of kcat/KM with meso-DAP and pyruvic acid, almost complete loss of catalytic ability 763313 1.4.1.16 V68M mutation decreases the kcat/KM value with meso-DAP, the catalytic efficiency toward pyruvic acid does not change 763313 1.4.1.16 W121L mutant accepts substrates D-2-phenylglycine and D-homophenylalanine 762879 1.4.1.16 W121L/H227I mutant sow improved enzyme activities towards various 2-oxoacids including sterically bulky substrates. The substrate binding cavity of the mutant enzyme is reshaped to accommodate these bulky substrates, thus leading to higher enzyme activity 762879