EC Number   |
Protein Variants |
Reference |
|---|
    1.1.2.3 | Y143F |
- |
348072 |
| 1.1.2.3 | Y254F |
- |
348069 |
| 1.1.2.3 | analysis |
a L-lactate-selective microbial biosensor is developed using permeabilized cells of gene-engineered thermotolerant methylotrophic yeast Hansenula polymorpha, over-producing FCb2. The HpCYB2 gene, encoding FCb2, under the control of the strong Hansenula polymorpha alcohol oxidase promoter in the frame of a plasmid for multicopy integration is transformed to the recipient strain Hansenula polymorpha C-105 (gcr1 catX) impaired in glucose repression and devoid of catalase activity. The biosensor based on recombinant yeast cells exhibit a higher Km value (Km: 3.02 mM) and hence expanded linear range toward l-lactate as compared to a similar sensor based on the initial cells of Hansenula polymorpha C-105 (Km: 0.33 mM) |
685700 |
| 1.1.2.3 | more |
active site structures of wild-type and mutant enzymes, detailed overview |
743420 |
| 1.1.2.3 | D72A |
activity and inhibitory profile similar to wild type |
654520 |
| 1.1.2.3 | K73A |
activity and inhibitory profile similar to wild type |
654520 |
| 1.1.2.3 | R38E |
activity and inhibitory profile similar to wild type |
654520 |
| 1.1.2.3 | Y74F |
activity and inhibitory profile similar to wild type |
654520 |
| 1.1.2.3 | more |
construction of a lactate-selective microbial sensor based on flavocytochrome b2-enriched yeast cells. Combination of recombinant technology and nanotechnology by recombinant overexpression of the enzyme in Hansenula polymorpha yeast cells additionally enriched by recombinant enzyme bound with gold nanoparticles, FC b2-nAu. The FC b2-nAu-enriched livingand permeabilized yeast cells are used for construction of a bioselective membrane of microbial L-lactate-selective amperometric biosensor. Phenazine methosulfate serves as a free defusing electron transfer mediator which provides effective electron transfer from the reduced enzyme to the electrode surface. The output to L-lactate of FC b2-nAu-enriched permeabilized yeast cells is 2.5fold higher compared to control cells. For intact cells, the treatment with FC b2-nAu results in 1.56fold increased enzymatic activity. For permeabilized cells, the enrichment efficacy is 2.33fold increased. Method evaluation |
743862 |
| 1.1.2.3 | A198G/L230A |
crystallization data |
654766 |
