| abstract |
L-pantolactone dehydrogenase, a mutant thereof, coexpression engineering bacteria and application thereof. The invention relates to the field of gene engineering, in particular to a novel L-pantolactone dehydrogenase (RhoLPLDH) and a mutant thereof, a coding gene, a recombinant vector containing the mutant gene, a co-expression engineering bacterium and application. The amino acid sequence of the L-pantolactone dehydrogenase is shown as SEQ ID NO.1, and the coding gene is shown as SEQ ID NO. 2. The invention provides a high catalytic activity L-pantoate lactone dehydrogenase RhoLPLDH and a mutant thereof, wherein the rhoLPLDH mutant and a molecular chaperone pGro7 are co-expressed to further improve the soluble expression of a target protein. Wherein when the mutant RhoLPLDH-V241I-pGro7 catalyzes 100mM substrate, the product concentration gradually increases along with the time, the reaction can be completed within 8h, the substrate conversion rate is more than 99 percent, and no intermediate product ketopantoate lactone is generated. When the mutant RhoLPLDH-L254I-pGro7 catalyzes 100mM of substrate, the product concentration gradually increases along with the passage of time, the substrate conversion rate reaches 93% at 8h, and the substrate conversion rate is more than 99% at 16 h. |