An L-glucitol oxidizing dehydrogenase from Bradyrhizobium japonicum USDA 110 for production of D-sorbose with enzymatic or electrochemical cofactor regeneration.
A gene in Bradyrhizobium japonicum USDA 110, annotated as a ribitol dehydrogenase (RDH), had 87 % sequence identity (97 % positives) to the N-terminal 31 amino acids of an L-glucitol dehydrogenase from Stenotrophomonas maltophilia DSMZ 14322. The 729-bp long RDH gene coded for a protein consisting of 242 amino acids with a molecular mass of 26.1 kDa. The heterologously expressed protein not only exhibited the main enantio selective activity with D-glucitol oxidation to D-fructose but also converted L-glucitol to D-sorbose with enzymatic cofactor regeneration and a yield of 90 %. The temperature stability and the apparent K m value for L-glucitol oxidation let the enzyme appear as a promising subject for further improvement by enzyme evolution. We propose to rename the enzyme from the annotated RDH gene (locus tag bll6662) from B. japonicum USDA as a D-sorbitol dehydrogenase (EC 1.1.1.14).
Microbiology, Campus Bldg. A1.5, Saarland University, 66123, Saarbrücken, Germany.
Sabrina
S
Gauer
Zhijie
Z
Wang
Harm
H
Otten
Mathieu
M
Etienne
Morten Jannik
MJ
Bjerrum
Leila
L
Lo Leggio
Alain
A
Walcarius
Friedrich
F
Giffhorn
Gert-Wieland
GW
Kohring
D001711:Biotransformation; D020369:Bradyrhizobium / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D003067:Coenzymes / Q000378:metabolism*; D004795:Enzyme Stability; D005632:Fructose / Q000378:metabolism; D015870:Gene Expression; D007700:Kinetics; D008970:Molecular Weight; D010084:Oxidation-Reduction; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013012:Sorbitol / Q000378:metabolism*; D013013:Sorbose / Q000378:metabolism*; D020615:Stenotrophomonas maltophilia / Q000201:enzymology / Q000235:genetics; D013401:Sugar Alcohol Dehydrogenases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013696:Temperature
D003067:Coenzymes; D011994:Recombinant Proteins; D005632:Fructose; D013012:Sorbitol; D013401:Sugar Alcohol Dehydrogenases; C022529:ribitol 2-dehydrogenase; D013013:Sorbose