L-glucitol oxidizing dehydrogenase from Bradyrhizobium japonicum USDA 110 for production of D-sorbose with enzymatic or electrochemical cofactor regeneration

• Published in: Applied microbiology and biotechnology Vol. 98; no. 7; pp. 3023 - 3032
• Main Authors: Gauer, Sabrina, Wang, Zhijie, Otten, Harm, Etienne, Mathieu, Bjerrum, Morten Jannik, Lo Leggio, Leila, Walcarius, Alain, Giffhorn, Friedrich, Kohring, Gert-Wieland
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer-Verlag 01.04.2014; Springer Berlin Heidelberg; Springer; Springer Nature B.V; Springer Verlag
• Subjects: Amino acids; Analysis; Bacteria; Biomedical and Life Sciences; Biosynthesis; Biotechnologically Relevant Enzymes and Proteins; Biotechnology; Biotransformation; Bradyrhizobium; Bradyrhizobium - enzymology; Bradyrhizobium - genetics; Bradyrhizobium japonicum; Chemical properties; Chemical Sciences; chemistry; Chromatography; Cloning; Cloning, Molecular; Coenzymes; Coenzymes - metabolism; Dehydrogenase; Dehydrogenases; E coli; Electrochemical reactions; Electrochemistry; Enzyme Stability; Enzymes; enzymology; Fructose; Fructose - metabolism; Gene Expression; genes; genetics; Kinetics; L-iditol dehydrogenase; Life Sciences; loci; metabolism; Microbial Genetics and Genomics; Microbiology; Molecular Weight; Oxidation; Oxidation-Reduction; Oxidoreductases; Recombinant Proteins; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; sequence analysis; Sequence Homology, Amino Acid; Sorbitol; Sorbitol - metabolism; Sorbose; Sorbose - metabolism; Stenotrophomonas maltophilia; Stenotrophomonas maltophilia - enzymology; Stenotrophomonas maltophilia - genetics; Studies; Sugar; Sugar Alcohol Dehydrogenases; Sugar Alcohol Dehydrogenases - chemistry; Sugar Alcohol Dehydrogenases - genetics; Sugar Alcohol Dehydrogenases - metabolism; Temperature; Germany; Electrochemistry; Dehydrogenase; Cofactor regeneration; glucitol; sorbose; D-sorbose; L-glucitol
• ISSN: 0175-7598; 1432-0614; 1432-0614
• DOI: 10.1007/s00253-013-5180-7

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).