title: Biochemical retrosynthesis of 2'-deoxyribonucleosides from glucose, acetaldehyde, and a nucleobase.
abstract: 2'-Deoxyribonucleosides are important as building blocks for the synthesis of antisense drugs, antiviral nucleosides, and 2'-deoxyribonucleotides for polymerase chain reaction. The microbial production of 2'-deoxyribonucleosides from simple materials, glucose, acetaldehyde, and a nucleobase, through the reverse reactions of 2'-deoxyribonucleoside degradation and the glycolytic pathway, was investigated. The glycolytic pathway of baker's yeast yielded fructose 1,6-diphosphate from glucose using the energy of adenosine 5'-triphosphate generated from adenosine 5'-monophosphate through alcoholic fermentation with the yeast. Fructose 1,6-diphosphate was further transformed to 2-deoxyribose 5-phosphate in the presence of acetaldehyde by deoxyriboaldolase-expressing Escherichia coli cells via D-glyceraldehyde 3-phosphate. E. coli transformants expressing phosphopentomutase and nucleoside phosphorylase produced 2'-deoxyribonucleosides from 2-deoxyribose 5-phosphate and a nucleobase via 2-deoxyribose 1-phosphate through the reverse reactions of 2'-deoxyribonucleoside degradation. Coupling of the glycolytic pathway and deoxyriboaldolase-catalyzing reaction efficiently supplied 2-deoxyribose 5-phosphate, which is a key intermediate for 2'-deoxyribonucleoside synthesis. 2'-Deoxyinosine (9.9 mM) was produced from glucose, acetaldehyde, and adenine through three-step reactions via fructose 1,6-diphosphate and then 2-deoxyribose 5-phosphate, the molar yield as to glucose being 17.8%.
authors.affiliation: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, Kyoto 606-8502, Japan.
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Horinouchi
authors.forename: Jun
authors.initials: J
authors.lastname: Ogawa
authors.forename: Takako
authors.initials: T
authors.lastname: Kawano
authors.forename: Takafumi
authors.initials: T
authors.lastname: Sakai
authors.forename: Kyota
authors.initials: K
authors.lastname: Saito
authors.forename: Seiichiro
authors.initials: S
authors.lastname: Matsumoto
authors.forename: Mie
authors.initials: M
authors.lastname: Sasaki
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Mikami
authors.forename: Sakayu
authors.initials: S
authors.lastname: Shimizu
mesh_terms: D000079:Acetaldehyde / Q000378:metabolism*; D000225:Adenine / Q000378:metabolism*; D001709:Biotechnology / Q000379:methods*; D003853:Deoxyribonucleosides / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005947:Glucose / Q000378:metabolism*; D006019:Glycolysis; D007288:Inosine / Q000031:analogs & derivatives / Q000378:metabolism; D012267:Ribosemonophosphates / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000254:growth & development / Q000378:metabolism
chemical_list: D003853:Deoxyribonucleosides; D012267:Ribosemonophosphates; D007288:Inosine; C058245:2-deoxyribose 5-phosphate; D000079:Acetaldehyde; C012271:deoxyinosine; D005947:Glucose; D000225:Adenine