One-pot microbial synthesis of 2'-deoxyribonucleoside from glucose, acetaldehyde, and a nucleobase.
A one-pot enzymatic synthesis of 2'-deoxyribonucleoside from glucose, acetaldehyde, and a nucleobase was established. Glycolysis by baker's yeast (Saccharomyces cerevisiae) generated ATP which was used to produce D: -glyceraldehyde 3-phosphate production from glucose via fructose 1,6-diphosphate. The D: -glyceraldehyde 3-phosphate produced was transformed to 2'-deoxyribonucleoside via 2-deoxyribose 5-phosphate and then 2-deoxyribose 1-phosphate in the presence of acetaldehyde and a nucleobase by deoxyriboaldolase, phosphopentomutase expressed in Escherichia coli, and a commercial nucleoside phosphorylase. About 33 mM 2'-deoxyinosine was produced from 600 mM glucose, 333 mM acetaldehyde and 100 mM adenine in 24 h. 2'-Deoxyinosine was produced from adenine due to the adenosine deaminase activity of E. coli transformants.
Horinouchi
Nobuyuki
N
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.
Ogawa
Jun
J
Kawano
Takako
T
Sakai
Takafumi
T
Saito
Kyota
K
Matsumoto
Seiichiro
S
Sasaki
Mie
M
Mikami
Yoichi
Y
Shimizu
Sakayu
S
D000079:Acetaldehyde / Q000378:metabolism; D001709:Biotechnology / Q000379:methods; D003853:Deoxyribonucleosides / Q000096:biosynthesis*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism; D010430:Pentosyltransferases / Q000494:pharmacology; D010770:Phosphotransferases / Q000378:metabolism; D012267:Ribosemonophosphates / Q000378:metabolism; D012440:Saccharomyces / Q000201:enzymology*; D014169:Transformation, Bacterial / Q000235:genetics* / Q000502:physiology
D003853:Deoxyribonucleosides; D012267:Ribosemonophosphates; C058245:2-deoxyribose 5-phosphate; D010430:Pentosyltransferases; C067995:nucleoside phosphorylase; D010770:Phosphotransferases; C022375:phosphopentomutase; D000079:Acetaldehyde; D005947:Glucose