Efficient production of 2-deoxyribose 5-phosphate from glucose and acetaldehyde by coupling of the alcoholic fermentation system of Baker's yeast and deoxyriboaldolase-expressing Escherichia coli.
2-Deoxyribose 5-phosphate production through coupling of the alcoholic fermentation system of baker's yeast and deoxyriboaldolase-expressing Escherichia coli was investigated. In this process, baker's yeast generates fructose 1,6-diphosphate from glucose and inorganic phosphate, and then the E. coli convert the fructose 1,6-diphosphate into 2-deoxyribose 5-phosphate via D-glyceraldehyde 3-phosphate. Under the optimized conditions with toluene-treated yeast cells, 356 mM (121 g/l) fructose 1,6-diphosphate was produced from 1,111 mM glucose and 750 mM potassium phosphate buffer (pH 6.4) with a catalytic amount of AMP, and the reaction supernatant containing the fructose 1,6-diphosphate was used directly as substrate for 2-deoxyribose 5-phosphate production with the E. coli cells. With 178 mM enzymatically prepared fructose 1,6-diphosphate and 400 mM acetaldehyde as substrates, 246 mM (52.6 g/l) 2-deoxyribose 5-phosphate was produced. The molar yield of 2-deoxyribose 5-phosphate as to glucose through the total two step reaction was 22.1%. The 2-deoxyribose 5-phosphate produced was converted to 2-deoxyribose with a molar yield of 85% through endogenous or exogenous phosphatase activity.
Horinouchi
Nobuyuki
N
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University.
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 / Q000737:chemistry / Q000378:metabolism*; D000438:Alcohols / Q000378:metabolism*; D000446:Aldehyde-Lyases / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005285:Fermentation; D005635:Fructosediphosphates / Q000096:biosynthesis; D005947:Glucose / Q000737:chemistry / Q000378:metabolism*; D007711:Klebsiella pneumoniae / Q000201:enzymology / Q000235:genetics; D015394:Molecular Structure; D010710:Phosphates / Q000494:pharmacology; D012267:Ribosemonophosphates / Q000096:biosynthesis*; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000378:metabolism*; D013696:Temperature
D000438:Alcohols; D005635:Fructosediphosphates; D010710:Phosphates; D012267:Ribosemonophosphates; C058245:2-deoxyribose 5-phosphate; D000446:Aldehyde-Lyases; C020019:deoxyribose-phosphate aldolase; D000079:Acetaldehyde; D005947:Glucose; C029063:fructose-1,6-diphosphate