Construction of allitol synthesis pathway by multi-enzyme coexpression in Escherichia coli and its application in allitol production.
An engineered strain for the conversion of D-fructose to allitol was developed by constructing a multi-enzyme coupling pathway and cofactor recycling system in Escherichia coli. D-Psicose-3-epimerase from Ruminococcus sp. and ribitol dehydrogenase from Klebsiella oxytoca were coexpressed to form the multi-enzyme coupling pathway for allitol production. The cofactor recycling system was constructed using the formate dehydrogenase gene from Candida methylica for continuous NADH supply. The recombinant strain produced 10.62 g/l allitol from 100 mM D-fructose. To increase the intracellular concentration of the substrate, the glucose/fructose facilitator gene from Zymomonas mobilis was incorporated into the engineered strain. The results showed that the allitol yield was enhanced significantly to 16.53 g/l with a conversion rate of 92 %. Through optimizing conversion conditions, allitol was produced effectively on a large scale by the whole-cell biotransformation system; the yield reached 48.62 g/l when 500 mM D-fructose was used as the substrate.
National Engineering Laboratory for Industrial Enzymes, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Xiqi Road 32#, Tianjin Airport Economic Area, Tianjin, 300308, People's Republic of China.
Yueming
Y
Zhu
Hongyi
H
Li
Pingping
P
Liu
Jiangang
J
Yang
Xueli
X
Zhang
Yuanxia
Y
Sun
D019149:Bioreactors; D053898:Biosynthetic Pathways / Q000235:genetics*; D001711:Biotransformation; D002175:Candida / Q000201:enzymology / Q000235:genetics; D002238:Carbohydrate Epimerases / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005560:Formate Dehydrogenases / Q000235:genetics* / Q000378:metabolism; D005632:Fructose / Q000378:metabolism; D041121:Klebsiella oxytoca / Q000201:enzymology / Q000235:genetics; D060847:Metabolic Engineering; D009243:NAD / Q000378:metabolism; D019998:Racemases and Epimerases / Q000235:genetics / Q000378:metabolism; D045857:Ruminococcus / Q000201:enzymology / Q000235:genetics; D013401:Sugar Alcohol Dehydrogenases / Q000235:genetics* / Q000378:metabolism; D013402:Sugar Alcohols / Q000378:metabolism*; D017355:Zymomonas / Q000235:genetics
D013402:Sugar Alcohols; D009243:NAD; C003243:psicose; D005632:Fructose; C527823:allitol; D013401:Sugar Alcohol Dehydrogenases; C022529:ribitol 2-dehydrogenase; D005560:Formate Dehydrogenases; D019998:Racemases and Epimerases; D002238:Carbohydrate Epimerases