One-pot enzymatic reaction sequence for the syntheses of d-glyceraldehyde 3-phosphate and l-glycerol 3-phosphate

Publikationstyp
Journal Article
Date Issued
2016-02-01
Sprache
English
Author(s)
Molla, Getachew Shibabaw  
Wohlgemuth, Roland  
Liese, Andreas  orcid-logo
Institut
Technische Biokatalyse V-6  
TORE-URI
http://hdl.handle.net/11420/4883
Journal
Journal of molecular catalysis / B Enzymatic  
Volume
124
Start Page
77
End Page
82
Citation
Journal of Molecular Catalysis B: Enzymatic (124): 77-82 (2016-02-01)
Publisher DOI
10.1016/j.molcatb.2015.12.004
Scopus ID
2-s2.0-84950263283
A one-pot enzymatic reaction sequence for the synthesis of optically pure d-glyceraldehyde 3-phosphate (d-GAP) and l-glycerol 3-phosphate (sn-G3P) was designed using fructose-bisphosphate aldolase from rabbit muscle (RAMA), sn-glycerol 3-phosphate dehydrogenase (sn-G3PDH) and formate dehydrogenase from Candida boidinii (FDH). The reaction sequence significantly improves the aldol cleavage of d-fructose 1,6-bisphosphate (d-F16BP) catalyzed by RAMA and yields 100% conversion of d-F16BP by overcoming thermodynamic limitation. The degradation kinetics of d-GAP under reaction conditions was investigated and a reaction kinetics model defining the entire cascade was developed. Validation of the model shows 98.5% correlation between experimental data and numerically simulated data matrices. The evaluation of different types of reactor was performed by combining the reaction kinetics model, mass balances and kinetics of the non-enzymatic degradation of d-GAP. Batch-wise operation in a stirred tank reactor (STR) is the most convenient procedure for the one-pot enzymatic syntheses of d-GAP and sn-G3P. The separation of the two products d-GAP and sn-G3P has been achieved using polyethylenimine (PEI)-cellulose TLC.
Subjects
Downstream processing
One-pot reaction sequence
Phosphorylated metabolite
Reaction kinetics
Reactor simulation
DDC Class
600: Technology