Molla, Getachew ShibabawGetachew ShibabawMollaKinfu, Birhanu M.Birhanu M.KinfuChow, JenniferJenniferChowStreit, Wolfgang R.Wolfgang R.StreitWohlgemuth, RolandRolandWohlgemuthLiese, AndreasAndreasLiese2019-08-222019-08-222016-12-19Biotechnology Journal 3 (12): 1600625 (2017-03-01)http://hdl.handle.net/11420/3159Enantiopure L-glyceraldehyde-3-phosphate (L-GAP) is a useful building block in natural biological and synthetic processes. A biocatalytic process using glycerol kinase from Cellulomonas sp. (EC 2.7.1.30) catalyzed phosphorylation of L-glyceraldehyde (L-GA) by ATP is used for the synthesis of L-GAP. L-GAP has a half-life of 6.86 h under reaction conditions. The activity of this enzyme depends on the Mg2+ to ATP molar ratio showing maximum activity at the optimum molar ratio of 0.7. A kinetic model is developed and validated showing a 2D correlation of 99.9% between experimental and numerical data matrices. The enzyme exhibits inhibition by ADP, AMP, methylglyoxal and Ca2+, but not by L-GAP and inorganic orthophosphate. Moreover, equal amount of Ca2+ exerts a different degree of inhibition relative to the activity without the addition of Ca2+ depending on the Mg2+ to ATP molar ratio. If the Mg2+ to ATP molar ratio is set to be at the optimum value or less, inorganic hexametaphosphate (PPi6) suppresses the enzyme activity; otherwise PPi6 enhances the enzyme activity. Based on reaction engineering parameters such as conversion, selectivity and specific productivity, evaluation of different reactor types reveals that batchwise operation via stirred-tank reactor is the most efficient process for the synthesis of L-GAP.en1860-731420163Wiley-VCHGlycerol kinaseL-Glyceraldehyde-3-phosphate instabilityMg to ATP ratio 2+Reaction kineticsReactor simulationBiowissenschaften, BiologieIngenieurwissenschaftenJournal Article10.1002/biot.201600625Other