This manuscript is dedicated to Wolf-Dieter "Woody" Fessneront he occasion of his 60th birthday.Biohybrid nanoreactors operating in one-pot cascade reactions were designed by co-immobilizationofe nzymes in an inorganic layered matrix,n amely,l ayered double hydroxides.T hese biohybrid systems were devoted to prepare dihydroxyacetone phosphate (DHAP) and phosphorylated sugars throughs tereoselectiveC ÀCb ond formation. In the first system,t wo kinases were exploited for the in situ generation of DHAP.I ncreasing the complexity,t he second nano-bioreactorc ombined up to four enzymest ol ead to d-fructose-6-phosphate from the aldol-catalyzed addition of dihydroxyacetone to d-glyceraldehyde-3-phosphate generated in situ from DHAP.T he biohybrid catalysts howedt he same reactionr ate as that of the free enzymes and was reusable.Ta ndema nd cascade reactions were highlighted by Trost et al. in 2008 [1] as being of fundamentali mportance to develop greener syntheses. Biocatalysis takes am ajor place in this field, in particular enzymatic cascades,w hich create artificial pathways for the preparation of chiral or complex chemicals.[2] This emerging concept was recently termed "systemb iocatalysis" by Fessner.[3] Nature can be mimicked by designing artificial metabolic and non-naturalb iocatalytic pathways in vitro. In such multienzymatics ystems, enzymes are brought together in the same reactionv essel, in which the product of one enzymei st he substrate of the other.T he reactionp arameters (e.g.,p H, enzyme quantity,s ubstrate concentration, cosolvent, temperature, etc.) are easier to control than in in vivo systems and can be adjusted to reach high yields and purities.[4-6] To approachn aturals ystemsf or which the cooperating enzymes are organized in specific assemblies leading, in some cases, to ac hanneling effect,t he artificial metabolic pathways can be immobilized following different strategies. They have been nicely reviewed by Schoffelen et al.[5] The strategies are classified in order of increasing controlo ft he enzyme organization: random co-immobilization to as olid support (e.g.,a garose, [7] chitin, [8] nylon, [8] polystyrene, [9] polyvinyl chloride [10] )o rn anoparticles( e.g.,g oldn anoparticles, [11] sol-gelm atrix [12] ), sequential immobilization in microfluidic channels, positional immobilization on DNA nanostructures, site-specific enzymei mmobilization, and finally scaffold-free cross-linking.In this work, we aim to design cascader eactions for the enzymatics ynthesis of phosphorylated sugars followingt he random co-immobilizationo fe nzymes embedded in layered double hydroxide (LDH) nanoplatelets. Indeed LDHs, M II 1Àx M III x (OH) 2 X x/q ·n H 2 O( Supporting Information, Scheme S1), owing to their two-dimensional structure, physicochemical properties, and controllable nanoplateleta ggregation, [13] offer as uitable microenvironment that favorsahigh loading of enzymes, an increase in their stability, andp reservation of their activity. [14] In the course of our work on green by design,e fficie...

Tw ok inases and the key fructose-6-phosphate aldolase (FSA) were involved in ao ne-pot multienzymatic procedure to obtain rare l-sugars. Starting from simplea chiral substrates such as glycolaldehyde andf ormaldehyde, l-glyceraldehyde-3phosphate( l -G3P) was generated. When this latter compound was used as an FSA acceptor substrate along with four different donor compounds,f our terminally phosphorylated l-monosaccharides from C 5 to C 7 chain lengths were preparedi ng oodi solated yields.