Coimmobilization and colocalization of a glycosyltransferase and a sucrose synthase greatly improves the recycling of UDP-glucose: Glycosylation of resveratrol 3-O-β-D-glucoside

Glycosyltransferase Co‐Immobilization for Natural Product Glycosylation: Cascade Biosynthesis of the C‐Glucoside Nothofagin with Efficient Reuse of Enzymes

“…The rate enhancement for the co‐immobilized enzyme preparation can probably be ascribed to shortened diffusion paths when enzymes are co‐localized on the porous surface of the solid carrier. Our results are consistent with study of Rocha‐Martin and colleagues who showed 2.4‐fold benefit of glycosyltransferase co‐immobilization, compared to individual enzyme immobilization on separate carrier particles, on piceid glycosylation from sucrose‐derived UDP‐glucose [17] . The effect was dedicated to an enhanced efficiency of the UDP/UDP‐glucose shuttle when enzymes are co‐localized on the solid carrier.…”

“…Recently, Guisan, Rocha‐Martin and their co‐workers have shown immobilization of bacterial sucrose synthases on suitably activated agarose carriers [15d] . They also demonstrated glycosyltransferase co‐immobilization on agarose, [17] as already discussed. Their studies showed glycosyltransferase immobilization in good loading (several mg protein/g carrier), high yield (≥50%) and typically good effectiveness (≥44%).…”

“…Recent study suggests that immobilization of O ‐glycosyltransferase (from apple) and sucrose synthase (from Acidithiobacillus caldus ) to co‐localize the two enzyme activities on the same porous carrier benefits the synthetic efficiency of the bi‐enzymatic glycosylation cascade [17] . The rate of 5‐O‐β‐glycosylation of piceid (3‐O‐β‐ d ‐glucoside of resveratrol) was enhanced ∼2.4‐fold compared to reaction of the two enzymes immobilized individually on separate carriers [17] . Therefore, to perform the 3‐ C ‐β‐glycosylation of phloretin efficiently, we considered co‐immobilization of Os CGT and Gm SuSy to place the enzymes closely together under the confinement of the solid surface.…”

“…Glycosyltransferase co‐immobilization can be achieved in principle via immobilizing each enzyme according to its own, enzyme‐specific strategy. Guisan, Rocha‐Martin and coworkers have recently presented an important example of such “individualized” glycosyltransferase co‐immobilization, applied to a natural product glycosyltransferase and a sucrose synthase, as discussed later in context of the evidence from the current study [17] . Alternatively, glycosyltransferase co‐immobilization can be uniform in the strategy used for tethering the enzymes to the solid carrier surface.…”

Multi-Combilipases: Co-Immobilizing Lipases with Very Different Stabilities Combining Immobilization via Interfacial Activation and Ion Exchange. The Reuse of the Most Stable Co-Immobilized Enzymes after Inactivation of the Least Stable Ones

“…The paper shows, as many others [66,[113][114][115][116][117][118], how different immobilization protocols may alter enzyme specificity. Moreover, it gives some clues, which need to be confirmed, on how the order of the co-immobilization of enzymes may give biocatalysts with different activity properties, mainly when diffusional limitations may be raised and the activity of the co-immobilized enzymes differ each other with different substrates [112,128]. Another interesting question that is opened in this communication is to make a deeper analysis on the reasons that cause that the use of a very high ionic strength on these biocatalysts is not convenient to release the enzymes immobilized following the presented strategy, problem that did not exist using galactosidase.…”