Artificial multi-enzyme systems for the transformation of the prebiotic compound formaldehyde into stereodefined functional sugars by stepwise cascade biocatalysis.
Disaccharides are valuable oligosaccharides
with an increasing
demand in the food, cosmetic, and pharmaceutical industries. Disaccharides
can be manufactured by extraction from the acid hydrolysate of plant-derived
substrates, but this method has several issues, such as the difficulty
in accessing natural substrates, laborious product separation processes,
and troublesome wastewater treatment. A chemical synthesis using glucose
was developed for producing disaccharides, but this approach suffers
from a low product yield due to the low specificity and requires tedious
protection and deprotection processes. In this study, we adopted an
artificial strategy for producing a variety of value-added disaccharides
from low-cost starch through the construction of an in vitro synthetic enzymatic platform: two enzymes worked in parallel to
convert starch to glucose and glucose 1-phosphate, and these two intermediates
were subsequently condensed together to a disaccharide by a disaccharide
phosphorylase. Several disaccharides, such as laminaribiose, cellobiose,
trehalose, and sophorose, were produced successfully from starch with
the yields of more than 80% with the help of kinetic mathematical
models to predict the optimal reaction conditions, exhibiting great
potential in an industrial scale. This study provided a promising
alternative to reform the mode of disaccharide manufacturing.
Laminaribiose is a reducing disaccharide linked by a β‐1,3 glycosidic bond; it is also a precursor for building blocks in the pharmaceutical industry, a powerful germinating agent and antiseptic, as well as a potential prebiotic. In this study, an in vitro enzymatic biosystem composed of α‐glucan phosphorylase, laminaribiose phosphorylase, isoamylase, and 4‐glucanotransferase is designed for the one‐pot synthesis of laminaribiose from low‐cost maltodextrin and glucose. Through condition optimization, 51 mM laminaribiose is produced from 10 g L−1 maltodextrin (55.5 mM glucose equivalent) and 90 mM glucose. The product yield based on maltodextrin is 91.9%. To investigate the industrial potential of this in vitro enzymatic biosystem, the production of laminaribiose from high concentrations of substrates is also examined, and 179 mM laminaribiose is produced from 50 g L−1 of maltodextrin and 450 mM glucose. This in vitro enzymatic biosystem comprised of thermophilic enzymes can drastically decrease the manufacturing cost of laminaribiose and provide a green method for the production of other disaccharides using phosphorylases.
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