High Conversion of D-Fructose into D-Allulose by Enzymes Coupling with an ATP Regeneration System.

Abstract:

:D-Allulose is a rare monosaccharide that exists in extremely small quantities in nature, and it is also hard to prepare at a large scale via chemical or enzyme synthetic route due to low conversion and downstream separation complexity. Using D-psicose epimerase and L-rhamnulose kinase, a method enabling high conversion of D-allulose from D-fructose without the need for a tedious isomer separation step was established recently. However, this method requires expensive ATP to facilitate the reaction. In the present study, an ATP regenerate system was developed coupling with polyphosphate kinase. In our optimized reaction with purified enzymes, the conversion rate of 99% D-fructose was achieved at the concentrations of 2 mM ATP, 5 mM polyphosphate, 20 mM D-fructose, and 20 mM Mg2+ when incubated at 50 °C and at pH 7.5. ATP usage can be reduced to 10% of the theoretical amount compared to that without the ATP regeneration system. A fed-batch mode was also studied to minimize the inhibitory effect of polyphosphate. The biosynthetic system reported here offers a potential and promising platform for the conversion of D-fructose into D-allulose at reduced ATP cost.

journal_name

Mol Biotechnol

journal_title

Molecular biotechnology

authors

Xiao Q,Niu J,Liu H,Liu Y,Zhou X

doi

10.1007/s12033-019-00174-6

subject

Has Abstract

pub_date

2019-06-01 00:00:00

pages

432-441

issue

6

eissn

1073-6085

issn

1559-0305

pii

10.1007/s12033-019-00174-6

journal_volume

61

pub_type

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