Open AccessD-Allulose Production from D-Fructose by Permeabilized Recombinant Cells of Corynebacterium glutamicum Cells Expressing D-Allulose 3-Epimerase Flavonifractor plautii
Author(s) -
Chul-Soon Park,
Tae-Yong Kim,
Sunwook Hong,
KyungChul Shin,
Kyoung-Rok Kim,
DeokKun Oh
Publication year - 2016
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0160044
Subject(s) - corynebacterium glutamicum , recombinant dna , escherichia coli , biochemistry , enzyme kinetics , enzyme , fructose , chemistry , microbiology and biotechnology , biology , gene , active site
A d -allulose 3-epimerase from Flavonifractor plautii was cloned and expressed in Escherichia coli and Corynebacterium glutamicum . The maximum activity of the enzyme purified from recombinant E . coli cells was observed at pH 7.0, 65°C, and 1 mM Co 2+ with a half-life of 40 min at 65°C, K m of 162 mM, and k cat of 25280 1/s. For increased d -allulose production, recombinant C . glutamicum cells were permeabilized via combined treatments with 20 mg/L penicillin and 10% (v/v) toluene. Under optimized conditions, 10 g/L permeabilized cells produced 235 g/L d -allulose from 750 g/L d -fructose after 40 min, with a conversion rate of 31% (w/w) and volumetric productivity of 353 g/L/h, which were 1.4- and 2.1-fold higher than those obtained for nonpermeabilized cells, respectively.