Characterization of a d-lyxose isomerase from Bacillus velezensis and its application for the production of d-mannose and l-ribose

Abstract

d-Mannose and l-ribose are two important monosaccharides, which have attracted public attention recently because of their great application potentials in food, cosmetic and pharmaceutical industries. Sugar isomerases catalyze the sugar isomerization and therefore can be used as the biocatalysts for production of the high-value sugars from inexpensive sugars. l-arabinose isomerase catalyzes the conversion of l-arabinose to l-ribulose, while d-lyxose isomerase catalyzes l-ribulose and d-fructose to l-ribose and d-mannose, respectively. In this paper, a putative d-LI from Bacillus velezensis (BvLI) was identified, characterized and used to produce d-mannose and l-ribose from d-fructose and l-arabinose, respectively. The recombinant BvLI exhibited a maximum activity at 55 °C and pH 6.5, in the presence of 0.1 mM Co2+. Approximately 110.75 g/L d-mannose was obtained from 500 g/L d-fructose in 6 h by the recombinant BvLI, and approximately 105 g/L l-ribose was obtained from 500 g/L l-arabinose in 8 h by the successive biocatalysis of l-arabinose isomerase from Bacillus licheniformis (BlAI) and BvLI.

Recent advances in biotransformation, extraction and green production of D-mannose

“…The isomerases currently used in the production of D-mannose mainly include D-lyxose isomerase (LIase) ( Guo et al, 2019 ; Huang et al, ; Park et al, 2010 ), D-mannose isomerase (Mlase) ( Hu et al, 2016a , Hu et al, 2016b ), cellobiose 2-epimerase (CEase) ( Park et al, 2011 ) and mannose 2-epimerase ( Saburi et al, 2019 ), etc ( Fig. 2 ).…”

“…Compared with the above isomerases, D-LIase is an aldose-ketose enzyme with a greater application value. It has broad substrate specificity which catalyzes the reverse isomerization of many functional sugars ( Guo et al, 2019 ), such as D-xylulose and D-lyxose, D-fructose, D-mannose, L-ribulose, and L-ribose ( Huang et al, 2018 ). The genes of D-LIase are widely distributed in many microorganisms such as Aerogenes PRL-R3 ( Hirose et al, 2001 ), Bacillus licheniformis DSM13 ( Patel et al, 2011 ), Lactobacillus RI-39 ( Zhang et al, 2019 ), Dictyostelium dilatum DSM 6724 ( Choi et al, 2012 ), Escherichia coli O157:H7 ( van Staalduinen et al, 2010 ), P. stuartii KCTC 2568 ( Kwon et al, 2010 ) and S. proteamaculans KCTC 2936 ( Park et al, 2010 ), etc.…”

“…For example, Guo et al identified a D-LIase from B. velezensis CICC 24777, in the presence of 0.1 mM Co 2+ , the genetic engineered D-LIase obtained exhibit maximum activity at 55 °C and pH 6.5. When using 25 U/mL enzyme and 500 g/L D-fructose as a substrate for 6 h, the reaction basically reached equilibrium and production rate of D-Mannose is 18.46 g/L/h ( Guo et al, 2019 ). Hao et al identified and characterized a novel putative D-LIase from the thermophilic strain Caldanaerobius polysaccharolyticus ( Wu et al, 2020 ).…”

Engineering the thermostability of d-lyxose isomerase from Caldanaerobius polysaccharolyticus via multiple computer-aided rational design for efficient synthesis of d-mannose

“…Until now, many types of d -LIase have been identified and characterized from Thermofilum sp. [ 8 ], Thermoprotei archaeon [ 9 ], Caldanaerobius polysaccharolyticus [ 10 ], Thermoflavimicrobium dichotomicum [ 11 ], Bacillus velezensi s [ 12 ], Thermosediminibacter oceani [ 13 ], and so on. Of these, some D-LIases achieve higher catalytic efficiency, such as C. polysaccharolyticus (conversion rate 25.6% with 45% enzyme activity retained at 65 °C after incubation for 3 h) [ 10 ], T. dichotomicum (conversion rate 24.2% and half-life 1.45 h at 65 °C) [ 11 ], and S. proteamaculans (conversion rate 20% and half-life 0.09 h at 50 °C) [ 14 ].…”

Biochemical and Structural Characterisation of a Novel D-Lyxose Isomerase From the Hyperthermophilic Archaeon Thermofilum sp.

“…Several LIs, including the enzyme from Bacillus velezensis (Guo et al, 2019), have also been applied in the production of L-ribose, a common intermediate of nucleoside based pharmaceuticals (Okano, 2009). The LIs have been structurally classified as members of the cupin superfamily, which is one of the most functionally diverse classes of proteins.…”