Highly efficiency production of D-allulose from inulin using curli fiber multi-enzyme cascade catalysis

Yao Chen; Yujin Chen; Dengming Ming; Liying Zhu; Ling Jiang

doi:10.1016/j.ijbiomac.2023.124468

Highly efficiency production of D-allulose from inulin using curli fiber multi-enzyme cascade catalysis

Int J Biol Macromol. 2023 Jun 30:241:124468. doi: 10.1016/j.ijbiomac.2023.124468. Epub 2023 Apr 21.

Authors

Yao Chen¹, Yujin Chen¹, Dengming Ming², Liying Zhu³, Ling Jiang⁴

Affiliations

¹ State Key Laboratory of Materials-Oriented Chemical Engineering, College of Food Science and Light Industry, Nanjing Tech University, Nanjing 211816, China; College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, China.
² College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, China.
³ School of Chemistry and Molecular Engineering, Nanjing Tech University, Nanjing 211816, China.
⁴ State Key Laboratory of Materials-Oriented Chemical Engineering, College of Food Science and Light Industry, Nanjing Tech University, Nanjing 211816, China. Electronic address: jiangling@njtech.edu.cn.

PMID: 37088188
DOI: 10.1016/j.ijbiomac.2023.124468

Abstract

D-Allulose is a rare sugar with numerous physiological benefits and low caloric content, which can be obtained from inulin through enzymatic catalysis. In this study, we combined D-allulose 3-epimerase, exo- and endo-inulinases (EXINU and ENINU) with the NGTag/NGCatcher/CsgA system to accelerate D-allulose accumulation from inulin. Molecular dynamics simulations were used to screen linkers of appropriate length for ENINU. In vitro, we successfully observed the assembled NGCatcher_ENINU_CsgA, NGTag_EXINU, and DAERK fibers using fluorescent labelling with GFP, YFP, and mCherry. The optimal pH and temperature of the tagged variants were comparable to those of the wild-type, and the MD simulations showed that NGCatcher_ENINU_CsgA had improved stability in the working environment of EXINU. D-Allulose accumulation rate of the assembled enzymes cascade (NGCatcher_ENINU_CsgA/NGTag_EXINU_CsgA/DAERK) reached 0.25 g/L min^-1 (1.25 mg_D-allulose mg_DAERK^-1 min^-1) at an inulin concentration of 100 g/L. The assembled system greatly improves the high-valued productions of rare sugars from cheap biomass.

Keywords: CsgA; D-Allulose; Inulin; NGTag/NGCatcher; Self-assembly.

MeSH terms

Catalysis
Fructose* / chemistry
Hydrogen-Ion Concentration
Inulin*

Substances

psicose
Inulin
Fructose