Characterization of a recombinant sucrose isomerase and its application to enzymatic production of isomaltulose
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ORIGINAL RESEARCH PAPER
Characterization of a recombinant sucrose isomerase and its application to enzymatic production of isomaltulose Feng Zhang . Feng Cheng . Dong-Xu Jia . Yue-Hao Gu . Zhi-Qiang Liu Yu-Guo Zheng
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Received: 15 February 2020 / Accepted: 3 September 2020 Ó Springer Nature B.V. 2020
Abstract Objective To characterize a recombinant isomerase that can catalyze the isomerization of sucrose into isomaltulose and investigate its application for the enzymatic production of isomaltulose. Results A sucrose isomerase gene from Erwinia sp. Ejp617 was synthesized and expressed in Escherichia coli BL21(DE3). The enzymatic characterization revealed that the optimal pH and temperature of the purified sucrose isomerase were 6.0 and 40 °C, respectively. The enzyme activity was slightly activated by Mn2?and Mg2?, but partially inhibited by Ca2?, Ba2?, Cu2?, Zn2? and EDTA. The kinetic parameters of Km and Vmax for sucrose were 69.28 mM and 118.87 U/mg, respectively. The time course showed that 240.9 g/L of isomaltulose was produced from 300 g/L of sucrose, and the yield reached 80.3% after bioreaction for 180 min. F. Zhang F. Cheng D.-X. Jia Y.-H. Gu Z.-Q. Liu (&) Y.-G. Zheng Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, 18 Chaowang Road, Hangzhou 310014, People’s Republic of China e-mail: [email protected] F. Zhang F. Cheng D.-X. Jia Y.-H. Gu Z.-Q. Liu Y.-G. Zheng The National and Local Joint Engineering Research Center for Biomanufacturing of Chiral Chemicals, Zhejiang University of Technology, 18 Chaowang road, Hangzhou 310014, People’s Republic of China
Conclusions This recombinant enzyme showed excellent capability for biotransforming sucrose to isomaltulose at the substrate concentration of 300 g/L. Further investigations should be carried out focusing on selection of suitable heterologous expression system with the aim to improve its expression level. Keywords Sucrose isomerase Isomaltulose Biotransformation Purification Characterization
Introduction Isomaltulose (a-D-glucopyranosyl-1,6-fructose) is a natural isomer of sucrose (a-D-glucopyranosyl-1,2fructose), which has a similar appearance and taster to sucrose (Zhan et al. 2019). It is commonly used as a functional carbohydrate with superior properties, including lower glycemic index, slower digestion, lower insulin reaction, less cariogenicity and prolonged energy release (Zheng et al. 2019). Recently, isomaltulose has attracted wide attention of researchers because of its industrial applications either as an ideal sucrose substitute in food processing or as a starting material for producing surfactants and polymers (Mu et al. 2014; Watzlawick and Mattes 2009). Sucrose isomerase (EC 5.4.99.11, SIase) catalyzes the isomerization of sucrose into isomaltulose and trehalulose. In this bioconversion process, isomaltulose is the main product, and a little amount of
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