Turanose, a potential novel sweetener in food industry, can be synthesized by Neisseria polysaccharea amylosucrase (NpAS). However, the malt-oligosaccharide byproduct affects the yield. In this study, the NpAS mutant G396S, which was expected to interfer with the extension of glucan by increasing steric hindrance, was obtained. The NpAS and G396S were heterologously expressed in Bacillus subtilis and enzyme properties were analyzed. Results showed that the polymerization activity of G396S was decreased. In addition, the mutant was used in the preparation of turanose. When using 2 M sucrose as substrate, the turanose yield reached 410.4 g·L-1, an increase of 61 g·L-1 compared with that of NpAS. When fructose was added, the optimal fructose concentration for G396S decreased from 0.75 M to 0.5 M. The turanose production reached 523 g·L-1 with the conversion rate of 76.5%. This study contributes the use of turanose in food industry.
Keywords: Bacillus subtilis; Enzymatic conversion; Molecular modification; NpAS; Turanose production.
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