Production of branched glucan polymer by a novel thermostable branching enzyme of Bifidobacterium thermophilum via one-pot biosynthesis containing a dual enzyme system

Abstract

Glycogen-like particles (GLPs) are applied in food, pharmaceutical, and cosmetics. The large-scale production of GLPs is limited by their complicated multi-step enzymic processes. In this study, GLPs were produced in a one-pot dual-enzyme system using Bifidobacterium thermophilum branching enzyme (BtBE) and Neisseria polysaccharea amylosucrase (NpAS). BtBE showed excellent thermal stability (half-life of 1732.9 h at 50 °C). Substrate concentration was the most influential factor during GLPs production in this system: GLPs yield and [sucrose]_ini decreased from 42.4 % to 17.4 % and 0.3 to 1.0 M, respectively. Molecular weight and apparent density of GLPs decreased significantly with increasing [sucrose]_ini. Regardless of the [sucrose]_ini, the DP 6 of branch chain length was predominantly occupied. GLP digestibility increased with increasing [sucrose]_ini, indicating that the degree of GLP hydrolysis may be negatively related to its apparent density. This one-pot biosynthesis of GLPs using a dual-enzyme system could be useful for the development of industrial processes.

Keywords: Amylosucrase; Apparent density; Bifidobacterium thermophilum; Branching enzyme; Digestibility; Glycogen-like particle.