Development of novel starch-based dietary fiber using microbial glucanotransferase

In this study, a glucanotransferase from prokaryotic Azotobacter chroococcum NCIMB 8003 was recombinantly expressed and its biochemical characteristics and bioconversion ability for starch were investigated. The purified enzyme has the optimum activity at 55 °C and pH 6.5-7.0, as well as a melting temperature of 62 °C. The double-charged ion Ca2+ stimulated the activity of the enzyme by approximately 2.4 times. The kinetic parameters and specificity analysis revealed that this glucanotransferase had a higher affinity for high-amylose starch. During the transglycosylation reaction, the starch molecule was converted into a relatively small polymer with a narrow size distribution. For the enzyme modification of high-amylose starch for 72 h, the amount of α-1,6 linkages increased from 1.9% to 22.7% and the content of resistant starch (RS) increased from 3.18% to 17.83%. In addition, the fine structure displayed the reuteran-like highly branched glucan linked by single linear α-1,6 linkages and α-1,4/6 branching points. These results revealed that a promising prebiotic dietary fiber was synthesized from starch with glucanotransferase modification.