Huan Jiang

Interaction mechanism between green tea extract and human α-amylase for reducing starch digestion.

This study evaluated the inhibitory effects of the green tea extract on human pancreatic α-amylase activity and its molecular mechanism. The green tea extract was composed of epicatechin (59.2%), epigallocatechin gallate (14.6%) and epicatechin gallate (26.2%) as determined by HPLC analysis. Enzyme activity measurement showed that % inhibition and IC50 of the green tea extract (10%, based on starch) were 63.5% and 2.07 mg/ml, respectively. The Michaelis-Menten constant remained unchanged but the maximal velocity decreased from 0.43 (control) to 0.07 mg/(ml × min) (4 mg/ml of the green tea extract), indicating that the green tea extract was an effective inhibitor against α-amylase with a non-competitive mode. The fluorescence data revealed that the green tea extract bound with α-amylase to form a new complex with static quenching mechanism. Docking study showed the epicatechin gallate in the green tea extract presented stronger affinity than epigallocatechin gallate, with more number of amino acid residues involved in amylase binding with hydrogen bonds and Van der Waals forces. Thus, the green tea extract could be used to manipulate starch digestion for potential health benefits.

Phytonutrients for controlling starch digestion: Evaluation of grape skin extract

The objective of this work was to evaluate the structure-function relationship between grape skin extract and human α-amylase. The grape skin extract was characterised as resveratrol-3-O-glucoside by RP-HPLC-ESI-MS, which showed strong inhibition towards α-amylase and the IC50 value was 1.35 mg/ml. The kinetic results demonstrated grape skin extract obeyed the non-competitive mode against amylase. Fluorescence data revealed the ability of grape skin binding to amylase belonged to static quenching mechanism with a complex formation and there was only one binding site in α-amylase for grape skin extract. Docking study showed a best pose with total energy value of -118.3 kJ/mol and grape skin extract interacted with side chain of Asp300 with hydrogen bonds and Van der Waals forces. This preliminary observation provides the basis for further evaluation of the suitability of grape skin extract as natural inhibitor with potential health benefits.

Enzymatic modification of corn starch with 4-α-glucanotransferase results in increasing slow digestible and resistant starch.

In this study, partial 4-α-glucanotransferase (4αGT) treatment was used to modulate the fine structure responsible for the slow digestion and resistant property of starch. Normal corn starch modified using 4αGT for 4h showed an increase of slowly digestible starch from 9.40% to 20.92%, and resistant starch from 10.52 to 17.63%, respectively. The 4αGT treatment decreased the content of amylose from 32.6% to 26.8%. The molecular weight distribution and chain length distribution of 4αGT-treated starch showed a reduction of molecular weight and a great number of short (DP<13) and long (DP>30) chains through cleaving and reorganization of starch molecules. Both the short and long chain fractions of modified amylopectin were attributed to the low in vitro digestibility. The viscosity was inversely related to the digestibility of the 4αGT-treated starch. These results suggested that the 4αGT modified starch synthesized the novel amylopectin clusters with slow digestible and resistant character.

Partial branching enzyme treatment increases the low glycaemic property and α-1,6 branching ratio of maize starch.

Partial branching enzyme treatment was used to modulate the starch fine chain structure responsible for a high content of slowly digestible starch fraction. Normal maize starch modified using branching enzyme for 4h showed a maximum slowly digestible starch content of 23.90%. The branching enzyme hydrolysis decreased the amylose content from 32.8% to 12.8%. The molecular weight distribution of enzyme-treated starches showed a larger proportion of low molecular weight fractions appeared in the enzyme treated starch sample compare to native starch. The number of shorter chains (DP<13) increased from 18.71% to 28.23.1%, accompanied by a reduction of longer chains (DP>30) from 20.11% to 11.95%. (1)H NMR spectra showed an increase of α-1,6 branching ratio from 4.7% to 9.4% during enzyme treatment. The increase in the amount of shorter chains and more α-1,6 linkages likely contribute to their slow digestion property. These results suggest that starches treated with partial branching enzyme synthesis a novel branched structure with slowly digestible character.

Improved the slow digestion property of maize starch using partially β-amylolysis.

In this study, partial β-amylolysis was used to modulate the starch fine structure responsible for the slow digestion property of starch. Normal maize starch modified using β-amylase for 2 h showed an increase of slowly digestible starch from 11.16% to 24.38%. The β-amylase treatment increased the amylose content from 28.4% to 32.5%, decreased the molecular weight from 32.5×10(7) to 3.8×10(5) g/mol and increased the number of shorter chains (DP<13) from 25.5% to 41.1%, accompanied by a reduction of longer chains (DP>13). (1)H NMR spectra showed an increase of α-1,6 linkages from 7.4% to 10.1% in the enzyme treated starches. Both the increase in the amount of shorter chains and the increase in α-1,6 linkages were attributed to the slow digestion property of starch. These results suggest that starches treated with partial β-amylolysis retain a branched structure and slow digestibility.