Hydrolysis of Maltoheptaose in Flow through Silicon Wafer Microreactors Containing Immobilised α‐Amylase and Glycoamylase

Abstract

In this study a silicon micro immobilised enzyme reactor (mIMER) has been applied for hydrolysis of maltoheptaose as a model maltodextrin and starch using immobilised aamylase (from Aspergillus oryzae) and glycoamylase (from Aspergillus niger). The influence of several parameters was investigated such as immobilisation chemistry, buffer constituents, pH, temperature, flow rate and substrate concentration. The conversion efficiency profile of the substrate was measured and the long-term stability of the reactor was tested. For separation and detection of the formed hydrolysis products, high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) was used. The results show that the mIMERs can also be used for hydrolysis of starch and also additionally be connected directly on-line with, e.g., liquid chromatography, making it possible to perform on-line characterisation and analysis of starch hydrolysis products.

Immobilization of α‐amylase on reactive modified fiber and its application for continuous starch hydrolysis in a packed bed bioreactor

“…However, for the following steps, a flow rate of 1.0 mL/min was chosen for practical reasons. Melander et al was reported that the hydrolysis efficiency decreases rapidly with increasing a flow rate for α ‐amylase and glucoamylase immobilized on silicon wafer. The immobilized inulinase on chitosan has been reported to hydrolyse inulin in continuous process.…”

Immobilization of alcohol dehydrogenase on ceramic silicon carbide membranes for enzymatic CH3OH production

“…For activation, 10 mL of 2.5% (v/v) GA was filtered through the membrane without applied pressure followed by a rinse with 50 mL of Milli‐Q water at 0.2 barg. This concentration of GA is commonly used for protein immobilization using functionalization with APTES or PEI (below) . Enzyme immobilization was completed by the filtration of ADH and subsequent wash as described for physical adsorption above.…”

Surface treatments and functionalization of metal‐ceramic membranes for improved enzyme immobilization performance

“…For activation, 10 mL of 2.5% ( v / v ) GA was filtered through the membrane without applied pressure followed by a rinse with 50 mL of Milli‐Q water at 0.2 barg. This concentration of GA is commonly used for protein immobilization using functionalization with APTES or PEI (below) . Enzyme immobilization was completed as described for physical adsorption.…”

“…PEI functionalization was carried out by filtration of 10 mL 0.1% (v/v) PEI solution with no applied pressure; PEI concentrations in this range are commonly used for enzyme immobilization. 25,26 Excess PEI was removed by filtration of 50 mL of Milli-Q water at 0.2 barg. Activation with GA, enzyme immobilization, and the subsequent wash were carried out as described for APTES-GA earlier.…”

“…This concentration of GA is commonly used for protein immobilization using functionalization with APTES or PEI (below). [25][26][27] Enzyme immobilization was completed as described for physical adsorption. This method is denoted the APTES-GA method.…”