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Monitoring of enzymatic hydrolysis of starch by microdialysis sampling coupled on‐line to anion exchange chromatography and integrated pulsed electrochemical detection using post‐column switching
Author(s) -
Torto N.,
Gorton L.,
MarkoVarga G.,
Emnéus J.,
Åkerberg C.,
Zacchi G.,
Laurell T.
Publication year - 1997
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/(sici)1097-0290(19971205)56:5<546::aid-bit8>3.0.co;2-i
Subject(s) - chemistry , chromatography , hydrolysis , hydrolysate , starch , ion chromatography , enzymatic hydrolysis , microdialysis , biochemistry , extracellular
A quantitative evaluation of the hydrolysis of wheat starch using Termamyl, a thermostable α‐amylase (endo‐1,4‐α‐d‐glucan, glucanohydrolase; EC 3.2.1.78), is reported. Data from the monitoring of the hydrolysis of wheat starch indicated that, after 1 h, glucose and maltooligosaccharides up to DP 7 were the main hydrolysis products and thus enabled optimization of a liquefication step during the production of L ‐lactic acid. The monitoring system used, both in the on‐ and off‐line mode, was based on continuous flow microdialysis sampling (CFMS) coupled to anion exchange chromatography and integrated pulsed electrochemical detection (IPED). A microdialysis probe equipped with a 5‐mm polysulfone (SPS 4005) membrane, with a molecular‐weight cut‐off of 5 kDa, was used to sample the hydrolysis products of native wheat starch at 90°C. Characteristic fingerprint separations were achieved by anion exchange chromatography after enzymatic hydrolysis. Post‐column switching improved the detection and, consequently, also quantification of the hydrolysates as fouling of the electrode could be reduced. Maltooligosaccharide standards were used for quantification and to verify the elution of the hydrolysates by spiking the off‐line samples. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 56: 546–554, 1997.