Kinetic modelling of enzymatic starch hydrolysis

Kinetic modelling of enzymatic starch hydrolysis – a summary K.A. Bednarska The dissertation entitled ‘Kinetic modelling of enzymatic starch hydrolysis’ describes the enzymatic hydrolysis and kinetic modelling of liquefaction and saccharification of wheat starch. After the background information about the enzymes, the substrate and the basics of the model in the first chapter, we describe a model predicting the outcome of wheat starch liquefaction by α-amylase from Bacillus licheniformis at 50°C in chapter 2. We demonstrate the ability of the model to predict starch hydrolysis products larger than the oligosaccharides considered in the existing models. The model in its extended version follows all the products of wheat starch hydrolysis separately, and despite the quantitative differences, the qualitative predictions are satisfactory. We also show that the difference between the experimental and computed data might stem from the inaccuracy of the subsite map. In the following chapters the model is used to find a better description of the hydrolysis data at two temperatures (50°C and 80°C), by varying the energy values of the subsite map and evaluating the inhibition. We hypothesize that a subsite map that is based on the cleavage patterns of linear, short molecules does not account for the complexity of hydrolysis of amylopectin. The branched structure of amylopectin molecules influences the composition of the hydrolysis products by restricting the access to some of the bonds. The presence of branches creates steric obstacles for the enzyme. The used α-amylase has difficulties hydrolysing and accommodating α-(1,6)-glycosidic bonds, which imposes on the hydrolysis of the α-(1,4)-glycosidic bonds located in its proximity. On this basis, we analyse the subsite maps in detail and suggest which of the subsites are crucial when making predictions about the product composition of starch hydrolysates. On top of that we propose new subsite maps that allow a quantitative description of the experimental data. After the model was shown to work at different experimental conditions, we also test it at increased the dry matter content during wheat starch hydrolysis. We follow both the liquefaction by BLA and the saccharification process by glucoamylase from Aspergillus niger at low moisture content. The liquefaction model, is used to predict all of the products of wheat starch hydrolysis at higher dry matter contents (30-60 w/w%). The liquefaction model also creates the substrate matrices representing maltodextrins to be used in the saccharification model. The saccharification of liquefacts to glucose is followed with a new mechanistic model, also using the assumptions of the subsite theory. The saccharification model predicts all of the reaction products using the subsite maps of glucoamylase available in literature. The findings described in the thesis are summarized and put in context in the general discussion. We demonstrate how the parameters of the liquefaction model at low moisture contents were chosen. The outcomes of the model are also compared with the experimental data at 30-60 w/w%. Next, we test our liquefaction model with starch hydrolysis data at 5 and 60 w/w% taken from literature, to verify both the approach we used and the validity of the parameters we obtained in previous chapters. The method used to improve the subsite maps is also tested on another enzyme, Bacillus amyloliquefaciens α-amylase. After discussing the factors that influence saccharification at high dry matter contents, we conclude the chapter with describing the potential of stochastic modelling and its practical use.

Bibliographic Details

Main Author:	Bednarska, K.A.
Other Authors:	van Boekel, Tiny
Format:	Doctoral thesis biblioteca
Language:	English
Published:	Wageningen University
Subjects:	enzymes, hydrolysis, processing, starch, stochastic models, enzymen, hydrolyse, stochastische modellen, verwerking, zetmeel,
Online Access:	https://research.wur.nl/en/publications/kinetic-modelling-of-enzymatic-starch-hydrolysis
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