Enzymatic production of purified isomaltooligosaccharides prebiotic preparations and their full qualitative and quantitative characterization

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COMMUNAUTE FRANCAISE DE BELGIQUE ACADEMIE UNIVERSITAIRE WALLONIE-EUROPE GEMBLOUX AGRO-BIO TECH, UNIVERSITE DE LIEGE

ENZYMATIC PRODUCTION OF PURIFIED ISOMALTOOLIGOSACCHARIDES PREBIOTIC PREPARATIONS AND THEIR FULL QUALITATIVE AND QUANTITATIVE CHARACTERIZATION Dorothée GOFFIN

Dissertation submitted in fulfilment of the requirements for the degree of Doctor in Agricultural Sciences andBiological Engineering

Supervisor : Prof. Michel Paquot Co-supervisor : Prof. Christophe Blecker

2010

COMMUNAUTE FRANCAISE DE BELGIQUE ACADEMIE UNIVERSITAIRE WALLONIE-EUROPE GEMBLOUX AGRO-BIO TECH, UNIVERSITE DE LIEGE

ENZYMATIC PRODUCTION OF PURIFIED ISOMALTOOLIGOSACCHARIDES PREBIOTIC PREPARATIONS AND THEIR FULL QUALITATIVE AND QUANTITATIVE CHARACTERIZATION Dorothée GOFFIN

Dissertationsubmitted in fulfilment of the requirements for the degree of Doctor in Agricultural Sciences and Biological Engineering

Supervisor : Prof. Michel Paquot Co-supervisor : Prof. Christophe Blecker

2010

Copyright Aux termes de la loi belge du 22 mars 1886, sur le droit d’auteur, seul l’auteur a le droit de reproduire cet ouvrage ou d’en autoriser la reproduction de quelque manière et sousquelque forme que ce soit. Toute photocopie ou reproduction sous autre forme est donc faite en violation de la loi

GOFFIN Dorothée (2010) Enzymatic production of purified Isomaltooligosaccharide prebiotic preparations and their full qualitative and quantitative characterization. (PhD Thesis). Gembloux, Université de Liège, Gembloux Agro-Bio Tech Prebiotics are non-digestible food ingredients thatbeneficially affect the host health by stimulating the growth and/or activity of one or a limited number of bacteria in the colon, and moreover can have systemic effects on metabolism regulation. Recent studies have pointed out the certain links between their structure (DP, types of linkage, sequence) and the specificity and intensity of their effects on host health. Therefore, the knowledge ofthe exact composition of prebiotic preparations has become a crucial step in the development of new products. Amongst these prebiotics, isomaltooligosaccharides (IMO), produced enzymatically from hydrolyzed starch, are the most developed in Asia thanks to their many favourable properties for application in food industry. However, their full characterization hasn’t been achieved yet. In the presentthesis, an HPAEC-PAD method has been set up in order to quantify the IMO present in syrups. Moreover, this step-forward analytical method permitted us to point out the presence of unknown IMO. Two structural determination methods were then set up and applied. PGC-LC-ESI-ITMS2 including a chromatographic separation on a porous graphitized carbon column and producing MS2 fragment ion profilesspecific to the linkage position proved to be promising but delicate for certain combination of linkage, while 1D and 2D NMR experiments give unambiguous structural determinations but need a preliminary chromatographic preparation step. Moreover, two enzymes from different families (a glucosyl-transferase and an α-glucosidase) were tested on maltose individually or in combination. The IMO kinetics offormation and mixtures obtained were compared with a view to produce IMO preparations with singular profiles. Finally, a new original method was set up to convert deleterious digestible saccharides, present in IMO preparations, into gluconic acid which presents various advantageous techno-functional properties as well as nutritional properties, in particular, prebiotic. GOFFIN Dorothée (2010) Laproduction enzymatique de préparations purifiées d’Isomaltooligosaccharides prébiotiques ainsi que leur caractérisation qualitative et quantitative complète. (Thèse de doctorat). Gembloux, Université de Liège, Gembloux Agro-Bio Tech Les prébiotiques sont des ingrédients alimentaires non digestibles qui produisent des effets bénéfiques pour la santé de l’hôte en stimulant la croissance ou l’activité...
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