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See detailDéveloppement d’un procédé biotechnologique de production de molécules à note verte à partir de feuilles de betterave
Gigot, Cédric ULg

Doctoral thesis (2011)

La grande richesse des feuilles de betterave en 13-HPL (entre 10 et 12 UE1/g de poids frais), une enzyme produisant le (3Z)-hexénol, le (2E)-hexénal et le n-hexanal, fait de ce substrat un candidat ... [more ▼]

La grande richesse des feuilles de betterave en 13-HPL (entre 10 et 12 UE1/g de poids frais), une enzyme produisant le (3Z)-hexénol, le (2E)-hexénal et le n-hexanal, fait de ce substrat un candidat potentiel pour le développement d’un procédé biotechnologique de production d’arômes alimentaires. Maillon essentiel du système de défense des plantes, l’enzyme 13-HPL est inductible par des stimuli externes et son expression dépend de l’âge et de l’état physiologique de la plante. Au cours de ce travail, l’optimisation de l’expression de cette enzyme au sein du végétal a été étudiée préalablement au développement d’un procédé industriel visant à exploiter ce substrat. Ce procédé comprend successivement quatre opérations chimiques : une hydrolyse d’huile végétale riche en acides gras insaturés, une oxydation de ces acides, un clivage par les 13-HPLs et une purification des aldéhydes et des alcools synthétisés. Selon nos résultats, l’étape limitante de ce procédé est la synthèse d’aldéhydes qui présente, dans des conditions optimales de réaction, une capacité de production maximale de 0,502mM à l’échelle pilote (100L). L’inhibition suicide par le substrat et l’instabilité combinée de l’enzyme et de ces produits de réaction réduisent le taux de clivage des aldéhydes. La mise au point d’un système d’extraction des arômes volatils par flux gazeux et d’une alimentation fedbatch en substrat permet de réduire l’impact limitant de ces facteurs et d’accroître la capacité de production à 1,37mM. Afin de mieux comprendre les mécanismes réactionnels, l’enzyme 13-HPL de la betterave a été séquencée et clonée dans un vecteur procaryote permettant de disposer d’une source d’activité enzymatique indépendante des contraintes du milieu extérieur. L’utilisation de ces techniques a permis de développer un procédé bivalent (à la fois de source microbienne et végétale) de production d’arôme exploitable au sein de l’industrie alimentaire. [less ▲]

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See detailThe lipoxygenase metabolic pathway in plants: potential for industrial production of natural green leaf volatiles
Gigot, Cédric ULg; Ongena, Marc ULg; Fauconnier, Marie-Laure ULg et al

in Biotechnologie, Agronomie, Société et Environnement = Biotechnology, Agronomy, Society and Environment [=BASE] (2010), 14(3), 451-460

Lipoxygenase enzymatic pathway is a widely studied mechanism in the plant kingdom. Combined actions of three enzymes: lipase, lipoxygenase (LOX) and hydroperoxide lyase (HPL) convert lipidic substrates ... [more ▼]

Lipoxygenase enzymatic pathway is a widely studied mechanism in the plant kingdom. Combined actions of three enzymes: lipase, lipoxygenase (LOX) and hydroperoxide lyase (HPL) convert lipidic substrates such as C18:2 and C18:3 fatty acids into short chain volatiles. These reactions, triggered by cell membrane disruptions, produce compounds known as Green Leaf Volatiles (GLVs) which are C6 or C9-aldehydes and alcohols. These GLVs are commonly used as flavors to confer a fresh green odor of vegetable to food products. Therefore, competitive biocatalytic productions have been developed to meet the high demand in these natural flavors. Vegetable oils, chosen for their lipidic acid profile, are converted by soybean LOX and plant HPL into natural GLVs. However this second step of the bioconversion presents low yield due to the HPL instability and the inhibition by its substrate. This paper will shortly describe the different enzymes involved in this bioconversion with regards to their chemical and enzymatic properties. Biotechnological techniques to enhance their production potentialities will be discussed along with their implication in a complete bioprocess, from the lipid substrate to the corresponding aldehydic or alcoholic flavors. [less ▲]

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See detailPilot scale biotransformation of vegetal oil into natural green note flavor using sugar beet leaves as sources of hydroperoxide lyase
Gigot, Cédric ULg; Ongena, Marc ULg; Fauconnier, Marie-Laure ULg et al

Poster (2009, June 02)

Natural green note aromas (GLVs) are highly attractive flavors commonly used in the food industry. These are produced in extremely low levels upon physiological stress in plant organs of any sort. This ... [more ▼]

Natural green note aromas (GLVs) are highly attractive flavors commonly used in the food industry. These are produced in extremely low levels upon physiological stress in plant organs of any sort. This weak sporadic presence entails a very expensive extraction step to obtain pure GLVs. Therefore catalytic biotransformations of fatty acid sources, the initial substrate for GLVs, have been developed. Enzymatic defense pathways and particularly the LOX pathway produce the major part of GLVs. Unlike GLV molecules that are emitted in the atmosphere, the enzymes are extractible from the plant material. Thus, a combination of plant enzyme extracts and substrate preparations provides all the ingredients for GLV production. Besides, sugar beet leaves present high levels of hydroperoxide lyase among plant sources and are available in large amounts during three months. In this enzymatic pathway, fatty acids are successively transformed by lipase, lipoxygenase and hydroperoxide lyase into aldehydes and alcohols, final compounds of GLVs pathway. Limiting and problematic steps occur with the action of hydroperoxide lyase, when enzymatic catalysis is followed by an enzyme destabilization. Alternative substrates bind irreversibly to the heme group of the enzyme and end the reaction. This poster briefly describes the development of a complete bioprocess for natural GLV production, from hydrolysis to purification. A high level of biotransformation could be achieved using optimum experimental conditions and a cheap source of plant materials. [less ▲]

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See detailSugar beet leaves as new source of hydroperoxide lyase in a bioprocess producing green-note aldehydes.
Rabetafika, Holy-Nadia ULg; Gigot, Cédric ULg; Fauconnier, Marie-Laure ULg et al

in Biotechnology Letters (2008), 30(6), 1115-9

Hydroperoxide lyase activity was found in sugar beet leaves. Its optimum pH and temperature were, respectively, 6.7 and 22 degrees C. Under these conditions, conversion of linolenic acid 13-hydroperoxide ... [more ▼]

Hydroperoxide lyase activity was found in sugar beet leaves. Its optimum pH and temperature were, respectively, 6.7 and 22 degrees C. Under these conditions, conversion of linolenic acid 13-hydroperoxide to cis-3-hexenal with a maximum yield of 80% was reached after only 2 min. The stability of cis-3-hexenal was improved by acidifying the reaction medium. Based on these studies, a bioprocess producing green-note aldehydes in a laboratory-scale was achieved. [less ▲]

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See detailSugar beet : a source of biofuel an chemicals.
Destain, Jacqueline ULg; Gigot, Cédric ULg; Fauconnier, Marie-Laure ULg et al

Poster (2007, June)

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See detailSugar beet leaves as source of green notes.
Gigot, Cédric ULg; Rabetafika, Holy-Nadia ULg; Fauconnier, Marie-Laure ULg et al

Poster (2007, June)

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See detailSugar beet leaves as source of lyase in bioprocess producing green note aldehydes
Rabetafika, Holy-Nadia ULg; Gigot, Cédric ULg; Fauconnier, Marie-Laure ULg et al

Poster (2007)

Sugar beet leaves were used as source of lyase in a biocatalytic process for the production of C6-aldehydes. Their hydroperoxide lyase activity was revealed important. The optimum values of pH and ... [more ▼]

Sugar beet leaves were used as source of lyase in a biocatalytic process for the production of C6-aldehydes. Their hydroperoxide lyase activity was revealed important. The optimum values of pH and temperature for reactions were respectively 6.7 and 22 °C. A simple and fast process providing high molar conversion of hydroperoxides to aldehydes is proposed. The yield of the main product cis-3-hexenal reached 80% after only 2 min and was stabilised by acidifying the reaction medium to pH 2. [less ▲]

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