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Spray Drying of Probiotic Bacteria: from Molecular Mechanism to Pilot-Scale Productio Song Huang

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Spray Drying of Probiotic Bacteria: from Molecular Mechanism to Pilot-Scale Productio Song Huang Spray drying of probiotic bacteria: From molecular mechanism to pilot-scale productio Song Huang To cite this version: Song Huang. Spray drying of probiotic bacteria: From molecular mechanism to pilot-scale productio. Food and Nutrition. AGROCAMPUS OUEST, 2017. English. tel-02791240 HAL Id: tel-02791240 https://hal.inrae.fr/tel-02791240 Submitted on 5 Jun 2020 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Distributed under a Creative Commons Attribution - ShareAlike| 4.0 International License RÉSUMÉ ABSTRACT Séchage par atomisation des bactéries probiotiques : des méca- Spray drying of probiotic bacteria: From molecular mechanism nismes de protection à la production à l’échelle pilote to pilot-scale production Les probiotiques sont des microorganismes vivants qui, ingérés Probiotics are live microorganisms that, when administered in en quantité suffi sante, exercent des effets positifs sur la santé. adequate amounts, confer a health benefi t on the host. Freeze La lyophilisation est aujourd’hui questionnée quant à sa consom- drying, the reference drying method, is currently challenged mation d’énergie et son caractère discontinu. S’il offre une al- because of its low energy-effi ciency and productivity. Therefore, ternative pour produire massivement des poudres probiotiques spray drying is expected to be an alternative and sustainable à faible coût, le séchage par atomisation induit quant à lui des method for producing probiotic powders. The issue remains stress thermiques et oxydatifs conduisant à des pertes de viabi- in the considerable inactivation of probiotics caused by high lité rédhibitoires. temperature and dehydration during the process. Dans ce travail, un procédé innovant de séchage par atomisation In this work, a novel spray-drying process for continuous est proposé. Du lactosérum doux concentré (jusqu’à 30% p/p) production of probiotics was challenged. Concentrated sweet est utilisé à la fois comme support de culture et de séchage de whey (up to 30% w/w dry matter) was used to both culture P. freudenreichii et L. casei. Ce procédé élimine les étapes inter- and spray dry P. freudenreichii ITG P20 and L. casei BL23. This médiaires à risque de contamination élevé, accroît la biomasse et process cuts down the steps between culturing and drying (e.g. améliore la viabilité des bactéries. harvesting, washing), increases the cell population after growth Les mécanismes sous-jacents ont été explorés au plan de la and improves spray drying productivity and probiotic viability. résistance bactérienne et des conditions de séchage. Le milieu The mechanisms were explored from bacterial physiology and drying concentré induit une osmoadaptation des bactéries par expres- process conditions. The hypertonic stress led to overexpression of sion de protéines de stress et accumulation de solutés compa- key stress proteins and accumulation of intracellular compatible tibles, conduisant à une tolérance accrue des probiotiques à solutes, which enhanced multistress tolerance. The presence of différents stress. La présence d’agrégats et la concentration en protein aggregates and optimal concentration of Mg2+ in matrix Mg2+ du milieu concentré pourraient également être impliquées. may also be involved. Le scale-up du procédé a été étudié : un schéma technologique The feasibility of scaling up this process was validated. A multi- semi-industriel impliquant séchage par atomisation, sur bande stage semi industrial drying process, coupling spray-drying with et en lit fl uidisé a permis d’atteindre une viabilité de 100% belt drying and fl uid-bed drying, was applied to further improve (> 109 CFU g-1). Par ailleurs, la fonctionnalité des poudres probio- the probiotic viability to 100% (> 109 CFU g-1). Moreover, the tiques a été évaluée in vitro and in vivo sur modèle porcelet. Ce functionality of these probiotic powders was investigated in vitro travail ouvre de nouvelles perspectives pour l’obtention à grande and in vivo with piglet model. This work opens new avenues échelle et selon un procédé sobre et durable de préparations pro- for the sustainable development of new starter and probiotic Song HUANG • 30 mai 2017 biotiques. preparations with enhanced robustness. Thèse AGROCAMPUS OUEST ÉCOLE DOCTORALE • Vie - Agro - Santé (VAS) Mots-clés : probiotiques ; séchage par atomisation ; osmorégula- Keywords: Probiotics; spray drying; osmoregulation; stress sous le label de l’Université Bretagne Loire LABORATOIRE D’ACCUEIL • UMR INRA - AGROCAMPUS OUEST tion ; tolérance ; viabilité tolerance; viability pour obtenir le grade de Science et Technologie du Lait et de l’Œuf (STLO) DOCTEUR D’AGROCAMPUS OUEST Spécialité Science de l’aliment THÈSE EN CO-TUTELLE • School of Chemical and Environmental Séchage par atomisation des bactéries probiotiques : des mécanismes de protection à la production à l’échelle pilote à l’échelle à la production de protection mécanismes : des probiotiques bactéries des atomisation par Séchage Engineering, Soochow University, Suzhou MURIEL THOMAS ong Séchage par atomisation Directrice de Recherche, INRA Micalis, France / présidente AGROCAMPUS OUEST • Institut supérieur des sciences agronomiques, CATHERINE STANTON agroalimentaires, horticoles et du paysage des bactéries probiotiques : 65 rue de Saint-Brieuc – CS84215 – F-35042 Rennes Cedex Professeure, University College Cork, Irlande / rapporteure Tél. : 02 23 48 50 00 S HUANG des mécanismes de protection à la CORDELIA SELOMULYA www.agrocampus-ouest.fr • Professeure, Monash University, Australie / rapporteure MAARTEN SCHUTYSER production à l’échelle pilote Professeur, Wageningen University, Pays-Bas / examinateur 2017-15 XIAO DONG CHEN — — Professeur, Soochow University, Chine / directeur de thèse ROMAIN JEANTET B-298 Professeur AGROCAMPUS OUEST, UMR INRA-AO STLO, France / directeur de thèse GWENAËL JAN Thèse Thèse Chargé de recherche, UMR INRA-AO STLO, France / co-directeur de thèse N° ordre : 2017-15 THESE / AGROCAMPUS OUEST N° Série : B-298 Sous le label de l’Université Européenne de Bretagne pour obtenir le diplôme de : DOCTEUR DE L'INSTITUT SUPERIEUR DES SCIENCES AGRONOMIQUES, AGRO-ALIMENTAIRES, HORTICOLES ET DU PAYSAGE Spécialité : Science de l'aliment Ecole Doctorale : VAS (Vie-Agro-Santé) présentée par : Song HUANG Spray drying of probiotic bacteria: From molecular mechanism to pilot-scale production Thèse préparée à l’UMR Sciences et Technologies du Lait et l’Œuf, INRA-Agrocampus Ouest, Rennes et School of Chemical and Environmental Engineering, Soochow University, Suzhou Soutenue le 30 May 2017 devant la commission d’examen Composition du jury : Catherine Stanton, Professeure, University College Cork, Irlande Rapporteur Cordelia Selomulya, Professeure, Monash University, Australie Rapporteur Muriel Thomas, Directrice de Recherche, INRA Micalis, France Présidente Maarten Schutyser, Professeur, Wageningen University, Pays-Bas Examinateur Xiao Dong Chen, Professeur, Soochow University, Chine Directeur de thèse Romain Jeantet, Professeur, Agrocampus Ouest, France Directeur de thèse Gwenaël Jan, Scientific Director, INRA (STLO), France Directeur de thèse This page is intentionally left blank. Foreword 欲穷千里目，更上一层楼。 - 王之涣 《登鹳雀楼》 Désirant embrasser toute l’étendue de ce paysage, je gravis un étage supplémentaire de la tour. De Zhihuan WANG (688-742) Acknowledgement I Acknowledgement Three years have gone by since the first time I arrived Rennes in 2014. France (Bretagne) is the first country (region) I reach abroad, and Rennes is the first city I live out of China. It seems not so far away from the first time I saw the amazingly huge green farms and clean blue sky here. As the first and only child who got the “college-level” education in my big family, it is hard to imagine this experience of doing a PhD in Europe. I am very grateful to this joint PhD project supported by Soochow University and Agrocampus Ouest. I also thank the joint research unit (STLO) between INRA and Agrocampus Ouest, and the Suzhou Key Lab of Green Chemical Engineering for providing the world-class laboratories. I would like to firstly thank Prof. Xiao Dong Chen, who teaches and helps me so much during the 7 years as my supervisor and mentor. Because of you, I fell in love with scientific research. You brought me to this “new world”, and because of this, everything happens now. Thank you so much for changing my life and making my dream of studying oversea comes true. Prof. Romain Jeantet, my dear “father” in France. Thank you so much for taking care of me in both study and life here. I appreciate this experience of working with you here, and the memories at beautiful Plonéour and also in China. I admire and also appreciate the great friendship between you and Dong. Because of it, I had this opportunity to come in France, to see this big big world. Thank you so much for everything you did for my PhD project. Dr. Gwénaël Jan, the super “Yoda”. I would like to thank you for your great supervison, showing me (your young padawan) the magic of microbiology and molecular world. I admire your enthusiasm in not only research,
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