microPAT: A protocol for direct in vitro NMR observation of lactic acid bacteria fermentations
Parvaneh Ebrahimi,a Flemming Hofmann Larsen,a Henrik Max Jensenb andSøren Balling Engelsena* a Chemometrics and Analytical Technology, Department of Food Science, Faculty of Science, University of Copenhagen, Rolighedsvej 30, 1958 Frederiksberg, Denmark b DuPont Nutrition Biosciences ApS, Edwin Rahrs vej 38, 8220 Brabrand, Denmark * Corresponding Author: [email protected]
Lactic acid bacteria (LAB) are widely used in the food industry as starter cultures to manufacturefermented food, and more recently there has been an increasing interest in using LAB for biopreservation of food products. The study of the metabolism of these bacteria is therefore of great interest, and NMR is the only omics analytical technology that hasthe potential to measure their metabolome non-destructively. In vivo NMR can provide valuable information about the dynamic metabolism of microorganisms, for example for studying the adaptive responses of microorganism to differentenvironmental and stress factors such as substrate, cohabitation, deleterious metabolites, as well as temperature and pH. This paper describes a protocol for in vitro NMR measurement of the fermentation of lactic acid bacteria. We call itmicroPAT (microbial process analytical technology)  as it monitors in real time the microbial processes of the fermentation. Some of the experimental issues that were encountered during the process of developing the protocol as well as the selectedapproach to overcome them are presented.