GROWTH RESPONSE OF LACTOBACILLUS RHAMNOSUS TO MICROAEROPHILIC INCUBATION AND ADDITION OF REDUCTANTS IN A RIBOFLAVIN MICROTITER PLATE ASSAY

Abstract

ABSTRACT Given the importance of riboflavin in human diets, a rapid riboflavin assay is needed by the food industry to allow simultaneous determination of riboflavin in numerous food sources. The growth responses of Lactobacillus rhamnosus to microaerophilic conditions and three reducing agents were examined as an attempt to reduce riboflavin assay time. Ascorbic acid, dithiothreitol and thioglycollate were applied at three concentrations. None of the concentrations of reducing agents caused a further increase of bacterial optical density (OD) or had a significant effect on the growth rates. More optimal microaerophilic conditions were achieved through the use of an anaerobic chamber. The growth rate of L. rhamnosus under microaerophilic conditions (0.30 ± 0.05/h) was not significantly different (P > 0.05) from the growth rate of the bacteria under aerobic conditions (0.34 ± 0.07/h). However, after 14 h of bacterial growth, the OD of the bacterial culture under anaerobic hood incubation (0.71 ± 0.09) was significantly higher (P < 0.05) than under aerobic incubation conditions (0.58 ± 0.05). Additional research is needed involving more controlled atmospheres for incubation during growth and a wider range of reductants to optimize and shorten the time required for riboflavin microtiter plate assay.PRACTICAL APPLICATIONSQuantifying riboflavin in foods and food ingredients is important to the food industry. Lactobacillus rhamnosus growth response to varying levels of riboflavin is used to estimate the amount of this vitamin. In an effort to optimize growth on microtiter plates and decrease assay time, microaerophilic conditions were examined. Although rate of growth was not altered, cell concentration measured as optical density (OD) did increase. Microaerophilic incubation of microtiter plates represents a potential approach to decrease assay time and provide the food industry with more timely results.