Antioxidative effect of Bacteroides thetaiotaomicron extracts: superoxide dismutase identification

Abstract

Bacteroides thetaiotaomicron, a bowel anaerobic commensal, seems to release enzymes detoxifying reactive oxygen species according to our recent work. This opportunistic pathogen would be beneficial in the case of an inflammatory process. To explore its role after an oxidative or nutritive stress, six to seven separate experiments were performed. The bacteria were grown on media restricted in growth factors or supplemented with bile. Their viability was checked after surface protein extraction. The extracts underwent 2D electrophoresis. Gel images were statistically analysed to construct "master" gels. Proteins were identified (peptide-mass fingerprinting technique). The effect of each extract on superoxide anions was evaluated (spectrophotometric method). Superoxide dismutase was identified and a major superoxide anion inhibition was shown by extracts obtained after a nutritive and oxidative stress without significant bacterial death. So, a therapeutic antioxidant potential is firmly hoped for. [figure: see text]

Interactions between Kluyveromyces marxianus from cheese origin and the intestinal symbiont Bacteroides thetaiotaomicron: Impressive antioxidative effects

“…However, superoxide dismutase is not detected in these extracts. As previously described (Hochart-Behra et al, 2008), chaperonin proteins were also found such as chaperone protein DnaK (c1), SCO1/SenC chaperone. Glutamate decarboxylase (s1), is another stress protein previously widely described in our extracts (Hochart-Behra et al, 2008).…”

“…The same process was applied to B. thetaiotaomicron ATCC 29741 and K. marxianus S-02-5 grown concomitantly and separately, in the same conditions as for their co-culture. After centrifugation, followed by three successive washes in Ringer medium ¼, the pellets of pure B. thetaiotaomicron ATCC 29741 and of pure K. marxianus S-02-5 and of B. thetaiotaomicron ATCC 29741 co-culturing with K. marxianus S-02-5 were placed in agitation in mild detergent solution containing 7 mM 3-[(3-cholamidopropyl) dimethylammonio]-1 propanesulfonate, 0.5 mM tris(hydroxymethyl) aminomethane hydrochloride at pH 6.8 (Serva, Heidelberg, Germany) -a cholic acid derivative -overnight at 4 C, as previously described for B. thetaiotaomicron (Hochart-Behra et al, 2008 at a rate of 4 ml of extraction solution per g of microorganisms. The suspensions were centrifuged and the supernatant was filtered 0.22 m (Millipore corporation, Bedford, MA, USA), and assayed for protein content (Hochart-Behra et al, 2008).…”

“…This bacterium has also received Orphan Designation for paediatric Crohn's disease by the US Food and Drug Administration (2013). Previously, we established that under defined conditions, B. thetaiotaomicron displayed antioxidant and anti-inflammatory properties and antioxidant enzyme/proteins, chaperonin proteins produced by this bacterium were identified (Hochart-Behra, Drobecq, Tourret, Dubreuil, & Behra-Miellet, 2014;Hochart-Behra et al, 2008). To gain more insights, we investigated the interactions between B. thetaiotaomicron ATCC 29741 and Kluyveromyces marxianus S-02-5 recently isolated from cheese and characterized for its beneficial attributes (Ceugniez, Drider, Jacques, & Coucheney, 2015).…”

“…The present work aimed at studying, under anaerobic conditions, the interactions between B. thetaiotaomicron ATCC 29741 and K. marxianus S-02-5. To this end, they were placed into a mild detergent mixture containing a cholic acid derivative preserving viability, in order to detect if these microorganisms are able to reject antioxidant elements from their surface at a higher level than previously described for B. thetaiotaomicron grown alone (Hochart-Behra et al, 2008. The activities against superoxide anion and hydrogen peroxide were measured in vitro.…”

“…The activities against superoxide anion and hydrogen peroxide were measured in vitro. These reactive oxygen species (ROS) are the first described in the oxidative cascade of phagocytes (Hochart-Behra et al, 2008. Two-dimensional (2D) gels and matrix-assisted laser desorption ionization time of flight e Time of flight (MALDI TOF/TOF) were used to separate and identify any antioxidant enzymes/proteins in the compounds secreted both microorganisms.…”