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. 2016 Jan 29:7:47.
doi: 10.3389/fmicb.2016.00047. eCollection 2016.

Ethanol Production by Selected Intestinal Microorganisms and Lactic Acid Bacteria Growing under Different Nutritional Conditions

Fouad M F Elshaghabee  1 , Wilhelm Bockelmann  2 , Diana Meske  2 , Michael de Vrese  2 , Hans-Georg Walte  3 , Juergen Schrezenmeir  4 , Knut J Heller  2
Affiliations

Ethanol Production by Selected Intestinal Microorganisms and Lactic Acid Bacteria Growing under Different Nutritional Conditions

Fouad M F Elshaghabee et al. Front Microbiol. .

Abstract

To gain some specific insight into the roles microorganisms might play in non-alcoholic fatty liver disease (NAFLD), some intestinal and lactic acid bacteria and one yeast (Anaerostipes caccae, Bacteroides thetaiotaomicron, Bifidobacterium longum, Enterococcus fecalis, Escherichia coli, Lactobacillus acidophilus, Lactobacillus fermentum, Lactobacillus plantarum, Weissella confusa, Saccharomyces cerevisiae) were characterized by high performance liquid chromatography for production of ethanol when grown on different carbohydrates: hexoses (glucose and fructose), pentoses (arabinose and ribose), disaccharides (lactose and lactulose), and inulin. Highest amounts of ethanol were produced by S. cerevisiae, L. fermentum, and W. confusa on glucose and by S. cerevisiae and W. confusa on fructose. Due to mannitol-dehydrogenase expressed in L. fermentum, ethanol production on fructose was significantly (P < 0.05) reduced. Pyruvate and citrate, two potential electron acceptors for regeneration of NAD(+)/NADP(+), drastically reduced ethanol production with acetate produced instead in L. fermentum grown on glucose and W. confusa grown on glucose and fructose, respectively. In fecal slurries prepared from feces of four overweight volunteers, ethanol was found to be produced upon addition of fructose. Addition of A. caccae, L. acidophilus, L. fermentum, as well as citrate and pyruvate, respectively, abolished ethanol production. However, addition of W. confusa resulted in significantly (P < 0.05) increased production of ethanol. These results indicate that microorganisms like W. confusa, a hetero-fermentative, mannitol-dehydrogenase negative lactic acid bacterium, may promote NAFLD through ethanol produced from sugar fermentation, while other intestinal bacteria and homo- and hetero-fermentative but mannitol-dehydrogenase positive lactic acid bacteria may not promote NAFLD. Also, our studies indicate that dietary factors interfering with gastrointestinal microbiota and microbial metabolism may be important in preventing or promoting NAFLD.

Keywords: Weissella confusa; arabinose; ethanol; fecal slurries; fructose; inulin; lactulose; non-alcoholic fatty liver disease.

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Figures

Figure 1
Figure 1
Separation of selected sugars, acids, and ethanol on a Metacarb 87H column (7.5 × 300 mm) with 0.0085 N H2SO4 at a flow rate of 0.3 ml/min and 65°C.
Figure 2
Figure 2
Ethanol production from glucose and fructose, respectively, of microorganisms grown anaerobically in MCB-medium at 37°C. Black, L. fermentum 92294; pink, W. confusa NRRL-B-14171; blue, S. cerevisiae 56101.
Figure 3
Figure 3
Metabolites profiles and colony forming units (cfu) of microorganisms grown anaerobically in MCB medium at 37°C with different supplements: Glu, glucose; Fru, fructose; Cit, citrate; Pyr, pyruvate. (A) L. fermentum 92294; (B) W. confusa NRRL-B-14171; (C) S. cerevisiae 56101.
Figure 4
Figure 4
Microbial alcohol dehydrogenase activity (ADH, mU/mL) of differently treated fecal slurries after 24 h incubation at 37°C in MCB containing fructose. A box-and whisker plot is shown, with the median indicated by the border of the black and gray box and the whiskers indicating maximal and minimal values. Slurry, without added bacterial strain; Ac, A. caccae DSM 14662; La, L. acidophilus NRRL-B-4495; Lf, L. fermentum 92294; Wc, W. confusa NRRL-B-14171. Different letters (a and b) indicate significant differences (P < 0.05) in post-hoc Tuckey test.
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