Journal: Applied and Environmental Microbiology

Article Title: H 2 O 2 Production in Species of the Lactobacillus acidophilus Group: a Central Role for a Novel NADH-Dependent Flavin Reductase

doi: 10.1128/AEM.04272-13

Figure Lengend Snippet: List of strains and plasmids used in this study

Article Snippet: NCC 533 , Lactobacillus johnsonii strain from the Nestec Culture Collection.

Techniques: Plasmid Preparation, Construct, Expressing

Journal: Applied and Environmental Microbiology

Article Title: H 2 O 2 Production in Species of the Lactobacillus acidophilus Group: a Central Role for a Novel NADH-Dependent Flavin Reductase

doi: 10.1128/AEM.04272-13

Figure Lengend Snippet: Growth and hydrogen peroxide concentration of L. johnsonii NCC 533 in LAPTg medium under anaerobic conditions (square symbols), aerobic conditions alone (circular symbols), or aerobic conditions with 0.5 mg ml−1 catalase added to the medium (triangular symbols). Culture densities were determined by optical density measurement at 600 nm (A), and H2O2 concentrations were determined by the phenol red enzymatic assay (B). The data represent duplicate experiments ± standard errors of the means.

Article Snippet: NCC 533 , Lactobacillus johnsonii strain from the Nestec Culture Collection.

Techniques: Concentration Assay, Enzymatic Assay

Journal: Applied and Environmental Microbiology

Article Title: H 2 O 2 Production in Species of the Lactobacillus acidophilus Group: a Central Role for a Novel NADH-Dependent Flavin Reductase

doi: 10.1128/AEM.04272-13

Figure Lengend Snippet: H2O2 production of L. johnsonii NCC 533 (wild type [wt]) and the derivatives NCC 9333 (Δpox), NCC 9334 (Δlox), and NCC 9337 (Δnox). Anaerobic logarithmic-phase cultures were transferred to a shake flask and incubated at 37°C. After 1 h (open bars) and 2 h (closed bars), cells were removed by centrifugation, and H2O2 concentrations were determined in the culture medium, using the phenol red-peroxidase enzymatic assay. Data represent the average of three independent experiments.

Article Snippet: NCC 533 , Lactobacillus johnsonii strain from the Nestec Culture Collection.

Techniques: Incubation, Centrifugation, Enzymatic Assay

Journal: Applied and Environmental Microbiology

Article Title: H 2 O 2 Production in Species of the Lactobacillus acidophilus Group: a Central Role for a Novel NADH-Dependent Flavin Reductase

doi: 10.1128/AEM.04272-13

Figure Lengend Snippet: Typical NADH-dependent flavin reductase activity in L. johnsonii cell extract. NADH consumption rates were measured by absorption at 340 nm (A). Endpoint H2O2 concentrations were determined using the phenol red assay (B). Either 500 μM NADH was used, replaced by 500 μM NADPH where indicated (A), or 250 μM NADH was replaced by 250 μM NADPH where indicated (B). A 250 μM or 25 μM concentration of FAD, FMN, or riboflavin was added as a flavin source. The protein concentration in the cell-free extracts was determined by the MicroBCA assay. Data represent the average of technical triplicates ± standard deviation and are representative of cell extracts derived in comparable experiments.

Article Snippet: NCC 533 , Lactobacillus johnsonii strain from the Nestec Culture Collection.

Techniques: Activity Assay, Concentration Assay, Protein Concentration, Standard Deviation, Derivative Assay

Journal: Applied and Environmental Microbiology

Article Title: H 2 O 2 Production in Species of the Lactobacillus acidophilus Group: a Central Role for a Novel NADH-Dependent Flavin Reductase

doi: 10.1128/AEM.04272-13

Figure Lengend Snippet: Typical NADH-dependent flavin reductase activity in cell extracts of wild-type L. johnsonii NCC 533 and its LJ_0548 LJ_0549 (NCC 9359) mutant derivative, with and without complementation by plasmid-borne expression of one (pDP1016 and pDP1017) or both (pDP1019) of the deleted genes. Standard assay conditions were employed, containing 500 μM NADH and 25 μM FMN (NADH consumption rates) and 250 μM NADH and 25 μM FMN (to determine H2O2 concentration). NADH consumption rates were measured by absorption at 340 nm (A). H2O2 concentrations were determined after 10 min of reaction using the phenol red assay (panel B). Protein concentrations in the cell extracts were determined by MicroBCA assay. Data represent the average of three technical replicates ± standard deviation and are representative for activity measured in multiple (>3) cell extracts.

Article Snippet: NCC 533 , Lactobacillus johnsonii strain from the Nestec Culture Collection.

Techniques: Activity Assay, Mutagenesis, Plasmid Preparation, Expressing, Concentration Assay, Standard Deviation

Journal: Applied and Environmental Microbiology

Article Title: H 2 O 2 Production in Species of the Lactobacillus acidophilus Group: a Central Role for a Novel NADH-Dependent Flavin Reductase

doi: 10.1128/AEM.04272-13

Figure Lengend Snippet: H2O2 production in L. johnsonii NCC 533 and NCC 9359, with or without complementation of LJ_0548 and LJ_0549 under aerobic conditions. Anaerobic logarithmic-phase cultures of wild-type Lactobacillus johnsonii and its deletion derivative were transferred to a shake flask and incubated at 37°C. After 1 h, cells were removed by centrifugation, and H2O2 concentrations were determined in the culture medium, using the phenol red assay. Data represent the average of three independent experiments ± standard deviation.

Article Snippet: NCC 533 , Lactobacillus johnsonii strain from the Nestec Culture Collection.

Techniques: Incubation, Centrifugation, Standard Deviation