For ribotyping, all the isolates were grown in Luria Bertani (LB) broth and total genomic DNA was extracted.
For sporulation assays in broth, the total counts of vegetative cells and spores were compared (in CFU/mL, mean of 3 biological replicates).
Furthermore, compared to the control growing in AYE broth, the total RNA extracted from Lp exposed to water dropped markedly after 2 h and reduced to only one tenth of the control after 24 h (data not shown).
(c) Butyraldehyde concentration in broth (square) and total butanol concentration (circle).
Application of a differential method in principle requires metabolite measurements in total broth and filtrate for each measurement.
To avoid metabolite leakage during quenching, metabolite quantification (glycolytic and TCA-cycle intermediates and nucleotides) was carried out with a differential method, whereby the amounts measured in the filtrate were subtracted from the amounts measured in total broth.
Except for the one without YE supplement (YE 0.00 %), maximum final broth pH, minimum total SL, lactonic SL and Lac SL/tot SL ratio of 4.77, 45.83 ± 2.00, 18.12 ± 0.48 g/L, and 39.53 % were obtained with 0.30 % of YE addition, respectively.
The culture medium was inoculated with a 24 h culture broth containing a total viable cell count (TVC) of 8.7 × 106 cfu/mL of the selected isolate and incubated at 35 °C for 48 h in a rotary shaker incubator.
Membrane filters (0.45 µm cellulose acetate, Millipore MF type HA) were placed on a pad with 2.5 ml of m-Endo broth MF for total coliforms, M-FC broth for fecal coliforms, KF Streptococcus agar for streptococci and/or enterococci, and thiosulfate citrate bile salts sucrose agar (TCBS) to isolate bacteria other than Vibrio, although this medium was designed for Vibrio spp. isolation [20].
A single colony was used to inoculate 100 mL of overnight 2×YT culture, and 5 mL of overnight culture was used to inoculate 1 L of Terrific Broth (7 L total).
