Api 50ch test

Metabolic characterization was assessed for a representative group of all the strains of the three species of photobacteria. A total of 14 strains of P. phosphoreum, 16 strains of P. carnosum, and 3 strains of P. iliopiscarium were assessed for carbohydrate acid production and enzymatic activities. Production of acid from different carbon sources was assessed by the API 50CH test (bioMérieux, Marcy-l’Étoile, France). Several enzymatic activities were tested with the API ZYM test (bioMérieux, Marcy-l’Étoile, France) according to the instructions from the manufacturer. Both procedures were performed according to the methodology followed by Hilgarth et al. (2018b) (link) and data for P. carnosum TMW2.2021/2.2022/2.2029/2.2030, P. phosphoreum DSM15556^T, and P. iliopiscarium DSM9896^T were taken from this study.

To determine hydrolysis of starch, urea, Tween 20, 40, 60, and 80, the isolate was cultured on TSA at 30°C for a week, as described by Cowan and Steel [37 ]. The enzyme activity was evaluated using API ZYM kit, API 20NE kit (bioMérieux, France), and acid production was tested API 50CH test (bioMérieux, France) according to the manufacturer’s instructions at 30°C for 2 days. The type strains, M. bovistercoris NEAU-LLE^T and M. pseudoresistence CC-5209^T, which are related to KUDC0405^T, were analyzed under the same conditions. The cell wall peptidoglycan was analyzed using an amino acid analyzer (L-8900; Hitachi, Japan). To analyze the polar lipids, two-dimensional thin layer chromatography (TLC) analysis were used according to Minnikin et al. [38 (link)]. The fatty acid composition analysis was performed using the Microbial Identification System from cells of the strain KUDC0405^T, and reference strains were incubated on TSA at 30°C for 7 days. To determine siderophore production by strain KUDC0405^T, chrome azurol S (CAS) media were used as previously described [39 (link), 40 (link)].

Fermentation patterns of 49 sugars under anaerobic conditions were determined using the API 50 CH test (BioMerieux, Marcy l’Etolie, France) as specified by the manufacturer and recorded after 24, 48 and 72 h of incubation at 30 °C. Growth tests were performed in M17 medium (Oxoid, Hampshire, United Kingdom) supplemented with 1% cellobiose (cel-M17), 1% glucose (glu-M17) or 1% mannose (man-M17), using Microbiology Reader Analyser (Bioscreen C; Oy Growth Curves Ab Ltd, Helsinki, Finland). Optical density at 600 nm was monitored during 72 h of incubation at 30 °C in 1-h intervals. Assays were performed in triplicate. Exponential growth rates were determined as previously described²⁷ . The ability to use mannose as a single carbon source was determined by growing bacteria in CDM medium supplemented with 1% mannose (man-CDM).