Despite being vintners of old, Çobo's forefathers were prohibited from making wine during the communist era: he's making up for lost time by experimenting with several 100-bottle "micro-fermentations" each year.
The 72 deletants identified as being protracted in fermentation in a high sugar medium (CDGJM_200) were screened as part of the original 336 in a micro-fermentation of low sugar medium (CDGJM_20) containing 20 g L−1 (Additional files 1 and 2).
After we selected the top candidate strains by micro-fermentation analysis, we needed to further optimize the medium and process conditions (e.g., T, pH, DO, substrate feed) for a given strain to maximize its EPA production and minimize by-product formation (e.g., organic acids), in order to reduce the cost of manufacture (COM) under commercially achievable conditions.
Micro-fermentations (210 μl) were carried out at 30°C and 200 rpm in a humidity shaker.
Differential growth kinetics was also observed during wine micro-fermentations in vessels with 0.45 L of Tempranillo grape must.
Micro-fermentations were carried out as mentioned above and, after centrifugation, different volumes of supernatant (1 30 μL) were transferred to new plates.
The assays were validated using fresh supernatants from the micro-fermentations of S. cerevisiae transformed cells secreting laccase (in 96-well plate format).
In order to eliminate the possibility that incomplete fermentation by these 336 deletants was due to impaired growth, micro-fermentations were repeated with the additional estimation of growth (OD 600 nm) over the duration of the fermentation.
This comparative transcriptomic study of S. cerevisiae and S. kudriavzevii was carried out during wine micro-fermentations in 500 mL vessels of Tempranillo grape must at 12°C and 28°C with wine yeast strain T73 and IFO1802.
There have been several different types of micro-fermenters reported in fermentation research work (Doig et al. 2005; Betts et al. 2006; Gilla et al. 2008).
The flasks were sealed with plastic wrap to result in micro-aerobic fermentation conditions and transferred to an orbital shaker set at 28°C and 200 rpm.
