2.3 Initial tolerance test of C. pasteurianum C8 to crude
glycerol
To evaluate the inhibition of crude glycerol, initial tolerance test was
performed in batch culture. A cyro stock culture stored in -80°C was
first grown in anaerobic bottle with the pre-culture medium described
above which contained 30 g/L pure glycerol. Then the culture in
exponential growth phase was inoculated into the pre-culture medium
containing various concentrations of crude glycerol (10, 15, 20, 25, 30,
40, 60, 80 g/L); a pre-culture medium with pure glycerol (30 or 80 g/L)
was also inoculated and used as control. After anaerobic cultivation at
35°C for 12 h, OD600 of the culture was determined and
compared. All tests were conducted in triplicates.