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ชื่อเรื่องว่าRepeated-batch ethanol fermentation from sweet
sorghum juice by free cells of Saccharomyces
cerevisiae NP 01

MATERIALS AND METHODS
Microorganism, inoculum preparation, inoculum preparation
(IP) medium
S. cerevisiae NP 01 isolated from Loog-pang (Chinese yeast cake)
from Nakorn Panom province, Thailand, was inoculated into a 250
ml Erlenmeyer flask containing 150 ml of YM medium as the first
medium. The medium contained (in g l-1) YE 3, malt extract (ME) 3,
peptone (PE) 5 and glucose 10. The flask was incubated on a
rotating shaker at 150 rev min-1, 30°C for 15 h.
To increase cell concentration, the yeasts (10% inoculum size)
were transferred into 500 ml Erlenmeyer flasks with 350 ml of
modified IP media as shown in Table 1. The flasks were further
incubated under the conditions previously mentioned and the viable
yeast cell concentration was measured every 3 h for 24 h. Specific
growth rate (μ) of the yeasts in the IP media was calculated. The IP
medium giving the highest μ was used for inoculum preparation for
ethanol production

Ethanol production medium
The concentrated SSJ was diluted with distilled water to the total
soluble solids of 24°Bx and used as an ethanol production medium.
The ethanol production medium was transferred into a 500 ml airlocked
Erlenmeyer flask (working volume of 400 ml) or a 2 L
bioreactor (working volume of 1500 ml) and autoclaved at 110°C for
15 (for the 500-ml flask) or 28 min

Repeated-batch fermentation system
The repeated-batch system in the 500 ml flask was first carried out
in batch mode as described above. When the total residual sugars
in the broth had dropped slowly as found in the batch fermentation
system, the fermented broth was withdrawn at 75 and 50% of the
working volume and the same amount of the fresh juice was
immediately replaced to initiate the next batch. This method was
called “fill and drain technique” (Anastassiadis and Rehm, 2006).
Eight successive batches were performed. The fill and drain volume
giving the maximum ethanol production was selected for the
fermentation in the 2 L bioreactor.
In the 2 L bioreactor, the fermentation was operated as in the
flask, but the medium was agitated at 100 rev min-1. Eight
successive batches were performed.
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