Title:
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Production of alpha-amylase by bacterial fermentation.
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An industrial-type fermentation for the production of alphaamylase
was studied at 3-litre scale, using a complex medium and
Bacillus subtilis B. 20, a high-yielding strain.
It was found that both the peak titre and the production
rate continued to increase up to the highest mass-transfer rates
used. Using an impeller of half the fermenter diameter at speeds
around 14 Hz (840 r. p. m. ) and aeration at 0.57 v. v. m., a batch
fermentation of 60-70 hours gave at least 9 u/ml (about 900
S. K. B. u/ml, measured at pH 6.0, or 36 000 Novo u/ml). Rapid
production began around hour 10, when growth was about half complete.
The volumetric production rate was almost constant for
40-50 hours, giving a constant specific productivity of about
0.75 x 10-11 u/cell/h after the completion of growth, until an
abrupt final decline. Rapid synthesis coincided with growth and
the rate was not directly related to population, the rates of
growth and mass-transfer or any other measured variable.
The termination of synthesis in simple batch operation was
apparently due to carbohydrate depletion. Thus, feeding increased
production by about 25%. Corrected for dilution, the production
rate was unchanged so that the specific productivity fell in
proportion to the increase in population during feeding.
It is suggested that the rate of synthesis was controlled
by a pool of a very stable regulator-inducer complex accumulated
before exhaustion of the inducer in the early stages of the fermentation.
There appears to be considerable potential for further
development along the lines followed in this work. The limits of
the stimulatory effects of mass transfer and feeding were not
passed. Neither was there evidence of depletion of a pool of
complexed regulator.
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