PRODUCTION AND PROPERTIES OF 2,3-BUTANEDIOL: XXXIII. AUTOMATIC pH CONTROL IN THE DISSIMILATION OF SUCROSE BY BACILLUS POLYMYXA

An electronic control circuit was used to maintain pH within ±0.02 units between successive additions of soluble alkali. Carbon balances show the effects of a series of pH levels and of a range of sucrose concentrations on the proportional yields of end products. There is a marked change in the chemical balance of this fermentation at about pH 7.0, correlated with a suppression of the acetoin enzyme system. Above pH 6.8 a sharp increase in acid production is correlated with decreased formation of diol and carbon dioxide: below pH 6.8 the yield of organic acids decreases steadily. Most efficient conversion to diol occurs from pH 6.0 to 6.4. Several reasons are advanced for selecting pH 6.2 as the optimum. Under anaerobic conditions the fermentation rate is increased over that under aerobic conditions. Diol yields increase and ethanol yields decrease steadily with increasing sucrose concentrations. The increases in diol are accompanied by decreasing yields of organic acids, and not by changes in carbon dioxide production, which remains relatively uniform. The sucrose concentration most efficient for conversion to diol is about 8%, which is dissimilated anaerobically in 30 hr. at pH 6.2 to yield 65 mM. (millimoles) of diol per 100 mM. of invert sugar fermented. Under aerobic conditions the diol–ethanol ratios show a marked increase, and reach a maximum of about 11 at 10% sucrose. This is due largely to increased acetoin and decreased ethanol formation. The dissimilation of 6% sucrose reaches 98% in 71 hr. under aerobic conditions and yields 82 mM. of diol plus acetoin per 100 mM. of invert sugar fermented. The use of either sodium or potassium hydroxide in place of ammonium hydroxide increases five times the period for complete dissimilation of 5% sucrose. Advantages of controlling the reaction by addition of ammonium hydroxide are reviewed.