Patent GB-415312-A (GB)
Title: Improvements in or relating to the production of butyl alcohol by fermentation
IPC: C12P7/28
Filed: 1933-12-27
Abstract: Acetone and normal butyl, isopropyl, and ethyl alcohols are produced by fermentation of mashes comprising monose sugars and degraded protein nitrogen with bacteria of a group named Clostridium propyl butylicum at a temperature between 23 and 38 DEG  C. and under conditions such that the final pH of the mash is between 5,0 and 6,5. The bacteria employed are principally characterized by: (a) inability to liquefy gelatine or utilize undegraded protein in fermentation; (b) ability to utilize degraded protein, including ammonia; (c) giving small yields from starch, sucrose or uninverted molasses mashes, but good yields with mashes comprising glucose or inverted molasses.  The pH control is effected preferably by addition of calcium, barium or iron carbonate in slightly greater proportion than that required to neutralize acidity.  Inverted molasses, wood sugar, hydrol, or hydrolyzed whey may be employed as the source of sugar, and ammonia, ammonium salts, urea, yeast water, or steep water as the source of nitrogen.  Examples: (1) Cuban molasses of 20 per cent sugar content is inverted by heating with sulphuric acid under pressure, ammonia (0,7 per cent of the sugar) is then added followed by finely divided calcium carbonate sufficient to neutralize the acid and leave an excess equal to 6 per cent of the sugar, and the mash is then diluted to 5 per cent sugar concentration, sterilized, and inoculated with 3 per cent of a culture from rotted wood and incubated at 30--32 DEG  C. for seventy-two hours; a yield is obtained equal to 28,8 per cent of the sugar and comprising 66,3 per cent butyl alcohol, 22,9 per cent isopropyl and ethyl alcohol and 10,8 per cent of acetone; (2) a mash comprising 5 per cent of corn sugar, 0,04 per cent ammonia as ammonium sulphate and 0,5 per cent calcium carbonate is inoculated and fermented as above; the yield comprises 27,3 per cent of the sugar and is constituted by 75,6 per cent butyl alcohol, 20,8 per cent isopropyl and ethyl alcohols and 3,6 per cent acetone.ALSO:Normal butyl, isopropyl, and ethyl alcohols, and acetone, are produced by fermentation of mashes comprising monose sugars and degraded protein nitrogen with bacteria of a group named Clostridium propyl butylicum at a temperature between 23 and 38 DEG  C. and under conditions such that the final pH of the mash is between 5,0 and 6,5.  The bacteria employed are principally characterized by:-(a) inability to liquefy gelatine or utilize undergraded protein in fermentation, (b) ability to utilize degraded protein, including ammonia, (c) giving small yields from starch, sucrose or uninverted molasses mashes, but good yields with mashes comprising glucose or inverted molasses.  The pH control is effected preferably by addition of calcium, barium or iron carbonate in slightly greater proportion than that required to neutralize acidity.  Inverted molasses, wood sugar, hydrol, or hydrolyzed whey may be employed as the source of sugar, and ammonia, ammonium salts, urea, yeast water, or steep water as the source of nitrogen. Examples:-(1) Cuban molasses of 20 per cent sugar content is inverted by heating with sulphuric acid under pressure, ammonia (0,7 per cent of the sugar) is then added followed by finely divided calcium carbonate sufficient to neutralize the acid and leave an excess equal to 6 per cent of the sugar, and the mash is then diluted to 5 per cent sugar concentration, sterilized, and inoculated with 3 per cent of a culture from rotted wood and incubated at 30-32 DEG  C. for seventy-two hours; a yield is obtained equal to 28,8 per cent of the sugar and comprising 66,3 per cent butyl alcohol, 22,9 per cent isopropyl and ethyl alcohol and 10,8 per cent of acetone: (2) a mash comprising 5 per cent of corn sugar, 0,04 per cent ammonia as ammonium sulphate and 0,5 per cent calcium carbonate is inoculated and fermented as above; the yield comprises 27,3 per cent of the sugar, and is constituted by 75,6 per cent butyl alcohol, 20,8 per cent isopropyl and ethyl alcohols, and 3,6 per cent acetone. Bacteria, cultivating.-The organisms used are obtainable from soil, mud, corn, corn stalks or rotted wood, for example by adding samples of these to the medium of example (1) at 80-85 DEG  C., or to a medium comprising 3,0 per cent glucose, 0,1 per cent ammonium sulphate, 0,15 per cent diammonium hydrogen phosphate, 0,05 per cent ammonium chloride, 0,05 per cent magnesium sulphate, and 0,3 per cent calcium carbondioxide adjusted to pH p  6,0.  The cultures are held at the temperature for one to three minutes, cooled to 32 DEG  C. and incubated for 48 hours, those strongly fermenting being allowed to sporulate at 32 DEG  C. for five days and being then transferred to flasks of the first of the above media at 95-100 DEG  C. which are subsequently cooled and incubated at 32 DEG  C.  After several repetitions of this procedure active cultures are tested for yield on the medium of example (1) and on a corresponding medium containing only uninverted molasses, those showing good yields on the former and poor yields on the latter being selected for further tests, or being plated and the best colonies selected one or more times.  The cultures may be stored indefinitely on dried sterile soil or a highly buffered medium, if stored otherwise they should be transferred every ten days to the medium of example (1) and allowed to germinate.