( 1208. ) 
the succession is then: a fat layer, gelatin, gelatin and calcium- 
carbonate gelatin, gelatin and fatty acid, gelatin, the gelatin consisting 
of broth-gelatin and 5°/, glucose. 
On this medium, about 0.5 em. thick, a fat-splitting microbe is 
cultivated, which forms acid from glucose, for example b. lipolyticum a, 
and one which does not do so, such as B. lipolyticum B. After two 
weeks’ cultivation fat-splitting is to be observed under B. lipolyticum a, 
whilst under B. lipolyticum B no splitting is seen. Hence, the acid- 
lipase under B. lipolyticum a diffuses through the fatty-acid layer, is 
neutralised by the carbonate layer and the further diffusing lipase 
decomposes the fat. The lipase of B. lipolytieum 3 combines with 
the fatty acid and diffuses no further. 
But if now one of these acids is transported to the place where 
B. lipolyticnm 8 has grown, it diffuses through the gelatin, forms 
with the lipase which is combined with fatty acid an acid-lipase, 
which diffuses and is decomposed in the carbonate layer, the further 
diffusing lipase decomposing the fat. It needs no explanation that 
lipase is likewise freed when the fatty acid is neutralised by addition 
of alkalis so that then also fields of decomposed fat arise. 
Hence, microbic lipase behaves in all these experiments as a feeble 
alkali forming compounds with acids and again is freed from these 
by alkalis. 
Influence of oxygen and light on the decomposition of fat by 
lipase produced by microbes. 
Presence of oxygen and light favour the decomposition of fat by 
the action of lipase. 
The favourable influence of oxygen on the decomposition of fat 
is very simply shown by means of fatted tubes filled with a lipase- 
containing gelatin. If some tubes are quite filled with the gelatin 
and some others thinly coated with it as is done at the preparation 
of roll-eultures, it is found that in the full tubes, where thus no 
oxygen can enter, the decomposition of fat proceeds much more 
slowly than in the other tubes; still there is more lipase in the 
former than in the latter although in the full tubes a more vigorous 
decomposition of the fat was to be expected. 
It is evident that together with the fat-splitting, oxidation of the 
fatty acids takes place. 
The special products resulting at the culture method of Eykman 
are fatty acids, soaps, oxidation products of fatty acids, and compounds 
of these acids with lipase. 
