The yeast is allowed to liquify for at least a month in a fairly warm
room without stirring. At the end of this time the surface is removed
and any supernatant liquid poured away. The lower sedimentary part is
thrown on a quick-acting filter and allowed to drain for two days.
To the filtrate, alcohol of specific gravity 0.87 is gradually added
to the extent of one and a half times its volume, with continued and
vigorous stirring. The process of adding the alcohol and stirring
should require about half an hour, after which the mixture is allowed
to stand for twenty-four hours to allow the precipitated invertase
to settle. The supernatant liquid is poured away and the precipitate
washed several times on successive days by decantation with alcohol of
0.92 specific gravity. When the washings become nearly colorless the
precipitate is thrown on a filter, allowed to drain, and immediately
removed and mixed with a large bulk of alcohol of 0.92 specific
gravity. The precipitate is again collected, mixed thoroughly with its
own bulk of water, and some alcohol of 0.97 specific gravity, allowed
to stand for a few hours and thrown on a filter. The filtrate contains
the invertase.
=97. Determination of Activity of Invertase.=—The activity of a
solution of invertase, prepared as above, is measured by the number
of minutes required for it to reduce to zero the optical power of a
solution of 100 times its weight of cane sugar at a temperature of
15°.5. In order to facilitate the action of the invertase, a trace of
sulfuric acid is added to the solution. The manipulation is as follows:
Fifty grams of sucrose are dissolved in water and made to a volume of
nearly a quarter of a liter and placed in a bath maintained at 15°.5.
Half a gram of the invertase is added, the time noted, the solution
immediately made up to a quarter of a liter and well shaken. The
contents of the flask are poured rapidly into five beakers; the actual
quantity in each beaker is not necessarily the same. To each of these
beakers, in succession, are added the following amounts of decinormal
sulfuric acid, _viz._, one-tenth, three-tenths, six-tenths, one, and
one and four-tenths cubic centimeters. After an hour a small quantity
of the solution is taken from beaker No. 3 and the reaction of the
invertase stopped by adding a few drops of strong potassium hydroxid
and the time of adding this reagent noted. This solution is then read
in the polariscope and the percentage of sugar inverted is calculated
from the formula C₁₂H₂₂O₁₁ + H₂O = C₆H₁₂O₆ + C₆H₁₂O₆.
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