Advances in Protein Chemistry, 8 by M L Anson; J T Edsall

By M L Anson; J T Edsall


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The insoluble impurities are precipitated in aqueous acid solution, and then removed. 0; the butanol solution is then extracted by ammonia, NATURALLY OCCURRING PEPTIDES 35 and the addition of methanol removes further impurities. CF is adsorbed on a Dowex 1 column, then eluted. The solution thus obtained is treated with carbon, which adsorbs the active substance. It i s eluted and adsorbed once again on a column of alumina from a water-ethanol solution. Finally, the product is purified by fractional crystallization as its barium salt.

The L . casei factor is csterified, and €he esterified product is extracted by butanol. After removal of butanol, the substance is dissolved in methanol and chromatographed on a column of Superfiltrol. It is then eluted with 70 % aqueous acetone. The active esterified product thus eluted is pure, and on hydrolysis yields the free acid which crystallizes. The latter is identical to the vitamin B, of Pfiffner et al. (459). Mitchell et al. (421) independently, isolated from spinach leaves a substance possessing biological properties closely related to those of PGA.

0 for 48 hours, to remove vitamin B,. The substances extracted by butanol are then treated by methanol. Ba(0H)z is added to the methanol extract. The precipitate is removed, and the vitamin is precipitated from the filtrate by zinc acetate. The salt thus formed is then decomposed by ammonium oxalate. 8 and the vitamin immediately precipitates. It crystallizes on cooling from a hot aqueous solution of the vitamin. The yield is about 60 mg. from 1000 lb. of liver. The same technique applies to the preparation of vitamin B, from horse liver, which is even richer than hog liver in this vitamin (459).

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