Glucoamylase purification

The purification was by chromatography on DEAE cellulose. Electrophoresis of the purified enzyme and the original filtrate are shown in Fig. (27) [71]. Rabbits were immunized with the pure glucoamylase [72]. Antibodies in the serum were purified by affinity chromatography. The affinity pattern for glucoamylase purification is shown in Fig. (28). Electrofocusing and assay by the coupled method for antibodies is shown in Fig. (29). The glucoamylase antibodies consist of 8 isomers [73],... 

Several variations on these production methods also exist, such as the use of immobilized CGTase (2i, 22), continuous ultrafiltration 23) and the use of isoamylase to increase CD yield. Variations of purification methods include the addition of glucoamylase to degrade non-CD starch hydrolysates to simply separation and the use of various S3mthetic ion exchange resins in chromatographic separations 24-26) and acuity columns (27),... 

In early experiments, glucoamylase was purified by ammonium sulfate precipitation, followed by an acid treatment to eliminate traces of alp/ia-amylase, but gel filtration and ion-exchange chromatography have now been successfully applied to purification of the A. niger, R. delemar, and gamma-amylases. Crystallization of the enzymes from A. niger and R. delemar has also been reported. ... 

ELUTION VOLUME(ml) Fig. (28). Affinity purification of anti-glucoamylase antibodies. 

Cellulose activated with epichlorohydrin has been treated with glycine and then coupled with a-amylase via a carbodi-imide mediator. Glucoamylase was also immobilized by a carbodi-imide-mediated reaction with epichlorohydrin-activated cellulose, which had previously been treated with 6-aminohexanoic acid, or by reaction with diazotized benzidine previously coupled to epichloro-hydrin-activated cellulose. Immobilized 4-chloromercuribenzoate was prepared by reaction with 1,6-diaminohexane coupled to epichlorohydrin-activated cellulose, and used for the affinity chromatographic purification of a-amylase and glucoamylase. 

Sephadexes have been used as affinity matrices in the purification of an exo-amylase from Pseudomonas stutzeri, mammalian glucoamylase ( glucoamylase-... 

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