Abstract
Crude porcine lipase (triacylglycerol lipase, EC 3.1.1.3) was purified in a single-stage Chromatographic process. The purification was accomplished in a batch, as well as in a continuous system. Two types of sizeexclusion packing materials (Sephadex and Sephacryl) were used. The average x-fold increase in purity, and the average recovered activity in the batch Sephadex and Sephacryl experiments were 13.6 and 89.7%, and 34.2 and 98.8%, respectively. The average x-fold increase in purity and the average activity recovered in the continuous Sephadex and Sephacryl experiments were 27.1 and 82.5% and 16.2 and 89%, respectively. Flow visualization experiments were carried out by tagging the protein to be separated with a fluorescent dye. The results from these experiments are also reported in this article.
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Abbreviations
- F:
-
flow rate, mL/min
- H:
-
plate height, cm
- h i :
-
absorbance, dimensionless
- L:
-
packing height, cm
- N:
-
number of plates, dimensionless
- t R :
-
peak retention time, min
- R s :
-
resolution, dimensionless
- V i :
-
elution volume, mL
- V R :
-
peak retention volume, mL
- σ:
-
standard deviation, mL.
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Genest, P.W., Field, T.G., Vasudevan, P.T. et al. Continuous purification of porcine lipase by rotating annular size-exclusion cnromatography. Appl Biochem Biotechnol 73, 215–230 (1998). https://doi.org/10.1007/BF02785657
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DOI: https://doi.org/10.1007/BF02785657