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Separation and purification of enzymes by continuous pH‐parametric pumping
Author(s) -
Huang Shih Yow,
Lin Ching Kuan,
Juang Long Yih
Publication year - 1985
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260271009
Subject(s) - chromatography , parametric statistics , chemistry , enzyme , separation (statistics) , biochemistry , mathematics , statistics
Trypsin and chymotrypsin were separated from porcine pancreas extract by continuous pH‐parametric pumping. CHOM (chicken ovomucoid) was convalently bound to laboratory‐prepared crab chitin with glutaraldehyde to form an affinity adsorbent of trypsin. The pH levels of top and bottom feeds were 8.0 and 2.5, respectively. Similar inhibitor, DKOM (duck ovomucoid), and pH levels 8.0 and 2.0 for top and bottom feeds, respectively, were used for separation and purification of chymotrypsin. ε‐Amino caproyl‐D‐tryptophan methyl ester was coupled to chitosan to form an affinity adsorbent for stem bromelain. The pH levels were 8.7 and 3.0. Separation continued fairly well with high yield, e.g., 95% recovery of trypsin after continuous pumping of 10 cycles. Optimum operational conditions for concentration and purification of these enzymes were investigated. The results showed that the continuous pH‐parametric pumping coupled with affinity chromatography is effective for concentration and purification of enzymes.