Purification of Aspergillus carbonarius polygalacturonase using polymeric membranes
IR@CFTRI: CSIR-Central Food Technological Research Institute, Mysore
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Relation |
http://ir.cftri.com/8694/
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Title |
Purification of Aspergillus carbonarius polygalacturonase using polymeric membranes
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Creator |
Nakkeeran, E.
Venkatesh, K. S. Subramanian, R. Umesh Kumar, S. |
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Subject |
05 Enzymes
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Description |
BACKGROUND: Microfiltration (MF: 70–450nm) and ultrafiltration (UF: 10–500 kDa) membranes were used to
eliminate carbohydrates and other non-protein impurities from Aspergillus carbonarius culture broth containing
polygalacturonase enzyme (EC 3.2.1.15) that would otherwise interfere with the purification processes and lead to
enzyme loss. Further, diafiltration was attempted to improve the elimination of impurities as well as recovery of
enzymes.
RESULTS: MF resulted in removal of 2–25% carbohydrates with an enzyme recovery of 69–82% from the
crude culture broth owing to the secondary layer formation. UF with 10kDa membrane eliminated most of the
carbohydrates (96%), phosphate salts and total acids with a recovery of 96% polygalacturonase and resulted in
greater productivity. Using the above procedure, the enzyme was concentrated nearly 10-fold while the purity
improved from 4.6 to 49.4Umg−1 of dry matter.
CONCLUSIONS: The results of this study focused on the elimination of carbohydrates and other non-protein
impurities showed that UF could be used efficiently as a primary purification step during downstream processing
of microbial culture broths containing enzymes. The present approach will ensure complete elimination of
non-protein impurities thereby reducing the losses and difficulties in the subsequent purification steps.
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Date |
2008
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Type |
Article
PeerReviewed |
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Format |
application/pdf
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Language |
en
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Identifier |
http://ir.cftri.com/8694/1/J_Chem_Technol_Biotechnol_%282008%29_83%287%29_957.pdf
Nakkeeran, E. and Venkatesh, K. S. and Subramanian, R. and Umesh Kumar, S. (2008) Purification of Aspergillus carbonarius polygalacturonase using polymeric membranes. Journal of Chemical Technology and Biotechnology, 83. pp. 957-964. ISSN 0268–2575 |
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