One-step, inexpensive high yield strategy for Candida antarctica lipase A isolation using hydroxyapatite
Lipase A from Candida antarctica (CAL A) was purified to apparent homogeneity in a single step using hydroxyapatite (HAP) chromatography. CAL A bound to HAP was eluted with 10 mM Na-phosphate buffer, pH 7.0 containing 0.5% Triton X-100. The protocol resulted in a 3.74-fold purification with 94.7% fi...
Published in: | Bioresource Technology |
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Main Authors: | , , , , , |
Format: | Article in Journal/Newspaper |
Language: | unknown |
Published: |
Elsevier Sci Ltd, Oxford
2012
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Subjects: | |
Online Access: | http://TechnoRep.tmf.bg.ac.rs/handle/123456789/2201 https://doi.org/10.1016/j.biortech.2011.11.077 |
Summary: | Lipase A from Candida antarctica (CAL A) was purified to apparent homogeneity in a single step using hydroxyapatite (HAP) chromatography. CAL A bound to HAP was eluted with 10 mM Na-phosphate buffer, pH 7.0 containing 0.5% Triton X-100. The protocol resulted in a 3.74-fold purification with 94.7% final recovery and 400.83 U/mg specific activity. Silver staining after SDS-PAGE revealed the presence a single band of 45 kDa. The enzyme exhibited a temperature optimum of 60 degrees C, was unaffected by monovalent metal ions, but was destabilized by divalent metal ions (Zn2+, Ca2+, Mg2+, Cu2+, Mn2+) and stimulated by 50 mM Fe2+. Detergents at 0.1% concentrations did not affect lipase activity. Except for Triton X-100, detergent concentrations of 1% had a destabilizing effect. |
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