One-step, inexpensive high yield strategy for Candida antarctica lipase A isolation using hydroxyapatite
Authorized Users Only
2012
Authors
Dimitrijević, AleksandraVeličković, Dušan
Bihelović, Filip
Bezbradica, Dejan
Jankov, Ratko
Milosavić, Nenad
Article (Published version)
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Show full item recordAbstract
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.
Keywords:
Candida antarctica / Lipase / Isolation / Purification / HydroxyapatiteSource:
Bioresource Technology, 2012, 107, 358-362Publisher:
- Elsevier Sci Ltd, Oxford
Funding / projects:
- Reinforcement of the Faculty of Chemistry, University of Belgrade, towards becoming a Center of Excellence in the region of WB for Molecular Biotechnology and Food research (EU-256716)
- Allergens, antibodies, enzymes and small physiologically important molecules: design, structure, function and relevance (RS-172049)
- Novel encapsulation and enzyme technologies for designing of new biocatalysts and biologically active compounds targeting enhancement of food quality, safety and competitiveness (RS-46010)
DOI: 10.1016/j.biortech.2011.11.077
ISSN: 0960-8524
PubMed: 22209131
WoS: 000301620600050
Scopus: 2-s2.0-84856573973
Institution/Community
Tehnološko-metalurški fakultetTY - JOUR AU - Dimitrijević, Aleksandra AU - Veličković, Dušan AU - Bihelović, Filip AU - Bezbradica, Dejan AU - Jankov, Ratko AU - Milosavić, Nenad PY - 2012 UR - http://TechnoRep.tmf.bg.ac.rs/handle/123456789/2201 AB - 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. PB - Elsevier Sci Ltd, Oxford T2 - Bioresource Technology T1 - One-step, inexpensive high yield strategy for Candida antarctica lipase A isolation using hydroxyapatite EP - 362 SP - 358 VL - 107 DO - 10.1016/j.biortech.2011.11.077 ER -
@article{ author = "Dimitrijević, Aleksandra and Veličković, Dušan and Bihelović, Filip and Bezbradica, Dejan and Jankov, Ratko and Milosavić, Nenad", year = "2012", abstract = "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.", publisher = "Elsevier Sci Ltd, Oxford", journal = "Bioresource Technology", title = "One-step, inexpensive high yield strategy for Candida antarctica lipase A isolation using hydroxyapatite", pages = "362-358", volume = "107", doi = "10.1016/j.biortech.2011.11.077" }
Dimitrijević, A., Veličković, D., Bihelović, F., Bezbradica, D., Jankov, R.,& Milosavić, N.. (2012). One-step, inexpensive high yield strategy for Candida antarctica lipase A isolation using hydroxyapatite. in Bioresource Technology Elsevier Sci Ltd, Oxford., 107, 358-362. https://doi.org/10.1016/j.biortech.2011.11.077
Dimitrijević A, Veličković D, Bihelović F, Bezbradica D, Jankov R, Milosavić N. One-step, inexpensive high yield strategy for Candida antarctica lipase A isolation using hydroxyapatite. in Bioresource Technology. 2012;107:358-362. doi:10.1016/j.biortech.2011.11.077 .
Dimitrijević, Aleksandra, Veličković, Dušan, Bihelović, Filip, Bezbradica, Dejan, Jankov, Ratko, Milosavić, Nenad, "One-step, inexpensive high yield strategy for Candida antarctica lipase A isolation using hydroxyapatite" in Bioresource Technology, 107 (2012):358-362, https://doi.org/10.1016/j.biortech.2011.11.077 . .