Metal chelate affinity precipitation: purification of (His)6-tagged lactate dehydrogenase using poly(vinylimidazole-co-N-isopropylacrylamide)copolymers
(2003) In Enzyme and Microbial Technology 33(1). p.113-117- Abstract
- Affinity precipitation is a highly selective bioseparation method, which has been successfully used in the purification of various enzymes/proteins. The separation of (His)6-tagged lactate dehydrogenase ((His)6-LDH) was carried through affinity precipitation technique using thermoresponsive copolymer poly(vinylimidazole-co-N-isopropyl-acrylamide) (poly(VI-NIPAM)). Nearly quantitative recovery of (His)6-LDH has been achieved using Cu(II)- and Ni(II)-loaded poly(VI-NIPAM) with Cu(II)-poly(VI-NIPAM) being more efficient in enzyme precipitation than Ni(II)-poly(VI-NIPAM). Optimal precipitation of (His)6-LDH was achieved at pH 6 and pH 6.5–7.0 with Cu(II)-poly(VI-NIPAM) and Ni(II)-poly(VI-NIPAM), respectively. Furthermore, the enzyme was... (More)
- Affinity precipitation is a highly selective bioseparation method, which has been successfully used in the purification of various enzymes/proteins. The separation of (His)6-tagged lactate dehydrogenase ((His)6-LDH) was carried through affinity precipitation technique using thermoresponsive copolymer poly(vinylimidazole-co-N-isopropyl-acrylamide) (poly(VI-NIPAM)). Nearly quantitative recovery of (His)6-LDH has been achieved using Cu(II)- and Ni(II)-loaded poly(VI-NIPAM) with Cu(II)-poly(VI-NIPAM) being more efficient in enzyme precipitation than Ni(II)-poly(VI-NIPAM). Optimal precipitation of (His)6-LDH was achieved at pH 6 and pH 6.5–7.0 with Cu(II)-poly(VI-NIPAM) and Ni(II)-poly(VI-NIPAM), respectively. Furthermore, the enzyme was purified by affinity precipitation upto 3.5- and 3.1-fold with recoveries of 95 and 82% using Cu(II)- and Ni(II)-loaded copolymers, respectively. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/129156
- author
- Kumar, Ashok LU ; Khalil, Ashraf LU ; Galaev, Igor LU and Mattiasson, Bo LU
- organization
- publishing date
- 2003
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Affinity precipitation, Lactate dehydrogenase, Thermoresponsive copolymers, Metal affinity
- in
- Enzyme and Microbial Technology
- volume
- 33
- issue
- 1
- pages
- 113 - 117
- publisher
- Elsevier
- external identifiers
-
- wos:000183967800016
- scopus:0038072717
- ISSN
- 0141-0229
- DOI
- 10.1016/S0141-0229(03)00092-9
- language
- English
- LU publication?
- yes
- id
- ea5de19d-0e9e-4ed9-9c49-d217f2cfee62 (old id 129156)
- date added to LUP
- 2016-04-01 12:34:30
- date last changed
- 2023-08-15 13:30:54
@article{ea5de19d-0e9e-4ed9-9c49-d217f2cfee62, abstract = {{Affinity precipitation is a highly selective bioseparation method, which has been successfully used in the purification of various enzymes/proteins. The separation of (His)6-tagged lactate dehydrogenase ((His)6-LDH) was carried through affinity precipitation technique using thermoresponsive copolymer poly(vinylimidazole-co-N-isopropyl-acrylamide) (poly(VI-NIPAM)). Nearly quantitative recovery of (His)6-LDH has been achieved using Cu(II)- and Ni(II)-loaded poly(VI-NIPAM) with Cu(II)-poly(VI-NIPAM) being more efficient in enzyme precipitation than Ni(II)-poly(VI-NIPAM). Optimal precipitation of (His)6-LDH was achieved at pH 6 and pH 6.5–7.0 with Cu(II)-poly(VI-NIPAM) and Ni(II)-poly(VI-NIPAM), respectively. Furthermore, the enzyme was purified by affinity precipitation upto 3.5- and 3.1-fold with recoveries of 95 and 82% using Cu(II)- and Ni(II)-loaded copolymers, respectively.}}, author = {{Kumar, Ashok and Khalil, Ashraf and Galaev, Igor and Mattiasson, Bo}}, issn = {{0141-0229}}, keywords = {{Affinity precipitation; Lactate dehydrogenase; Thermoresponsive copolymers; Metal affinity}}, language = {{eng}}, number = {{1}}, pages = {{113--117}}, publisher = {{Elsevier}}, series = {{Enzyme and Microbial Technology}}, title = {{Metal chelate affinity precipitation: purification of (His)6-tagged lactate dehydrogenase using poly(vinylimidazole-co-N-isopropylacrylamide)copolymers}}, url = {{http://dx.doi.org/10.1016/S0141-0229(03)00092-9}}, doi = {{10.1016/S0141-0229(03)00092-9}}, volume = {{33}}, year = {{2003}}, }