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Metal chelate affinity precipitation: purification of (His)6-tagged lactate dehydrogenase using poly(vinylimidazole-co-N-isopropylacrylamide)copolymers

Kumar, Ashok LU ; Khalil, Ashraf LU ; Galaev, Igor LU and Mattiasson, Bo LU (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)
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author
organization
publishing date
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
2007-07-02 12:44:59
date last changed
2018-05-29 09:44:46
@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},
  keyword      = {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},
  volume       = {33},
  year         = {2003},
}