Purification of hyperthermophilic archaeal amylolytic enzyme (MJA1) using thermoseparating aqueous two-phase systems

Mian Li; Tonya L. Peeples

doi:10.1016/j.jchromb.2004.03.030

Purification of hyperthermophilic archaeal amylolytic enzyme (MJA1) using thermoseparating aqueous two-phase systems

Mian Li, Tonya L. Peeples

Research output: Contribution to journal › Conference article › peer-review

40 Scopus citations

Abstract

Purification of a recombinant, thermostable α-amylase (MJA1) from the hyperthermophile, Methanococcus jannaschii, was investigated in the ethylene oxide-propylene oxide random copolymer (PEO-PPO)/(NH₄) ₂SO₄, and poly(ethylene glycol) (PEG)/(NH ₄)₂SO₄ aqueous two-phase systems. MJA1 partitioned in the top polymer-rich phase, while the remainder of proteins partitioned in the bottom salt-rich phase. It was found that enzyme recovery of up to 90% with a purification factor of 3.31 was achieved using a single aqueous two-phase extraction step. In addition, the partition behavior of pure amyloglucosidase in polymer/salt aqueous two-phase systems was also evaluated. All of the studied enzymes partitioned unevenly in these polymer/salt systems. This work is the first reported application of thermoseparating polymer aqueous two-phase systems for the purification of extremophile enzymes.

Original language	English (US)
Pages (from-to)	69-74
Number of pages	6
Journal	Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume	807
Issue number	1
DOIs	https://doi.org/10.1016/j.jchromb.2004.03.030
State	Published - Jul 25 2004
Event	12th International Conference on Biopartitioning and Purif. - Vancouver,BC, Canada Duration: Jun 1 2003 → Jun 1 2003

All Science Journal Classification (ASJC) codes

Analytical Chemistry
Biochemistry
Clinical Biochemistry
Cell Biology

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title = "Purification of hyperthermophilic archaeal amylolytic enzyme (MJA1) using thermoseparating aqueous two-phase systems",

abstract = "Purification of a recombinant, thermostable α-amylase (MJA1) from the hyperthermophile, Methanococcus jannaschii, was investigated in the ethylene oxide-propylene oxide random copolymer (PEO-PPO)/(NH4) 2SO4, and poly(ethylene glycol) (PEG)/(NH 4)2SO4 aqueous two-phase systems. MJA1 partitioned in the top polymer-rich phase, while the remainder of proteins partitioned in the bottom salt-rich phase. It was found that enzyme recovery of up to 90% with a purification factor of 3.31 was achieved using a single aqueous two-phase extraction step. In addition, the partition behavior of pure amyloglucosidase in polymer/salt aqueous two-phase systems was also evaluated. All of the studied enzymes partitioned unevenly in these polymer/salt systems. This work is the first reported application of thermoseparating polymer aqueous two-phase systems for the purification of extremophile enzymes.",

author = "Mian Li and Peeples, {Tonya L.}",

note = "Funding Information: This work was supported in part by NSF Career grant BES 9702588 and by NASA NAG8-1593. ; 12th International Conference on Biopartitioning and Purif. ; Conference date: 01-06-2003 Through 01-06-2003",

year = "2004",

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doi = "10.1016/j.jchromb.2004.03.030",

language = "English (US)",

volume = "807",

pages = "69--74",

journal = "Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences",

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TY - JOUR

T1 - Purification of hyperthermophilic archaeal amylolytic enzyme (MJA1) using thermoseparating aqueous two-phase systems

AU - Li, Mian

AU - Peeples, Tonya L.

N1 - Funding Information: This work was supported in part by NSF Career grant BES 9702588 and by NASA NAG8-1593.

PY - 2004/7/25

Y1 - 2004/7/25

N2 - Purification of a recombinant, thermostable α-amylase (MJA1) from the hyperthermophile, Methanococcus jannaschii, was investigated in the ethylene oxide-propylene oxide random copolymer (PEO-PPO)/(NH4) 2SO4, and poly(ethylene glycol) (PEG)/(NH 4)2SO4 aqueous two-phase systems. MJA1 partitioned in the top polymer-rich phase, while the remainder of proteins partitioned in the bottom salt-rich phase. It was found that enzyme recovery of up to 90% with a purification factor of 3.31 was achieved using a single aqueous two-phase extraction step. In addition, the partition behavior of pure amyloglucosidase in polymer/salt aqueous two-phase systems was also evaluated. All of the studied enzymes partitioned unevenly in these polymer/salt systems. This work is the first reported application of thermoseparating polymer aqueous two-phase systems for the purification of extremophile enzymes.

AB - Purification of a recombinant, thermostable α-amylase (MJA1) from the hyperthermophile, Methanococcus jannaschii, was investigated in the ethylene oxide-propylene oxide random copolymer (PEO-PPO)/(NH4) 2SO4, and poly(ethylene glycol) (PEG)/(NH 4)2SO4 aqueous two-phase systems. MJA1 partitioned in the top polymer-rich phase, while the remainder of proteins partitioned in the bottom salt-rich phase. It was found that enzyme recovery of up to 90% with a purification factor of 3.31 was achieved using a single aqueous two-phase extraction step. In addition, the partition behavior of pure amyloglucosidase in polymer/salt aqueous two-phase systems was also evaluated. All of the studied enzymes partitioned unevenly in these polymer/salt systems. This work is the first reported application of thermoseparating polymer aqueous two-phase systems for the purification of extremophile enzymes.

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DO - 10.1016/j.jchromb.2004.03.030

M3 - Conference article

C2 - 15177162

AN - SCOPUS:2642572488

SN - 1570-0232

VL - 807

SP - 69

EP - 74

JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

JF - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

IS - 1

T2 - 12th International Conference on Biopartitioning and Purif.

Y2 - 1 June 2003 through 1 June 2003

ER -

Purification of hyperthermophilic archaeal amylolytic enzyme (MJA1) using thermoseparating aqueous two-phase systems

Abstract

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