A semi-pilot-scale procedure for isolating and purifying soybean (Glycine max) lectin

Yewande O. Fasina; Harold E. Swaisgood; Jim D. Garlich; Henry L. Classen

doi:10.1021/jf021125l

A semi-pilot-scale procedure for isolating and purifying soybean (Glycine max) lectin

Yewande O. Fasina, Harold E. Swaisgood, Jim D. Garlich, Henry L. Classen

Research output: Contribution to journal › Article › peer-review

Abstract

Availability of gram quantities of purified soybean lectin (SBL) to scientists will foster discovery of novel biomedical applications of the lectin and provide the opportunity to investigate the antinutritional effects of SBL in soybean-consuming food animals and poultry. Therefore, a semi-pilot-scale procedure for isolating and purifying SBL was designed. Defatted soyflour was extracted overnight with 0.9% NaCl at 4 °C. The extract obtained was filtered (0.45 μm membrane) and subjected to affinity chromatography using a column containing N-acetyl-D-galactosamine resin that is specific for SBL. Bound SBL was eluted off the column with 0.14 M galactose solution. The eluent was ultrafiltered (30 kDa), and the resulting solution (SBL and water) was freeze-dried. Electrophoretic analysis and hemagglutination assay revealed that the freeze-dried SBL was similar to Sigma-grade SBL in purity and activity (35 and 33 HU/mg protein, respectively). The procedure yielded 141 mg of SBL/100 g of soyflour.

Original language	English
Pages (from-to)	4532-4538
Number of pages	7
Journal	Journal of Agricultural and Food Chemistry
Volume	51
Issue number	16
DOIs	https://doi.org/10.1021/jf021125l
State	Published - Jul 30 2003
Externally published	Yes

Keywords

Affinity chromatography
Glycine max
Lectin purification
SDS- and native-PAGE
Scale-up
Soybean lectin (SBL)

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10.1021/jf021125l

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@article{02968ece98e84159846e1920c7ad07e8,

title = "A semi-pilot-scale procedure for isolating and purifying soybean (Glycine max) lectin",

abstract = "Availability of gram quantities of purified soybean lectin (SBL) to scientists will foster discovery of novel biomedical applications of the lectin and provide the opportunity to investigate the antinutritional effects of SBL in soybean-consuming food animals and poultry. Therefore, a semi-pilot-scale procedure for isolating and purifying SBL was designed. Defatted soyflour was extracted overnight with 0.9\% NaCl at 4 °C. The extract obtained was filtered (0.45 μm membrane) and subjected to affinity chromatography using a column containing N-acetyl-D-galactosamine resin that is specific for SBL. Bound SBL was eluted off the column with 0.14 M galactose solution. The eluent was ultrafiltered (30 kDa), and the resulting solution (SBL and water) was freeze-dried. Electrophoretic analysis and hemagglutination assay revealed that the freeze-dried SBL was similar to Sigma-grade SBL in purity and activity (35 and 33 HU/mg protein, respectively). The procedure yielded 141 mg of SBL/100 g of soyflour.",

keywords = "Affinity chromatography, Glycine max, Lectin purification, SDS- and native-PAGE, Scale-up, Soybean lectin (SBL)",

author = "Fasina, \{Yewande O.\} and Swaisgood, \{Harold E.\} and Garlich, \{Jim D.\} and Classen, \{Henry L.\}",

year = "2003",

month = jul,

day = "30",

doi = "10.1021/jf021125l",

language = "English",

volume = "51",

pages = "4532--4538",

journal = "Journal of Agricultural and Food Chemistry",

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T1 - A semi-pilot-scale procedure for isolating and purifying soybean (Glycine max) lectin

AU - Fasina, Yewande O.

AU - Swaisgood, Harold E.

AU - Garlich, Jim D.

AU - Classen, Henry L.

PY - 2003/7/30

Y1 - 2003/7/30

N2 - Availability of gram quantities of purified soybean lectin (SBL) to scientists will foster discovery of novel biomedical applications of the lectin and provide the opportunity to investigate the antinutritional effects of SBL in soybean-consuming food animals and poultry. Therefore, a semi-pilot-scale procedure for isolating and purifying SBL was designed. Defatted soyflour was extracted overnight with 0.9% NaCl at 4 °C. The extract obtained was filtered (0.45 μm membrane) and subjected to affinity chromatography using a column containing N-acetyl-D-galactosamine resin that is specific for SBL. Bound SBL was eluted off the column with 0.14 M galactose solution. The eluent was ultrafiltered (30 kDa), and the resulting solution (SBL and water) was freeze-dried. Electrophoretic analysis and hemagglutination assay revealed that the freeze-dried SBL was similar to Sigma-grade SBL in purity and activity (35 and 33 HU/mg protein, respectively). The procedure yielded 141 mg of SBL/100 g of soyflour.

AB - Availability of gram quantities of purified soybean lectin (SBL) to scientists will foster discovery of novel biomedical applications of the lectin and provide the opportunity to investigate the antinutritional effects of SBL in soybean-consuming food animals and poultry. Therefore, a semi-pilot-scale procedure for isolating and purifying SBL was designed. Defatted soyflour was extracted overnight with 0.9% NaCl at 4 °C. The extract obtained was filtered (0.45 μm membrane) and subjected to affinity chromatography using a column containing N-acetyl-D-galactosamine resin that is specific for SBL. Bound SBL was eluted off the column with 0.14 M galactose solution. The eluent was ultrafiltered (30 kDa), and the resulting solution (SBL and water) was freeze-dried. Electrophoretic analysis and hemagglutination assay revealed that the freeze-dried SBL was similar to Sigma-grade SBL in purity and activity (35 and 33 HU/mg protein, respectively). The procedure yielded 141 mg of SBL/100 g of soyflour.

KW - Affinity chromatography

KW - Glycine max

KW - Lectin purification

KW - SDS- and native-PAGE

KW - Scale-up

KW - Soybean lectin (SBL)

UR - https://www.scopus.com/pages/publications/0041302298

U2 - 10.1021/jf021125l

DO - 10.1021/jf021125l

M3 - Article

SN - 0021-8561

VL - 51

SP - 4532

EP - 4538

JO - Journal of Agricultural and Food Chemistry

JF - Journal of Agricultural and Food Chemistry

IS - 16

ER -

A semi-pilot-scale procedure for isolating and purifying soybean (Glycine max) lectin

Abstract

Keywords

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