Lysozyme purification from tobacco extract by polyelectrolyte precipitation

Chenming Zhang¹, Raymond Lillie, Jackie Cotter

¹Department of Biological Systems Engineering, Virginia Polytechnic Institute and State University, 210 Seitz Hall, Blacksburg, VA 24061, USA. cmzhang@vt.edu

|April 23, 2005

Summary

Isoelectric precipitation effectively removes native tobacco proteins. Poly(acrylic) acid then efficiently recovers recombinant lysozyme from tobacco extract, improving downstream processing for plant-based protein production.

Area of Science:

Plant biotechnology
Biochemistry
Protein purification

Background:

Tobacco is a model for transgenic protein production.
Downstream processing of large biomass quantities presents challenges.
Efficient recovery methods are needed for recombinant proteins from plants.

Purpose of the Study:

To investigate isoelectric precipitation for native tobacco protein removal.
To evaluate polyelectrolyte precipitation for recombinant protein recovery from tobacco extract.
To improve downstream processing for plant-derived recombinant proteins.

Main Methods:

Isoelectric precipitation using hydrochloric acid (HCl) at pH 4.
Polyelectrolyte precipitation using poly(acrylic) acid (PAA) on lysozyme.
Analysis of protein precipitation and co-precipitation in tobacco extract.

Main Results:

Hydrochloric acid precipitated 60% of native tobacco protein at pH 4.
Isoelectric precipitation at pH 5 followed by PAA treatment precipitated 85% of soluble lysozyme.
Minimal co-precipitation of native tobacco proteins was observed with PAA.

Conclusions:

Isoelectric precipitation is effective for removing native tobacco proteins.
Poly(acrylic) acid significantly enhances recombinant lysozyme recovery from tobacco extract.
This two-step method improves downstream processing efficiency for plant-based protein production.

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Lysozyme purification from tobacco extract by polyelectrolyte precipitation

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Related Experiment Videos

Lysozyme purification from tobacco extract by polyelectrolyte precipitation

Area of Science:

Background:

Purpose of the Study:

Main Methods:

Main Results:

Conclusions:

Related Concept Videos

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