Recombinant bacterial expression of the lysozyme from the tobacco-hornworm Manduca sexta with activity at low temperatures

A gene coding for lysozyme from the insect Manduca sexta (Ms-lyz) was expressed in Escherichia coli. The protein was produced as an insoluble cytoplasmic inclusion body which was denatured in 8 M: guanidine-HCl, renatured and purified by affinity and ion-exchange chromatography. The N-terminal seque...

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Bibliographic Details
Published in:Biotechnology letters 2005-08, Vol.27 (15), p.1075-1080
Main Authors: GARCIA-OROZCO, Karina D, LOPEZ-ZAVALA, Alonso A, PUENTES-CAMACHO, Daniel, CALDERON-DE-LA-BARCA, Ana Maria, SOTELO-MUNDO, Rogerio R
Format: Article
Language:English
Subjects:
Animals
Aspergillus - enzymology
Bacteria
Bacteria - metabolism
Biochemistry - methods
Biological and medical sciences
Bioreactors
Biotechnology
Chromatography
Chromatography, Affinity
Chromatography, Ion Exchange
Cytoplasm - metabolism
Disulfides
E coli
Electrophoresis, Polyacrylamide Gel
Escherichia coli
Escherichia coli - metabolism
Fundamental and applied biological sciences. Psychology
Guanidine - chemistry
Insecta
Insects
Low temperature
Manduca - enzymology
Manduca sexta
Micrococcus luteus
Muramidase - chemistry
Muramidase - metabolism
Protein Folding
Protein Structure, Tertiary
Proteins
Recombinant Proteins - chemistry
Temperature
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Summary:A gene coding for lysozyme from the insect Manduca sexta (Ms-lyz) was expressed in Escherichia coli. The protein was produced as an insoluble cytoplasmic inclusion body which was denatured in 8 M: guanidine-HCl, renatured and purified by affinity and ion-exchange chromatography. The N-terminal sequence and the activity of the recombinant protein against Micrococcus luteus confirmed that correct expression had occurred. When Ms-lyz activity was compared to hen egg white lysozyme, the insect lysozyme was active at lower temperatures. These results demonstrate the feasibility of producing a disulfide-bonded lysozyme enzyme in bacteria and suggest that the insect Ms-lyz is an interesting system for further development of an antibacterial functional at low temperatures.
ISSN:0141-5492
1573-6776
DOI:10.1007/s10529-005-8452-1