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

• Published in: Biotechnology letters Vol. 27; no. 15; pp. 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: Journal Article
• Language: English
• Published: Dordrecht Springer 01.08.2005; Springer Nature B.V
• 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; Manduca - enzymology; Manduca sexta; Micrococcus luteus; Muramidase - chemistry; Muramidase - metabolism; Protein Folding; Protein Structure, Tertiary; Proteins; Recombinant Proteins - chemistry; Temperature; Enzyme; Insecta; Refolding; bacterial over-expression; Gene overexpression; Sphingidae; Affinity chromatography; Arthropoda; Glycosidases; Lepidoptera; Manduca sexta; Hydrolases; Bacteria; Invertebrata; O-Glycosidases; Lysozyme; Low temperature
• ISSN: 0141-5492; 1573-6776
• DOI: 10.1007/s10529-005-8452-1

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.