title: Recombinant bacterial expression of the lysozyme from the tobacco-hornworm Manduca sexta with activity at low temperatures.
abstract: 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.
authors.affiliation: Aquatic Molecular Biology Laboratory, Centro de Investigación en Alimentación y Desarrollo, A.C. P.O. Box 1735, 83000, Hermosillo, Sonora, México.
authors.forename: Karina D
authors.initials: KD
authors.lastname: García-Orozco
authors.forename: Alonso A
authors.initials: AA
authors.lastname: López-Zavala
authors.forename: Daniel
authors.initials: D
authors.lastname: Puentes-Camacho
authors.forename: Ana Maria
authors.initials: AM
authors.lastname: Calderón-de-la-Barca
authors.forename: Rogerio R
authors.initials: RR
authors.lastname: Sotelo-Mundo
mesh_terms: D000818:Animals; D001230:Aspergillus / Q000201:enzymology; D001419:Bacteria / Q000378:metabolism*; D001671:Biochemistry / Q000379:methods; D019149:Bioreactors; D002845:Chromatography; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D003593:Cytoplasm / Q000378:metabolism; D004220:Disulfides; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D019791:Guanidine / Q000737:chemistry; D007313:Insecta; D018402:Manduca / Q000201:enzymology*; D009113:Muramidase / Q000737:chemistry / Q000378:metabolism*; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry*; D013696:Temperature
chemical_list: D004220:Disulfides; D011994:Recombinant Proteins; D009113:Muramidase; D019791:Guanidine