Expression of the Human Lysozyme Gene in Kluyveromyces lactis
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Abstract:
Human lysozyme, an enzyme with natural broad-spectrum antimicrobial activity, has potential applications in the food and pharmaceutical industries. To obtain human lysozyme preparations with high activity, a Kluyveromyces lactis yeast expression system was used for soluble expression of the human lysozyme gene (hLYZ) with optimized codons. An artificial hLYZ gene was cloned into the K. lactis expression vector pKLAC1, resulting in the recombinant vector pKLAC1-hLZY, which was then linearized with SacII and transformed into K. lactis GG799 by electroporation. Following whole-cell PCR, a multicopy recombinant strain (named hLYZ1) was obtained. The engineered strain secreted a target protein of approximately 14 ku in molecular weight, which agreed with that of the expected product. The maximum activity of hLZY in the culture supernatant reached 1430 U/mL after 128 h of culture. Antibacterial activity testing confirmed that the recombinant human lysozyme was effective against Micrococcus luteus, Escherichia coli, and Bacillus subtilis. In conclusion, a recombinant hLYZ with high enzyme activity was successfully expressed in K. lactis GG799, thereby providing a basis for studies on the large-scale production of recombinant human lysozyme using the K. lactis expression system.
