Tsuchiya Y, Morioka K, Shirai J, Yoshida K; International Conference on AIDS (15th : 2004 : Bangkok, Thailand).

Int Conf AIDS. 2004 Jul 11-16; 15: abstract no. WePeA5628.

National Institute of Animal Health, Tokyo, Japan

Background: Although human lysozyme is known as the antibacterial medicine for many years, it turns out that it is effective also in the medical treatment of AIDS in recent years. However, a mass production system for it is not established yet and its clinical application to AIDS is not made. In order to spread human lysozyme through the medical treatment of AIDS, it is necessary to establish the mass production system and to lower manufacture cost. Methods: Human lysozyme (HLY) gene connected with human lysozyme signal peptide (HLSP) gene was used for the secretion of recombinant HLY (rHLY) from yeast cells. HLSP portion in this gene was substituted with chicken lysozyme signal peptide (CLSP) gene or bovine lysozyme signal peptide (BLSP) gene, or the amino acid sequence near the n-terminal was altered, and effect of mutations on the secretion of HLY was estimated. The secretion amount of recombinant HLY in the medium was measured by the lytic activity against bacterial cell wall. Results: As the results of expression of each genes in yeast, rHLYs were secreted as follows; 18.1 mg/L in HLSP-HLY, 16.2 mg/L in BLSP-HLY, 21.5 mg/L in CLSP-HLY. These results shows that the CLSP is excellent than wild-type (HLSP). So next, n-terminal region of CLSP gene was altered and expressed in yeast. Deletion of arginine (Arg) residue decreased the HLY secretion (7.3 mg/L), but additioin of Arg increased the secretion (44.2 mg/L). Conclusions: The ability of signal peptides for secretion of rHLY are as follows: CLSP >wild-type (HLSP) >BLSP. By the addition of 2 or 3 Arg residues in the n-terminal region of CLSP, rHLY was successfully produced in large quantities (more than 50 mg/L). This indicates that the positive charge of this region in CLSP plays an important role in yeast as well as prokaryotes. The mass production system of rHLY was established here.

Publication Types:

• Meeting Abstracts

Keywords:

• Amino Acid Sequence
• Animals
• Cattle
• Chickens
• Gene Expression
• Gene Expression Regulation, Fungal
• Humans
• Muramidase
• Mutation
• Pharmaceutical Preparations
• Protein Sorting Signals
• Saccharomyces cerevisiae
• genetics

Other ID:

• GWAIDS0039346

UI: 102283562

From Meeting Abstracts