title: Use of Bacillus brevis for efficient synthesis and secretion of human epidermal growth factor.
abstract: Using previously isolated Bacillus brevis strains that secrete large amounts of proteins but little protease into the medium, we have developed a host-vector system for very efficient synthesis and secretion of heterologous proteins. The multiple promoters and the signal-peptide-coding region of the MWP gene, a structural gene for one of the major cell wall proteins of B. brevis strain 47, were used to construct expression-secretion vectors. With this system, a synthetic gene for human epidermal growth factor (hEGF) was expressed efficiently and a large amount (0.24 g per liter of culture) of mature hEGF was secreted into the medium. hEGF purified from the culture supernatant had the same NH2-terminal amino acid sequence, COOH-terminal amino acid, and amino acid composition as natural hEGF, and it was fully active in biological assays. These results, in combination with previous results, showed that mammalian proteins can be produced in active form 10-100 times more efficiently in B. brevis than has been reported in other systems.
authors: Department of Food Science and Technology, Faculty of Agriculture, Nagoya University, Japan.
authors: H
authors: H
authors: Yamagata
authors: K
authors: K
authors: Nakahama
authors: Y
authors: Y
authors: Suzuki
authors: A
authors: A
authors: Kakinuma
authors: N
authors: N
authors: Tsukagoshi
authors: S
authors: S
authors: Udaka
mesh_terms: D000595:Amino Acid Sequence; D001407:Bacillus / Q000502:physiology*; D001483:Base Sequence; D002451:Cell Compartmentation; D003001:Cloning, Molecular / Q000379:methods*; D004815:Epidermal Growth Factor / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000302:isolation & purification / Q000502:physiology; D013329:Structure-Activity Relationship; D013997:Time Factors
chemical_list: D011994:Recombinant Proteins; D004815:Epidermal Growth Factor