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Biosci Biotechnol Biochem

Title:		Human lysozyme secretion increased by alpha-factor pro-sequence in Pichia pastoris
Author(s):		Oka C; Tanaka M; Muraki M; Harata K; Suzuki K; Jigami Y;
Address:		"Chiba Prefectural Industrial Research Institute, Chiba, Japan. coka@iri.pref.chiba.jp"
Journal Title:		Biosci Biotechnol Biochem
Year:		1999
Volume:		63
Issue:		11
Page Number:		1977 - 1983
DOI:		10.1271/bbb.63.1977
ISSN/ISBN:		0916-8451 (Print) 0916-8451 (Linking)
Abstract:		"To get high level secretion of human lysozyme in Pichia pastoris, the following three signal sequences and one prepro sequence were evaluated: chicken lysozyme signal peptide, leucine-rich artificial signal peptide, Saccharomyces invertase signal peptide, and Saccharomyces prepro sequence of alpha factor (MF-alpha Prepro). Transformants harboring a lysozyme gene with MF-alpha Prepro secreted 20-fold more lysozyme than those harboring the lysozyme gene with any one of the other three signal sequences. Three mutant leader sequences derived from MF-alpha Prepro were constructed to discover the function of the pro region. The secretion was dramatically decreased by eliminating the pro region of MF-alpha Prepro. In contrast, MF-alpha Prepro with the EAEAEA sequence directed the secretion of an equivalent level of lysozyme having the extra amino acids (EAEAEA) in its N-terminus. For the effective secretion of native human lysozyme, MF-alpha Prepro without any spacer sequences was most suitable. The secreted protein by MF-alpha Prepro construct was identical with the authentic human lysozyme, judging from N-terminal amino acid sequencing and molecular mass spectrometric and crystallographic analysis"
Keywords:		"Amino Acid Sequence Animals Base Sequence Chickens Cloning, Molecular/methods DNA Primers Glycoside Hydrolases/genetics Humans Mating Factor Molecular Sequence Data Muramidase/*biosynthesis/chemistry/*genetics Peptides/*genetics Pheromones/genetics *Pichi;"
Notes:		"MedlineOka, C Tanaka, M Muraki, M Harata, K Suzuki, K Jigami, Y eng England 2000/01/15 Biosci Biotechnol Biochem. 1999 Nov; 63(11):1977-83. doi: 10.1271/bbb.63.1977"