High-level secretory production of phospholipase A1 by Saccharomyces cerevisiae and Aspergillus oryzae

Phospholipase A 1 (PLA 1 ) is a hydrolytic enzyme that catalyzes the removal of the acyl group from position 1 of lecithin to form lysolecithin. The PLA 1 gene, which had been cloned from Aspergillus oryzae, was expressed in Saccharomyces cerevisiae and A. oryzae. Through the modification of the med...

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Bibliographic Details
Published in:Bioscience, biotechnology, and biochemistry biotechnology, and biochemistry, 2001-01, Vol.65 (1), p.94-101
Main Authors: Shiba, Y. (Sankyo Co. Ltd., Iwaki, Fukushima (Japan). Biomedical Research Labs.), Ono, C, Fukui, F, Watanabe, I, Serizawa, N, Gomi, K, Yoshikawa, H
Format: Article
Language:English
Subjects:
ASPERGILLUS ORYZAE
Aspergillus oryzae - enzymology
Aspergillus oryzae - genetics
Biological and medical sciences
Biotechnology
Blotting, Southern
CPY
Culture Media - chemistry
Enzyme engineering
ESTERASES
Ethanol - metabolism
Fermentation
Fundamental and applied biological sciences. Psychology
Gene Dosage
GENE EXPRESSION
Genes, Fungal
Genetic engineering
Genetic technics
GENETICALLY MODIFIED MICROORGANISMS
Immunoblotting
Industrial Microbiology - methods
Lysophosphatidylcholines - chemical synthesis
Lysophosphatidylcholines - metabolism
Methods. Procedures. Technologies
Modification of gene expression level
mycelial morphology
MYCELIUM
phospholipase A
Phospholipases A - biosynthesis
Phospholipases A - genetics
Phospholipases A1
Plasmids - genetics
Plasmids - metabolism
PRODUCTION
Production of selected enzymes
Promoter Regions, Genetic - genetics
Protein Sorting Signals
Recombinant Proteins - biosynthesis
SACCHAROMYCES CEREVISIAE
Saccharomyces cerevisiae - enzymology
Saccharomyces cerevisiae - genetics
SECRETION
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Description
Summary:Phospholipase A 1 (PLA 1 ) is a hydrolytic enzyme that catalyzes the removal of the acyl group from position 1 of lecithin to form lysolecithin. The PLA 1 gene, which had been cloned from Aspergillus oryzae, was expressed in Saccharomyces cerevisiae and A. oryzae. Through the modification of the medium composition and the feeding conditions of substrate, the production level of PLA 1 by S. cerevisiae was increased to a level fivefold higher than that indicated in a previous report. In the case of A. oryzae, introduction of multicopies of PLA 1 expression units, and the morphological change from the pellet form to the filamentous form were effective for the enhancement of PLA 1 production. We succeeded in producing 3,500 U/ml of PLA 1 using an industrial-scale fermentor.
ISSN:0916-8451
1347-6947
DOI:10.1271/bbb.65.94