High-level secretory production of phospholipase A1 by Saccharomyces cerevisiae and Aspergillus oryzae.
Phospholipase A1 (PLA1) is a hydrolytic enzyme that catalyzes the removal of the acyl group from position 1 of lecithin to form lysolecithin. The PLA1 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 PLA1 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 PLA1 expression units, and the morphological change from the pellet form to the filamentous form were effective for the enhancement of PLA1 production. We succeeded in producing 3,500 U/ml of PLA1 using an industrial-scale fermentor.
Lead Discovery Research Laboratories, Sankyo Co. Ltd.,, Iwaki, Fukushima, Japan. shibay@ona.sankyo.co.jp
Y
Y
Shiba
C
C
Ono
F
F
Fukui
I
I
Watanabe
N
N
Serizawa
K
K
Gomi
H
H
Yoshikawa
D001236:Aspergillus oryzae / Q000201:enzymology* / Q000235:genetics; D015139:Blotting, Southern; D003470:Culture Media / Q000737:chemistry; D000431:Ethanol / Q000378:metabolism; D005285:Fermentation; D018628:Gene Dosage; D005800:Genes, Fungal; D015151:Immunoblotting; D007218:Industrial Microbiology / Q000379:methods*; D008244:Lysophosphatidylcholines / Q000138:chemical synthesis / Q000378:metabolism; D010741:Phospholipases A / Q000096:biosynthesis* / Q000235:genetics; D054466:Phospholipases A1; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D021382:Protein Sorting Signals; D011994:Recombinant Proteins / Q000096:biosynthesis; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*
D003470:Culture Media; D008244:Lysophosphatidylcholines; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D000431:Ethanol; D010741:Phospholipases A; D054466:Phospholipases A1