Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Pivot Concepts:
Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Target Concepts:
Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Query: UNIPROT:P00750 (
PLA
)
16,800
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
Phospholipase A
2
(
PLA
2
) has found extensive use in industry. However, recombinant
PLA
2
production in different expression systems is a difficult task because of its toxicity to cell membranes. We report here the development of an effective method for production of highly active
PLA
2
from Streptomyces violaceoruber strain A-2688 in the yeast Saccharomyces cerevisiae. The method is based on the use of the
PRP8
mini-intein (from Penicillium chrysogenum) inserted into the phospholipase sequence with the purpose of temporal inactivation of the enzyme and its subsequent delayed autoactivation. We demonstrate that the most effective site for intein insertion is Ser76 of the mature phospholipase. As a result of intein-containing precursor secretion from yeast cells and its subsequent autocatalytic splicing, highly active enzyme accumulated in the yeast culture fluid. The properties of the obtained recombinant phospholipase A
2
protein were similar to those of the native Streptomyces violaceoruber
PLA
2
protein. A possible evolutionary role of delayed autoactivation of intein-containing proteins is also discussed.
...
PMID:Expression of Highly Active Bacterial Phospholipase A
2
in Yeast Using Intein-Mediated Delayed Protein Autoactivation. 3238 5
