Purification and Characterization of Phospholipase B fromCandida utilis
| Autor: | Satoko Akaboshi, Tomonari Fujita, Shuji Fujino, Yasuo Watanabe, Youichi Tamai, Daigo Akiyama |
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| Rok vydání: | 2006 |
| Předmět: |
Molecular Sequence Data
Applied Microbiology and Biotechnology Biochemistry Substrate Specificity Analytical Chemistry Phospholipase A2 Cations Catalytic triad Consensus sequence Amino Acid Sequence Lipase Molecular Biology Peptide sequence Conserved Sequence Candida chemistry.chemical_classification Kluyveromyces lactis Phospholipase B biology Organic Chemistry Temperature General Medicine Hydrogen-Ion Concentration biology.organism_classification Molecular biology Amino acid Molecular Weight chemistry Metals biology.protein Electrophoresis Polyacrylamide Gel Lysophospholipase Sequence Alignment Acyltransferases Biotechnology |
| Zdroj: | Bioscience, Biotechnology, and Biochemistry. 70:377-386 |
| ISSN: | 1347-6947 0916-8451 |
| Popis: | Phospholipase B (PLB) from the asporogenous yeast Candida utilis was purified to homogeneity from a culture broth. The apparent molecular mass was 90-110 kDa by SDS-PAGE. The enzyme had two pH optima, one acidic (pH 3.0) and the other alkaline (pH 7.5). At acidic pH the enzyme hydrolyzed all phospholipids tested without metal ions. On the other hand, the PLB showed substrate specificity and required metal ions for alkaline activity. The cDNA sequence of the PLB was analyzed by a combination of several PCR procedures. The PLB encoded a protein consisting of 643 amino acids. The amino acid sequence contained a lipase consensus sequence (GxSxG) and catalytic arginine and aspartic acid motifs which were identified as the catalytic triad in the PLB from Kluyveromyces lactis, suggesting that the catalytic mechanism of the PLB is similar to that of cytosolic phospholipase A(2) (cPLA(2)), found in mammalian tissues. |
| Databáze: | OpenAIRE |
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