Purification and characterisation of an Aspergillus niger invertase and its DNA sequence
| Autor: | Thierry Bergès, D. J. Ballance, Melanie J. Dobson, L. M. Boddy, John F. Peberdy, Christian Barreau, Marilene Henning Vainstein |
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| Rok vydání: | 1993 |
| Předmět: |
Glycoside Hydrolases
Size-exclusion chromatography Molecular Sequence Data Restriction Mapping Substrate Specificity chemistry.chemical_compound Hydrolysis Genetics Amino Acid Sequence Raffinose DNA Fungal Polyacrylamide gel electrophoresis biology Molecular mass Base Sequence beta-Fructofuranosidase Aspergillus niger Nucleic acid sequence Temperature General Medicine Hydrogen-Ion Concentration biology.organism_classification Chromatography Ion Exchange Blotting Southern Invertase Biochemistry chemistry Protein Biosynthesis Chromatography Gel Electrophoresis Polyacrylamide Gel |
| Zdroj: | Current genetics. 24(1-2) |
| ISSN: | 0172-8083 |
| Popis: | A secreted invertase was purified 23-fold by ultrafiltration, ion-exchange, and gel filtration chromatography from the culture supernatant of 18 h sucrose-grown cultures of Aspergillus niger. The purified enzyme hydrolysed sucrose and raffinose but there was no detectable hydrolysis of inulin, melezitose or PNPG. Invertase activity was optimal at pH 5.5 and 50 degrees C. The molecular mass of reduced invertase was 115 kDa, as determined by SDS gel electrophoresis. The native molecular weight of between 225 kDa and 250 kDa, estimated by electrophoresis under non-denaturing conditions, suggests that the protein is a dimer of identical subunits. The suc1 gene encoding this protein was completely-sequenced. The translated sequence yields a protein of 566 amino acids with a calculated molecular mass of 61 kDa, suggesting that carbohydrates represent about 50% of the mass of the protein. |
| Databáze: | OpenAIRE |
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