Aspergillus fumigatus Produces Two Arabinofuranosidases From Glycosyl Hydrolase Family 62: Comparative Properties of the Recombinant Enzymes
| Autor: | Rodrigo Sciaraffia Pérez, Jaime Eyzaguirre |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Glycoside Hydrolases Bioengineering Applied Microbiology and Biotechnology Biochemistry Pichia Substrate Specificity Pichia pastoris 03 medical and health sciences chemistry.chemical_compound Enzyme Stability Arabinoxylan Hydrolase Glycoside hydrolase Amino Acid Sequence Cloning Molecular Molecular Biology Peptide sequence Thermostability chemistry.chemical_classification biology Aspergillus fumigatus Temperature General Medicine biology.organism_classification Arabinose Recombinant Proteins Kinetics 030104 developmental biology Enzyme chemistry Xylans Biotechnology |
| Zdroj: | Applied Biochemistry and Biotechnology. 179:143-154 |
| ISSN: | 1559-0291 0273-2289 |
| Popis: | The genes of two α-L-arabinofuranosidases (AbfI and II) from family GH 62 have been identified in the genome of Aspergillus fumigatus wmo. Both genes have been expressed in Pichia pastoris and the enzymes have been purified and characterized. AbfI is composed of 999 bp, does not contain introns and codes for a protein (ABFI) of 332 amino acid residues. abfII has 1246 bp, including an intron of 51 bp; the protein ABFII has 396 amino acid residues; it includes a family 1 carbohydrate-binding module (CBM) in the N-terminal region, followed by a catalytic module. The sequence of ABFI and the catalytic module of ABFII show a 79 % identity. Both enzymes are active on p-nitrophenyl α-L-arabinofuranoside (pNPAra) with KM of 94.2 and 3.9 mM for ABFI and II, respectively. Optimal temperature for ABFI is 37 °C and for ABFII 42 °C, while the pH optimum is about 4.5 to 5 for both enzymes. ABFII shows a higher thermostability. When assayed using natural substrates, both show higher activity over rye arabinoxylan as compared to wheat arabinoxylan. ABFII only is active on sugar beet pulp arabinan and both are inactive towards debranched arabinan. The higher thermostability, higher affinity for pNPAra and wider activity over natural substrates shown by ABFII may be related to the presence of a CBM. The availability of the recombinant enzymes may be useful in biotechnological applications for the production of arabinose. |
| Databáze: | OpenAIRE |
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