Produção e caracterização das Tanases do fungo filamentoso Aspergillus carbonarius
Nowadays, biotechnology is accompanied by functional, structural and application studies, mainly in industry, of microbial enzymes, which have aroused great interest in researchers around the world. Specially filamentous fungi have been highlighted as the major enzymes producers, mostly Aspergillus...
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| Format: | master thesis |
| Status: | Published version |
| Publication Date: | 2014 |
| Country: | Brasil |
| Institution: | Universidade Estadual Paulista (UNESP) |
| Repository: | Repositório Institucional da UNESP |
| Language: | Portuguese |
| OAI Identifier: | oai:repositorio.unesp.br:11449/124410 |
| Online Access: | http://hdl.handle.net/11449/124410 |
| Access Level: | Open access |
| Keyword: | Biotecnologia Enzimas Fungos filamentosos Fermentação Aspergillus Fermentation |
| Summary: | Nowadays, biotechnology is accompanied by functional, structural and application studies, mainly in industry, of microbial enzymes, which have aroused great interest in researchers around the world. Specially filamentous fungi have been highlighted as the major enzymes producers, mostly Aspergillus genera, an ascomycete. Among the enzymes of biotechnological interest we can found the tannin acyl hydrolase (EC 3.1.1.20), also known as tannase that can be produced by filamentous fungi, yeasts and bacterias. Acoording to the this objective of this work was to study the production of tannase by Aspergillus carbonarius standardizing the best physico-chemical conditions for the microorganism growth, in order to obtain high levels of tannase, purifying and characterizing them biochemically. The higher enzymes levels in SSF were obtained when it was used green tea leaves as carbon source moistured with tap water (1: 1 w / v) at 30° C for 3 days. Tannase was purified 11,3 fold with 98% of recover after two chromatographic steps: DEAE-celulose and Sepharose CL-6B. The enzyme has a molecular weight of 134,89 kDa with 50% of carbohydrates. The optimal temperature of activity was 60ºC and the optimal pH was 5.0. Tannase showed quite stability to temperatures between 40ºC and 65ºC and under acid pH. The enzyme activity was strongly inhibited by Mg+, Fe2+, Zn2+, Al3+ e Cu2+. The kinetic parameters were analyzed and the enzyme showed higher affinity to the substrate methyl gallate (Km 1.42mM) in it compared to tannic acid (Km 2.2mM). Therefore it is concluded that the tannase produced by Aspergillus carbonarius has great biotechnological potential and it is promising for industrial use. |
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