Purificação de quitosanases produzidas por Bacillus cereus utilizando cromatografía líquida rápida de proteínas
Rio Grande do Norte is a historical major producer and consumer of shrimp, thus being a major generator of the residues coming from this product processing. The shrimp residue, its exoskeleton, is rich in chitin. This is raw material for obtaining chitosan, which is generated by chitin deacetylation...
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| Formato: | bachelorThesis |
| Idioma: | pt_BR |
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Universidade Federal do Rio Grande do Norte
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| Endereço do item: | https://repositorio.ufrn.br/handle/123456789/38815 |
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| Resumo: | Rio Grande do Norte is a historical major producer and consumer of shrimp, thus being a major generator of the residues coming from this product processing. The shrimp residue, its exoskeleton, is rich in chitin. This is raw material for obtaining chitosan, which is generated by chitin deacetylation. Chitosan is rich in biological properties, but it is difficult to manipulate due to its low solubility in water. The chitosan hydrolysis generates chitooligosaccharides (QOS), which have several biological properties as antimicrobial, antitumor, anti-inflammatory and anti-cytotoxic. Obtaining QOS by enzymatic hydrolysis has several industrial advantages when compared, for example, to acid hydrolysis. However, for the use of enzymes in products for medicinal and pharmaceutical purposes, it is necessary that they possess a high purification factor. So, the development of low cost and efficient purification methodologies for these enzymes is important. Purification is a complex step and involves high industrial cost, so it is important that it is going to have few steps and high yield. Thus, this work was carried out to evaluate the performance of the AKTA Plus chromatography system to purify chitosanases produced from Bacillus cereus. Our research group has already carried out work of purification of this enzyme, but in bench chromatography. A fixed bed column with anionic ion exchange resin (Streamline DEAE) was used, and a linear gradient of 0 to 1 M NaCl was applied in the elution step of proteins from the adsorbent matrix. In general, the AKTA Plus system showed a good performance in the enzyme purification, obtaining the best elution result with 0.25 M NaCl buffer solution, reaching a purification factor of 9.54. In the overall evaluation of the elution process, the first integrated fraction of the elution resulted in a purification factor of 2.7. The individual point of best purification factor had a yield of 7.27% and the integrated fraction of improved elution performance had a yield of 18.96%. The purification of this enzyme using this system brought a considerable purification factor increase comparing with the methodology initially employed by our group. |
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