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Master's Dissertation
DOI
https://doi.org/10.11606/D.9.2005.tde-24112008-135527
Document
Author
Full name
Cristina de Simone Carlos Iglesias Pascual
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2005
Supervisor
Committee
Finardi Filho, Flavio (President)
Iguti, Antonia Miwa
Marquez, Úrsula Maria Lanfer
Title in Portuguese
Caracterização físico-química e purificação de enzimas amilolíticas de mandioca (Manihot esculenta Crantz) cv. Zolhudinha
Keywords in Portuguese
Manihot esculenta
Alfa-amilase
Caracterização enzimática
Mandioca
Pós-colheita
Purificação
Abstract in Portuguese
A mandioca (Manihot esculenta Crantz) é uma raiz originária e cultivada na América do Sul, com alta perecibilidade no período pós-colheita. Seus principais processos de deterioração envolvem reações enzimáticas, oxidativas e microbiológicas. Neste trabalho foram estudadas raízes de mandioca da variedade Zolhudinha catalogada pela EMBRAPA como IM-158, provenientes da região amazônica, que se destacam pela alta atividade amilolítica. Foram analisadas as condições físico-químicas junto com o isolamento e purificação da α-amilase da raiz e a possível participação desta enzima no processo deteriorativo pós-colheita. Por ser uma variedade de mandioca não comercial, o tempo de cocção foi em média de 4,30 h, teor de umidade em tomo de 64 % e porcentagem de amido de cerca de 30 %. A atividade amilásica decai em 1/3 de sua intensidade no quinto dia pós-colheita, em contraponto a formação de açúcares redutores, cuja concentração aumenta cinco vezes. A purificação foi obtida com duas etapas cromatográficas, com DEAE-celulose e Sephacryl S-200, revelando duas isoenzimas de α-amilase treze vezes mais purificadas, com recuperação protéica de 7,5 % e com pesos moleculares entre 14 e 19 kDa.
Title in English
Characterization physical-chemistry and purification of amylolytic enzymes from cassava (Manihot esculenta Crantz) cv.Zolhudinha
Keywords in English
Manihot esculenta
Alpha-amylase
Cassava
Enzyme characterization
Post-harvest
Purification
Abstract in English
Cassava (Manihot esculenta Crantz) is a root from a native plant, cultivated in South América, that is hightly perish on the post-harvest time. The mechanisms of the root deterioration are due to enzymatic and oxidative reactions as well as the microbiological attack. In this work were studied roots of Zolhudinha variety, EMBRAPA - IM 158, cultivated in Amazonian area, which distinguishes from others varieties by its higher amylase activity. Physicochemical properties were analyzed during the post-harvest time, the purification of α-amylase were performed to establish a possible involvement on the deteriorative process. As a non-commercial variety, the cooking time of the roots was 4.30 hours on average, with 64 % of water content and 30 % of starch. The amylase activity during the post-harvest decrease 1/3 from the original at the day 5, that matches with the reducing sugar in roots by increase of five times. The purification was achieved by two chromatography steps on DEAE-cellulose and Sephacryl S-200, providing two isozymes of α-amylase, thirteen times more purified with a recovery of 7.5 % of the protein fraction, the estimated molecular weights were between 14 and 19 kDa.
 
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Publishing Date
2008-11-25
 
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