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26

3.2.2 ANALISE DE AMID°

Os teores de amido foram determinados enzimaticamente pelo metodo

descrito por Areas e Lajolo (1981). 0 amido foi extraido corn hidr6xido de sOdio

0,5N, precipitado corn etanol 80%, hidrolisado coal amiloglicosidase e a glicose

liberada determinada pelo sistema glicose oxidase/peroxidase/ABTS, segundo

metodo de Bergmeyer (1974).

3.2.3 DETERMINACA 0 DE ACOCARES SOLUVEIS

Os acOcares solOveis (glicose, frutose e sacarose) foram extraidos corn

etanol 80% a 80°C por tres vezes, os sobrenadantes foram combinados, o etanol

evaporado por sistema a vàcuo a baixas temperaturas (speed vac) e o volume

reconstituido corn âgua.

Os acOcares foram estimados por HPLC por cromatografia de troca i6nica

utilizando urn cromat6grafo da DIONEX modelo DX500 acoplado corn detector

amperomêtrico de puiso, modelo ED40. A coluna utilizada foi a CARBOPAC PA 1

(4x250 mm). A fase mOvel foi constituida 100% de NaOH 18 mM ern fluxo de 1,0

ml/min.

3.2.4 PREPARO DOS EXTRATOS ENZIMATICOS

Os extratos enzimàticos, para doseamento das atividades da SS e SPS,

foram feitos seguindo metodologia otimizada por Cordenunsi (1989), onde cerca

de 0,5g de polpa de banana foi homogeneizada em 2,0 ml de meio extrator, em

homogeneizador Brinkman (tipo Turrax), por cerca de 30 segundos em velocidade

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IL

O amadurecimento é uma etapa exclusiva do estádio de desenvolvimento

dos frutos, que envolve uma série de transformações metabólicas a partir de

diferentes fontes de energia. Ele é mediado por um dinâmico complexo

enzimático, resultando em sínteseldegradação e conversão de cornpostos que

tornarão o fruto aceitável para o consumo. Dependendo do tipo de fruto, essa

fonte de energia pode ser na forma de ácidos orgânicos, sacarose vinda da

própria planta e na forma de amido. A banana (Musa acuminata) e uma fruta de

comportamento climatérico que utiliza como principal fonte de carbono o amido,

que é reduzido durante o climatério de teores que variam de 12 a 20% a menos de

1 %. Concomitante a esta degradação, o teor de sacarose pode atingir até 15%,

dependendo da cultivar. O ácido giberélico (GA3) é um fitohormônio da família das

giberelinas que tem sido muito estudado em cereais por aumentar a transcrição

gênica das a-amilase. Em frutos, ele é responsável por manter a textura firme e o

teor de sólidos solúveis, e atrasar o amadurecimento. Ao estudar a influência do

GA3 no metabolismo arnido-sacarose em fatias de banana, observou-se neste

trabalho que o fitohormônio não alterou o pico respiratório nem a síntese de

etileno. Entretanto, atrasou a degradação do amido e o acúmulo de açúcares

solúveis por três dias. Este atraso foi acorriparihado pela dirriinuiçãolatraso na

atividade das enzimas que degradam o amido e sintetizam sacarose, a e

p-amilase e sacarose fosfato sintase, respectivamente, sendo que não foi

observada uma diferença no aumento de expressão gênica da sacarose fostato

sintase e das fosforilases.

ABSTRACT

The ripening is an exclusive stage of fruit development, that involves a serie

of metabolic transformation from different energy source. It is mediated by a

dynarriic enzymatic complex, resulting in formationldegradation of different

coumpouds that will render fruit acceptable for the consumption. Depending on the

type of fruit, this energy source can be in the form of organic acid, sucrose of the

plant or starch. Banana (Musa acuminata) is a climateric fruit that uses starch as

main carbon source, which is reduced during banana ripening of levels that vary

.from 12 to 20% to less than 1 %. Concomitant to the this degradation, the levels of

sucrose can reach up to 15%, depending of the cultivar. The gibberellic acid, GA3-

mediated is a plant growth regulation of the giberellins family, that has been

studied in cereals because of their enhancing effect of gene expression of a-

amylase. In fruits, it is responsible for keeping the texture firm and the soluble

solid levels, and deiaying the ripening. The influence of the GA3 in the starch-

sucrose metabolism in banana slices, were observed. That the phytohormone did

not modify the respiratory peak nor the synthesis of ethylene. However, it delayed

the starch degradation and the soluble sugars accumulation for about three days.

This delay was followed by decrease andlor delay in the activity of the enzymes

related to starch degradation: the a and p-amylases; and sucrose synthesis the

sucrose phosphate synthase (SPS). However, was not observed a difference in the

increase of gene expression of SPS and phosphorilase