Studies on the purification of banana polyphenoloxidase

Studies on the extractability of polyphenoloxidase (PPO) from the pulp of five banana cultivars revealed a varietal difference in the nature of binding of the PPO in the cell, with the enzyme being entirely in the soluble fraction in one and partly associated with the cell wall in others, necessitating use of a detergent to release it from the latter. Partial purification by acetone precipitation and chromatography using a DEAE-cellulose column yielded two major fractions DE-I and DE-II with purifications of 4- and 16·3-fold and activity recoveries of 38·2 and 43·3% respectively. Further gel filtration of the two fractions on a Sephadex G-100 column improved the purifications to 44- and 50-fold respectively with full activity recovery. Polyacrylamide gel electrophoretic studies showed the two fractions to be composed of isoenzymes differing in pattern. The purified enzyme showed maximum absorption at 275 nm.

POLYPHENOLOXIDASE FROM ATEMOYA FRUIT (ANNONA CHERIMOLA MILL. ANNONA SQUAMOSA L.)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Some Biochemical properties of polyphenoloxidase from spearmint (Mentha arvensis)

Polifenoloxidase (PPO, EC 1.14.18.1) extraída de folhas de Mentha arvensis foi isolada por precipitação com (NH4)2SO4 e diálise extensiva. Seu pH e temperatura ótimos variaram com o substrato. A PPO apresentou atividade com vários difenóis. Valores de Km foram 0,825; 0,928 e 7,41 mM para ácido caféico, 4-metilcatecol e catecol, respectivamente. Na inativação térmica, 50% da enzima foi inativada após 60 e 15 segundos a 70 e 75ºC, respectivamente. A medida de atividade residual mostrou um efeito estabilizante de sacarose a várias temperaturas e uma energia de ativação (Ea) para inativação aumentando com a concentração de sacarose de 0 a 40% (p/p). Valores de energias de ativação de 78,13; 80,37; 82,79 and 81,00 kJ/Mol foram encontradas para 0, 15, 30 e 40% de sacarose, respectivamente. A PPO foi inibida pelos ácidos ascórbico, benzóico, cinamico, ferulico, p-cumárico, protocatéquico, além de metabisulfito de sódio, resorcinol e pirogalol. Os valores de Ki mostram que o ácido ascórbico foi o mais efetivo inibidor. O tipo de inibição foi determinado para cada inibidor.

Isolation and characterization of latent and active polyphenoloxidase in BRS Clara (CNPUV 154-147 x Centennial seedless) and BRS Morena (Marroo seedless x Centennial seedless) seedless table grapes

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Polyphenoloxidase activity in peaches after vibration

The vibration during transportation is one of the main causes of fruit damage. The actual methodologies used for damage evaluation in fruits seem to be subjective, since most of them are based on visual evaluation. In this study the effect of vibration, for one and two hours, on polyphenoloxidase (PPO) activity in peach pulp was investigated. The relation among PPO activity variation, postharvest time and resting time were also considered. Results showed that vibration affects PPO activity. However, it was not possible to correlate PPO activity variation and vibration.

Soluble proteins and polyphenoloxidase activity in bud flowers, flowers and leaves of cold stored lisianthus

This study evaluated the activity of the enzyme polyphenol oxidase (PPO) and the content of soluble protein present in lisianthus bud flowers, flowers and leaves in room temperature (24±2°C) and pre-exposure cold chamber at 9±2°C for 24 h, in order to examine a possible correlation between these parameters and postharvest longevity of lisianthus flowers. After treatments, flowers were kept in pots with water, stored at room temperature and evaluated every three days until the end of their decorative life for biochemical analyzes. During the experimental period the enzymatic activity increased with the aging of the material, directly related to the high concentration of phenolics that were accumulated in injured tissue, providing browning, while soluble protein content slightly decreased. Thus, PPO enzyme activity can be applied for plant senescence evaluation, acting as a biochemical marker for product visual quality.

花椒叶和土壤浸提液对土壤微生物、土壤酶及土壤化学性质的影响

花椒（Zanthoxylum bungeanum Maxim.）是川西地区重要的经济植物，化感作用是花椒连作障碍的原因之一，而花椒凋落物和根系分泌物对土壤质量的影响是花椒化感作用的一个重要方面。系统研究花椒如何影响土壤有助于深入理解和解决花椒连作障碍。本文主要以大红袍（10a生）花椒叶和种植过花椒的土壤的浸提液浇灌花椒幼苗进行试验，分析叶浸提液与土壤浸提液对非花椒生长土壤中土壤微生物、土壤酶及土壤化学性质的影响。主要结果如下： 1.花椒叶浸提液和土壤浸提液减少了土壤中微生物的种类、组成和数量。本试验中未施加浸提液的土样中根际微生物明显高于非根际区，在经过花椒叶浸提液处理后，根际细菌、真菌和放线菌数量以及微生物总数都有所减少，这样将会导致土壤中的有效养分的供给减少，进而可能影响植物的生长。 2．施加花椒叶浸提液和土壤浸提液，以及花椒幼苗的栽种，对不同土样中的土壤酶各有促进和抑制作用。在浸提液处理下，水解酶之间及氧化还原酶之间各存在相互促进作用。 3.施加花椒叶浸提液和土壤浸提液均抑制了根际土中全氮和有机质含量，叶浸提液还抑制了无苗土中全磷含量，土壤浸提液还抑制了无苗土中全氮含量与根际土全磷、有机质含量。但两种浸提液均促进了根际土中有效磷和水解性氮含量、根外土中全磷含量，叶浸提液促进了根际土中全磷含量，土壤浸提液促进了根外土中有效磷含量。全氮和有机质含量的下降可能对植物生长发育不利。 4．土壤化学性质与土壤酶活性在不同土样中有不同的相关性。全氮含量在施加叶浸提液的土样中与蛋白酶活性呈正相关。水解性氮含量在施加叶浸提液的土样中与蛋白酶活性、蔗糖酶活性呈正相关。全磷含量在施加叶浸提液的土样中与多酚氧化酶活性呈正相关；在施加土壤浸提液的土样中与蛋白酶活性、蔗糖酶活性呈正相关，与多酚氧化酶活性呈负相关。有效磷含量在施加叶浸提液的土样中与多酚氧化酶活性呈正相关，与蛋白酶活性呈负相关；在施加土壤浸提液的土样中与蛋白酶活性、过氧化氢酶活性呈正相关。有机质含量在施加叶浸提液的土样中与蛋白酶活性、蔗糖酶活性呈正相关。 Zanthoxylum bungeanum is one of the most important cash crops in Eastern Tibetan Plateau. Allelopathic effects could be one of reasons for Z. bungeanum’s continuous cropping impediment. The effects of secretion of leaf and root of Z. bungeanum on soil quality is a important way of Z. bungeanum’s allelopathic effects. However, allelopathic effect of Z.bungeanum on soil microbes, enzyme activities and chemical property were seldom studied. In this study, leaf and soil extracts of Da Hongpao(DHP), the most common varieties of Z.bungeanum in this area, were used to assess allelopathic effect of Z. bungeanum on soil biology and biochemistry by pot experiments . The main results showed that: 1. The irrigation of two kinds of extracts reduced the species, component and quantity of soil microbes. In rhizosphere soil which irrigated by distilled water, the quantity of soil microbes is significantly different from exoroot soil. In rhizosphere soil which irrigated by leaf extracts, the quantity of bacterial, fungi, actionmycete and gross of microbes were decreased, it may resulted in reduce of Available nutrient in soil, and influenced the growth of plants. 2．The irrigation of two kind of extracts reduced or enhanced the enzyme activities in different soils. Interaction between hydrolytic ferments and redoxases were promoted each other. 3. The irrigation of two kinds of extracts reduced the total N and organic matter in rhizosphere soil. Leaf extracts also reduced the total P in soil without seedling. Soil extracts reduced total N in soil without seedling and total P, organic matter in rhizosphere soil. But both extracts also enhanced available P and hydrolysable N in rhizosphere soil, total P in exoroot soil. Leaf extracts enhanced total P in rhizosphere soil. Soil extracts enhanced available P in exoroot soil. The reduction of total N and organic matter may influence growth of plants. 4．Positive correlations between total N and prolease, hydrolysable N and prolease, hydrolysable N and saccharase, total P and polyphenol oxidase, available P and polyphenoloxidase, organic matter and prolease, organic matter and saccharase, were studied in soil irrigated by leaf extracts. In soil irrigated by soil extracts, there are positive correlations between total P and prolease, total P and saccharase, available P and prolease, available P and catalase, while negative correlation between total P and polyphenoloxidase, available P and prolease, available P and catalase was found.

Técnicas de análise multivariada na avaliação de injúrias pelo frio em mangas cv. Tommy Atkins

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Chilling Injury in Avocados and Its Prevention with Thermal Treatment

The combinations of temperature and time which can cause chilling injuries in avocados 'Geada', 'Quintal' and 'Fortuna' were determined. The binomial 4 degrees C/ 28 days was selected to determine the activity of peroxidase (POD), polyphenoloxidase (PPO), polygalacturonase (PG) and methylesterase pectin (PME). The respiratory activity was also evaluated. The fruits were stored at this condition until being transferred to ambient conditions (22 degrees C and 77% RH) until maturity, when they were compared to fruits stored permanently at this environment, after being cleaned (control). In the second part of this work, different hydrothermal treatments were tested to prevent or minimize chilling injuries. Avocados 'Geada', 'Quintal' and 'Fortuna' were treated at 38 degrees C for 0, 30, 60 and 90 minutes before storage at 4 degrees C for 28 days. It was observed that the activity of enzymes associated to browning, POD and PPO, and to maturation, PG and PME, had become greater in fruits stored at 4 degrees C, or when they were transferred to environmental conditions. Fruits subjected to refrigeration, after transferred to environment, presented lower respiratory peak intensity and it occurred earlier than the others. Treatments using 38 degrees C for 60 and 90 min minimized the symptoms of chilling injury in avocados 'Geada' while for 'Quintal' the most efficient was 38 degrees C for 60 min. In 'Fortuna' these treatments did not minimize the damage by cold.

Armazenamento refrigerado de abacates 'Hass'

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Caracterização físico-química, bioquímica e funcional da jabuticaba armazenada sob diferentes temperaturas

O presente trabalho teve como objetivo avaliar a qualidade pós-colheita de jabuticabas submetidas a diferentes temperaturas de armazenamento refrigerado (AR). Após a colheita, os frutos fisiologicamente maduros foram acondicionados em bandejas de poliestireno expandido (EPS), revestidas por filme plástico de polietileno de baixa densidade (PEBD) e armazenados sob refrigeração a 0; 3; 6; 9 e 12 ± 1ºC e U.R. 87 ± 2%, sendo avaliados a cada 5 dias. Os frutos foram avaliados quanto à perda de massa, atividade respiratória, pH, acidez titulável, sólidos solúveis, ácido ascórbico, textura, pectina total e solúvel, atividade da enzima polifenoloxidase (PFO), compostos fenólicos e atividade antioxidante. Para frutos refrigerados a 9 e 12ºC, o pico respiratório atrasou em relação aos demais tratamentos, além de apresentarem as menores taxas respiratórias. O teor de sólidos solúveis aumentou com o tempo de armazenamento para todas as temperaturas, contudo, em 9 e 12ºC, esse aumento foi em menor proporção. A firmeza e o teor de ácido ascórbico também foram superiores nos frutos armazenados a 9 e 12ºC, enquanto os contéudos de pectina solúvel foram menores. Observou-se a diminuição da atividade da enzima PFO ao longo dos 30 dias do AR, independentemente da temperatura utilizada; entretanto, os menores valores foram encontrados nos frutos mantidos a 9 e 12ºC. Os frutos armazenados a 12ºC apresentaram os maiores conteúdos de compostos fenólicos totais e a maior atividade antioxidante ao final do experimento. Nesse sentido, a temperatura de 12ºC foi a mais efetiva na manutenção da qualidade pós-colheita das jabuticabas.

Capacidade antioxidante e qualidade pós-colheita de abacate 'Fuerte'

A capacidade antioxidante e a qualidade pós-colheita do abacate 'Fuerte' foram avaliadas. Os frutos foram mantidos sob temperatura ambiente (24 ± 1 ºC) e sob refrigeração em 10 ± 1 ºC e 90±5% UR e avaliados durante 15 dias. Determinaram-se a perda de massa e a taxa respiratória. As características físico-químicas avaliadas foram acidez total titulável (AT), sólidos solúveis (SS), ratio, pH, firmeza, atividade da enzima polifenoloxidase (PPO), cor (L, a* e b*), fenólicos totais e atividade antioxidante (AA) por DPPH·. Nas duas condições de armazenamento, o pico respiratório ocorreu no 9º dia.Os frutos mantidos sob temperatura refrigerada apresentaram menor perda de massa e maior firmeza em relação àqueles em temperatura ambiente. O comportamento respiratório refletiu na redução dos valores de AT e no conteúdo de SS que diminuíram a partir do pico de respiração. Os valores de pH aumentaram como consequência da diminuição da AT. Os valores de luminosidade mantiveram-se elevados, observando-se diminuição do componente de cor verde na polpa com o armazenamento. A atividade da PPO diminuiu com o armazenamento. Os valores de AA variaram de 17,6 a 68,7%. O conteúdo de fenólicos totais diminuiu após o pico respiratório, e valores crescentes de AA foram observados mesmo após o 9º dia.

Ineficiência de acibenzolar-S-methyl na indução de resistência de feijoeiro comum à murcha-de-Curtobacterium

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)