Mango starch degradation. II. The binding of alpha-amylase and beta-amylase to the starch granule

During mango ripening, soluble sugars that account for mango sweetening are accumulated through carbon supplied by both photosynthesis and starch degradation. The cultivar Keitt has a characteristic dependence on sugar accumulation during starch degradation, which takes place during ripening, only a few days after detachment from the tree. Most knowledge about starch degradation is based on seeds and leaves currently used as models. However, information about the mango fruit is scarce. This work presents the evaluation of alpha- and beta-amylases in the starch granule surface during fruit development and ripening. Extractable proteins were assayed for amylase activity and detected by immunofluorescence microscopy and correlated to gene expression. The results suggest that both amylases are involved in starch degradation during mango ripening, probably under the dependence of another signal triggered by the detachment from the mother-plant.     

Chromoplast morphology and beta-carotene accumulation during postharvest ripening of Mango Cv. 'Tommy Atkins'

Accumulation of beta-carotene and trans-cis isomerization of ripening mango mesocarp were investigated as to concomitant ultrastructural changes. Proceeding postharvest ripening was shown by relevant starch degradation, tissue softening, and a rising sugar/acid ratio, resulting in a linear decrease (R (2) = 0.89) of a ripening index (RPI(KS)) with increasing ripening time. A modest accumulation of all-trans-beta-carotene and its cis isomers resulted in a slight pigmentation of the mango chromoplasts, because ambient temperatures of 18.2-19.5 degrees C provided suboptimal ripening conditions, affecting color development and beta-carotene biosynthesis. The ultrastructures of chromoplasts from mango mesocarp and carrot roots were comparatively studied by means of light and transmission electron microscopy. Irrespective of the ripening stage, mango chromoplasts showed numerous plastoglobuli varying in size and electron density. They comprised the main part of carotenoids, thus supporting the partial solubilization of the pigments in lipid droplets. However, because different pigment-carrying tubular membrane structures were also observed, mango chromoplasts were assigned to the globular and reticulotubular types, whereas the crystalline type was confirmed for carrot chromoplasts. The large portions of naturally occurring cis-beta-carotene in mango fruits contrasted with the predominance of the all-trans isomer characteristic of carrots, indicating that the nature of the structure where carotenoids are deposited and the physical state of the pigments are crucial for the stability of the all-trans configuration.     

Plantain and banana starches: granule structural characteristics explain the differences in their starch degradation patterns

Different banana cultivars were used to investigate the influences of starch granule structure and hydrolases on degradation. The highest degrees of starch degradation were observed in dessert bananas during ripening. Scanning electron microscopy images revealed smooth granule surface in the green stage in all cultivars, except for Mysore. The small and round granules were preferentially degraded in all of the cultivars. Terra demonstrated a higher degree of crystallinity and a short amylopectin chain length distribution, resulting in high starch content in the ripe stage. Amylose content and the crystallinity index were more strongly correlated than the distribution of amylopectin branch chain lengths in banana starches. α- and β-amylase activities were found in both forms, soluble in the pulp and associated with the starch granule. Starch-phosphorylase was not found in Mysore. On the basis of the profile of α-amylase in vitro digestion and the structural characteristics, it could be concluded that the starch of plantains has an arrangement of granules more resistant to enzymes than the starch of dessert bananas.     

Olive fruit cell wall: degradation of pectic polysaccharides during ripening

Olive fruits at three stages of ripening (green, cherry, and black) have been studied. After cell wall isolation, the compositions of the cell wall and that of the phosphate-soluble polysaccharides were determined. In cell walls, decreases in arabinose, xylose, glucose, and uronic acid levels were observed, together with a slight increase in mannose on ripening. At the beginning of ripening, fragments of pectic polymers were the major constituents of the phosphate-soluble fraction, with the hemicellulosic ones increasing toward the end of the process. The molecular weight of the fragments solubilized was approximately 6 kDa. After cell wall fractionation, the pectic polysaccharides soluble in imidazole and sodium carbonate were also studied. In both fractions, between the green and cherry stages of ripening, a significant loss of homogalacturonans took place. Between the cherry and black stages of ripening, rhamnogalacturonan side chains were also released in addition to homogalacturonans. In any of the pectic fractions, changes in apparent molecular weight were quantified.     

几种化学药物混合物熏气催熟杧果的综合效应

为解决果后熟过程中存在的黄熟程度不均匀、易感炭疽病和蒂腐病等问题,进行几种化学药物的混合物所产生气体催熟果的试验。结果表明:最佳的处理(处理Ⅱ),果黄熟仅需4 d,防炭疽病效果可达93%,防蒂腐病效果可达100%,可溶性固形物含量提高1%以上,轻耗仅为自然后熟的17.38%,果皮色泽鲜艳,果肉硬,风味佳,外观整洁,商品率高。该处理方法简单易行。     

Olive fruit cell wall: degradation of cellulosic and hemicellulosic polysaccharides during ripening

Cellulose and hemicelluloses obtained from the cell walls of partially depectinated olives have been studied at three stages of ripening (green, cherry, and black). Hemicelluloses were fractionated into two groups, the amounts of which diminished during ripening: those soluble in 4% KOH diminished between the cherry and black stages, whereas those soluble in 24% KOH did so between the green and cherry stages. Arabinoxylans, xyloglucans, and homo- and/or rhamnogalacturonans to a lesser extent were present in these fractions. After ion exchange and size exclusion chromatographies, decreases in the molecular weights of hemicelluloses, mainly in the neutral fractions, were observed. The amount of cellulose also decreased, but at the second stage of the ripening process. Approximately 2 mg/fruit of glucose was lost from cellulose, and the amount of uronic acids increased (0.23 mg/fruit).     

苹果果实成熟期间电特性的研究

为了了解果实成熟期间电特性的变化规律,该文以红富士苹果为对象,研究了测试信号的频率和电压对成熟期苹果电参数的影响,果实生长中电参数和生理生化参数的变化,并分析了电参数变化的原因。研究结果表明：频率对苹果电参数的影响规律与苹果成熟度无关,但电压对苹果电参数的影响规律与苹果成熟度有关。苹果成熟期间,其呼吸强度持续下降;果实成熟前期,相对介电常数和pH值逐渐下降,后期逐渐增大;而电阻率和可溶性固形物含量在前期逐渐增大,后期减小。这些参数的转折点皆出现在同一时间,说明电参数与苹果内部品质之间存在必然的联系,可以用电参数反映内部物质成分的变化。     

Soil degradation: I. Basic processes

This paper examines soil and land degradation. It describes basic processes and factors responsible for degradation, illustrates the cause-effect relationships and differentiates between natural and man-induced regressive effects. The ‘critical limit’ concept is described in terms of properties beyond which the soil will not support an economically-viable agriculture. This paper is not an exhaustive literature review but emphasizes the scientific principles involved and highlights natural against man-induced processes. Important natural processes are: laterization, hard-setting, fragipan and clay pan formation, and geologic erosion. In comparison, man-induced processes consist of: soil compaction, accelerated erosion, desertification, salt accumulation and leaching and acidification. One of the principal constraints is the problem of data reliability. A reliable database and precise criteria are definitely lacking and hinder the assessment of the extent, type and degree of soil degradation and establishing the cause-effect scenario. Improving our database is, therefore, of a high priority if we are to adopt land use policy for sustainable soil management and long-range resource management. Also outlined, are vital research and development strategies. Judicious resource management policy should emphasize managing prime agricultural land to produce to its maximum potential so that there is no need to cultivate marginal and easily-degraded fragile ecosystems. A strict code of conduct is needed for utilizing marginal/fragile lands. Methods of restoring the productivity of degraded lands must be researched so as to minimize the need to clear and develop new lands.     

芒果动态运输试验考核

使用汽车、火车等运输工具,对经过不同保鲜处理的芒果进行了远距离、长时间的生产型动态运输考核,较系统地观测了整个运输过程中环境因子的变化,根据实际考核筛选出了最佳动态保鲜技术方案。     

Functional characterization of FaCCD1: a carotenoid cleavage dioxygenase from strawberry involved in lutein degradation during fruit ripening

A gene encoding a carotenoid cleavage dioxygenase class 1 enzyme (FaCCD1) was identified among a strawberry fruit expressed sequence tag collection. The full-length cDNA was isolated, and the expression profiles along fruit receptacle development and ripening, determined by quantitative real time polymerase chain reaction, showed that FaCCD1 is a ripening-related gene that reaches its maximal level of expression in the red fully ripe stage. FaCCD1 was expressed in Escherichia coli, and the products formed by the recombinant protein through oxidative cleavage of carotenoids were identified by liquid chromatography-mass spectrometry and gas chromatography-mass spectrometry analyses. The FaCCD1 protein cleaves zeaxanthin, lutein, and beta-apo-8'-carotenal in vitro. Although beta-carotene is not a good substrate for FaCCD1 in vitro, the expression of FaCCD1 in an engineered carotenoid-producing E. coli strain caused the degradation of beta-carotene in vivo. Additionally, the carotenoid profile in strawberry was analyzed by high-performance liquid chromatography-photodiode detection, and a correlation between the increase of the expression level of FaCCD1 during ripening and the decrease of the lutein content suggests that lutein could constitute the main natural substrate of FaCCD1 activity in vivo.     

Milling of rice grains. The degradation on three structural levels of starch in rice flour can be independently controlled during grinding

Whole polished rice grains were ground using cryogenic and hammer milling to understand the mechanisms of degradation of starch granule structure, whole (branched) molecular structure, and individual branches of the molecules during particle size reduction (grinding). Hammer milling caused greater degradation to starch granules than cryogenic milling when the grains were ground to a similar volume-median diameter. Molecular degradation of starch was not evident in the cryogenically milled flours, but it was observed in the hammer-milled flours with preferential cleavage of longer (amylose) branches. This can be attributed to the increased grain brittleness and fracturability at cryogenic temperatures, reducing the mechanical energy required to diminish the grain size and thus reducing the probability of chain scission. The results indicate, for the first time, that branching, whole molecule, and granule structures of starch can be independently altered by varying grinding conditions, such as grinding force and temperature.     

Molecular degradation rate of rice and corn starches during acid-methanol treatment and its relation to the molecular structure of starch

The degradation rates of rice and corn starches with different contents of amylose treated in methanol containing 0.36% HCl at 25 degrees C for 1-15 days were evaluated by monitoring the weight average degree of polymerization of starch. A two-stage degradation pattern during acid-methanol treatment was found for the starches studied, which were the slow (first) and the rapid (second) degradation stages. Waxy starches showed a shorter time period of the first stage than that of nonwaxy starch. Rice starch showed a shorter time period of the first stage and a higher degradation rate of the second stage than the counterpart corn starch with similar amylose content. Despite the botanic source and amylose content of starch, the degradation rate of starch in the second stage significantly (p < 0.05) correlated to the S/L ratio (r = -0.886) and polydispersity (r = 0.859) of amylopectin branch chains of native starch.     

Granule structure and distribution of allomorphs in C-type high-amylose rice starch granule modified by antisense RNA inhibition of starch branching enzyme

C-type starch, which is a combination of both A-type and B-type crystal starch, is usually found in legumes and rhizomes. We have developed a high-amylose transgenic line of rice (TRS) by antisense RNA inhibition of starch branching enzymes. The starch in the endosperm of this TRS was identified as typical C-type crystalline starch, but its fine granular structure and allomorph distribution remained unclear. In this study, we conducted morphological and spectroscopic studies on this TRS starch during acid hydrolysis to determine the distribution of A- and B-type allomorphs. The morphology of starch granules after various durations of acid hydrolysis was compared by optical microscopy, scanning electron microscopy, and transmission electron microscopy. The results showed that amorphous regions were located at the center part of TRS starch subgranules. During acid hydrolysis, starch was degraded from the interior of the subgranule to the outer surface, while the peripheral part of the subgranules and the surrounding band of the starch granule were highly resistant to acid hydrolysis. The spectroscopic changes detected by X-ray powder diffraction, 13C cross-polarization magic-angle spinning NMR, and attenuated total reflectance Fourier transform infrared showed that the A-type allomorph was hydrolyzed more rapidly than the B-type, and that the X-ray diffraction profile gradually changed from a native C-type to a CB-type with increasing hydrolysis time. Our results showed that, in TRS starch, the A-type allomorph was located around the amorphous region, and was surrounded by the B-type allomorph located in the peripheral region of the subgranules and the surrounding band of the starch granule. Thus, the positions of A- and B-type allomorphs in the TRS C-type starch granule differ markedly from those in C-type legume and rhizome starch.     

Effect of temperature during ripening on grain quality of rice

Temperature during grain ripening has been shown to affect amylose content and gelatinization temperature of rice starch (1–6). These studies demonstrated that high ambient temperature results in lower amylose content and higher gelatinization temperature of the starch. Rice starches obtained from rice grains that matured at lower temperature had higher iodine blue values and were more susceptible to alkali digestion than those that ripened at higher temperature.     

Evolution of the taste of a bitter Camembert cheese during ripening: characterization of a matrix effect.

The objective of this study was to characterize the effect of ripening on the taste of a typically bitter Camembert cheese. The first step was to select a typically bitter cheese among several products obtained by different processes supposed to enhance this taste defect. Second, the evolution of cheese taste during ripening was characterized from a sensory point of view. Finally, the relative impact of fat, proteins, and water-soluble molecules on cheese taste was determined by using omission tests performed on a reconstituted cheese. These omission tests showed that cheese taste resulted mainly from the gustatory properties of water-soluble molecules but was modulated by a matrix effect due to fat, proteins, and cheese structure. The evolution of this matrix effect during ripening was discussed for each taste characteristic.     

Distribution and characterization of enzymes causing starch degradation in rice (Oryza sativa cv. koshihikari)

The thermal dependency and stability of enzymes producing reducing sugar (RS) were examined in bran, the exterior 13% part (outer endosperm), and the remaining inner endosperm of rice grains. RS-producing enzymes in the inner endosperm showed a higher optimum temperature than those in other parts of the rice grain. Diethylaminoethyl-Sephacel chromatography of crude extracts revealed two peaks of RS-producing activity with different optimum temperatures (60 and 37 degrees C) in all three parts. alpha-Glucosidase (EC 3.2.1.20) and alpha-amylase (EC 3.2.1.1) isoform G were thought to be major components of the RS-producing activities with high and low optimum temperatures, respectively. The peak with a high optimum temperature was a more abundant component in the inner endosperm, compared with other parts of the rice grain. Thus, different parts of rice were found to have distinct enzyme sets having different thermal dependency and to be involved in starch degradation to various sugars.     

Changes in chloroplast pigments of olive varieties during fruit ripening

Changes in chlorophyll and carotenoid pigments of five olive (Olea europaea L.) varieties destined for milling were investigated at six consecutive ripening stages. There was a manifest dependence between olive variety, moment of picking, and chloroplast pigment composition of the fruits. Although the content of chlorophylls and carotenoids differed with fruit variety, ripening always involved their gradual loss, which becames more pronounced with increased presence of anthocyanin compounds. The relative rates of disappearance of chlorophylls and carotenoids were markedly different between varieties, implying that the catabolism of these pigments takes place at a relative rate inherent to each variety. The varieties less rich in pigments showed the most extreme behavior. The highest relative rate of disappearance was observed in fruits of the Blanqueta variety, and the lowest was observed in those of Arbequina. The chlorophyll a/chlorophyll b ratio remained practically constant during ripening, with a value very similar for Hojiblanca, Picual, Cornicabra, and Blanqueta, but much higher for Arbequina, implying that the structure of the photosynthetic apparatus is different in the latter variety. In the five varieties studied, lutein was the slowest carotenoid to be degraded, so that its percentage in the fruits increased with ripening, whereas beta-carotene was the fastest to disappear. In ripe fruits covered with anthocyanins, chloroplast pigments were retained in both skin and pulp, with the rate of disappearance being much higher in the latter.     

Mozzarella干酪成熟中蛋白水解与功能特性的变化

为控制干酪的质量,对Mozzarella干酪成熟过程中蛋白质的水解(测定SDS凝胶电泳和可溶性氮)和未融化干酪的质构变化以及融化干酪功能特性变化进行了研究,干酪成熟过程中由于凝乳酶和乳酸菌酶的作用使蛋白水解,从而使pH 4.6可溶性氮(SN)和12% TCA SN逐渐增加;凝乳酶主要影响酪蛋白的水解范围,乳酸菌及其酶,不但影响酪蛋白的水解范围,而且主要影响酪蛋白的水解深度。干酪中的残留凝乳酶和乳酸菌酶使酪蛋白水解为大分子量的肽段,而乳酸菌酶还可将大分子量的肽段进一步降解为小分子量的肽段和游离氨基酸。由于酪蛋白的水解,使干酪的硬度和弹性下降,融化性和油脂析出性增加,随着小分子量肽和游离氨基酸的增加,干酪的褐变性提高。     

Protein hydrolysis and proteinase activity during the ripening of salted anchovy (Engraulis encrasicholus l.). A microassay method for determining the protein hydrolysis.

The protein hydrolysis and proteinase activity during the ripening of salted anchovy were studied. A rapid, simple, and inexpensive microassay method for determining the protein hydrolysis by trinitrobenzenesulfonic acid (TNBS) has been developed. A linear relationship was observed between proteolysis determination by the TNBS method and ripening time in the fish muscle and in the brine (r = 0.99). A linear relationship was also observed between the ratio nonprotein nitrogen and total nitrogen (NPN/TN) and ripening time (r = 0.98). Proteolysis by the TNBS method and NPN/TN determination could be considered as objective methods to follow and assess the ripening process of an anchovy. A value of proteolysis by the TNBS method of 240 mM leucine in the fish muscle and/or 200 mM leucine in the brine would indicate the ripening point. The crude enzyme prepared of fish muscle and brine showed that alkaline proteinases dominate.     

Modification of rice starch by selective degradation of amylose using alkalophilic Bacillus cyclomaltodextrinase

A cyclomaltodextrinase (CDase) isolated from alkalophilic Bacillus sp. I-5 (CDase I-5) exists in a dodecameric form, an assembly of six dimers, each catalytic site of which is located in a narrow groove at the interface of the dimeric unit. Because of the unique geometric shape of the catalytic site, the enzyme has the ability to discriminate the molecular size of substrates. An analysis of the hydrolysis reaction of the enzyme revealed that its kcat/Km value on amylose was 14.6 s(-1) (mg/mL)(-1), whereas that for amylopectin was 0.92 s(-1) (mg/mL)(-1), showing an exceptionally high preference toward amylose. CDase I-5 was applied to modify the starch structure to produce low-amylose starch products by incubating rice starch with this enzyme. We found that the amylose content of rice starch decreased from 28.5 to 9%, while the amylopectin content remained almost constant with no significant change in the side chain length distribution. When the CDase I-5-treated rice starch was stored at 4 degrees C for 7 days, the retrogradation rate was significantly retarded as compared to that in the control sample.