Purification and characterisation of carrot (Daucus carota L) pectinesterase

A pectinesterase isoform with an alkaline isoelectric point of over 8.66 was detected in crude extracts of carrot. The enzyme was purified by ion exchange and molecular exclusion chromatography. The molecular weight of the isoform was 25 kDa, determined in native conditions by filtration through Sephadex G‐75 SF. The enzyme showed a high affinity for its substrate, with K_m and V_max values of 0.031 mg ml^?1 and 6.77 units respectively for apple pectin. The pectinesterase activity exhibited an optimum around pH 7.4 and was activated by metallic ions, with optimum activities at NaCl concentrations between 130 and 330 mM and at CaCl₂ concentrations between 15 and 50 mM . The enzyme was activated most by Ca²⁺ and exhibited a greater tolerance of high concentrations of Na⁺. Comparison of its heat stability with other pectinesterases of vegetable origin indicated that the purified isoform was very thermolabile, being rendered inactive by heating for 5 min at 70 °C. The enzyme was inhibited by high concentrations of polygalacturonic acid and competitively inhibited by D ‐galacturonic acid, with a K_i value of 1 mM . Copyright © 2003 Society of Chemical Industry     

Evaluating eggplant (Solanum melongena L) genotypes for bioactive properties: A chemometric approach

Thirty four genotypes of eggplant and its wild relatives were evaluated for their total phenolics, flavonoid content and antioxidant capacity. Free radical scavenging was evaluated using four in-vitro assays, viz. FRAP (Ferric reducing antioxidant power), CUPRAC (cupric reducing antioxidant capacity), TEAC (Trolox equivalent antioxidant capacity) and DPPH (2, 2-diphenylpicrylhydrazyl). Total phenolics in eggplant showed a wide variation, ranging from 22.62 to 234.46 mg gallic acid equivalents (GAE)/100 g fw (244.28 to 2990.64 mg GAE/100 g dw). With the exception of Solanum aethiopicum and its accessions (Ac-1, Ac-2 and Ac-3), all wild relatives had significantly (p < 0.05) higher total phenolics and flavonoid content than cultivated ones. Total antioxidant capacity (FRAP) in wild genotypes ranged from 1.13 to 8.04 μmol Trolox (TE)/g. The hierarchy in decreasing order was Solanum khasianum > Solanum torvum > Solanum sisymbriifolium > Solanum incanum > Solanum integrifolium > S. aethiopicum. The antioxidant capacity correlated high with total phenolics in all assays. In cultivated group, JBR-99 and RCMBL-3 possessed high antioxidant capacity than the rest. Principal component analysis (PCA) and Hierarchical cluster analysis (HCA) revealed distinct similarity between JBR-99 (green), S. sisymbriifolium, S. khasianum and S. torvum. Overall results indicate that the wild species of S. torvum, S. incanum and S. sisymbriifolium are potential candidates for improving the functional quality of cultivated eggplant.     

Modeling Wound-induced Respiration of Fresh-cut Carrots (Daucus carota L.)

ABSTRACT A mechanistic model based on synthesis and inactivation of enzymes, associated to the respiration pathway, was proposed to describe wound-induced respiration of fresh-cut produce. Carrots ( Daucus carota L.) were cut into slices, sticks, a combination of both, and shreds to obtain different wounding intensities, defined by the ratio of new area created to tissue weight (A/W). Respiration rates obtained at 10°C showed a typical increase, a maximum peak, and a decrease until reaching steady state. A 2-phase exponential decay equation described the process, with the obtained key parameters relating to A/W. This model can be used to understand wound-induced respiration and the possible implications in produce quality changes and packaging design.     

Color evolution of aqueous solutions obtained by thermal processing of carrot (Daucus carota L.) roots: influence of light

ABSTRACT:  The color of aqueous solutions obtained by heating carrot ( Daucus carota L.) roots in water ("stocks") is different when the thermal treatment is applied with or without exposure to light. CIE L *, a *, and b * scale values of stocks processed for different times were recorded and 4 patterns were initially observed. To explain the 1st part of this evolution (patterns 1 and 2), pectin extraction and β-elimination in stocks were studied. Light dependence was investigated to explain patterns 3 and 4. A model with 2 compounds is proposed to explain all the color variations.     

Optimization of enzymatic process parameters for increased juice yield from carrot (Daucus carota L.) using response surface methodology

The effects of enzyme concentration (50–650 mg/kg grated carrot), pectolytic and cellulolytic enzyme ratio (3:7–7:3), incubation time (30–150 min) and temperature (25–65 °C) on juice recovery and viscosity from grated carrot were studied. A central composite rotatable design (CCRD) was used in designing the treatment combinations of four variables at five levels. The process involved in treating the blanched grated carrot with mixture of crude pectolytic enzyme from Aspergillus foetidus and crude cellulolytic enzyme from Trichoderma ressi, keeping the samples at the desired time, followed by extraction of juice. Enzyme-treated grated carrot sample showed increased juice recovery as compared to control. A second-order response surface model adequately fitted the data. All the variables affected juice recovery and viscosity significantly. Enzyme concentration, pectolytic and cellulolytic enzyme ratio, incubation time and temperature had total and combined effect at linear, square and interactive level on both responses. The optimum condition was enzyme concentration, 210.7 mg/kg of grated carrot; pectolytic and cellulolytic enzyme ratio, 3.84:6.16; incubation time, 130 min and incubation temperature 47 °C. Under the optimal conditions, juice extracted from enzyme-treated grated carrot was 74.3% having juice viscosity 1.07 cP, corresponding to the increase in yield by 13.95% and decrease in viscosity by 0.45 cP.     

Degradation of carrot (Daucus carota) fibres with cell-wall polysaccharide-degrading enzymes

Carrot Daucus carota L fibres were degraded with two enzyme preparations, SP249 from Aspergillus aculeatus and Celluclast from Trichoderma reesei. The enzymic activities of these complexes indicate that SP249 was particularly active on pectic polymers, and Celluclast could degrade amorphous and crystalline cellulose. A combination of both preparations degraded carrot fibres with a synergistic effect and led to the solubilisation of 95% of the cell-wall polysaccharides. The kinetics of solubilisation of sugars and gel-permeation chromatography of the soluble products show that pectic polymers were rapidly solubilised and then, in a second stage, degraded mainly to monomers, whereas cellulose was more slowly hydrolysed to cellobiose and glucose. Part (67%) of the polysaccharides were saccharified, the residual soluble material being rhamnogalacturonans containing arabinose residues. Residual insoluble fibres (10% of initial weight) were not liquefied and were composed mainly of lignin, proteins and polymers of glucose, xylose and galacturonic acid.     

Low Frequency Ultrasonics for Texture Measurements in Cooked Carrots (Daucus carota L.)

The use of low frequency ultrasound for texture evaluation of carrots cooked at 100°C for 0 to 15 min was examined. Uniaxial compression, microscopy and analyses of density and dry matter were used to provide comparative results for evaluating relationships between texture and ultrasonic parameters. During the first minutes of cooking the sound velocity (v) decreased and a positive correlation was found between compressive Young's modulus (E_cy) and sound velocity (v). During extended heating, sound velocity (v) increased and attenuation (α) decreased as a result of changes in mechanical properties and air and water content of the tissue.     

Activity,localisation and thermal inactivation of deteriorative enzymes in Australian carrot (Daucus carota L) varieties

Four Australian carrot varieties have been selected for a study of deteriorative enzymes. The varieties included those commonly used for table and also for processing purposes (Red Hot Original, Red Count, Top Pak and Red Hot Carotene 100). The activities of three deteriorative enzymes (peroxidase, catechol oxidase and pectinesterase) have been assayed in juices prepared from whole carrot as well as superficial tissues, core, root tip and stem end. The levels and relative distribution vary for the different enzymes and varieties studied. Thermal inactivation was assessed over a range of temperatures. Catechol oxidase was found to be the least stable and pectinesterase the most stable in each of the varieties. In most cases, effective inactivation was achieved within 2 min at 85 °C. The enzymes of Top Pak variety showed greater thermal stability. In this variety, pectinesterase required treatment at 90 °C to ensure rapid inactivation. It is concluded that pectinesterase should be used as the indicator enzyme in the assessment of blanching sufficiency for processing of carrots. © 1999 Society of Chemical Industry     

Effects of limonene on the growth and physiology of cabbage (Brassica oleracea L) and carrot (Daucus carota L) plants

Monoterpenes are plant‐based alternatives for pest control in sustainable agriculture. This study investigated the phytotoxic effects of the monoterpene limonene on white cabbage (cv Rinda and Lennox) and carrot (cv Parano and Splendid) plants. The effects of increasing concentrations (from 10 to 120 ml l^?1) of limonene on visible leaf injuries, growth, chlorophyll fluorescence, net photosynthesis and stomatal conductance were studied. Limonene concentrations of 90 and 120 ml l^?1 were significantly phytotoxic in both cabbage and carrot, depending on the cultivar. However, there were major differences in limonene responses between the cultivars in both species. In carrot, lower shoot and root biomass was related to lower chlorophyll fluorescence (indicating lower efficiency of the photochemical processes) in cv Splendid, whereas in cv Parano it was related to lower stomatal conductance (indicating reduced gas exchange), suggesting that the mechanism behind the phytotoxicity may be different in different cultivars. In cabbage, better limonene tolerance and faster recovery processes in cv Lennox compared with cv Rinda were evident through increasing efficiency of photochemistry (chlorophyll fluorescence), and in the higher stomatal conductance and net photosynthesis rates, providing energy for defence and repair action. Since direct phytotoxic effects were detected on carrot and cabbage with limonene, we conclude that critical threshold concentrations for cultivated plants should be determined before monoterpenes are widely used as repellents and deterrents. These thresholds should, as far as possible, be species‐ and cultivar‐specific in order to provide full protection of plants. Copyright © 2004 Society of Chemical Industry     

微波提取对胡萝卜中的多糖基本结构特征及固体形貌的影响

研究微波提取对胡萝卜中的多糖基本结构特征及固体形貌的影响。采用沸水浴、微波两种方式提取胡萝卜中的多糖(分别记为WEP和MEP),通过基本理化性质测定、高效体积排阻色谱(high performance size exclusion chromatography,HPSEC)和单糖组成分析等方法比较了两者的基本结构特征,并采用扫描电子显微镜对两者的固体形貌进行表征。结果显示,WEP和MEP的中性糖、糖醛酸和总酚含量存在差异,两者均由半乳糖醛酸、半乳糖、葡萄糖、阿拉伯糖、鼠李糖组成,但MEP总糖含量更高并含有更多果胶类单糖;WEP和MEP的HPSEC图有一定相似性,但WEP中含有重均分子质量高达7.77×105 D的组分;WEP表现较为聚集的片状结构,而MEP组分则表现较为分散且主要为小片段状态。上述结果表明,两种不同提取方法所得到的多糖具有显著差异。     

Effect of carrot (Daucus carota) antifreeze proteins on the fermentation capacity of frozen dough

Carrot (Daucus carota) antifreeze proteins have strong anti-recrystallization ability and potential uses in food industry. In this study, the effects of carrot concentrated protein supplementation containing 18.3% carrot antifreeze protein on the fermentation capacity of frozen dough were evaluated. Adding carrot concentrated protein to frozen dough increased the retention capacity of the dough, decreased mortality of the yeast, caused a thermal-hysteresis phenomenon during freezing–thawing cycle, and retarded the crystal forming. The application of carrot concentrated protein in frozen dough was proved to be a promising improver for the frozen dough fermentation.     

Quality attributes of shredded carrot (Daucus carota L. cv. Nantes) as affected by alternative decontamination processes to chlorine

The effects of alternative decontamination processes to chlorine: ozonated-water (1 ppm/5 min), hot water (100 °C/45 s) and ultrasonication (45 kHz/1 min), applied pre- or post-cut in the technological diagram of minimal processing of carrots were tested. Ultrasonication in chlorinated-water and thermo-ultrasonication as combined processes applied just in pre-cut carrot were also tested. The initial microbial load reduction, soluble solids content, pH and sensorial attributes of shredded carrot just after processing were evaluated. Decontamination processes applied on pre-cut carrot provided maintenance of fresh-like sensorial quality, regardless the type of treatment, due to diminished leaching phenomena which is critical for shredded carrot. Chlorination, ozonization and ultrasonication achieved ca. 1 Log₁₀ reduction of initial microbial load. No additional decontamination effect in combined processes was observed. The use of heat in pre-cut carrot proved to be the most efficient process regarding microbial reduction (3 Log₁₀ units) providing, as well, an acceptable fresh-like quality product.Industrial relevanceThe major constraint for marketability of minimally processed shredded carrot is its limited shelf-life due to rapid microbial growth and colour loss (decrease of orange intensity and/or whitening of the shreds). These questions arise from the practical experience of a fresh-cut industry directly involved in the R&D research project which supported this study. Chlorine solutions have been widely used to sanitize fruit and vegetables in the fresh-cut industry. However, reduced microbiological efficiency allied to the eventual formation of carcinogenic chlorinated compounds pointed out the need for alternative methodologies. The present work aimed the evaluation of clean alternative decontamination processes applied both to pre- and post-cut carrot for the production of shredded carrot, operating under conditions of industrial practice at pilot-plant scale.     

On the induction of cold acclimation in carrots (Daucus carota L.) and its influence on storage performance

We investigated the role of cold acclimation in carrot plants with respect to its influence on the storage performance of the harvested taproots. The induction of cold acclimation was followed in plants cultivated in a growth chamber under strict climate control and in taproots harvested from two separate field cultivations where the plants had been exposed to the natural variations in climate. Under controlled growth conditions, levels of antifreeze protein (AFP) mRNA were used as a marker for cold acclimation in carrot taproot tissue. Expression of this gene was induced by cold in discs excised from harvested taproots and this induction was clearly affected by the growth temperature of the plants from which the taproots were taken. These in vitro data were consistent with those from field-grown plants. In the cell wall of taproots harvested in year 2000, where the intact plants had frequently been exposed to temperatures below 6 °C, a 36 kDa AFP accumulated to higher levels during storage than in the taproots harvested from plants grown in year 2001, where cultivation temperatures had rarely dropped below 6 °C. The taproots from 2001 exhibited poor storage performance as shown by an earlier increase in relative electrolyte leakage and decrease in dry matter compared to taproots harvested in 2000. The capacity of the AFP to accumulate during storage was consistent with a high storage performance.     

Effect of Sodium Chloride, Acetic Acid, and Enzymes on Carotene Extraction in Carrots (Daucus carota L.)

ABSTRACT: Carrot root cores were cut off longitudinally and treated with NaCl (0.6 and 1.2 M ) and/or acetic acid (1.33%, 2.67%, and 4%) solutions. The extractability of the carotenes was estimated. Similarly, carrot cores were also treated with some degrading enzymes (carbohydrases, lipases, and proteases) alone or in combination to study the effect of the tissue rupture or the hydrolysis of possible complexes or interactions between carotenes and other components on the carotene extractability. The results showed that acetic acid increased the extractability of α- and β carotenes up to 99.8% and 94.6%, respectively, at a 4% acid concentration compared with the samples without any treatment. This increase was directly proportional to the acid concentration. An increase in extractability was also observed for NaCl, although the increases were not as high as in the previous case with values of 49% and 41.4% for α- and β-carotenes respectively at a 0.6 M concentration. The study of microstructural changes and extractability revealed that the enzymatic treatments could have broken some carotene complexes and interactions and altered the carbohydrate matrix structure, increasing to a certain extent the extractability of carotenes. It can be concluded then that pickling with 4% acetic acid is a good method to increase the extractability of α- and β-carotenes.     

The survival of E. coli O157:H7, S. Typhimurium and L. monocytogenes in black carrot (Daucus carota) juice

In this study the survival and growth patterns of Escherichia coli O157:H7, Salmonella Typhimurium and Listeria monocytogenes in various concentrations of black carrot juice were investigated during incubation period at 4 °C and 37 °C for 7 days. Several parameters, such as juice concentration (%), pH, incubation temperature (°C) and time (days) were found effective on the survival of pathogens tested. Although L. monocytogenes has been found to be the less resistant microorganism to the variable conditions, there were only ca. 1 and 2 log reductions in the number of the cells in the juice samples incubated at 4 °C for 2 and 7 days, respectively. Incubating at low temperature (4 °C) enhanced the survival of test microorganisms.