Changes in texture qualities and pectin characteristics of fermented minced pepper during natural and inoculated fermentation process

Texture qualities and pectin characteristics in fermented minced pepper (FMP) prepared by natural fermentation (NF) and inoculated fermentation (IF) were analysed during fermentation. The results showed variation in texture qualities and pectin characteristics was similar during NF and IF process. The hardness, cell wall material, sodium carbonate-soluble pectin (SSP) and chelate-soluble pectin (CSP) content, and CSP esterification degree decreased, while water-soluble pectin (WSP) content significantly (P < 0.05) increased after fermentation. The rhamnose (Rha) molar ratio in three pectins increased, but arabinose (Ara) and galactose (Gal) molar ratios in most pectins decreased after fermentation. Changes in Ara/Gal and (Gal + Ara)/Rha ratios represented the backbone and branched chains of rhamnogalacturonan-I in three pectins depolymerised during fermentation. The decrease of molecular weight (M_w) in CSP was more obvious than that in WSP and SSP, and it was extensively depolymerised into low-M_w pectin after fermentation. Pearson's correlation analysis showed FMP hardness was extremely (P < 0.01) positively correlated with CSP content and significantly (P < 0.05) positively correlated with SSP content and CSP M_w. Hence, CSP was the main pectin to affect texture compared with WSP and SSP, and its characteristics played a crucial role for regulating FMP texture during NF and IF process.     

Firmness and Cell Wall Characteristics of Pasteurized Jalapeño Pepper Rings Affected by Calcium Chloride and Acetic Acid

Pepper rings packed in brine containing CaCl₂ were firmer, had higher bound calcium, chelator soluble pectin and pectin DE, and less water-soluble pectin (WSP) than peppers packed in brine containing no CaCl₂. Pepper rings packed in low acid brines (1% and 1.2% acetic acid) were firmer and had less WSP than those packed in high acid brine. Those samples (4% acetic acid) resulted in softening and pectin solubilization, but CaCl₂ resulted in less softening. Monosaccharide composition of cell walls was not affected by CaCl₂ or acetic acid. Firmness retention in CaCl₂ treated samples was probably due to greater association between calcium ions and pectic substances, which resisted acid hydrolysis.     

Textural properties of gelling system of low-methoxy pectins produced by demethoxylating reaction of pectin methyl esterase

ABSTRACT:  After deesterification of commercial pectins with a pectin methyl esterase (PME), their gelling properties were characterized using instrumental texture analysis. The final degree of esterification (DE) of the high- and low-methoxy pectins reached approximately 6% after the PME treatment, while deesterification of low-methoxy amidated pectin stopped at 18% DE. Furthermore, DE of high-methoxy pectin was tailored to be 40%, which is equivalent to the DE of commercial low-methoxy pectin. As a result, significant changes in molecular weight (Mw) distribution were observed in the PME-treated pectins. The texture profile analysis showed that PME modification drastically increased hardness, gumminess, and chewiness, while decreasing cohesiveness and adhesiveness of the pectin gels ( P < 0.05). The pectin gel with relatively high peak molecular weight (Mp, 3.5 × 10⁵) and low DE (6), which was produced from high-methoxy pectin, exhibited the greatest hardness, gumminess, chewiness, and resilience. The hardness of low-methoxy amidated pectin increased over 300% after PME deesterification, suggesting that the effects of amide substitution could be reinforced when DE is even lower. The partial least square regression analysis indicated that the Mw and DE of the pectin molecule are the most crucial factors for hardness, chewiness, gumminess, and resilience of gel matrix.     

PECTIC SUBSTANCES AND FIRMNESS OF CUCUMBER PICKLES AS INFLUENCED BY CACL2, NACL AND BRINE STORAGE1

The effects of brine treatments (CaCl₂ and NaCl) and storage on pectic substances and texture of cucumber pickles were examined. Firmness of cucumber pickles was closely associated with the solubility characteristics and degree of esterification (DE) of pectic substances. Brining, storage, time of CaCl₂ addition and concentration of NaCl and CaCl₂ were all observed to influence the characteristics of pectic substances. Important to preventing softening was maintenance of pectic substances not extractable (NXP) by conventional methods, i.e., by water (WSP), the chelator sodium hex-ametaphosphate (CSP), and dilute alkali (OHSP). Erosion of NXP consistently resulted in increased levels of CSP and reduced firmness. Reducing the amount of demethylation of pectins was also associated with maintaining firmness. Although the DE declined rapidly during brining, tissues from treatments that enhanced firmness had pectic substances with the highest DE. CaCl₂ added to brine at the beginning of fermentation was most effective in preventing the demethylation of pectins and solubilization of NXP. In contrast, delayed addition of CaCl₂ and storage in low NaCl concentration (5% or less) caused greater pectin demethylation, erosion of NXP with concomitant increases in CSP and tissue softening.     

Quality attributes and cell wall properties of strawberries (Fragaria annanassa Duch.) under calcium chloride treatment

Effects of CaCl₂ (0%, 1% and 4%) treatment on quality attributes and cell wall pectins of strawberry fruits stored at 4 °C for 15 d were investigated. Strawberry firmness was not significantly affected by CaCl₂ treatment. Compared to the other groups, the 1% CaCl₂ group had better quality attributes, including decay rate, weight loss and soluble solids content. The treatment with 4% CaCl₂ inhibited weight loss but caused phytotoxicity. During storage, the chain widths and lengths of water-soluble pectin (WSP), chelate-soluble pectin (CSP) and sodium carbonate-soluble pectin (SSP) decreased. Strawberry softening seemed to be due to modifications of CSP and SSP, especially the side chains. CaCl₂ treatment significantly slowed the breakdown of CSP and SSP chains by strengthening the ionic crosslinkages among these pectin molecules. These results illustrate the fundamental CaCl₂ effects and will help improve the application of CaCl₂ to postharvest fruits.     

Freezing of strawberries by immersion in CaCl2 solutions

The present investigation studied the freezing of strawberries by immersion in CaCl₂ solutions, analysing drip loss, pectin content and the degree of esterification of the pectins, total and cell-wall bonded calcium contents, the ratio bonded calcium/total calcium, and textural parameters. In addition, the effect of immersion in pectin methylesterase (PME) solutions prior to immersion freezing (IF) was analysed. The firmness of thawed fruit decreased by approximately 74% with respect to fresh strawberries, and neither IF-CA (freezing by immersion in CaCl₂ solution) nor PME–IF-CA (immersion in PME solution + freezing by immersion in CaCl₂ solution) provided significant benefit in maintaining firmness when compared to slow freezing (SF). However, IF provided a significant benefit in reducing drip loss of thawed strawberries when compared to SF, but pre-treatment with PME did not provide any additional benefit.     

Extraction, thermal stability and kinetic behavior of pectinmethylesterase from hawthorn (Crataegus pubescens) fruit

Pectinmethylesterase (PME) extracted from hawthorn (Crataegus pubescens) fruit was evaluated for its thermal stability and kinetic behavior. The enzyme extraction process was established after studying different NaCl concentrations (0.5-3.0 moles/L). A maximum PME extraction of 51.61 units/mg protein was obtained using 2 moles/L NaCl. Kinetic parameters (K_m and V_max) were determined using a commercial citrus pectin and C. pubescens pectin as substrates. The effects of NaCl concentration, pH and temperature on PME activity were investigated. PME showed higher affinity for C. pubescens pectin (K_m and V_max of 2.84 mg/mL protein, and 64.10 units/mg protein, respectively) than for citrus pectin. C. pubescens PME extract showed maximum activity at 0.4 moles/L NaCl, pH 7.5, and 55 °C. The E_a and Q₁₀ for thermal activation were 36.27 kJ/mol and 2.01 (20-30 °C), respectively. About 50% of the activity still remained after heating for 25 min at 60 °C, and it was completely inactivated by incubation at 80 °C for 10 min. The Q₁₀ and E_a values for thermal inactivation reaction were 20.06 (70-80 °C) and 146.16 kJ/mol, respectively. These results provide useful information about the factors that affect the activity of C. pubescens PME, and might be used as a starting point for texture control during post-harvest handling and processing of this fruit.     

Thermal and Calcium Pretreatment Affects Texture, Pectinesterase and Pectic Substances of Frozen Sweet Cherries

Freezing causes loss of turgidity and firmness in sweet cherries. Thermal pretreatment at 50°C for 10 min followed by immersion in 100 mM CaCl₂ and thermal pretreatment at 70°C/2 min with or without immersion in 100 mM CaCl₂ prevented freezing-induced loss of firmness. Thermal pretreatments increased the pectin fraction soluble in EDTA, reduced the degree of pectin esterification, and increased both the concentration of divalent cations in the cell wall and the pectinesterase activity bounded to the cell wall. Immersion in CaCl₂ increased the concentration of Ca²⁺ cations in the cell wall and enhanced the effect of thermal pretreatments on pectinesterase activity.     

Pectin methyl esterase treatment on high‐methoxy pectin for making fruit jam with reduced sugar content

BACKGROUND: Pectin methyl esterase (PME) has been postulated to catalyse the transacylation reaction between pectin molecules. The present study aimed to prove the occurrence of this reaction. The feasibility of applying PME‐catalysed transacylation between high‐methoxy pectin molecules in making fruit jam with reduced sugar content was also investigated. RESULTS: PME treatment increased the turbidity and particle size in pectin solution and the molecular weight of pectin, while it decreased the number of methoxy ester linkages and the intensity of the CH₃ absorption peak in the Fourier transform infrared spectrum without changes in the number of total ester linkages in pectin molecules. These findings support the occurrence of PME‐catalysed transacylation between pectin molecules. Higher values of hardness, gumminess and chewiness were found in a jam containing PME‐treated citrus pectin (10 g L^?1) and sugar (350 g L^?1) as compared with either a jam containing untreated citrus pectin (10 g L^?1) and sugar (350 g L^?1) or strawberry jam containing pectin (10 g L^?1) from the fruit and sugar (650 g L^?1). CONCLUSION: The demand for sugar in jam making can be greatly reduced by the use of PME‐treated high‐methoxy pectin. Copyright © 2012 Society of Chemical Industry     

Constitutive expression,purification and characterisation of pectin methylesterase from Aspergillus niger in Pichia pastoris for potential application in the fruit juice industry

BACKGROUND: Pectin methylesterase (PME) catalyses the hydrolysis of the methyl ester of pectin, yielding free carboxyl groups and methanol. PME is widely used in the food, cosmetic and pharmaceutical industries. RESULTS: PME from Aspergillus niger was constitutively expressed to a high level in the yeast Pichia pastoris. The recombinant PME was purified by a combination of ammonium sulfate fractionation and ion exchange chromatography, giving an overall yield of 28.0%. It appeared as a single band in sodium dodecyl sulfate polyacrylamide gel electrophoresis, with a molecular mass of about 45 kDa. Optimal activity of the enzyme occurred at a temperature of 50 °C and a pH of 4.7. The K_m, V_max and k_cat values of the enzyme with respect to pectin were 8.6 mmol L^?1 [ ], 1.376 mmol min^?1 mg^?1 and 8.26 × 10² s^?1 respectively. Cations such as K⁺, Mg²⁺, Ni²⁺, Mn²⁺ and Co²⁺ slightly inhibited its activity, whereas Na⁺ had no effect. CONCLUSION: PME from A. niger was constitutively expressed to a high level in P. pastoris without methanol induction. The recombinant PME was purified and characterised and shown to be a good candidate for potential application in the fruit juice industry. Copyright © 2012 Society of Chemical Industry     

Physicochemical and molecular analysis of cell wall metabolism between two navel oranges (Citrus sinensis) with different mastication traits

BACKGROUND: FJ72‐1 navel orange and its bud mutant FJWC exhibit differences in melting texture character which are influenced mostly by cell wall metabolism. Here we compared the contents of water soluble pectin (WSP), protopectin, total pectin (TP), cellulose, and hemicellulose, activities of polygalaturonase (PG), pectin methylesterase (PME), pectate lyases (PL), cellulase (Cel) and gene expression levels of PG, PME, PL and Cel between the two cultivars. RESULTS: The content of cellulose and hemicellulose decreased progressively during fruit ripening. At the harvest time (230 DAF), the content of cellulose and hemicellulose in FJWC were obviously higher than those in FJ72‐1; the WSP content, PG activities and its gene expression level in FJWC was lower than those in FJ72‐1. Moreover, gene expression levels of PME and Cel in FJWC were only one‐quarter of those in FJ72‐1 at 230 DAF. CONCLUSION: The present work showed that the inferior melting character of FJWC attributed to the lower WSP, higher TP or protopectins, higher cellulose and hemicellulose than those in the pulp of FJ72‐1 at harvest time. Lower expression levels of PG, PME and Cel at harvest time might be associated with the inferior melting character. Copyright © 2010 Society of Chemical Industry     

Valencia PME isozymes create charge modified pectins with distinct calcium sensitivity and rheological properties

Pectin was de-esterified by U-PME or B-PME, pectinesterases that differed in cationic character and peptide composition. Pectin fractions of unmodified (O-Pec) and Valencia PME modified pectins (B-Pec and U-Pec) were separated by ion exchange (IEX) chromatography and characterized for calcium sensitivity by ζ-potential, texture profile analysis (TPA), and viscoelastic properties. U- and B-Pec had more negative ζ-potential than O-Pec, and became increasingly negative at higher pH values. Without CaCl_2, viscosity of O-Pec was higher than B- and U-Pec. With 35 mM CaCl₂, B- and U-Pec gelled, but O-Pec did not gel. TPA values were not significantly different between B-Pec and U-Pec. The G' values of O-Pec was 20–50-fold lower than G' for B-Pec or U-Pec. After separation with ion exchange chromatography, later eluting fractions formed stronger gels than earlier eluting fractions. However, pectin fractions had lower viscosity and G' values than unfractionated pectin, which is likely related to molecular weight differences. Results show that U-Pec forms stronger gels than B-Pec that is related to length of charge block and numbers of de-esterified pectin molecules.     

Comparison of cell wall metabolism in the pulp of three cultivars of ‘Nanfeng’ tangerine differing in mastication trait

BACKGROUND: Like sweet orange (Citrus sinensis), tangerine (Citrus reticulata) is another citrus crop grown widely throughout the world. However, whether it shares a common mechanism with sweet orange in forming a given mastication trait is still unclear. In this study, three ‘Nanfeng’ tangerine cultivars, ‘Yangxiao‐26’ (‘YX‐26’) with inferior mastication trait, elite ‘YX‐26’ with moderate mastication trait and ‘Miguang’ (‘MG’) with superior mastication trait, were selected to investigate the formation mechanism of mastication trait. RESULTS: ‘MG’ had the lowest contents of total pectin, protopectin and lignin and the highest gene expression levels of citrus polygalacturonase (PG) and pectin methylesterase (PME) at the end of fruit ripening, whereas ‘YX‐26’ had the lowest water‐soluble pectin (WSP) content, the highest lignin content and the lowest PG and PME expression levels. The contents of cellulose and hemicellulose were similar among the three tangerines. CONCLUSION: The fruit mastication trait of C. reticulata was determined by the proportions of WSP and protopectin as well as lignin content, not by cellulose and hemicellulose contents. Pectin content could be a major contribution to the feeling of mastication trait, while PG and PME exhibited an important role in forming a given mastication trait according to the present results as well as previous results for C. sinensis. Copyright © 2011 Society of Chemical Industry     

ACEROLA's CLONES OF INDUSTRIAL INTEREST

In order to know which clone of acerola is better for acerola industrialization, we studied the pectin methylesterase (PME) specific activity, pectin content and vitamin C content in five different clones of acerola. The pectin yield varied from 1.37 to 2.99% and the highest content of pectin occurred in clones 3 and 5. Ascorbic acid varied significantly from 1157.5 to 1735.5 mg/100 g of pulp in the five clones. The highest content of vitamin C occurred in clone 4. The PME specific activity varied from 0.79 to 2.92 units g ^?1 /g of pulp and the highest values occurred in clone 2. We also studied the optimum temperature and the optimum pH of this enzyme. Clones 1, 2, 4 and 5 showed optimum temperature at 90C. Clone 3 showed practically the same specific activity at all temperatures studied. Clones 1 and 4 showed an optimum pH of 9.0 and clone numbers 2, 3 and 5 showed a pH optimum at 8.5.     

雪里蕻腌菜的质构与果胶组分含量的关系

对雪里蕻腌菜中果胶组分的含量与质构变化进行了检测分析,雪里蕻腌菜在正常腌渍过程中总果胶含量趋于稳定,而水溶性果胶增加至一稳定值,六偏磷酸钠溶解性果胶下降至一稳定值.在腐烂过程中,总果胶含量显著下降,同时水溶性果胶组分增加和六偏磷酸钠溶解性果胶下降明显.研究结果表明,可用检测雪里蕻腌菜醇不溶物中果胶组分含量的变化来监测腌菜质构变化.     

促进植物内源性果胶甲酯酶催化作用的因素与机制及其在果蔬加工中的应用

植物内源性果胶甲酯酶（pectin methylesterase,PME）存在于天然植物组织中,可以催化高酯果胶脱甲基酯化生成低酯果胶,进而影响果蔬硬度、出汁率等质构及加工品质。一方面,为了避免贮藏期间出现果蔬汁浑浊沉淀、果蔬罐头中果肉变软、调味酱（如辣椒酱）中皮肉分离等不良品质,会在加工过程中抑制PME活性;另一方面,为了提高果蔬汁出汁率、生产低酯果胶等,也可在食品中添加外源PME。目前,对于通过激活内源PME来改善食品加工的研究较少,因此,本综述重点讨论促进植物内源性PME催化作用的因素与机制及其在果蔬贮藏加工中的应用,为植物内源性PME在果蔬加工中的充分利用提供理论参考。     

Improved Firmness in Calcified Diced Tomatoes by Temperature Activation of Pectin Methylesterase

ABSTRACT: The effects of temperature and calcium on pectin methylesterase (PME) activity and texture in tomato pericarp material were examined. Heating thin slices of pericarp to temperatures between 50°C and 75°C led to the rapid evolution of methanol from the material, indicating an activation of PME. This activity was further stimulated when CaCl₂ (up to 2.0% w/v) was added. When applied to half-inch diced tomato pericarp, the same conditions that led to the activation of PME also improved firmness. Diced tomatoes treated for 5 min with 0.5% CaCl₂ at 70°C were 2.5 times firmer than diced tomatoes treated with CaCl₂ at room temperature. This improvement in texture by treating with CaCl₂ at elevated temperatures was only apparent when the tomatoes received a subsequent 100°C treatment. Heating tomatoes to 70°C either before or after the CaCl₂ treatment also improved firmness through a subsequent high-temperature treatment, but to a lesser extent than heating during the CaCl₂ treatment. These results are consistent with the model that heating to 70°C greatly increases PME activity, leading to extensive pectin de-esterification and increased calcium cross-linking of the pectins in the middle lamella. Production of thermally processed diced tomatoes with improved firmness should be possible by increasing the temperature during and after calcium treatment.     

Comparison of enzymatic de-esterification of strawberry and apple pectin at elevated pressure by fungal pectinmethylesterase

Water soluble pectin was isolated from strawberries. Sugar composition, degree of esterification and molar mass were compared with commercial apple pectin. Both pectins were subjected to enzymatic de-esterification by recombinant Aspergillus aculeatus pectinmethylesterase (PME). Rate of enzymatic de-esterification at elevated pressures (0.1–500 MPa) and temperatures (20–60 °C) was assayed by measuring the release of methanol as a function of time. Optimal activity was observed at 200–300 MPa combined with 45–55 °C. At all conditions investigated, both pectins were de-esterified at similar initial rates. However, after prolonged enzymatic treatment at atmospheric pressure and 30 °C, apple pectin was de-esterified to a significantly lower degree of esterification (7%) than strawberry pectin (32%). The mode of action of A. aculeatus PME was investigated by enzymatic fingerprinting of de-esterified pectin chains. The enzyme de-esterified according to a “multiple chain, multiple attack” mechanism, irrespective of the substrate.Industrial relevanceThis article demonstrates that both strawberry and apple pectin de-esterification by recombinant Aspergillus aculeatus PME is accelerated by high hydrostatic pressure. Since de-esterification of pectin in fruits gives rise to a texture improvement, this enzyme can be used to minimize texture damage during high-pressure processing of fruits and fruit-based products. It was also shown that this enzyme de-esterifies strawberry and apple pectin according to a “multiple chain, multiple attack” mechanism. The resulting pattern of esterification might have an influence on textural properties of fruits.     

Enzyme infusion and thermal processing of strawberries: Pectin conversions related to firmness evolution

Strawberries were infused with fungal pectinmethylesterase (PME) and/or calcium chloride with the aim of minimising tissue damage during subsequent thermal processing (95 °C). Firmness measurements and micrographs provided information on the extent of tissue damage. These observations were linked to the chemical structure of pectin. When PME was infused in absence of Ca²⁺, the degree of methoxylation of pectin was lowered, but chains remained water soluble, indicating that they were not crosslinked. Thermal processing of PME-infused strawberries resulted in pectin solubilisation and depolymerisation which was reflected in pronounced firmness decrease and tissue damage, comparable to non-infused processed strawberries. On the other hand, when a combination of both PME and Ca²⁺ was infused, an important decrease in processing-related tissue damage was perceived. This can be explained by increased crosslinking of pectin chains with low degree of methoxylation, rendering them insoluble and less susceptible to thermal depolymerisation.     

Effect of Phenolic Acid on Antioxidant Activity of Wine and Inhibition of Pectin Methyl Esterase

Antioxidant activity, malolactic fermentation and sensory evaluation of the grape must after fermentation in the presence of gallic acid and coumaric acid, as well as the inhibitory mechanism of gallic acid and coumaric acid on pectin methyl esterase (PME), were investigated. The content of malic acid and lactic acid increased 40.4% and 36.9% compared to the control when commercial pectic enzyme (CPE) was used. The increase in malic acid content was enhanced by 64.8% and 83.4%, compared to the control in the presence of CPE + Gallic acid and CPE + Coumaric acid respectively. Ferric reducing/antioxidant power (FRAP) increased in the samples with added CPE. In addition to an increase in the FRAP, antioxidant capacity was enhanced in the CPE + Gallic acid and CPE + Coumaric acid samples. No significant differences were found in the content of total anthocyanin and in the value of sensory characteristics. The content of total flavanols increased significantly in the samples with added CPE. Lineweaver‐Burk plots of PME, with gallic acid or coumaric acid, indicated that gallic acid and coumaric acid were mixed inhibitors of PME.