• Korean Journal of Food Science and Technology
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• v.27 no.5
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• pp.658-665
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• 1995

Low yield of a thermostable pectinesterase(TSPE) from citrus fruits has made its detailed study extremely tedious and difficult; therefore, maximizing TSPE extraction is desirable. It is assumed that TSPE is bound to the cell components via ionic linkage and covalent bonds. Therefore, in this study, variations in extraction time, pH, NaCl concentration and commercial enzyme preparations were used to increase the yield of TSPE from Valencia orange. The largest recovery of TSPE, obtained by heating extracted pectinesterase(PE) at $70^{\circ}C\;for\;5{\sim}10$ minute, was achieved using actate buffer(pH 4.14) with 1 M NaCl and 0.2% $Cytolase^{TM}$ 104(a mixture of cellulase, hemicellulase and pectinase; Genecor, Inc). The two aquous phase partitioning with 5.0% Triton X-114 could be used as a tool for separation of thermolabile pectinesterase(TLPE) and TSPE from crude PE. Also, water extraction and $0{\sim}0.3$ ammonium sulfate fractionation could be used for removing non-pectinesterase protein.

• Korean Journal of Food Science and Technology
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• v.27 no.5
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• pp.673-679
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• 1995

Pectinesterase(PE) has been definitively established as the causative agent for clarification of citrus juice and gelation of frozen concentrates. This enzyme is present in multiple forms in citrus fruit. Although representing a minor fraction of the total PE activity, thermostable pectinesterase(TSPE) accounts for the severe time/temperature processing treatment required to inactivate PE. This study was undertaken to separate and purify thermolabile pectinesterase(TLPE) and TSPE from the crude PE extracts of Valencia orange rag-pulp powder. The approach taken was to carry out the three kind of chromatographies of CM-Sephadex C-50, Phenyl Sepharose CL-4B and CM-Biogel A to the unheated crude PE or the heated crude PE. All of them used could increase the purity of PEs and, specially, Phenyl Sepharose CL-4B chromatography could separate crude PE as the mixture of PEs into two forms of TLPE and TSPE. The purified TLPE had specific activity of 1,005 units/mg, yield of 13.6% and purification of 35 fold, while the TSPE separated from the unheated crude PE showed specific activity of 3,115 units/mg, yield of 1.5% and purification of 100.5 fold, and another TSPE from the heated crude PE was found to be specific activity of 1,803 units/mg, yield of 15.4% and purification of 140 fold.

• Korean journal of food and cookery science
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• v.11 no.5
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• pp.542-547
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• 1995

Pectinesterase was isolated from the culture medium of Aspergillus japonicus and partially purified by DEAE-Sephadex batchwise, Sephadex G-75 gel filtration and ion exchange chromatography. The purified enzyme solution was completely free from polygalacturonase which depolymerizes pectin molecule. The ability of the pectinesterase to demethylate high methoxyl pectin was investigated. On 20 minute of incubation methoxyl content of low methoxyl pectin decreased from 88％ to 6.93%. In general gel prepared with the pectin containing lower methoxyl content showed the lower value of percent sag, and showed the hieher Bel strength. Textural characteristics of pectin gel determined by Rheometer showed that as the methoxyl content was lowered, hardness and resilience of the gel were increased and cohesiveness was decreased. Apple juice containing HMP and organic acids can be converted into low methoxyl pectin apple jelly by the action of pectinesterase and addition of calcium ion. The strength of low methoxyl pectin apple jelly increased when it stored at room temperature.

• Korean Journal of Food Science and Technology
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• v.32 no.5
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• pp.1079-1086
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• 2000

Pectinesterase inhibitor(PEI) of ripened kiwi fruit(Actinidia chinensis) was separated with affinity chromatography using CNBr-activated Sepharose 4B being covalently bound by orange pectinesterse(PE). The affinity resin strongly and selectively bound PEI, which could be eluted in high yield as a single peak by pH 9.5 without loss of inhibitory activity. The separated PEI had maintained almost inhibitory activity at $-25^{\circ}C$ and $5^{\circ}C$ during 30 days but lost it at room temperature in 4 weeks. The PEI possessed a molecular weight of 16.6 KDa, as estimated by 12.5% SDS-PAGE. PEI had optimum pH of 7.5, optimum temperature of below $10^{\circ}C$ and stability up to $70^{\circ}C$. Also, optimum inhibitory activity for PEI was obtained in 0.2 M NaCl of substrate solutions. The kind of inhibition on tomato pectinesterase was found to be noncompetitive, using citrus pectin as substrate. Fresh orange juice added with crude PEI extracts maintained almost the same cloud stability as pasteurized juice. In case of apple juice, the addition of crude PEI extracts to apple juice had decrease of L-ascorbic acid with nearly no effect on cloud loss.

• Korean Journal of Food Science and Technology
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• v.31 no.1
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• pp.263-266
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• 1999

Pectic substances are important to sustain the textural properties of kimchi during fermentation and distribution. Therefore proper control of pectin degrading enzyme activity is critical on quality control in kimchi industry. Pectin degrading enzymes of kimchi ingredients were assayed to improve the product quality. Among pectin degrading enzymes, polygalacturonase and pectinesterase were selected. The specific activity of polygalacturonase was the highest in salted and fermented anchovy, followed by chinese radish. Considering the amount of protein contents, salted and fermented anchovy and dried red pepper showed higher polygalacturonase activity than other ingredients. In terms of specific activity, chinese radish showed the highest pectinesterase activity, followed by salted and fermented anchovy. However, the total activity of salted and fermented anchovy was the highest. Chinese radish showed higher pectinesterase activity than any other ingredients.

• Korean Journal of Food Science and Technology
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• v.28 no.2
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• pp.393-395
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• 1996

This study was conducted to investigate the effect of infusion of pectinesterase (PE) along with preheating treatment in $CaCI_2$ solution on softening Kimchi tissue. Preheating treatment showed preventive effect on softening of Kimchi tissue. Especially, the preheating in the presence of P was more effective in preserving firmness of both stem and leaf. As for of effect of PE infusion by vacuum treatment, a maximum crispness was obtained from the vacuum-treated stems in 0.05 M $CaCI_2$ solution containing PE. The crispness of all the samples decreased during fermentation and the PE-infused samples by vacuum treatment showed higher crispness than the control until the fourth day of fermentation.

• Korean Journal of Food Science and Technology
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• v.27 no.5
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• pp.666-672
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• 1995

This study was carried out to characterize thermolabile pectinesterase (TLPE) and thermostable pectinesterase (TSPE) separated from crude PE of Valencia orange in order to investigate the preventive measures of cloudy juice clarification. The TLPE was observed to be mixture of several isoenzymes with the same molecular weight of 36 KD (37.5 KD) but different isoelectric point of pH 8.4, 8.7, 8.9, 9.8 and ${\geq}10$ which were unstable at $70^{\circ}C$, and the TSPE also was found to be mixture of two or three isoenzymes with the same molecular weight of 53 KD (50 KD) but different isoelectric point of pH 8.7, 9.2 and ${\geq}10$ which had slightly different stability from one another at $70^{\circ}C$. The TLPE and the TSPE had the optimum reaction pH of 7.0 and $7.0{\sim}8.5,\;appK_{M}$ of 1.1 and 1.7 mg/ml, appVmax of 0.53 and $1.01\;{\mu}mol/min/{\mu}g$, and the turnover number of 19.000 and 54,000 mol/mol/min toward Kodak pectin, respectively. The TSPE had higher storage stabiblity and cloud loss effect on orange juice than the TLPE. Above all, the crude PE was most effective on orange juice cloud loss among the PEs used.

• The Korean Journal of Food And Nutrition
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• v.33 no.1
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• pp.1-8
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• 2020

The purpose of this study was to prevent excessive softening of seasoned spinach by heating and freezing through the pre-heating treatment in calcium chloride solution. The pre-heating temperature was set at 60℃, which supports the highest activity of pectinesterase around 60℃. Pre-heating spinach in calcium solution does not affect the chromaticity characteristics of spinach leaves and significantly improves its texture compared to conventional blanching only and pre-heating in water. The improvement of the firmness is related to the formation of strongly cross-linkages between carboxyl groups and Ca²⁺ by the action of the pectinesterase in spinach. For reheated spinach, the firmness of calcium chloride treated stems were 37.80~44.44 kg, and the control was 28.73 kg. The firmness of calcium chloride treated leaves was 19.73~40.79 kg, and the control was 9.63 kg. Additionally, the total aerobic bacteria in the fresh samples were 3.25~3.99 log CFU/g and 1.97~2.72 log CFU/g in the reheated samples. And the total coliform was not detected in the reheated samples. Considering the color, texture, microbial characteristics and taste quality, the optimum pre-treatment conditions to prevent excessive softening of spinach were 3% calcium chloride solution and 15minutes of treatment.

• The Korean Journal of Food And Nutrition
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• v.9 no.1
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• pp.52-58
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• 1996

This study was carried out 19 investigate the characteristics of pectinesterase (PE), polygalacturonase (PG), lipoxygenase(LOX) and peroxidase (POD) in hot pepper to know the effect of hot pepper on food quality during food processing and storage. The results were as follows : 1. The optimum pH of PE was pH 7.5 and the activity of PE below pH 5.5 was revealed scarcely, The concentration of NaCl and $CaCl_2$ that showed the highest activity of PE were 0.2M and 0.05M, respectively. 2. The optimum pH of PG was pH 6.0 and the activity of PG in acidity was higher than that in alkalinity. The activity of PG was maximum at 0.3M NaCl and 0.2mM $CaCl_2$. Above the concentration of 0.5M NaCl and 0.5M $CaCl_2$, the activity of PG was lower than that of PG not adding these salts 3. The optimum pH of LOX was pH 7.0 and pH 8.5. 4. The optimum pH of POD was pH 6.0 and the activity of POD was higher in weak acidity and neutrality than in alkalinity. POD activity was slightly decreased by the increase of NaCl and $CaCl_2$ concentration.

• Korean Journal of Food Science and Technology
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• v.29 no.5
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• pp.974-981
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• 1997

A combined treatment of high hydrostatic pressure and carbonation was used to inactivate pectinesterase (PE) and sterilize microorganisms in Valencia orange juice without major changes in its nutritive components. Quality characteristics of Valencia orange juice, such as microorganisms, PE activity, vitamin C content and color, were evaluated after it was treated with pressure, carbonation-and-pressure, and heat. Quality changes during storage at $4^{\circ}C$ and $30^{\circ}C$ after the treatments were also investigated. Pressurized orange juice (pressurized at 600 MPa for 10 min at $20^{\circ}C$) showed 7.0% residual PE activity, while the carbonated-and-pressurized orange juice (207 $kPa-CO_2$ gas pressure, pressurized at 600 MPa for 10 min at $20^{\circ}C$) showed 0%. Pressurization at 400 MPa or higher decreased the population of microorganisms in the orange juice to less than 10 CFU/mL. Carbonated-and-pressurized orange juice showed slight decrease in vitamin C content when stored at both $4^{\circ}C\;and\;30^{\circ}C$. While heat-treated ($90^{\circ}C$ for 60 sec) orange juice showed 75% decrease in vitamin C content when stored at $30^{\circ}C$. L value (lightness) and b value (yellowness) of carbonated-and-pressurized orange juice were higher than those of heat-treated orange juice when they were stored at $4^{\circ}C$ for 30 days.