• Korean Journal of Microbiology
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• v.33 no.2
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• pp.131-135
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• 1997

Several microorganisms capable of producing protopectinase, which catalyzes dissociation of plant tissue to single cells, were isolated from soils. Crude enzymes prepared from culture supernatants of the strains released potato cells as a single cell from potato tissues. One of the isolated strains showing higher activity of protopectinase was selected and identified as Rhizopus sp. from the morphological characteristics. For preparing single cells from potato tissues, optimum enzyme activity of protopectinase, which was prepared from culture filtrate of Rhizopus sp. R2, was obescrved at $40^{\circ}C$ and pH 4.

• Journal of the Korean Society of Food Science and Nutrition
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• v.26 no.3
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• pp.430-435
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• 1997

Protopectinases are heterologous group of enzymes that degrades insoluble protopectin which consists of middle lamella between cells of plant tissues. Tissues of potato and carrot could be separated to single cells by addition of protopectinase isolated from Rhizopus sp. Color changes ofthe suspensions treated with protopectinase and mechanically macerated after 2 weeks at 4$^{\circ}C$, were investigated. Color change of the latter was very serious, however, that of the former was insignificant. Furthermore, after heat treatment at 121$^{\circ}C$ for 5min, the constituents of mechanically macerted carrot suspension were separated into two layers, but those of single celled carrot were not. Yields of carrot juices extracted from single celled suspensions and mechanical maceration were 93.6% and 56.0%, respectively. These results support that treatment of protopectinase can increase yield of juices extracted from plant, and manufacture high value-added products in food processing.

• Food Science and Biotechnology
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• v.18 no.2
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• pp.442-448
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• 2009

In this study Bacillus subtilis PTCC 1023 was used for the production of protopectinase using soybean based media. The use of isolated soybean protein (ISP) and soybean flour resulted in similar protopectinase production and growth rates. The effect of medium composition on protopectinase production was studied using central composite design (CCD) methodology. The change in the concentration of ISP (1-7%), glucose (0-10%), and phosphate (0.1-0.3 M) was found to affect the protopectinase activity (response variable) after 24 hr of cultivation. In the range studied, ISP and glucose had a negative effect on the response variable, whereas phosphate had a positive effect. A statistically significant interaction was identified between phosphate and ISP, suggesting that correct optimization of medium formulation in this case can only be obtained using factorial design of experiments. Protopectinase activity exceeding 215 U/mL was obtained in a medium containing 4% ISP, 0.3M phosphate, and no added sugar.

• Culinary science and hospitality research
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• v.18 no.2
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• pp.162-171
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• 2012

This study was conducted to investigate the changes of macerating properties from grape suspensions for which both were treated with protopectinase(PPase) and mechanical maceration stored at $4^{\circ}C$ for 28 days. Changes of macerating properties such as pH, total acidity, color, total polyphenol and antioxidant activity of suspensions after enzymatic hydrolysis were determined before and after heating, and microscopic observation made on suspensions containing single cells. With the PPase, grapes were enzymatically macerated to separate cells to primary cell wall without damage. Also, the changes of macerating properties in grape suspension treated with PPase and after heat treatment at $80^{\circ}C$ for 30 min were more stable than those of mechanical maceration, indicating the thermal stability. Thus, the PPase treatment for grapes could be a better choice for preparing highly valuable and functional processed food as well as for prolonging preservation periods.

• Korean Journal of Food Science and Technology
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• v.39 no.3
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• pp.276-282
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• 2007

In this study, attempts were made to develop a garlic juice extraction method that would result in minimum changes in quality. Protopectinase and a mutienzyme containing cellulase, pectinesterase, ${\beta}-glucanase$, etc. were applied to garlic residue after first extraction, and the yields of garlic juice and the flavor component changes of the juices were investigated. Enzyme concentrations of 0.04, 0.08, and 0.12% which were based on pulp weight before extraction were added and allowed to hydrolyze for 30, 60, 90 and 120 min,. respectively. Increase in the garlic juice yield was observed according to the amount of enzyme added and the reaction time until reaching a maximum point. When 0.12% protopectinase was applied to the garlic residue for 90 min, the yield increased by 13.8%. Under the same conditions, when multienzyme was applied to the garlic residue, the yield increased by 14.5%, which was considered the maximum. The flavor compounds decreased when compared with the total GC peak area of garlic juice prepared without enzymes(control). The volatile flavor compounds in garlic juice prepared with 0.12% protopectinase for 60 min decreased by 6%. The free sugars profile of the garlic juice prepared with 0.12% protopectinase for 60 min was similar to that of the control. The type of enzyme used did not affect the free amino acid profile of the garlic juice. These results indicate that the optimum conditions for extraction of garlic juice are hydrolyzing the residue with 0.12% protopectinase for 60 min, extracting garlic juice from the hydrolyzed reside, and then combining the extracted juice with the first extraction.

• Microbiology and Biotechnology Letters
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• v.10 no.1
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• pp.45-52
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• 1982

A fungus with the highest protopectinase productivity was selected among 205 strains isolated from the soil and identified as a Verticillium sp. The Verticillium sp. was cultivated on wheat bran and the crude extruct of its culture medium showed the highest protopectinase activity on the following conditions: 3 days of cultivation time, 27$^{\circ}C$ of cultivation temperature, 1.2 $m\ell$/g wheat bran of water content, and reinforcement of ammonium nitrate and calcium chloride at the concentration of 0.5 and 0.02%, respectively. The optimum conditions for pectin production from Citrus peel pulp by the protopectinase were consequently obtained as follows: 20$m\ell$/g of liquid volume-to-pulp weight ratio, 4$0^{\circ}C$ of reaction temperature, and 4 of reaction pH. The higher the enzyme concentration, the better the yield of pectin and the shorter the reaction time. Total 45.6mg of pectin/g peel was produced by 1 hour reaction at the enzyme concentration of 10.5 units/$m\ell$. Molecular weight of the pectin produced by the enzyme was estimated to be about 62,000 by Smit and Bryant's method.

• Journal of the Korean Chemical Society
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• v.3 no.1
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• pp.28-30
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• 1954

It was pointed out by Katagiri and Nakahama that the useful retling bacteria acted selectively on vegetable fiber materials and also they proposed that this selectivity was based on the specificity of protopectinase of the bacteria, and that the two characters might have certain parallel relation with each other. The result of the further experiments carried out by the author with mulberry-tree bark also confirmed the selectivity of some kinds of retling bacteria, but found no remarkably different action on protopectin from mulberry-tree bark. Accordingly no parallel relation between the selectivity of the retling bacteria and the specificity of the protopectinase was identified.

• Applied Biological Chemistry
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• v.42 no.1
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• pp.1-5
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• 1999

For effective utilization of apple pomace, protopectinase(PPase) produced from Bacillus subtilis IFO 12113 was used for pectin extraction from apple pomace. Optimal conditions for enzyme treatment were found at $60^{\circ}$, pH 7.8, 48 hours with 20 : 1 ratio of substrate to enzyme. The yield of extracted pectin from water-alcohol insoluble pectin by enzyme at optimal condition was 34.3%. The purity and methoxyl content of extracted pectin by PPase at optimal condition were measured as 52.9% and 2.75%, respectively. Intrinsic viscosity and average molecular weight of extracted pectin by enzymatic method were 0.178 ml/g and $4.9{\times}10^3$, respectively.

• Applied Biological Chemistry
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• v.40 no.2
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• pp.107-111
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• 1997

To recover protopectinase (PPase) secreted from Bacillus subtilis IFO 12113, culture filtrate of the microorganism was treated with acetone, methanol, and ethanol, respectively. In the case of treatment with acetone at a ratio of 1: 1 (culture filtrate: acetone, v/v), PPase was purified 1.7-fold with 59.2% recovery The recovery of PPase was increased by increasing the acetone concentration. PPase was purified 4-fold with 100% recovery when the culture filtrate was precipitated with methanol at a ratio of 1 : 2 (culture filtrate: methanol, v/v). However, recovery of PPase was decreased by increasing the methanol concentration. PPase was purified 13.5-fold resulting in 68% recovery by the addition of ethanol with the final ratio 1 : 1(culture filtrate: ethanol, v/v) to the supernatant, which was obtained after precipitation of the culture filtrate with ethanol at a ratio of 1 : 0.5. These results show that methanol treatment is better than other organic solvent treatments for the simple recovery of PPase, whereas fractionated treatment of ethanol can recover PPase with higher purification fold.

• Microbiology and Biotechnology Letters
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• v.10 no.3
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• pp.197-203
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• 1982

The protopectinase from the culture extract of a Verticillium sp. was purified about 1000 fold by ammonium sulfate fractionation, DEAE-Sephadex treatment and Sephadex G-75 column chromatography. The purified enzyme was homogeneous on electrophoresis and its molecular weight was estimated to be 38000 by Andrew's gel filtration, method. The enzyme was almost stable under the temperature of 4$0^{\circ}C$ and within the pH range of 3-5. Its optimum pH and temperature were 4 and 4$0^{\circ}C$, respectively. The activity was markedly inhibited by galacturonic acid. The purified enzyme was able to macerate various kinds of plant tissues, such as radish, cucumber, onion, carrot, and potato. It also reduced the viscosity of pectin solution more rapidly than that of pectic acid solution and showed no lyase or CMCase activity.