• 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.29 no.3
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• pp.176-180
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• 2001

In plant tissues intercellular cementing portion called as middle lamella consists of high proportion of protopectin that is water insoluble form of pectin on their backbone Protopectinase (PPase) a heterogeneous group of enzymes that hydrolyze or dissolve the insoluble protopectin in plant tissues by restricted depolymerization liberates water solu- ble pectin with the resultant separation of plant tissues that have been protected against environmental shock by rigid cel wall . Bacillus subtilis EKll was most effective for PPase Production For increasing of PPase productivity effects of glucose concentrations, pHs and aeration rates were studied in batch culture The most proper concentra- tion of glucose pH and air condition for PPase Production were 1% 8.0 and lvvm respectively In these condi- tion PPase productivity was $84,364 UL^{-1}$ $h^{-1}$ and increased about 15.6 times than flask fermentation.

• Microbiology and Biotechnology Letters
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• v.27 no.5
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• pp.378-384
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• 1999

Protopectinase (PPases) are heterologous group of enzymes that degrade pectin from the insoluble protopection which is constituent of the middle lamella and primary cell wall of higher plants by restricted depolymerization. From the previous report[6], enzymatically separated plant cells, which are produced from plant tissues by PPases treatment, showed well-conserved cellular components with their rigid cell wall and this characteristic is applicable to preparation of novel food material. The purpose of this study is to investigate the effect of medium composition of PPase production from Bacillus subtilis EK11 which was selected as a PPase producer. Various carbon sources and concentrations on PPase production were studied and corn starch at 0.7% was the most effective for production of PPase. Among the nitrogen sources, yeast extract was the most effective for PPase production and the effect of (NH4)2SO4 was notable as inotganic nitrogen source. Inorganic compounds such as KH2PO4, K2HPO4, Na3-citrate.2H2O and MgSO4 were optimized for PPase production. PPase activity was inhibited by the adition of Ba2+ or Zn2+. The optimal medium for PPase production was devised: 0.7% corn starch, 0.3% yeast extract, 1.4% KH2PO4, 0.6% K2HPO4, 0.1% Na3-citrate.2H2O and 0.02% MgSO4. PPase production by using the optimum medium was carried out with shaking cultivation at 37$^{\circ}C$. The maximum PPase activity of 256unit/ml could be obtained after the cultivation for 48hrs. The activity was increased about 2.2timesthan the activity, 112 unit/ml, in basal medium.

• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.3
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• pp.401-406
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• 2000

In development of the processed food, it is important not only to make the food delicious but to enhance its storage span and thermal stability without change of the food quality in color, which greatly affects the tastes of customers. Protopectinase (PPase) from Bacillus subtilis EK11 hydrolyses or dissolves protopectin in the middle lamella of plant tissues with the resultant separation of plant cells from each other, called enzymatic maceration. With the PPase, Kiwifruit was enzymatically macerated to separate cells to primary cell wall without damage. Yields of kiwifruit treated with PPase and mechanical maceration were 82% and 60%, respectively. Total and reducing sugars, crude protein and fat in the enzymatic maceration were well preserved as in the mechanical maceration. Importantly, over 95% of vitamin C, which is the most unstable component in application of the mechanical maceration, remained with intact form for one day after the enzymatic treatment. When the suspensions of kiwifruit macerated with both treatments had been stored at $4^{\circ}C for 6 days, the suspension of kiwifruit mechanically macerated was decolorized. whereas decolorization was not found in the enzymatically macerated kiwifruit. Moreover, the mechanically macerated kiwifruit was greatly deteriorated after heat treatment at $100^{\circ}C for 60 min ; the cell suspension of the exzymatically separated kiwifruit appeared to be stable, indicating the thermal stability. Thus, the PPase treatment could be a better choice for preparation of the highly valuable and functional processed food of kiwifruit as well as for prolonging the preservation period of the processed kiwifruit.

• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.1
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• pp.29-34
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• 2003

In development of the processed food, it is important not only to make the food delicious but to enhance its storage span and thermal stability without change in color, which greatly affects the tastes. Protopectinase (PPase) from Bacillus subtilis EK11 hydrolyses or dissolves protopectin in the middle lamella of plant tissues with the resultant separation of plant cells from each other, called enzymatic maceration. With the PPase, persimmon was enzymatically macerated to separate cells to primary cell wall without damage. Recovery rates of persimmon treated with PPase and mechanical maceration were 95% and 85%, respectively. Total and reducing sugars, crude protein and fat in the enzymatic maceration were well preserved as in the mechanical maceration. Importantly, over 50% of vitamin C, which is the most unstable component during the mechanical maceration, remained with an intact form for one day after the enzymatic treatment. When the suspensions of persimmon macerated with both treatments were stored at 4$^{\circ}C$ for 9 days, the mechanically macerated persimmon suspension was decolorized, whereas decolorization, was not found in the enzymatically macerated persimmon suspension. Moreover the mechanically macerated persimmon was greatly deteriorated after heat treatment at 10$0^{\circ}C$ for 60 min, whereas cells of the enzymatically separated persimmon suspension appeared to be stable, indicating increased thermal stability Thus, the PPase treatment of persimmon could be a better choice for preparation of highly valuable and functional processed food as well as for increase in preservation period.

• Korean Journal of Food Science and Technology
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• v.38 no.3
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• pp.369-377
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• 2006

Protopectinase (PPase) from Bacillus subtilis was used to investigate enzymatic maceration of vegetable tissues. Optimum concentration and pH of PPase were 0.75, 0.75, and 0.5%, and 5.0, 8.0, 7.0 for red pepper, garlic, and cucumber, respectively. Optimum shaking-rate, reaction time, and temperature of PPase were 250 rpm, 150 min, and $37^{\circ}C$, respectively. Yields of mechanically macerated red pepper, garlic, and cucumber were 45.8, 47.5, and 82.1%, whereas those treated with PPase were 81.8, 84, and 98%. Over 40% Vitamin C, the most unstable component during mechanical maceration, remained intact for 12 days after enzymatic treatment. Color differences $({\Delta}E)$ of mechanically macerated red pepper, garlic, and cucumber were 1.16, 2.86, and 3.27, whereas those of PPase-treated ones were 2.87, 7.68, and 5.22 after heat treatment at $100^{\circ}C$ for 20 min. Capsaicin content of mechanically macerated red pepper was 0.4 mg/100 g, whereas that treated with PPase was 1.32 mg/100 g. Viscosity of PPase-treated vegetable decreased slowly with increasing storage period, whereas that of mechanically macerated vegetable sharply decreased. These results indicate PPase treatment of vegetable could be better choice for preparation of high-values and functionally processed food and for extending preservation period.

• Journal of Microbiology and Biotechnology
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• v.16 no.7
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• pp.1144-1148
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• 2006

DNA can oxidatively be deaminated by ROS, which converts DNA base amino groups to keto groups and can trigger abnormal mutations, resulting in mutagenesis in organisms. In this study, a noncanonical purine dNTP pyrophosphatase (AfPPase) from a hyperthermophilic archaeon Archaeoglobus fulgidus, which hydrolyzes aberrant nucleoside triphosphates, was overexpressed in E. coli, purified, and characterized. The purified AfPPase showed remarkably high activity for XTP and dITP, suggesting that the 6-keto group of these nucleotides is critical for the reactivity. Under optimal reaction conditions, the reaction rate for these substrates was about 120 times that with dGTP. Therefore, AfPPase may play a significant role in DNA repair by hydrolysis of noncanonical nucleotides before they are misincorporated into DNA.

• 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.

• Journal of Plant Biotechnology
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• v.39 no.4
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• pp.253-260
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• 2012

On the basis of the reported agriculturally valuable phenotypes resulted from ectopic overexpression of Arabidopsis vacuolar $H^+$-PPase (AVP1), we generated the Chinese cabbage lines expressing AVP1 which then subjected to salt stress to determine the AVP1 expression if it consistently confers the capability for increasing biomass and enhancing tolerance to salinity in other species. Collectively, here we demonstrate that the transgenic young plants show more vigorous growth and higher tolerance to salt stress than wild-type ones. Increased biomass phenotype by AVP1 expression was supported by comparing fresh and dry weights of transgenic and wild type plants grown under normal condition, while higher salt tolerance trait was confirmed by tracing the kinetics of photosystem II quantum yield and DAB-staining under gradually intensified salt stress induced by MS salt or NaCl, followed by normal condition.

• Applied Microscopy
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• v.31 no.2
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• pp.199-206
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• 2001

In 1980s, the fragmentation or subdivision of protein deposits at the periphery of protein storage vacuole was suggested as the only route of PB development in pea cotyledon cells. Since then, other independant processes such as terminal dilation , transformation and de novo development have been discussed as alternative routes for PB development, and today, these multiple mechanisms of PB development are accepted as a result of active investigations. For analysis of the protein accumulations in the ER cisternae during seed development, immunocytochemical gold labellings were applyed on the single cells separated by enzymatic digestion from cotyledon tissue. Anti-legumin labellings at the early stage, and anti-vicilin labellings at the intermediate stage were observed on the protein-filled ER. The $\alpha-Tip$, which is the ER retention protein, was labelled somewhat at late stage, and PPase, a sort of tonoplast membrane protein, was labelled at early stage.