• The Korean Journal of Mycology
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• v.22 no.4
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• pp.298-308
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• 1994

The properties of polygalacturonase by Ganoderma lucidum in liquid culture were investigated. The enzyme was composed of an endo- and an exo-polygalacturonase. The endo- and exo-polygalacturonase were purified approximately 56 and 9.2-fold, respectively, through ammonium sulfate fractionation, gel filtration on Biogel P-100, anion exchange chromatography on DEAE-cellulose, gel chromatography on Sephadex G-150 and re-gel chromatography on Sephadex G-150. The endo- and exo-polygalacturonase had higher affinity for apple pectin than for citrus pectin or pectic acid. The Km values of the endo- and exo-polygalacturonase for apple pectin, determined on the Lineweaver-Burk plot, were 1.44 and 10.6 mg $ml^{-1}$ for apple pectin, respectively. Purified endo-polygalacturonase was found to be homogeneous electrophoretically and had a molecular weight of 54,000 estimated on SDS polyacrylamide gel. The optimal pH for the activity of the enzymes was 4.0. The endo- and exo-polygalacturonase were stable in the pH range of 4.0 to 6.0 and 3.5 to 5.5, respectively. The optimal temperatures of the endo- and exo-polygalacturonase were 40 and $60^{\circ}C$, respectively. The exo-polygalacturonase was more resistant to heat than the endo-polygalacturonase, requiring heating for 40 min at $80^{\circ}C$ for complete inactivation. The activity of the endo-polygalacturonase was increased by $Ca^{++}$ and $Mn^{++}\;ions$, while that of the exo-polygalacturonase was increased by $Ca^{++}\;ion$ only, and was not affected by $Mn^{++}\;ion$.

• Journal of the Korean Society of Food Science and Nutrition
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• v.26 no.2
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• pp.180-185
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• 1997

This work was carried out to investigate change of polygalacturonase activities and polygalacturonase patterns by gel filtration chromatography during softening of persimmon and jujube fruits. During softening of two kinds of fruit, polygalacturonase activities of water-soluble and salt-soluble proteins were increased, but that of cell wall-bound proteins was decreased. In water-soluble and salt-soluble proteins of persimmon fruits, two peaks of polygalacturonase activity were separated in mature stages, but one peak in soft stages. During softening of those fruits, the peaks of polygalacturonase activity in water-soluble and salt-soluble proteins appeared on the same fraction with the peaks of polygalacturonase activity in cell wall-bound proteins.

• Journal of the Korean Society of Food Science and Nutrition
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• v.19 no.6
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• pp.596-604
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• 1990

Polygalacturonase activity was not detected at turning stage but were 55.01 and 206.70 units/100g -fr. wt. in mature and soft persimmon, respectively. Polygalacturonase have two isoenzymes and its molecular weight was estimated to be 55,000 doltons by the method of gel filtration. Vmax and Km of polygalacturonase 1 were 0.195$\mu$ mole reducing-sugar/$m\ell$/30 min. and 3.50mg/$m\ell$, respectively. The optimum temperature and pH of the enzyme appeared to 4$0^{\circ}C$ and 3.5, respectively. Polygalacturonase I had Vmax of 0.110m mo1e reducing-sugar/$m\ell$/30min. and Km value of 2.50mg/$m\ell$, The optimum temperature and pH of polygalcturonase II appear 4$0^{\circ}C$ and 4.0, respectively. Polygalacturonase I was fairly stable at 6$0^{\circ}C$while polygalacturonase II appeared to be stable up to 4$0^{\circ}C$.

• Applied Biological Chemistry
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• v.32 no.4
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• pp.367-373
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• 1989

The effect of gelatin concentration, pH of fruit juice and other factors for the enzymatic clarification of fruit juice by the use of polygalacturonase was studied. The proper addition of gelatin was much effected on clarification by polygalacturonase and optimum concentrations of gelatin for the enzymatic clarification of Zugaru juice, Magnolia Gold juice, Golden Delicious juice, Jonagold juice, Jonathan juice, Campbell Early juice were 0.04, 0.01% 0.02%, 0.06% 0.01%, and 0.04%, respectively. Optimum pH for the clarification by the use of polygalacturonase and gelatin was $3.2{\sim}3.5$ contrary to optimum pH 4.8 of polygalacturonase for the hydrolysis of pectic acid. At the reaction temperature of $45^{\circ}C$ and polygalacturonase concentration of 0.01mg/ml, fruit juices were completely clarified by optimum addition of gelatin for 60min.

• The Korean Journal of Mycology
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• v.21 no.3
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• pp.195-199
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• 1993

This experiments was designed for development of Fusarium oxysporum strains with enhanced activity of polygalacturonase by means of mutants and protoplasts fusion. Six auxotrophic mutants of F. oxysporum were induced by treatment of MNNG. Protoplasts from mutants were formed from early exoponential mycelium after treatment with driselase(10 mg/ml) using 0.6 M KCl as osmotic stabilizer. Fusion experiments between protoplasts of several auxotrophic mutants were done using polyethyleneglycol 8,000 and $CaCl_2$. The frequency of fusion was $5.0{\times}10^{-3}$ as determined from several experiments. Activity of polygalacturonase was determined by the methods of modified DNS. Consequently, the polygalacturonase activities of mutants and fusant derived F. oxysporum were 1.4 to 3.5 times greater than that of the parental strain, and mutant Fx-2 seemed to be the best strain. Thus, the method we used seemed to be favorable for the improvement of strains.

• The Korean Journal of Mycology
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• v.22 no.4
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• pp.286-297
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• 1994

The optimum nutritional and cultural conditions of polygalacturonase by Ganoderma lucidum in liquid culture were studied. The optimal temperature, pH, and the duration of culture for production of the enzyme was $30^{\circ}C$, 5.5 and 14 days, respectively. The maximal production of the enzyme was obtained in a synthetic medium containing 10 g of pectin, 10 g of soluble starch, 1 g of yeast extract, 2 g of peptone, 1 g of phenylalanine, 2 g of $KH_2PO_4$, 0.2 g of $MgSO_4{\cdot}7H_2O$, 0.05 g of $CaCI_2$ and 100 g of $thiamin{\cdot}HCI$ in 1000 ml of distilled water.

• Journal of the Korean Society of Food Science and Nutrition
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• v.24 no.4
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• pp.570-577
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• 1995

Aspergillus sp. SB-2704 was selected for its strong polygalacturonase activity among various strain of mold found in soil. It was found that production of polygalacturonase reached to maximum when the wheat bran medium containing 1% polypepton, 1% glucose, and 0.2% FeSO4 were cultured for 3 days at 35$^{\circ}C$. Polygalacturonase was purified 20.90 fold from Aspergillus SB-2704. The purification procedures include ammonium sulfate treatment, gel filtration on Sephdex G-150 and DEAE-cellulose ion exchange chromatography. Yield of the enzyme purification was 4.34%. Purified enzyme was confirmed as a single band by the polyacrylamide gel electrophoresis. When the purified enzyme was applied to SDS-polyacrylamide gel electrophoresis, the molecular weight was estimated to be 36,000. The optimum pH for the enzyme activity was 5.5 and optimum temperature was 5$0^{\circ}C$. The enzyme is stable in acidic condition. The activity of purified enzyme was inhibited by Pb2+, Hg2+ and Ba2+, whereas activated by Cu2+, Mn2+, Mg2+ and Fe2+. The activity of polygalacturonase was inhibited by the treament wit maleic anhydride, iodine, and EDTA. The result indicate the possible involvement of histidine and metal ion at active site.

• Journal of the Korean Society of Food Science and Nutrition
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• v.24 no.2
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• pp.247-253
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• 1995

This paper was performed to investigate the changes of non-cellulosic neutral sugars composition in cell wall of persimmon fruit by treatment of cell wall degrading enzyme in vitro. Rhamnose, xylose and galactose in cell wall by polygalacturonase treatment, arabinose, galactose and rhamnose in cell wall by mixed enzyme treatment and arabinose and galactose in cell wall by ${\beta}-galactosidase$ treatment decreased, respectively. Noncellulosic neutral sugars of pectins extracted cell wall by enzyme treatments decreased and those by polygalacturonase treatment decreased remarkably. Rhamnose, arabinose and xylose in hemicellulose I of cell wall by polygalacturonase treatment were higher than those of untreated, and rhamnose and xylose in that by ${\beta}-galactosidase$ treatment were higher but arabinose, mnnose and galactose decreased. Xylose, mannose and glucose in that by mixed enzyme treatment were higher than those of untreatment and arabinose and galactose decreased. Contents of total non-cellulosic neutral sugars in hemicellulose of untreatment, and contents xylose, and glucose in hemicellulose II of cell wall by polygalacturonase treatmet decreased but those of other treatments were not changed.

• Korean Journal of Microbiology
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• v.35 no.2
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• pp.158-163
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• 1999

A ploygalacturonase-produchg yeast was isolated from Cheju soil by selective eivichment media. One strain which has the highesl activity of polygalacturonase was selected. The characle~ishcs of the strain CS-2 were as follows: CS-2 utilized xylose. sucrose, maltose, u.ehalose, cellobiose. melibiose, lactose, raffinose, inosiiol, dulicilol, and dextrose, but did not utilized galactose, nitrate. nit~te, and lysine. Growth of CS-2 was inhibited by cyclohexamide, 1% acetic acid, and high concenaation (over 50%) of glucose. It grew at $30^{\circ}C$ but did 'IIOL $35^{\circ}C$. The cell size ofthe strain CS-2 was 2.9 p ~ n in length and 1.3 $\mu$ in diameter. Vegetable reproductmn was multiple budding and ascospre was present I to 4. Pseudomycelia or true myceliua formation were not observed In any of the cullureq. These results suggest that strain CS-2 is most likely a strain related Cryptococcus spp. (Cryptococcu spp. CS-2). When polygalacturonase or ihe yeast was induced by addition of polygalactoronic acid, polygalacturonase activity was detected in culture supernatent. There was a peak of specific activity a1 he mid-stationary phase(3 days culture) of growth. Polygalacturonase specific activity of Crylmcoccus sp. CS-2 was 2.96 unitsling. The molecular weighl ol'polygalacturonase was showed to be 46 KDa by both SDS-PAGE and activity stailling.

• The Korean Journal of Food And Nutrition
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• v.5 no.2
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• pp.111-115
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• 1992

To examine ripening in peach types, cell wall contents and polygalacturonase activity were compared in Changbang, Daegubo and Yumyung peaches. Among peach types, the hardness of Daegubo was the lowest. Yumyung peach had the highest content of alcohol-insoluble substances and Changbang peach of cell wall. The contents of total and insoluble pectic substances were little different between Changbang and Yumyung peach, while the lowest in Daegubo. Daegubo peach had the highest activity of polygalacturonase, Changballg and Yumyung peach in succession.