• 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.

• 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 $Mn^{++}\;ion$.

• Korean Journal of Microbiology
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• v.21 no.1
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• pp.1-6
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• 1983

Effects of carbon sources, incubation time, incubation temperature, initial incubation pH, and vitamin B complex on the polygalacturonase activity of Byssochlamys fulva were studied to confirm the optimum conditions for the production of that enzyme. When pectin was used as carbon source, polygalacturonase activity reached to the maximum value of 0.50 units/ml. After 5 days of incubation, polygalacuturonase activity reached to its maximum of 0.48units/ml. Polygalacturonase activities were similar between $25^{\circ}C\;and\;45^{\circ}C$, however, decreased dramatically in the outside of this range. Polygalacturonase activity was not signicicantly influenced by the variation of initial incubation pH. However, at pH5.0, polygalacturonase activity was slightly through the addition of thiamine and riboflavin, and the optimum concentrations were $10^{-2}M$ in case of thiamine and $10^{-3}M$ in riboflavin.

• 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.

• 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.

• 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$0^{\circ}C$.

• 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.

• 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.

• Korean Journal Plant Pathology
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• v.11 no.4
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• pp.312-317
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• 1995

The polygalacturonase (PG) production in rotten apples by Botryosphaeria dothidea was purified by using gel filtration and ion exchange column chromatography, and the biochemical characters of PG were investigated. The purified PG appeared as a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with approximate molecular weight of 49 kilodalton (kDa). The molecular weight was equal to the native molecular weight estimated by gel filtration. The Km and Vmax values of PG were 0.51 mg/ml and 90.9 $\mu$M/min/ml, respectively. Optimum pH was 4.0~5.0, and the PG activity was stable from pH 5.0~10.0. Optimum temperature of the enzyme activity was 4$0^{\circ}C$. The PG activity was relatively stable at 2$0^{\circ}C$, but it was reduced 45% at 4$0^{\circ}C$ and completely inactivated at 8$0^{\circ}C$. The PG activity was considerably inhibited by Cu2+, Zn2+, SDS and EDTA, whereas it was not effected by Ca2+, K+, Mg2+ or Na+ ions.

• Korean Journal of Microbiology
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• v.21 no.2
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• pp.86-94
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• 1983

Polygalacturonase of Byssochlamys fulva was purified and characterized. Specific activity increased from 2.21 units/mg protein to 10.47 units/mg protein through $(NH_4)_2SO_4$ treatment, SephadexG-100 gel filtration, and DEAE-Sephadex ion exchange chromatography. Divalent cations, such as $Ca^{++}\;and\;Cu^{++}$, increased polygalacturonase activity greatly. Added as $10^{-3}M$ concentration, $Ca^{++}$ ion enhanced enzyme activity 9.8folds. Optimum temperature was $50^{\circ}C$ and optimum pH was 5.0. Activation energy of reaction was 8.69 Kcal/mole. Michaelis-Menten $constant(K_M)\;and\;V_{max}$ of reaction were $6.27{\times}10^{-3}mole/l\;and\;2.85{$\mu}moles/min$. Polygalacturonase of Byssochlamys fulva preferred polygalacturonic acid to pectin as substrate and was presumed as endo-type on the basis of the relationship between viscosity reduction and substrate degradation. Molecular weight of polygalacturonase was estimated as 55,000.