• 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 $Mn^{++}\;ion$.

• The Korean Journal of Mycology
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• v.26 no.2 s.85
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• pp.281-286
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• 1998

Ganoderma lucidum produced the potent pectolytic enzymes for clarifying cloudy fruit juice. Among the purified polygalacturonases (endo- and exo-polygalacturonase), endo-polygalacturonase had a good effect on juice clarification. The optimum temperature and concentration of endo-polygalacturonase for the juice clarification were $40^{\circ}C$ and 4 unit/5 ml juice, respectively. The apple juice was almost completely clarified at $40^{\circ}C$ for 60 min. It was suggested that culture filtrate of Ganoderma lucidum or it's ammonium sulfate fraction should be used as a good source of pectolytic enzyme for juice clarification.

• Korean Journal of Food Science and Technology
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• v.16 no.1
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• pp.41-46
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• 1984

The pectic enzymes produced from Aspergillus niger were separated into three fractions (F-A, F-I and F-II) by means of Sephadex and DEAF-Sephadex column chromatography. Each enzyme fraction was characterized by determining viscosity change and reducing surgar of the pectic acid-enzyme mixture and analyzing thin layer chromatogram of the reaction products. F-I rapidly reduced the viscosity of pectic acid solution and released reducing groups in a random manner so that appeared to be an endo-polygalacturonase (endo-PG). The optimum pH of endo-PG for viscosity reducing activity was 4.2 and that for releasing reducing surgar was 4.7. In the thermal inactivation of endo-PG of $30-45^{\circ}C$, the enthalpy of activation was 217.3 kj/mole and z-value was $7.5^{\circ}C$. F-II and F-A were determined as endo-polymethylgalacturonase and exo-polygalacturonase, respectively.

• The Korean Journal of Mycology
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• v.18 no.1
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• pp.7-12
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• 1990

Isolates of Penicillium expansum with reduced pathogenicity were arbitrarily selected among benomyl-resistant isolates in order to investigate relationship of their pectolytic enzyme acitivity with pathogenicity. In artificial medium, strongly pathogenic isolate $S_1$ and weakly pathogenic isolate $R_2$ produced considerable amonts of endo-polymethylgalacturonase, endo-polygalacturonase, pectin methyl-trans-eliminase, and polygalacturonate-trans-eliminase. No marked difference in enzyme activities was observed between two isolates. In apple medium, the activities of endo-polymethylgalacturonase and endo-polygalacturonase of isolate $S_1$ were over 6 times higher than those of isolate $R_2$. But pectin methyl-trans-eliminase and polygalacturonate-trans-­eliminase did not show a great difference. Activities of endo-polymethylgalacturonase and endo­polygalacturonase precipitated at 80-95% saturation of ammonium sulfate were highest, and addition of these enzyme solutions increased pathogenicity of weakly pathogenic isolates $R_{1-4}$.

• Applied Biological Chemistry
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• v.37 no.4
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• pp.243-247
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• 1994

Endo-polygalacturonase was purified from Jujube. The purification procedures included DEAE-cellulose ion exchange chromatography and gel filtration on Sephdex G-100. Enzyme was purified as a single protein band and purification yield was about 6%. When the purified enzyme was applied to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the molecular weight was estimated about 19,000. Purified enzyme formed hexagonal board type.

• Applied Biological Chemistry
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• v.33 no.1
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• pp.73-78
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• 1990

Endo-polygalacturonase was purified from tomato, Lycopersicon esculentum L. The purification procedures included gel filtration on Sephadex G-150 and DEAE-cellulose ion exchange chromatography. Yield of the enzyme purification was 12.74 %. Purified enzyme was confirmed as a active single band by the SDS-polyacrylamide gel electrophoresis. When the purified enzyme was applied to SDS-PAGE, the molecular weight was estimated about 50,000. The optimum pH for the enzyme activity was 5.0 and the range of its stability to the pH was 4.0 to 5.0. The optimum temperature was $50^{\circ}C$, while the enzyme was abruptly inactivated above $50^{\circ}C$. From the result of the study on the effects of metals ion, it was found that $Ag^+$, $Zn^{++}$ and $Mg^{++}$ increased on the enzyme activity. In contrast, $Ba^{++}$, $Hg^{++}$, $Pb^{++}$, $Ca^{++}$, $Mn{++}$, $Cu^{++}$, $Fe^{+++}$, $Na^+$ and $K^+$ decreased it. the reaction catalyzed by this enzyme followed typical Michaelis-Menten kinetics with the Km value of $1.43{\times}10^{-1}\;mol/l$.

• Applied Biological Chemistry
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• v.37 no.5
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• pp.356-360
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• 1994

The optimum pH and temperature for endo-polygalacturonase activity from Jujube were 5.0 and $50^{\circ}C$. The range of its stability to pH was 4.0 to 5.0. The enzyme was inactivated about 35% by treatment at $70^{\circ}C$ for 1 hr. It was found that $Ag^+$, $Zn^{++}$ and $Mg^{++}$ increased the enzyme activity. In contrast, $Ba^{++}$, $Hg^{++}$, $Pb^{++}$, $Ca^{++}$, $Mn^{++}$, $Cu^{++}$, $Fe^{+++}$, $Na^+$ and $K^+$ decreased it. The enzyme was inactivated by treatment with maleic anhydride, iodine and 2,4-dinitrophenol. The results indicate that active site is a imidazole group on the enzyme.

• The Korean Journal of Mycology
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• v.25 no.4 s.83
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• pp.330-339
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• 1997

Botrytis cinerea T91-1 has shown to produce at least four different polygalacturonases in a liquid medium containing citrus pectin as a carbon source. One of the enzymes, its molecular weight was estimated as 37 kDa by denatured polyacrylamide gel electrophoresis, was purified by a series of procedures including acetone precipitation, ion exchange, heparin affinity, and reverse phase column chromatographies. By viscometric analysis, the enzyme was revealed as an endo-polygalacturonase. The enzyme activity was inhibited by divalent cations such as $Ca^{2+}$, $Co^{2+}$, and $Cu^{2+}$. Km and Vmax for polygalacturonic acid hydrolysis were 0.33 mg/ml and 28.6 nM/min, respectively. The optimum temperature for enzymatic activity was $55^{\circ}C$ and the enzyme showed optimal pH values between 4.0 and 4.5. The enzyme was stable up to 12 hours in the range of pH 4 to 7 and at the temperature below $30^{\circ}C$. Amino acid sequence from N-terminal up to 6 amino acids determined by Edman degradation showed little homology with polygalacturonases from fungi and plants.

• Journal of Microbiology and Biotechnology
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• v.32 no.4
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• pp.464-472
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• 2022

An endo-polygalacturonase (endo-PGase) exhibiting excellent performance during acidic fruit juice production would be highly attractive to the fruit juice industry. However, candidate endo-PGases for this purpose have rarely been reported. In this study, we expressed a gene from Penicillium oxalicum in Pichia pastoris. The recombinant enzyme PoxaEnPG28C had an optimal enzyme activity at pH 4.5 and 45℃ and was stable at pH 3.0-6.5 and < 45℃. The enzyme had a specific activity of 4,377.65 ± 55.37 U/mg towards polygalacturonic acid, and the K_m and V_max values of PoxaEnPG28C were calculated as 1.64 g/l and 6127.45 U/mg, respectively. PoxaEnPG28C increased the light transmittance of orange, lemon, strawberry and hawthorn juice by 13.9 ± 0.3%, 29.4 ± 3.8%, 95.7 ± 10.2% and 79.8 ± 1.7%, respectively; it reduced the viscosity of the same juices by 25.7 ± 1.6%, 52.0 ± 4.5%, 48.2 ± 0.7% and 80.5 ± 2.3%, respectively, and it increased the yield of the juices by 24.5 ± 0.7%, 12.7 ± 2.2%, 48.5 ± 4.2% and 104.5 ± 6.4%, respectively. Thus, PoxaEnPG28C could be considered an excellent candidate enzyme for acidic fruit juice production. Remarkably, fruit juice production using hawthorn as an material was reported for the first time.

• Korean journal of applied entomology
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• v.13 no.1 s.18
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• pp.1-10
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• 1974

The activities of pectolytic and cellulolytic enzymes produced from slices of ginseng root infected with Cylindrocarpon destructains(Zins.) Scholtern were proportional to each concentration and reaction time. Activities of cellulase(Cx), endo-polygalacturonase(endo-PG), endo-polymethylg-alacturonase(endo-PMG), exo-polygalacturonase(exe-PG), and exe-polymethylgalacturonase(exo-PMG) were maximum on the 4th day after inoculation. No endo-PG and endo-PMG were detected at the first and second days, while exo-PG exo-PMG were active. On the 6th day, all pectic enzymes were completely lost, whereas Cx remained at a high concentration. pH optima of Cx, endo-PG, endo-PMG, exo-PG, and exo-PMG were 6.0, 5.5, 8.0, 7.0 to 7.5, and 8.5, respectively. Temperature optima of Cx, endo-PG, endo-PMG exo-PG, and exo-PMG were $66^{\circ}C\;53^{\circ}C\;41^{\circ}C\;37^{\circ}C\;and\;40^{\circ}C$, respectively. Cx was only inhibited by $0.05M\; Hg^{++}$ among 16 ions tested. Inhibitory effects of ions on pectolytic enzymes varied, however$M Fe^{+++}\;and\;0.05M\;Al^{+++}$ were the best in general. Among 8 fungicides, none of them inhibited all the enzymes studied at $0.1\%$, active ingredients. Exo-PG were highly inhibited by all of the fungicides, of which difolatan was the most inhibitory to all the pectic enzymes. $Ca^{++}\; at\; 0.02M\; and\;Fe^{+++}\;at\;0.02M$ completely inhibited all the pectolytic enzymes, and Cx was inhibited $30\%$ and $70\%$ at the same concentration, respectively Formalin almost inhibited exo-PG and exe-PMG at $0.8\%$ but not the other enzymes especially Cx. Difolatan at $0.8\%$ inhibited all the enzymes concerned above $80\%$. The cellulolytic and pectolytic enzymes of C. destructans must be closely associated with the ginseng root rot and should be inhibited to control the disease effectively.