• 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.

• Journal of Microbiology and Biotechnology
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• v.33 no.4
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• pp.533-542
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• 2023

Exo-polygalacturonase (exo-PG) hydrolyzes pectin acids and liberates mono-galacturonate, which plays an important role in juice extraction, and has rarely been reported. Exo-PG (AfumExoPG28A) from Aspergillus fumigatus belongs to the glycoside hydrolase 28 family. In this study, its gene was cloned and the protein was expressed and secreted in Pichia pastoris with a maximal activity of 4.44 U/ml. The optimal temperature and pH of AfumExoPG28A were 55℃ and 4.0, respectively. The enzyme exhibited activity over almost the entire acidic pH range (>20.0% activity at pH 2.5-6.5) and remained stable at pH 2.5-10.0 for 24 h. The K_m and V_max values of AfumExoPG28A were calculated by the substrate of polygalacturonic acid as 25.4 mg/ml and 23.6 U/mg, respectively. Addition of AfumExoPG28A (0.8 U/mg) increased the light transmittance and juice yield of plantain pulp by 11.7% and 9%, respectively. Combining AfumExoPG28A (0.8 U/mg) with an endo-PG (0.8 U/mg) from our laboratory, the enzymes increased the light transmittance and juice yield of plantain pulp by 45.7% and 10%, respectively. Thus, the enzyme's potential value in juice production was revealed by the remarkable acidic properties and catalytic activity in fruit pulp.

• Preventive Nutrition and Food Science
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• v.7 no.3
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• pp.317-319
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• 2002

Exo-polygalacturonase (EPG) from Rhizopus sp. was applied to the extraction of alginic acid from sea mustard to increase extraction yield. EPG digestion was examined under distinct conditions within temperatures from $25^{\circ}C$ to 5$0^{\circ}C$, pH 5 to 9, and treatment times from 0 to 36 hr. The optimal conditions fur alginic acid extraction with EPG were: pH 7.0 at 3$0^{\circ}C$ for 24 hrs. The EPG hot water extraction yield was 3.4 times higher yield than hot water extraction alone. Using EPG to extract alginic acid from sea mustard should be considered a viable alternative to conventional extraction, with the advantage of reducing hazardous wastes such as strong acid and alkali solutions.

• Korean Journal of Food Science and Technology
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• v.31 no.1
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• pp.68-73
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• 1999

In order to overcome same disadvantages of acidic solubilization of pectin from apple pomace, enzymatic selective solubilization with exo-polygalacturonase was developed. According to analyses of the effects of temperature, pH and reaction time on extraction yield, the maximum yield by the enzymatic method, shown under the extraction condition of $45^{\circ}C$, pH 7 and 60 hours, was 31.0%, whereas the yield from an acidic method was 8%. Also the quality of pectin extracted by the enzymatic method was to that from acidic solubilization. The purity and methoxyl content of enzyme-extracted pectin were 80.1% and 6.36%, respectively, which were higher than 75.7% and 2.44% of acid-solubilized pectin. Viscosity average molecular weights of enzymic extracted and acid solubilized pectins were $1.50{\times}10^4\;and\;7.66{\times}10^4$, respectively.

• Korean Journal Plant Pathology
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• v.13 no.4
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• pp.225-231
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• 1997

Activities of polygalacturonase, laccase, and intra- and extra-cellular $\beta$-glucosidase produced by 20 Botrytis cinerea isolates in liquid culture media containing cucumber cell was as a carbon source were measured and their relationships to the pathogenicity were analyzed. No significant correlations between these enzyme activities and the pathogenicity of B. cinerea were found. Mycelial growth rate on Bayendamm media, however, was higthly correlated with the pathogenicity (r=0.522) anong these isolates. Immuno-blot analysis of the culture filtrate using antibody against against exo-polygalacturonase revealed that only one band with molecular weight of 66 kDa was detected amone 34 tested isolates. It appears that these enzymes may not be primary factors in dermining the pathogenicity of B. cinerea.

• Journal of Microbiology
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• v.35 no.2
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• pp.134-140
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• 1997

Botrytis cinerea T91-1 has been shown to produce at least four different polygalacturonases into a liquid medium containing citrus pectin, a carbon sousrce. One of the enzymes, which had an apparent molecular weight of 66 kDa estimated by denatured polyacrylamide gel electrophoresis, was purified to electrophoretic homogeneity by a series of procedures including a cetone precipitation, ion exchange, heparin affinity, and reverse phase column chromatographies. The molecular weight of native enzyme was determined to be 64 kDa by gel permeation chromatography indicating the enzyme to be a single polypeptide chain. By viscometric analysis, the enzyme was revealed as exo-polygalacturonase. The enzyme activity was inhibited by divalent cations such as $Ca^{2+}$, $Mg^{2+}$, 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 5$0^{\circ}C$. And the enzyme showed optimal pH values between 4.0 and 5.0. The enzyme was stable upto 12 hours in the range of pH 3 to 8 and at temperature below 3$0^{\circ}C$.

• 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.

• 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.

• The Korean Journal of Mycology
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• v.21 no.2
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• pp.106-111
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• 1993

Pectolytic enzymes were extracted in apple fruits rotted by Botryosphaeria dothidea, and their activities and change of pectic substances were investigated. Exo-polygalacturonase(exo-PG), exo-polymethylgalacturonase(exe-PMG), polygalacturonate-trans-eliminase(PGTE) and pectin-methyl-trans-eliminase(PMTE) were produced by the pathogen. Activities of exo-PG and exe-PMG extracted from rotten apple fruits were high to 21.15 and 24.65 units/mg protein in specific activity at seven days after inoculation, respectively. Activities of PGTE and PMTE showed 5.60 and 7.90 units/mg protein, respectively, but they were lower than those of the exo-type enzymes. Water-soluble and versene-soluble pectins were 11.50 mg/100 mg-AIS and 7.31 mg/100 mg-AIS at 14 days after inoculation, namely, they were increased by 4.23 and 2.16 mg/100 mg-AIS over those of sound apples, respectively. Total soluble pectic substances of rotten apple were 72.4% of total pectic substances and it was higher by 24.8% than sound apple. Insoluble pectic substance was notably decreased from 15.32 to 7.16 mg/100 mg-AIS according to progress of decay while total pectic substances were not changed remarkably.