• Title/Summary/Keyword: exo-polygalacturonase

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Investigation of Reactive Conditions to Extract Pectin with Exo-polygalacturonase from Pear Pomace (배박에서 펙틴의 추출을 위한 Exo-polygalacturonase의 반응조건 검토)

  • Yuk, Hyun-Gyun;Choi, Jin-Ho;Cho, Yong-Jin;Ha, Jung-Uk;Hwang, Yong-Il;Lee, Seung-Cheol
    • Korean Journal of Food Science and Technology
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    • v.31 no.4
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    • pp.971-976
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    • 1999
  • Exo-polygalacturonase (EPG) from Rhizopus sp. was applied to the extraction of pectin from pear pomace because EPG produces pectin by solubilizing protopectin. The content of total galacturonic acid in water-alcohol insoluble pectin (WAIP) was determined as 34.6%. Pear pomace was solubilized by using EPG, with regarding reaction pH, temperature, time and ratio of enzyme to substrate in order to find optimum condition. While the yield by an acidic treatment was 6.2%, the maximum yield by an enzymatic treatment was 23.4% under the extraction condition of pH 7.8, $60^{\circ}C$, 36 hr and 1/10 of enzyme/substrate. At this condition, the purity and methoxyl content of enzyme-extracted pectin were, respectively, 34.7% and 0.7%, while those of acid-extracted pectin were, respectively, 71.1% and 5.0%. Meanwhile, the average molecular weight of pectin extracted by the enzymatic method was $2.5{\times}10^{3}$ while that of acid-solubilized pectin was $8.4{\times}10^{3}$.

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Separation and Characterization of Endo-Polygalacturonase from Aspergillus niger (Aspergillus niger가 생산(生産)하는 Endo-Polygalacturonase의 분리(分離)와 특성(特性))

  • Park, Kyong-Bin;Park, Kwan-Hwa
    • 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.

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Production of Pectolytic enzymes by Alternaria mali Roberts (Alternaria mali Roberts에 의(依)한 Pectin질(質) 분해효소(分解酵素)의 생산(生産))

  • Kim, Kee-Hong;Lee, Chang-Un
    • The Korean Journal of Mycology
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    • v.16 no.2
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    • pp.64-69
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    • 1988
  • Isoates with changed pathogenicity were selected among iprodione-resistant Alternaria mali to investigate any relationship between their pectolytie enzyme activity and pathogenicity. In artificial medium, strongly pathogenic isolates $S_1$, and $R_3$ showed higher enzyme activity than weakly pathogenic isolate $R_8$.Activity of endo-polymethylgalacturonase and end-o-polygalacturonase was more than 3 times. But activity of pectin methylesterase and pectin Iyase by isolaste $S_1$ was higher than those by $R_3$ and $R_8$ isolates. In apple medium dialyzed against distilled water, activity of enzyme by each isolate was increased but growth of each isolate was reduced. When iprodione was added to the medium, enzyme activity and growth of isolate $S_1$, were reduced but strongly pathogenic isolate .$R_3$ among iprodione-resistant ones showed increased enzyme activity except for exo-polygalacturonase in dialyzed apple medium.

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Phylogenetics and Gene Structure Dynamics of Polygalacturonase Genes in Aspergillus and Neurospora crassa

  • Hong, Jin-Sung;Ryu, Ki-Hyun;Kwon, Soon-Jae;Kim, Jin-Won;Kim, Kwang-Soo;Park, Kyong-Cheul
    • The Plant Pathology Journal
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    • v.29 no.3
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    • pp.234-241
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    • 2013
  • Polygalacturonase (PG) gene is a typical gene family present in eukaryotes. Forty-nine PGs were mined from the genomes of Neurospora crassa and five Aspergillus species. The PGs were classified into 3 clades such as clade 1 for rhamno-PGs, clade 2 for exo-PGs and clade 3 for exo- and endo-PGs, which were further grouped into 13 sub-clades based on the polypeptide sequence similarity. In gene structure analysis, a total of 124 introns were present in 44 genes and five genes lacked introns to give an average of 2.5 introns per gene. Intron phase distribution was 64.5% for phase 0, 21.8% for phase 1, and 13.7% for phase 2, respectively. The introns varied in their sequences and their lengths ranged from 20 bp to 424 bp with an average of 65.9 bp, which is approximately half the size of introns in other fungal genes. There were 29 homologous intron blocks and 26 of those were sub-clade specific. Intron losses were counted in 18 introns in which no obvious phase preference for intron loss was observed. Eighteen introns were placed at novel positions, which is considerably higher than those of plant PGs. In an evolutionary sense both intron loss and gain must have taken place for shaping the current PGs in these fungi. Together with the small intron size, low conservation of homologous intron blocks and higher number of novel introns, PGs of fungal species seem to have recently undergone highly dynamic evolution.

Production of Pectolytic Enzymes by Botryosphaeria dothidea (사과겹무늬썩음병균(病菌) Botryosphaeria dothidea에 의한 Pectin질(質) 분해효소(分解酵素)의 생산)

  • Park, Seok-Hee;Kim, Kee-Hong;Lee, Chang-Un
    • The Korean Journal of Mycology
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    • v.19 no.2
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    • pp.143-147
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    • 1991
  • Botryosphaeria dothidea causing apple fruit rot was cultured in pectin-polypectate mine­raI salts or apple mediumss, to investigate pectolytic enzyme production and activity. Exo-polygalactu­ronase(PG) and exo-polymethylgalacturonase (PMG) in apple medium showed maximum of activity to 6.4 and 7.2 units at six days of culture, respectively. Their maximum activity in pectin-polypectate mineral salts medium was 5.9 and 5.3 units at eight days of culture lower than in apple medium respectively. Endo-PG and endo-PMG in pecin-polypectate mineral salts medium were maximum of activity to 4.4 and 16.2 units at six and eight days of culture, respectively, but activities in apple medium were 3.2 and 6.7 units at eight days of culture. Activity of polygalacturonate-trans-­eliminase(PGTE) and pectinmethyl-trans-eliminase(PMTE) was higher in pectin-polypectate mineral salts medium than in apple medium. Fungal growth was maximum at six and eight days of culture in pectin-polypectate mineral salts and apple medium, respectively.

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Structural elucidation of immuno-stimulating polysaccharide, galactomannan isolated from Colocasia esculenta (토란으로부터 분리한 면역활성 다당 galactomannan의 구조적 특성 규명)

  • Lee, Hee-Won;Shin, Kwang-Soon
    • Korean Journal of Food Science and Technology
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    • v.52 no.6
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    • pp.595-603
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    • 2020
  • To elucidate the structure-function relationship of polysaccharides obtained from Colocasia esculenta, the immuno-stimulating polysaccharide, CE-4a was purified to homogeneity from the crude polysaccharide (CE) extracted from the corms of C. esculenta by two subsequent column chromatographies using DEAE-Sepharose FF and Sephadex G-100, and analysis of their immuno-stimulatory activities and structure were conducted. CE-4a showed an increase in anti-complementary activity in a dose-dependent fashion. The molecular mass was estimated to be 182.4 kDa, which mainly consisted of galactose (43.5%) and mannose (18.2%). Methylation analysis indicated that CE-4a comprised at least 10 different glycosyl linkages, such as terminal Galp, 3-linked Galp, and 4-linked Manp, as well as a characteristic linkage, 2,4,6-branched Manp residue. To analyze the fine structure of CE-4a, it was sequentially digested using endo-α-(1→4)-polygalacturonase, exo-α-galactosidase and endo-β-1,4-D-mannanase. These analyses suggested that CE-4a is to be a highly branched galactomannan with a (1→4)-mannan backbone and galactopyranosyl oligosaccharide side chains.

Changes in Pectin and Pectin Degrading Enzymes Activity during Storage of Kiyomi Tangor Produced in Jeju (제주산 만감류 청견의 저장 중 펙틴 및 펙틴분해효소 활성의 변화)

  • 강문장;임자훈;고정삼
    • Food Science and Preservation
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    • v.8 no.2
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    • pp.131-139
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    • 2001
  • Kiyomi tangor(Citus unshiu x sinensis) was stored at 3$\^{C}$ and 85% relative humidity, and the changes in firmness, pectin degrading enzymes activity and other physicochemical properties of citrus fruits during storage were investigated. Decay ratio and weight loss during 180 days’ storage were increased gradually to 13.0% and 12.9%, respectively. Firmness of fruits with 2 mm probe was decreased gradually from 808.7 g-force to 406.4 g-force, and moisture of peel and flesh were decreased from 76.5% to from 89.6% to 87.6% during storage, respectively. Exo-polygalacturonase activity of peel after 150 days’ storage were increased gradually to 558.09 units/100g. Pectin methylesterase activity of peel and flesh were increased from 14.7 units/g to 2.3 units/g, and from 9.4 units/ml to 2.7 units/ml at 150days’ storage, respectively. Endo-polygalacturonase activities were not changed notably during storage. Alcohol-insoluble solid(AIS) of peel was not changed notably. During storage of the fruits water soluble pectin(WSP) of peel and flesh were increased from 474.49 mg/100g to 614.29mg/100g, and from 66.91mg/100g to 92.74mg/100g as wet basis, respectively. Hexameta-phosphate soluble pectin(HMP) of peel were decreased from 405.5mg/100g to 270.43mg/100g, hydochloric acid soluble pectin(HSP) of peel was also decreased from 544.02mg/100g to 412.64mg/100g during storage. Total pectin substance(TPS) of peel and flesh were decreased from 1,424.01mg/100g to 1,297.36mg/100g, and from 165.51mg/100g to 171.54mg/100g, respectively. Composition ratio of pectin was in order of WSP > HSP > HMP.

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Production of Endo-Polygalacturonase of a Mutant of Aspergillus niger (Aspergillus niger의 변이주(變異株)에 의(依)한 Endo-polygalacturonase의 생산(生産))

  • Park, Yoon Joong;Shon, Cheon Bae
    • Korean Journal of Agricultural Science
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    • v.12 no.2
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    • pp.324-332
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    • 1985
  • Aspergillus niger B-15 with strong Endo-polygalacturonase (Endo-PG) activities was selected out from a total of 1,573 fungal strains isolated from various testing materials. A mutant strain, U-46, was obtained from the Aspergillus niger B-15 by repeated irradition of ultra-violet light. The objectives of the study were to investigate the fungal properties of the parental and mutant strains obtained and to study the condition of enzyme production and reaction. The results obtained are summarized as follows: 1. The size of conidial head of the U-46 mutant was smaller than that of the parental strains, B-15 and the length of the conidiophore was also shorter than that of the parental strains. 2. The optimum conditions for the Endo-PG production of the parental B-15 strain in the wheat bran Koji were obtained when 40% of water was added to the wheat bran and the temperature was 30 to $35^{\circ}C$. However, the best condition for the mutant U-46 strain was attained when 60 to 70% of water was added and the temperature was $35^{\circ}C$. The optimum growing periods were two to three days for both parental and mutant strains. 3. Under the optimum producing conditions of each strains, the enzymatic activity of the mutant U-46 was 20 times higher than the Endo-PG of the parental strain, B-15. 4. When both strains were cultured in the wheat bran Koji containing 60% of water at $35^{\circ}C$ for three days, the mutant strain. U-46, was about 46 times higher in the Endo-PG activity and about 18 times greater in Exo-PG activity than the parental strain, B-15. The activities of cellulase, $\alpha$-amylase, and glucoamylase were also highly increased in the mutant strain. 5. The mutant strain, U-46, increased its Endo-PG activity up to 20% over that of ordinary case when 1.2 to 1.5% of ammonium sulphate was added to the wheat bran. 6. The optimum condition for Endo-PG activity of crude enzyme of the mutant strain, U-46, was attained when pH of reaction solution was 4.0 to 4.5 and the temperature was $50^{\circ}C$.

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Changes in Pectin-degrading Enzymes activity during Storage of Satsuma Mandarin (온주밀감의 저장 중 성분과 펙틴분해효소의 변화)

  • Kang, Moon-Jang;Kim, Ji-Yong;Koh, Jeong-Sam
    • Applied Biological Chemistry
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    • v.43 no.3
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    • pp.179-183
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    • 2000
  • Satsuma mandarin(Citrus unshiu Marc. var. miyagawa) was stored at $3^{\circ}C$ and 85% relative humidity, and then the changes of firmness, pectin- degrading enzymes activity and other physicochemical properties of citrus fruits during storage were investigated. Firmness of fruits with 2 m probe was decreased quickly from 1,176.8 g-force to 503.6 g-force, and moisture of peel and flesh were decreased from 75.3% to 74.9%, and from 91.8% to 90.7% during maturation, respectively. Decay ratio was increased to 18.75% after 90 days' storage, and after then it was increased rapidly. Weight loss was increased gradually to 24.5% during long-term storage. Firmness with 2 mm probe were decreased from 538.9 g-force to 336.9 g-force gradually during storage. Peel moisture was decreased from 75.8% to 72.6%, and flesh moisture was also decreased gradually from 90.3% to 88.3% during storage. Exopoly-galacturonase activity of peel and flesh were increased from 326.0 units/100 g to 534.9 units/100 g, and from 63.1 units/100 g to 81.0 units/100 g at 90 day's storage, respectively. After then, He enzyme activities were decreased from 394.0 units/100 g and 38.0 units/100 g, respectively. Pectinesterase activity of peel and flesh were increased from $14.4\;{\mu}mol$ to $38.8{\mu}mol$, and from $26.0{\mu}mol$ to $39.0{\mu}mol$ at 60 days' storage, respectively. After then, the enzyme activities were decreased to $6.0{\mu}mol$ and $8.2{\mu}mol$, respectively.

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