• Title/Summary/Keyword: cell wall-degrading enzyme

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Cloning of celC, Third Cellulase Gene, from Pectobacterium carotovorum subsp. carotovorum LY34 and its Comparison to Those of Pectobacterium sp.

  • LIM WOO JIN;RYU SUNG KEE;PARK SANG RYEOL;KIM MIN KEUN;AN CHANG LONG;HONG SU YOUNG;SHIN EUN CHULE;LEE JONG YEOUL;LIM YONG PYO;YUN HAN DAE
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.302-309
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    • 2005
  • Phytopathogenic Pectobacterium carotovorum subsp. carotovorum (Pcc) LY34 secretes multiple isozymes of the plant cell wall degrading enzyme endoglucanases. We have cloned a third cel gene encoding CMCase from Pcc LY34. The structural organization of the celC gene (AY188753) consisted of an open reading frame (ORP) of 1,116 bp encoding 371 amino acid residues with a signal peptide of 22 amino acids within the NH$_2$-terminal region of pre-CelC. The predicted amino acid sequence of CelC was similar to that of Peetobaeterium ehrysanthemi Cel8Y (AF282321). The CelC has the conserved region of the glycoside hydrolase family 8. The apparent molecular mass of CelC was calculated to be 39 kDa by CMC-SDS-PAGE. The cellulase­minus mutant of Pee LY34 was as virulent as the wild-type in pathogenicity tests on tubers of potato. The results suggest that the CelC of Pce LY34 is a minor factor for the pathogenesis of soft-rot.

Screening of White Rot Fungi with Selective Delignification Capacity for Biopulping (백색목재부후균중 Biopulping에 이용가능한 선택적 리그닌분해균의 스크리닝)

  • Lee, Jong-Kyu;Oh, Eun-Sung
    • The Korean Journal of Mycology
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    • v.26 no.2 s.85
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    • pp.144-152
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    • 1998
  • To obtain white rot fungi which have selective delignification capacity and can be used in biopulping processes, 94 different wood rotting fungi were screened and the capabilities of selected species were evaluated on deciduous and coniferous wood blocks. White rot fungi, first of all, were selected by simple enzyme tests, i.e., cellulase activity test; phenol oxidase activity test; laccase and peroxidase activity test. Most organisms that gave a positive Bavendamm gave a strongly positive laccase test with syringaldazine whereas most of those that gave a negative Bavendamm test also negative test for laccase and peroxidase, even if some exceptions were noted. Wood decay experiement were carried out to select fungal species with selective lignin-degrading ability by inoculating selected fungi to both wood blocks of Populus tomentiglandulosa and Larix leptolepis. After 12 weeks of incubation, weight losses, lignin losses, and morphological characteristics of the decayed wood were investigated. Almost all fungi tested caused 2 or more times of weight losses in P. tomentiglandulosa than in L. leptolepis, while no weight losses were detected from the un-inoculated wood blocks. Ceriporiopsis subvermispora and Phanerochaete chrysosporium were the best delignifiers for both hardwood and softwood. P. chrysosporium, however, was less effective than C. subvermispora. Bjerkandera adusta and two unidentified spp. caused delignification for only P. tomentiglandulosa. B. adusta caused simultaneous rot of all cell wall components, resulted in thinning of the secondary cell wall layers. Other fungi caused selective delignification resulting in the removal of lignin from middle lamella and separation of cells from each other.

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Selection of parental monokaryons from Korean Hypsizigus marmoreus by protoplast regeneration (원형질체 재생을 통한 느티만가닥버섯 단핵균주 선발)

  • Oh, Youn-Lee;Kong, Won-Sik;Jang, Kab-Yeul;Shin, Pyung-Gyun;Kim, Eun-Sun;Oh, Min ji;Choi, In-Geol
    • Journal of Mushroom
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    • v.13 no.3
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    • pp.270-273
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    • 2015
  • Hypsizigus marmoreus is commercially the most important edible mushroom in Japan. This mushroom is usually cultivated for a longer period (about 85~120 days) than other mushroom. In order to develop a new cultivar that has a shortened cultivation period, the genome analysis of this strain has been considered. This study aims to obtain parental monokaryotic strains reproducing 'Haemi' cultivar in Hypsizigus marmoreus for reference genome sequencing. The mycelia were cultured in MCM and MYG media for various incubation periods. Homogenized mycelia were treated with commercial cell wall degrading enzymes to maximize protoplasts production yield from Hypsizigus marmoreus. The greatest number of protoplasts was obtained from mycelia cultured in MCM media for 3 days using Novozyme enzyme. The isolated protoplasts were grown in regeneration agar media after two weeks. Regenerated colonies were picked and moved on separated dishes for microscopic observation. Neohaplonts regenerated from dikayotic strains were identified by the absence of clamp connections. We confirmed that one of monokaryotic strains is a parental strain by crossing with an original compatible strain of 'Haemi' cultivar. This parental strain will be used for reference genome sequence analysis.

Changes in chemical characteristics of cellulase-treated wheat germ extract (효소 처리 밀 배아 추출물의 화학적 특성 변화)

  • Lee, Jae-Kang;Jang, Davin;Kang, Dongwoo;Lee, Jeonghoon;Kum, Hyeim;Choi, Yonghyoun;Kang, Hee;Choi, Yong-Seok;Kim, Dae-Ok
    • Korean Journal of Food Science and Technology
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    • v.51 no.2
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    • pp.97-102
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    • 2019
  • Wheat germ, which is rich in nutrients and phytochemicals, is a by-product during the milling process of wheat kernel. In this study, we aimed to increase the amount of bioactive 2,6-dimethoxy-1,4-benzoquinone (2,6-DMBQ) in wheat germ using the cell-wall-degrading enzyme cellulase (Celluclast 1.5L). The amounts of organic acids, free sugars, and 2,6-DMBQ in wheat germ treated with Celluclast 1.5L were evaluated at various reaction times and temperatures. The results of reversed-phase high-performance liquid chromatography of Celluclast 1.5L-treated wheat germ revealed 2,6-DMBQ, four organic acids (tartaric, acetic, lactic, and succinic acids), and three free sugars (sucrose, fructose, and glucose). As reaction time and temperature of the mixture of wheat germ and Celluclast 1.5L increased, the contents of four organic acids, glucose, fructose, and 2,6-DMBQ increased, but that of sucrose decreased. Taken together, these results suggest that Celluclast 1.5L-treated wheat germ containing increased amounts of 2,6-DMBQ serves as a source of functional ingredients in food industry.

Study on the Genetic Characteristics of Waterlogging Tolerant Pepper (Capsicum annuum L.) for Breeding Tolerant Varieties against Flooding Stress (내습성 고추 품종 육성을 위한 선발계통의 유전적 특성 구명)

  • Yang, Eun Young;Chae, Soo-Young;Hong, Jong-Pil;Lee, Hye-Eun;Park, Eun Joon;Moon, Ji-hye;Park, Tae-Sung;Roh, Mi-Young;Kim, Ok Rye;Kim, Sang Gyu;Kim, Dae Young;Lee, Sun Yi;Cho, Myeong Cheoul
    • Journal of Life Science
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    • v.27 no.10
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    • pp.1111-1120
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    • 2017
  • This study was conducted to select pepper lines that were tolerant to excessive water injury among the pepper germplasm and investigate the genetic characteristics of those lines to contribute to the breeding of pepper cultivars with stable productivity in abnormal weather. Each of the tolerant and susceptible lines went through immersion treatment, and differentially expressed genes between them were analyzed. The tolerant line showed increased expression of the CA02g26670 gene, which is involved in the CONSTANS protein pathway and regulation of flowering by day length, but it exhibited decreased expressions of CA01g21450, CA01g22480, CA01g34470, CA02g00370 and CA02g00380. The susceptible line showed increased gene expressions of CA02g09720, CA02g21290, CA03g16520, CA07g 02110, and CA12g17910, which are involved in the inhibition of proteolytic enzyme activity, DNA binding, inhibition of cell wall-degrading enzyme, and inhibition of nodulin, respectively. Meanwhile the expressions of CA02g02820, CA03g21390, CA06g17700 and CA07g18230 decreased in the susceptible line, in relation to calcium-ion binding, high temperature, synthesis of phosphocholine and cold stress, respectively. The expressions of genes related to apoptosis and peroxidase increased, while that of CA02g16990, which functions as a nucleoside transporter, decreased in both the tolerant and susceptible lines. Based on the different gene expressions between the tolerant and susceptible lines, further studies are needed on breeding abiotic stress-tolerant lines.