• 제목/요약/키워드: cell wall degrading enzymes

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다양한 다당류를 분해하는 세균 Microbulbifer agarilyticus GP101의 완전한 유전체 서열 (Complete genome sequence of Microbulbifer agarilyticus GP101 possessing genes coding for diverse polysaccharide-degrading enzymes)

  • 정재준;배승섭;정다운;백경화
    • 미생물학회지
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    • 제54권3호
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    • pp.299-301
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    • 2018
  • Microbulbifer agarilyticus GP101은 소라(Turbo cornutus)의 내장에서 분리되었으며 해조류 유래 다당류인 한천, 알긴산, ${\kappa}$-카라기난을 분해하는 특징이 있다. GP101 균주의 유전체는 4,255,625 bp 크기로 3,458개의 코딩 서열을 포함하며 55.4%의 GC 함량을 가진다. BLASTP 분석 결과 7개의 agarase, 5개의 alginate lyase, 10개의 glucanase, 4개의 chitinase, 2개의 xylanases, 1개의 ${\kappa}$-carrageenase, 1개의 laminarinase의 존재를 확인하였다. M. agarilyticus GP101의 유전체 정보는 다당류의 생물전환 공정에 이용할 수 있는 유전 정보를 제공할 수 있을 것이다.

Molecular characterization of yeast Snf1 homologue (sucrose non-fermenting gene) from Magnaporthe grisea

  • Yi, Mi-Hwa;Jeong, Jae-Gyu;Kim, Bong-Gyu;Ahn, Joong-Hoon;Lee, Yong-Hwan
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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    • pp.84.2-85
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    • 2003
  • Magnaporthee grisea causes the devastating blast disease of rice. Entensive research has been conducted on infection mechanisms, particularly on appressorium formation and penetration, of this fungus during the last decade. However, the role(s) of cell-wall-degrading enzymes (CWDEs) on pathogenesis is not clearly demonstrated at molecular level. Many CWDES in plant pathogenic fungi including M. grisea are redundant; that is, there are multiple genes encoding enzymes with a similar or overlapping spectrum of activities. It is laborious to isolate all of the genes encoding related enzymes and to construct mutants lacking all 9f them. Thus, we considered alternative strategies to address the role of CWDEs in pathogenesis. Since expression of CWDE genes Is repressed by a simple sugar, as the first step, we cloned a Snfl (sucrose non-fermenting) gene (MgSnf1) from M. grisea. The predicted amino acid sequence showed a high identity with other Snf1 genes from various fungi. To elucidate molecular function of MgSnf1, a transformant lacking MgSnf1 was created by targeted gene replacement. En glucose, sucrose, and xylan the MgSnf1 mutant grew normally but in pectin and complex media, it grew slower than wild type. Expression of various CWDEs in MgSnf1 mutant was investigated and found that expression of some CWDEs is repressed. However, no significant difference was observed in conidial germination, appressorium formation, and pathogenicity in MgSnf1 mutant. However, MgSnf1 functionally complemented a yeast MgSnf1 mutant. These results suggest that MgSnf1 is involved in regulation of CWDEs and MgSnf1 is dispensable in pathogenicity of M. grisea.

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먹물버섯 Coprinellus congregatus에서 분열자를 사용한 형질전환 (Genetic Transformation of a Mushroom Forming Fungus Coprinellus congregatus to an Antibiotic Resistance Using Oidia Instead of Protoplast Generation)

  • 박남미;김동식;최형태
    • 미생물학회지
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    • 제42권1호
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    • pp.59-61
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    • 2006
  • 먹물버섯의 하나인 Coprinellus congregatus를 대상으로 유전자의 도입을 위한 형질전환실험에서 원형질체를 생성하지 않고 분열자 (oidium)를 사용하는 방법을 확립하였다. 분열자는 20일 이상된 CKMM 한천배지에서 생성되며, 이를 밤샘배양으로 발아를 촉진시킨 상태에서 전기천공방법으로 항생물질 basta에 대한 형질전환을 수행한 결과 10-20 형질전환체/${\mu}g$ DNA의 수율로 형질전환체를 확보할 수 있었다. 이 형질전환체들은 도입된 벡터가 염색체 상에 삽입되어 유전적으로 안정된 상태를 유지하였다.

Genetic Diversity of Pectobacterium carotovorum subsp. brasiliensis Isolated in Korea

  • Lee, Dong Hwan;Kim, Jin-Beom;Lim, Jeong-A;Han, Sang-Wook;Heu, Sunggi
    • The Plant Pathology Journal
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    • 제30권2호
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    • pp.117-124
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    • 2014
  • The plant pathogenic bacterial genus Pectobacteirum consists of heterogeneous strains. The P. carotovorum species is a complex strain showing divergent characteristics, and a new subspecies named P. carotovorum subsp. brasiliensis has been identified recently. In this paper, we re-identified the P. carotovorum subsp. brasiliensis isolates from those classified under the subspecies carotovorum and newly isolated P. carotovorum subsp. brasiliensis strains. All isolates were able to produce plant cell-wall degrading enzymes such as pectate lyase, polygalacturonase, cellulase and protease. We used genetic and biochemical methods to examine the diversity of P. carotovorum subsp. brasiliensis isolates, and found genetic diversity within the brasiliensis subsp. isolates in Korea. The restriction fragment length polymorphism analysis based on the recA gene revealed a unique pattern for the brasiliensis subspecies. The Korean brasiliensis subsp. isolates were divided into four clades based on pulsed-field gel electrophoresis. However, correlations between clades and isolated hosts or year could not be found, suggesting that diverse brasiliensis subsp. isolates existed.

키위 나무에서 분리한 Pectobacterium carotovorum subsp. actinidiae KKH3 균주의 유전체 분석 및 이를 통한 생물전환 소재로서의 가능성 연구 (The draft genome sequence of Pectobacterium carotovorum subsp. actinidiae KKH3 that infects kiwi plant and potential bioconversion applications)

  • 이동환;임정아;고영진;허성기;노은정
    • 미생물학회지
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    • 제53권4호
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    • pp.323-325
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    • 2017
  • Pectobacterium carotovorum subsp. actinidiae KKH3는 Enterobacteriaceae에 속하는 세균으로서, 키위 나무에 동고병과 같은 병을 일으키는 병원성 균주이다. 이 균주는 목본에서 분리되었으며 다양한 식물 세포벽 분해 효소를 가지고 있다. 따라서, 본 연구에서 제공하는 유전체 정보는 KKH3 균주의 병원성 기작을 이해하는 것뿐만 아니라 bioconversion 연구를 위한 토대로 활용될 수 있다.

Disruption of the metC Gene Affects Methionine Biosynthesis in Pectobacterium carotovorum subsp. carotovorum Pcc21 and Reduces Soft-Rot Disease

  • Seonmi, Yu;Jihee, Kang;Eui-Hwan, Chung;Yunho, Lee
    • The Plant Pathology Journal
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    • 제39권1호
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    • pp.62-74
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    • 2023
  • Plant pathogenic Pectobacterium species cause severe soft rot/blackleg diseases in many economically important crops worldwide. Pectobacterium utilizes plant cell wall degrading enzymes (PCWDEs) as the main virulence determinants for its pathogenicity. In this study, we screened a random mutant, M29 is a transposon insertion mutation in the metC gene encoding cystathionine β-lyase that catalyzes cystathionine to homocysteine at the penultimate step in methionine biosynthesis. M29 became a methionine auxotroph and resulted in growth defects in methionine-limited conditions. Impaired growth was restored with exogenous methionine or homocysteine rather than cystathionine. The mutant exhibited reduced soft rot symptoms in Chinese cabbages and potato tubers, maintaining activities of PCWDEs and swimming motility. The mutant was unable to proliferate in both Chinese cabbages and potato tubers. The reduced virulence was partially restored by a complemented strain or 100 µM of methionine, whereas it was fully restored by the extremely high concentration (1 mM). Our transcriptomic analysis showed that genes involved in methionine biosynthesis or transporter were downregulated in the mutant. Our results demonstrate that MetC is important for methionine biosynthesis and transporter and influences its virulence through Pcc21 multiplication in plant hosts.

Practical significance of plant growth-promoting rhizobacteria in sustainable agriculture: a review

  • Subhashini Wijeysingha;Buddhi C. Walpola;Yun-Gu Kang;Min-Ho Yoon;Taek-Keun Oh
    • 농업과학연구
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    • 제50권4호
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    • pp.759-771
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    • 2023
  • Plant growth-promoting rhizobacteria (PGPR) are naturally occurring bacteria that intensively colonize plant roots and are crucial in promoting the crop growth. These beneficial microorganisms have garnered considerable attention as potential bio-inoculants for sustainable agriculture. PGPR directly interacts with plants by providing essential nutrients through nitrogen fixation and phosphate solubilization and accelerating the accessibility of other trace elements such as Cu, Zn, and Fe. Additionally, they produce plant growth-promoting phytohormones, such as indole acetic acids (IAA), indole butyric acids (IBA), gibberellins, and cytokinins.PGPR interacts with plants indirectly by protecting them from diseases and infections by producing antibiotics, siderophores, hydrogen cyanide, and fungal cell wall-degrading enzymes such as glucanases, chitinases, and proteases. Furthermore, PGPR protects plants against abiotic stresses such as drought and salinity by producing 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase and modulating plant stress markers. Bacteria belonging to genera such as Bacillus, Pseudomonas, Burkholderia, Pantoa, and Enterobacter exhibit multiple plant growth-promoting traits, that can enhance plant growth directly, indirectly, or through synergetic effects. This comprehensive review emphasizes how PGPR influences plant growth promotion and presents promising prospects for its application in sustainable agriculture.

세포벽 분해 효소 처리 및 Saccharomyces cerevisiae 발효가 녹차 잎의 항산화 및 항균 활성에 미치는 영향 (Influence of cell-wall degrading enzyme treatment and Saccharomyces cerevisiae fermentation on the antioxidant and antibacterial activities of green tea leaf)

  • 임동욱;이가양;정민정;김병목;전준영
    • 한국식품저장유통학회지
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    • 제30권6호
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    • pp.1043-1055
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    • 2023
  • 본 연구에서는 항산화와 항균 활성 등이 우수한 녹차 잎을 대상으로 관련 물질 회수에 적합한 추출 용매의 농도 설정과 세포벽 분해 효소 처리 및 효모 발효가 활성을 높이는데 도움을 줄 수 있는지 살펴보았다. 입도 약 4-10 ㎛의 녹차 분말을 사용하여 추출물을 제조할 때, 다양한 에탄올 농도 가운데 50% 에탄올을 추출 용매로 하고 가열 처리 (121℃, 15 min)를 진행할 경우 높은 수율과 DPPH 라디칼 소거능 및 항균 활성을 나타내었다. 이 추출물은 B. cereus, B. licheniformis, S. aureus subsp. aureus 및 A. hydrophila subsp. hydrophila 에서 항균 활성을 나타내었다. 녹차 잎 분말에 효소 처리 및 효모 발효 진행 시 최종 녹차 잎 추출물의 항산화와 항균 활성에 미치는 영향을 조사하기 위해, 효소 처리에는 cellulase와 pectinase를 혼합(2.5% + 2.5%)하여 사용하였고, 효모 발효에는 S. cerevisiae 가 사용되었다. 녹차 잎을 효소 처리할 경우 추출물의 수율은 증가되었으나, 50% 에탄올 추출물(대조구)에 비해 항산화와 항균 활성은 유의적으로 감소되었다(p<0.05). 그에 반해 효모 발효를 단독으로 진행할 경우 최종 추출물의 수율 증가는 없었지만, 총페놀화합물과 플라보노이드 함량을 높여 항산화와 항균 활성을 높이는데 긍정적으로 작용하였다.

NADP+-Dependent Dehydrogenase SCO3486 and Cycloisomerase SCO3480: Key Enzymes for 3,6-Anhydro-ʟ-Galactose Catabolism in Streptomyces coelicolor A3(2)

  • Tsevelkhorloo, Maral;Kim, Sang Hoon;Kang, Dae-Kyung;Lee, Chang-Ro;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • 제31권5호
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    • pp.756-763
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    • 2021
  • Agarose is a linear polysaccharide composed of ᴅ-galactose and 3,6-anhydro-ʟ-galactose (AHG). It is a major component of the red algal cell wall and is gaining attention as an abundant marine biomass. However, the inability to ferment AHG is considered an obstacle in the large-scale use of agarose and could be addressed by understanding AHG catabolism in agarolytic microorganisms. Since AHG catabolism was uniquely confirmed in Vibrio sp. EJY3, a gram-negative marine bacterial species, we investigated AHG metabolism in Streptomyces coelicolor A3(2), an agarolytic gram-positive soil bacterium. Based on genomic data, the SCO3486 protein (492 amino acids) and the SCO3480 protein (361 amino acids) of S. coelicolor A3(2) showed identity with H2IFE7.1 (40% identity) encoding AHG dehydrogenase and H2IFX0.1 (42% identity) encoding 3,6-anhydro-ʟ-galactonate cycloisomerase, respectively, which are involved in the initial catabolism of AHG in Vibrio sp. EJY3. Thin layer chromatography and mass spectrometry of the bioconversion products catalyzed by recombinant SCO3486 and SCO3480 proteins, revealed that SCO3486 is an AHG dehydrogenase that oxidizes AHG to 3,6-anhydro-ʟ-galactonate, and SCO3480 is a 3,6-anhydro-ʟ-galactonate cycloisomerase that converts 3,6-anhydro-ʟ-galactonate to 2-keto-3-deoxygalactonate. SCO3486 showed maximum activity at pH 6.0 at 50℃, increased activity in the presence of iron ions, and activity against various aldehyde substrates, which is quite distinct from AHG-specific H2IFE7.1 in Vibrio sp. EJY3. Therefore, the catabolic pathway of AHG seems to be similar in most agar-degrading microorganisms, but the enzymes involved appear to be very diverse.

Pathogenicity of Bacillus Strains to Cotton Seedlings and Their Effects on Some Biochemical Components of the Infected Seedlings

  • Aly, Aly A.;El-Mahdy, Omima M.;Habeb, Marian M.;Elhakem, Abeer;Asran, Amal A.;Youssef, Maryan M.;Mohamed, Heba I.;Hanafy, Rania S.
    • The Plant Pathology Journal
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    • 제38권2호
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    • pp.90-101
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    • 2022
  • Pathogenicity of eight Bacillus strains to seedlings of four cotton cultivars was evaluated under greenhouse conditions. Each of the tested cultivars was individually treated with powdered inoculum of each bacterial strain. Untreated seeds were planted as control treatments in autoclaved soil. Effects of the tested strains on levels and activities of some biochemical components of the infected seedlings were also assayed. The biochemical components included total soluble sugars, total soluble proteins, total free amino acids, peroxidase, polyphenol oxidase, phenols, and lipid peroxidation. ANOVA showed that Bacillus strain (B) was a very highly significant source of variation in damping-off and dry weight. Cotton cultivar (V) was a nonsignificant source of variation in damping-off while it was a significant source of variation in dry weight. B × V interaction was a significant source of variation in damping-off and a nonsignificant source of variation in dry weight. Bacillus strain was the most important source of variation as it accounted for 59.36 and 64.99% of the explained (model) variation in damping-off and dry weight, respectively. The lack of significant correlation between levels and activities of the assayed biochemical components and incidence of damping-off clearly demonstrated that these biochemical components were not involved in the pathogenicity of the tested strains. Therefore, it was hypothesized that the pathogenicity of the tested strains could be due to the effect of cell wall degrading enzymes of pathogenic toxins. Based on the results of the present study, Bacillus strains should be considered in studying the etiology of cotton seedling damping-off.