• Title/Summary/Keyword: isolation of enzymes

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Isolation and Characteristics of Trichoderma harzianum FJI Producing Cellulases and Xylanase

  • Kim, Kyoung-Cheol;Yoo, Seung-Soo;Oh, Young-A;Kim, Seong-Jun
    • Journal of Microbiology and Biotechnology
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    • v.13 no.1
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    • pp.1-8
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    • 2003
  • Strain FJI, a filamentous fungus isolated from rotten wood, showed high ability to hydrolyze cellulosic materials. To identify the strain FJI, ITS sequencing analysis and morphological observation were performed. The strain FJI was identified as Trichoderma harzianum. The strain produced a large amount of CMCase, xylanase, ${\beta}-glucosidase$, and avicelase. Optimal culture conditions for the production of the enzymes, such as pH, temperature, and inoculation concentration, were initial pH 6.0-7.0,$25-30^{\circ}C$, and $10^4$ ea-spores/ml in Mandel's medium, respectively. T.hanzianum FJI utilized various cellulosic materials and organic nitrogen sources to produce cellulases and xylanase, and also considerably a crystalline and/or insoluble material like Avicel and rice straw. The highest levels of CMCase and xylanase were 41.2 and 65.6 U/ml in 7 days of cultivation using 2.5% of carbon source (Avicel+CMC) and 0.5% of nitrogen source (peptone), respectively.

Isolation and Characterization of an Antifungal and Plant Growth-Promoting Microbe

  • Park, Se Won;Yang, Hee-Jong;Seo, Ji Won;Kim, Jinwon;Jeong, Su-ji;Ha, Gwangsu;Ryu, Myeong Seon;Yang, Hee Gun;Jeong, Do-Youn;Lee, Hyang Burm
    • The Korean Journal of Mycology
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    • v.49 no.4
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    • pp.441-454
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    • 2021
  • Fungal diseases including anthracnose, stem rot, blight, wilting, and root rot of crops are caused by phytopathogens such as Colletotrichum species, Sclerotinia sclerotiorum, Phytophthora species, and Fusarium oxysporum and F. solani which threaten the production of chili pepper. In this study, to identify biological control agents (BCAs) of phytopathogenic fungi, potentially useful Bacillus species were isolated from the field soils. We screened out five Bacillus strains with antagonistic capacity that are efficiently inhibiting the growth of phytopathogenic fungi. Bacillus species were characterized by the production of extracellular enzymes, siderophores, and indole-3-acetic acid (IAA). Furthermore, the influence of bacterial strains on the plant growth promoting activity and seedling vigor index were assessed using Brassica juncea as a model plant. Inoculation with Bacillus subtilis SRCM 121379 significantly increased the length of B. juncea shoots and roots by 45.6% and 52.0%, respectively. Among the bacterial isolates, Bacillus subtilis SRCM 121379 showed the superior enzyme activities, antagonistic capacity and plant growth promoting effects. Based on the experimental results, Bacillus subtilis SRCM 121379 (GenBank accession no. NR027552) was finally selected as a BCA candidate.

Plant Regeneration of B.juncea Through Plant Tissue and Protoplast Culture

  • Lian, Yu-Ji;Lim, Hak-Tae
    • Journal of Plant Biotechnology
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    • v.3 no.1
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    • pp.27-31
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    • 2001
  • New types of cytoplasmic male sterility in Brassica species would be very useful for the production of F$_1$, hybrid seeds. Leaves and stems of rapid cycling stock of B.juncea (CrGC4-3) containing Anand CMS were used as experimental materials for plant regeneration from protoplast culture. Very high plant regeneration rate (85%) was found in the Kao & Michayluk medium supplemented with 2 mg/L zeatin, 0.5 mg/L BAP, and 1 mg/L NAA when only leaf, not stem, segments were cultured. Protoplasts were isolated from leaves using mixtures of enzymes (1% Cellulycin, 0.5% Macerozyme) in 0.4 M mannitol and 50 mM $CaCl_2$.$2H_2$O. Mcrocalli induced from protoplasts were transferred to the shoot regeneration medium containing 2 mg/L BAP, 2 mg/L zeatin, and 0.5 mg/L NAA. After 60 days of initial protoplast culture, regenerated plantlets were obtained, acclimatized, transplanted into the pots, and grown up to the flowering stage.

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Isolation and Purification of Protoplasts from PorPhyra tenera Thalli (김엽체에서의 원형질체분리와 정제)

  • SONG Sang Ho;CHUNG Gyu Hwa
    • Journal of Aquaculture
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    • v.1 no.1
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    • pp.103-108
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    • 1988
  • Protoplasts were isolated enzymatically from thalli of Porphyra tenera in relation to its utilization in breeding. As the concentration of the enzymes and incubation period increased, so did the yield of protoplasts. When a 250 mg fresh weight of the thalli was incubated in the $12\%$ abalone digestive enzyme mixture at $22^{\circ}C$ for 3 hours, about $2.5\times10^6$ protoplasts were released. The size of protoplasts ranged between 9$\mu$m and 25$\mu$m, with an average of 14.5$\mu$m.

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Isolation of Rhizopus sp. R2 Producing Protopectinase and Optimum Condition ofr Pre-paring Single Cells from Potato Tissues (Protopectinase 생산균주, Rhizopus sp. R2의 분리 및 감자조직의 단세포화를 위한 최적조건)

  • 이승철;고보성;김향미;김기운;황용일
    • Korean Journal of Microbiology
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    • v.33 no.2
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    • pp.131-135
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    • 1997
  • Several microorganisms capable of producing protopectinase, which catalyzes dissociation of plant tissue to single cells, were isolated from soils. Crude enzymes prepared from culture supernatants of the strains released potato cells as a single cell from potato tissues. One of the isolated strains showing higher activity of protopectinase was selected and identified as Rhizopus sp. from the morphological characteristics. For preparing single cells from potato tissues, optimum enzyme activity of protopectinase, which was prepared from culture filtrate of Rhizopus sp. R2, was obescrved at $40^{\circ}C$ and pH 4.

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Isolation, Cloning and Co-Expression of Lipase and Foldase Genes of Burkholderia territorii GP3 from Mount Papandayan Soil

  • Putra, Ludwinardo;Natadiputri, Griselda Herman;Meryandini, Anja;Suwanto, Antonius
    • Journal of Microbiology and Biotechnology
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    • v.29 no.6
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    • pp.944-951
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    • 2019
  • Lipases are industrial enzymes that catalyze both triglyceride hydrolysis and ester synthesis. The overexpression of lipase genes is considered one of the best approaches to increase the enzymatic production for industrial applications. Subfamily I.2. lipases require a chaperone or foldase in order to become a fully-activated enzyme. The goal of this research was to isolate, clone, and co-express genes that encode lipase and foldase from Burkholderia territorii GP3, a lipolytic bacterial isolate obtained from Mount Papandayan soil via growth on Soil Extract Rhodamine Agar. Genes that encode for lipase (lipBT) and foldase (lifBT) were successfully cloned from this isolate and co-expressed in the E. coli BL21 background. The highest expression was shown in E. coli BL21 (DE3) pLysS, using pET15b expression vector. LipBT was particulary unique as it showed highest activity with optimum temperature of $80^{\circ}C$ at pH 11.0. The optimum substrate for enzyme activity was $C_{10}$, which is highly stable in methanol solvent. The enzyme was strongly activated by $Ca^{2+}$, $Mg^{2+}$, and strongly inhibited by $Fe^{2+}$ and $Zn^{2+}$. In addition, the enzyme was stable and compatible in non-ionic surfactant, and was strongly incompatible in ionic surfactant.

Characterization of NAD-Dependent Formate Dehydrogenase from Trametes versicolor Using a Cell-Free Protein Expression System

  • LEE, Su-Yeon;JANG, Seokyoon;LEE, Soo-Min
    • Journal of the Korean Wood Science and Technology
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    • v.50 no.3
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    • pp.159-166
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    • 2022
  • CO2 emissions are the primary reason for global warming; hence, biological and chemical technologies for converting CO2 into useful compounds are being actively studied. Biological methods using enzymes can convert CO2 under mild conditions. Formate dehydrogenase (FDH) is a representative CO2 conversion enzyme. Its function was revealed after isolation from bacteria, yeast, and plants. In this study, we evaluated the CO2 conversion potential of FDH isolated from wood-rotting fungi. After isolating the FDH gene (TvFDH) from Trametes versicolor, we cloned the full-length FDH from T. versicolor and expressed it in a cell-free expression system. The gene encoding TvFDH was identified as 1,200 bp open reading frame (ORF) and the expected molecular weight of the protein was approximately 42 kDa. Overexpression of the recombinant crude protein including TvFDH was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Enzyme activities and metabolite analyses confirmed the efficiency of TvFDH for CO2 reduction.

Isolation, Characterization, and Metabolic Profiling of Ceratorhiza hydrophila from the Aquatic Plant Myriophyllum spicatum

  • Elsaba, Yasmin M.;Boroujerdi, Arezue;Abdelsalam, Asmaa
    • Mycobiology
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    • v.50 no.2
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    • pp.110-120
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    • 2022
  • The goal of the present study was to investigate the antibacterial properties, enzyme production, and metabolic profiling of a new Ceratorhiza hydrophila strain isolated from the submerged aquatic plant Myriophyllum spicatum. Furthermore, the fungus' morphological characterization and DNA sequencing have been described. The fungus has been identified and submitted to the GenBank as Ceratorhiza hydrophila isolate EG19 and the fungus ID is MK387081. The enzyme analyses showed its ability to produce protease and cellulase enzymes. According to the CSLI standard, the ethyl acetate extract of C. hydrophila showed intermediate antibacterial activity against Streptococcus pneumonia, Micrococcus luteus, and Staphylococcus aureus. Metabolic profiling has been carried out using 700 MHz NMR spectroscopy. Based on the 1H and 1H-13C heteronuclear single quantum coherence (HSQC) NMR data and NMR databases, 23 compounds have been identified. The identified metabolites include 31% amino acids, 9% sugars, 9% amines, 4% sugar alcohols, and 4% alkaloids. This is the first report for the metabolic characterization of C. hydrophila, which gave preliminary information about the fungus. It is expected that our findings not only will pave the way to other perspectives in enormous applications using C. hydrophila as a new promising source of antimicrobial agents and essential metabolites, but also it will be valuable in the classification and chemotaxonomy of the species.

Isolation of a Lignolytic Bacterium for Degradation and Utilization of Lignocellulose (Lignocellulose의 분해 및 이용을 위한 Lignin 분해 세균의 분리)

  • 김용균;김한수;김근기;손홍주;이영근
    • Journal of Life Science
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    • v.12 no.4
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    • pp.392-398
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    • 2002
  • 38 strains were isolated in order to utilize lignin degrading ability from soil and compost. A organism having high lignin degrading ability of the isolated strains determined morphologcal and biochemical characteristics. Enrichment technique yielded a lignin degrading bacterium characterized as Pseudomonas sp. LC-2. This strain was able to degrade lignin which are the true representatives of native lignin and transform lignin to a lot of aromatic compounds as HPLC analysis of culture. By polyacrylamide gel analysis, it was determined that peroxidase consisted of three enzymes, with only one, the lignin peroxidase having high activity.

Isolation and characterization of 4-chlorophenoxyacetic acid-degrading bacteria from agricultural soils

  • Chung, Min-Jae;Shin, Se-Young;Park, Yong-Keun;Min, Kyung-Hee;Ka, Jong-Ok
    • Journal of Microbiology
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    • v.35 no.2
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    • pp.117-122
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    • 1997
  • Several dominant 4-CPA-degrading bacteria were isoalted from agricultural soils. Most of the isolates were identified as Burkholderia species by fatty acid methyl ester (FAME) analysis, but they were idstinct in chromosomal patterns obtained by PCR amplification of repetitive extragenic palindromic (REP) sequences. These strains were generally restricted in their substrate utilization capabilities. The 4-CPA degradative enzymes were idnducible by 4-CPA and some isolates appeared to mineralize 4-CPA via formation of 4-chlorophenol and 4-chlorocatechol as intermediates during its biodegradation pathway. Plasmid DNAs were not detected from most of the isoaltes and their 4-CPA genes wer on the chromosomal DAN. The 4-CPA degradation patterns in axenic cultures and natural soils varied depending on the strains and soils. The inoculation of 4-CPA degraders much improved the removal of 4-CPA from the 4-CPA treated soils.

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