• Title/Summary/Keyword: species characterization

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Production, Purification and Characterization of a Melanin Bleaching Enzyme from Trametes velutina JS18 (Trametes velutina JS18 유래 멜라닌 탈색 효소의 생산, 정제 및 특성)

  • Jeon, Sung-Jong;Kim, Tae-Yun
    • Microbiology and Biotechnology Letters
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    • v.48 no.4
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    • pp.463-470
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    • 2020
  • The JS18 strain was isolated from an old tree forest and produced extracellular enzymes that decolorize synthetic melanin. Phylogenetic analysis, based on the internal transcribed spacer (ITS) sequence, indicate that JS18 belongs to the Trametes velutina species. JS18 demonstrated laccase activity but no manganese peroxidase or lignin peroxidase activity. Batch culture indicated that the melanin decolorization activity of JS18 strain originated from the laccase. Syringic acid and CuSO4 induced maximum laccase production, yielding 98 U/ml laccase activity after cultivation for 7 days at 25℃. T. velutina secretes an extracellular laccase in GYP medium, and this enzyme was purified using (NH4)2SO4 precipitation, Hi-trap Q Sepharose columns and gel filtration. The molecular weight of the purified enzyme was estimated to be 67 kDa using sodium dodecyl sulfate polyacrylamide gel electrophoresis. This enzyme produced 80% of its melanin decolorization activity within the first 24 h of evaluation in the presence of 1-hydroxybenzotriazole (HBT), while only about 4% of the melanin was decolorized in the absence of the mediator. The greatest decolorization was observed at 1.5 mM/l HBT, which decolorized 81% of the melanin within the first 24 h. The optimum pH and temperature for this decolorization were found to be 5.0 and 37℃, respectively. Our results suggest the possibility of applying HBT induced T. velutina JS18 laccase-catalyzed melanin decolorization.

Molecular Characterization of Vibrio parahaemolyticus Isolates from Seawater, Fish Tanks, and Distributed Fishery Products in Jeju (제주지역 해수, 수족관물, 유통수산물에서 분리된 장염비브리오균의 분자생물학적 특성)

  • Man Jae Cho;Eunok Kang;Ye-seul Heo;Eun A Koh
    • Journal of Food Hygiene and Safety
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    • v.38 no.4
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    • pp.246-254
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    • 2023
  • V. parahaemolyticus causes waterborne and foodborne disease such as acute diarrhea. In this study, V. parahaemolyticus isolates from seawater, fish tanks, and distributed fishery products in Jeju were investigated for potential toxin or species-specific genes (tdh, trh, tlh, and toxR) using RT-PCR and their genetic characteristics were analyzed using Pulsed-field gel electrophoresis (PFGE). Overall, V. parahaemolyticus of 90 strains (36.7%), including 33 strains from seawater, 8 strains from fish tanks, and 50 strains from fishery products, were isolated from 245 samples. All V. parahaemolyticus strains did not detect the tdh gene, whereas all strains detected tlh or toxR genes. In addition, trh genes were detected in 3 strains from seawater and 1 strain from fishery products. Monthly quantitative testing of seawater revealed that V. parahaemolyticus was positively correlated with water temperature. The 90 strains of V. parahemolyticus obtained in this study showed by gene homology between types, ranging from 64.0-97.3%. Among these, thirteen types showed 100% homology between genes. These results indicate that continuous monitoring is needed to facilitate food poisoning epidemiological investigations because some isolated V. parahaemolyticus strains harbored toxin genes and V. parahaemolyticus strains isolated from seawater, fish tanks, and distributed fishery products showed genetic similarity.

Breeding and characterization of a new white cultivar of Pleurotus ostreatus, 'Sena' (갓이 백색인 느타리 신품종 '세나'의 육성 및 특성)

  • Minji Oh;Min-Sik Kim;Ji-Hoon Im;Youn-Lee Oh
    • Journal of Mushroom
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    • v.21 no.3
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    • pp.179-184
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    • 2023
  • The development of automated bottle cultivation systems has facilitated the large-scale production of Pleurotus ostreatus, a commonly cultivated oyster mushroom species in South Korea. However, as the consumption of this product is decreasing and production quantities are exceeding demand, farmers are seeking various other mushroom types and cultivars. In response to this, we have developed a new oyster mushroom cultivar named 'Sena'. This high-yielding cultivar has a white pileus and excellent quality. The white oyster mushroom cultivars 'Goni' and 'Miso' were selected as parental strains from the genetic resources of the National Institute of Horticultural and Herbal Science's Mushroom Division. By crossing their monokaryons, hybrids were developed and subjected to cultivation trials and characteristic evaluations to select the superior cultivar. The optimal temperature for 'Sena' mycelial growth is 25-30℃, with inhibition occurring at temperatures above 30℃, whereas the temperature for mushroom growth is 14-18℃. The mushrooms grow in clusters, with the white pileus having a shallow funnel shape. Optimal mycelial growth occurs in malt extract agar medium. When cultivated in 1,100 cc bottles, the 'Sena' cultivar had 35 available individuals, surpassing the number 16 available from the control cultivar 'Goni'. The yield per bottle also increased by approximately 157 g, a 24% increase over the control cultivar amount. When 300 g samples of harvested mushrooms were packed and stored at 4℃ in a cold storage facility for 28 days, the weight loss rate of 'Sena' was approximately 4.22%, lower than that of 'Goni'. Moreover, the changes in pileus and stipe whiteness (measuring 6.99 and 8.33, respectively) were also lower than those of the control cultivar. Since the appearance of a white cap is crucial for quality assessment, the 'Sena' cultivar is superior to the 'Goni' cultivar in terms of both weight and quality after undergoing low-temperature storage.

Identification and Characterization of Polymorphic Microsatellite DNA Markers Using Next-generation Sequencing in Parapristipoma trilineatum (차세대 염기서열 분석법을 사용한 벤자리(Parapristipoma trilineatum)의 microsatellite 마커의 개발 및 유전학적 특성 분석)

  • Chun Mae Dong;Mi-Nan Lee;Jae Koo Noh;Jin Woo Park;Young-Ok Kim;Eun-Mi Kim
    • Journal of Life Science
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    • v.33 no.8
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    • pp.623-631
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    • 2023
  • This study was conducted to develop microsatellite markers in Parapristipoma trilineatum using next-generation sequencing. A total of 402,244,934 reads were generated on the Illumina Hiseq X Ten System, yielding 60,738,985,034 bp of sequences. The de novo assembly resulted in 1,320,995 contigs. A total of 952,326 contigs (0.016%) including 151 microsatellite loci were derived from the 1,320,995 contigs longer than 640 bp. A total of 34 primer sets were designed from the 151 microsatellite loci. As a result, 15 microsatellite loci were chosen and used for assuming population genetic parameters in the wild and farmed populations. The mean number of effective alleles was 12, ranging from 6 to 25. The observed heterozygosity (HO) and the expected heterozygosity (HE) ranged between 0.530 and 0.873, with an average of 0.750, and from 0.647 to 0.895, with an average of 0.793, respectively. According to these results, the developed set of 15 microsatellite markers is expected to be useful for the analysis of genetic characteristics in the population of P. trilineatum in Korea. There are requirements now for further genetic information, fishery resource management, breeding guidelines, support with the selection of breeds and studies on the effects of release, all of which will improve species conservation, and through future research, we aim to offer genetic foundational data with that goal.

Characterization of broad bean wilt virus 2 isolated from Perilla frutescens in Korea (국내 잎들깨에서 발생한 잠두위조바이러스2의 특성 구명)

  • Hyun-Sun Kim;Hee-Seong Byun;You-Ji Choi;Hyun-Yong Choi;Jang-Kyun Seo;Hong-Soo Choi;Bong-Choon Lee;Mikyeong Kim;Hae-Ryun Kwak
    • Korean Journal of Environmental Biology
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    • v.41 no.1
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    • pp.1-13
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    • 2023
  • Broad bean wilt virus 2 (BBWV2) is a species in the genus Fabavirus and family Secoviridae, which is transmitted by aphids and has a wide host range. The BBWV2 genome is composed of two single-stranded, positive-sense RNAs, RNA-1 and RNA-2. The representative symptoms of BBWV2 are mosaic, mottle, vein clearing, wilt, and stunting on leaves, and these symptoms cause economic damage to various crops. In 2019, Perilla fructescens leaves with mosaic and yellowing symptoms were found in Geumsan, South Korea. Reverse-transcription polymerase chain reaction (RT-PCR) was performed with specific primers for 10 reported viruses, including BBWV2, to identify the causal virus, and the results were positive for BBWV2. To characterize a BBWV2 isolate (BBWV2-GS-PF) from symptomatic P. fructescens, genetic analysis and pathogenicity tests were performed. The complete genomic sequences of RNA-1 and RNA-2 of BBWV2-GS-PF were phylogenetically distant to the previously reported BBWV2 isolates, with relatively low nucleotide sequence similarities of 76-80%. In the pathogenicity test, unlike most BBWV2 isolates with mild mosaic or mosaic symptoms in peppers, the BBWV2-GS-PF isolate showed typical ring spot symptoms. Considering these results, the BBWV2-GS-PF isolate from P. fructescens could be classified as a new strain of BBWV2.

Taxonomic characterization of novel Hymenobacter sp. B2 isolated from a freshwater environment (민물환경에서 분리된 novel Hymenobacter sp. B2의 분류학적 특성연구)

  • Young-Min Bae
    • Journal of the Korean Applied Science and Technology
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    • v.40 no.4
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    • pp.881-889
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    • 2023
  • The genus Hymenobacter, type genus of the family Hymenobacteraceae and a member of the phylum Bacteroidota includes gram-negative and red-pigmented rods. Those bacteria have been isolated from various environments of the earth. I isolated a red-pigmented, gram-negative rod from a pond in the campus of the Changwon University, Changwon, Kyeongnam and designated this bacterium as strain B2. Strain B2 was further analyzed phylogenetically and biochemically, and concluded as a member of genus Hymenobacter. BLAST search of the 16S rRNA gene sequence of strain B2 showed its homology lower than 98.7% with those sequences of the other bacteria whose 16S rRNA gene sequences have been reported. Fatty acid composition of the strain B2 was analyzed and its major fatty acids are summed feature 3(C16:1 ω7c and/or C16:1 ω6c, 22.8%), iso-C15:0 (16.2%), anteiso-C15:0(12.9%), C16:1ω5c(12.4%) and summed feature 4 (iso-C17:1 I/anteiso-C17:1)(9.5%) showing significant differences in fatty acid compositions between strain B2 and the other known Hymenobacter species. DNA sequence of 16S rRNA gene of strain B2 was deposited in genbank under accession number OQ318247.

Exploring the role and characterization of Burkholderia cepacia CD2: a promising eco-friendly microbial fertilizer isolated from long-term chemical fertilizer-free soil

  • HyunWoo Son;Justina Klingaite;Sihyun Park;Jae-Ho Shin
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.394-403
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    • 2023
  • In the pursuit of sustainable and environmentally-friendly agricultural practices, we conducted an extensive study on the rhizosphere bacteria inhabiting soils that have been devoid of chemical fertilizers for an extended period exceeding 40 years. Through this investigation, we isolated a total of 80 species of plant growth-promoting rhizosphere bacteria and assessed their potential to enhance plant growth. Among these isolates, Burkholderia cepacia CD2 displayed remarkable plant growth-promoting activity, making it an optimal candidate for further analysis. Burkholderia cepacia CD2 exhibited a range of beneficial characteristics conducive to plant growth, including phosphate solubilization, siderophore production, denitrification, nitrate utilization, and urease activity. These attributes are well-known to positively influence the growth and development of plants. To validate the taxonomic classification of the strain, 16S rRNA gene sequencing confirmed its placement within the Burkholderia genus, providing further insights into its phylogenetic relationship. To delve deeper into the potential mechanisms underlying its plant growth-promoting properties, we sought to confirm the presence of specific genes associated with plant growth promotion in CD2. To achieve this, whole genome sequencing (WGS) was performed by Plasmidsaurus Inc. (USA) utilizing Oxford Nanopore technology (Abingdon, UK). The WGS analysis of the genome of CD2 revealed the existence of a subsystem function, which is thought to be a pivotal factor contributing to improved plant growth. Based on these findings, it can be concluded that Burkholderia cepacia CD2 has the potential to serve as a microbial fertilizer, offering a sustainable alternative to chemical fertilizers.

Identification and molecular characterization of the chitinase gene, EaChi, from the midgut of the earthworm, Eisenia andrei (붉은줄지렁이 (Eisenia andrei) 중장에서 발현되는 chitinase 유전자, EaChi의 동정 및 분자생물학적 특성에 관한 연구)

  • Tak, Eun Sik;Kim, Dae hwan;Lee, Myung Sik;Ahn, Chi Hyun;Park, Soon Cheol
    • Journal of the Korea Organic Resources Recycling Association
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    • v.18 no.3
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    • pp.31-37
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    • 2010
  • Chitinases (EC 3.2.1.14) hydrolyze the ${\beta}$-1,4-linkages in chitin, the second most abundant polymer of N-acetyl-${\beta}$-D-glucosamine which is a structural component of protective biological matrices such as fungal cell walls and insect exoskeletons. The glycosyl hydrolases 18 family including chitinases is an ancient gene family widely expressed in archea, prokaryotes and eukaryotes. Since earthworms live in the soil with a lot of microbial activities and fungi are supposed to be a major component of the diet of earthworm, it has been reported that there would be appropriate immune system to protect themselves from microorganisms attacks. In this study, the novel chitinase, EaChi, from the midgut of earthworm, Eisenia andrei, were identified and characterized. To obtain full-length cDNA sequence of chitinase, RT-PCR and RACE-PCR analyses were carried out by using the previously identified EST sequence amongst cDNA library established from the midgut of E. andrei. EaChi, a partial chitinase gene, was composed of 927 nucleotides encoding 309 amino acids. By the multiple sequence alignments of amino acids with other different species, it was revealed that EaCHI is a member of glycosyl hydrolases 18 family, which has two highly conserved domains, substrate binding and catalytic domain.

Molecular Characterization of Hanwoo Glucose Transporter 4 Gene (한우 Glucose Transporter 4 유전자의 분자생물학적 해석)

  • Lee, S.M.;Jeong, Y.H.;Kim, H.M.;Park, H.Y.;Yoon, D.H.;Moon, S.J.;Chung, E.R.;Kang, M.J.
    • Journal of Animal Science and Technology
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    • v.47 no.6
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    • pp.1087-1094
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    • 2005
  • The uptake of glucose for metabolism and growth is essential to most animal cells and is mediated by glucose transport protein. In the glucose transport protein family, GLUT4 plays a key role in cellular glucose uptake stimulated by insulin in skeletal muscles and adipose tissue in rodents and human. In this studies, we reported the identification, characterization, and expression of Hanwoo GLUT4 gene. The Hanwoo GLUT4 cDNA includes a 1527 bp open reading frame encoding a protein of 509 amino acids. The GLUT4 amino acid sequences of the Hanwoo show strong conservation with the corresponding sequences reported in other species. The highest mRNA expression of GLUT4 was detected in heart and lower expression was detected in rib meat, sirloin, and colon. We confirmed the expression of GLUT4 in the subcutaneous and small intestinal adipose tissue using RT-PCR. To investigate the expression of GLUT4 in the bovine intramuscular adipose differentiation, fibroblast-like cells were isolated from the sirloin of Hanwoo bull aged 12 months by collagenase digestion of minced tissue and cultured with activators of PPAR gamma. We identified that GLUT4 mRNA expression decreased during differentiation of preadipocytes into adipocyte in Korean cattle. These results indicated that function of GLUT4 in bovine adipose tissue was different from that of mouse and human.

Characterization of Anti-Advanced Glycation End Products (AGEs) and Radical Scavenging Constituents from Ainsliaea acerifolia (단풍취의 최종당화산물 생성 저해 및 라디칼 소거 물질의 동정)

  • Jeong, Gyeng Han;Kim, Tae Hoon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.6
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    • pp.759-764
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    • 2017
  • Reactive oxygen species (ROS) and advanced glycation end products (AGEs) are valuable therapeutic targets for the regulation of diabetic complications. Activity-guided isolation of the ethylacetate (EtOAc)-soluble portion of 70% ethanolic extract from aerial parts of Ainsliaea acerifolia was performed, followed by AGE formation inhibition assay for the characterization of four dicaffeoylquinic acid derivatives of a previously known structure, methyl 3,5-di-O-caffeoyl-epi-quinate (1), 3,5-di-O-caffeoyl-epi-quinic acid (2), 4,5-di-O-caffeoyl-quinic acid (3), and methyl 4,5-di-O-caffeoyl-quinate (4). The structures of these compounds were confirmed by interpretation of nuclear magnetic resonance (NMR, $^1H-$, $^{13}C-NMR$, two-dimensional NMR) and mass spectroscopic data. Among the isolates, the major secondary metabolites, 3,5-di-O-caffeoyl-epi-quinic acid (2) and 4,5-di-O-caffeoyl-quinic acid (3) showed the most potent inhibitory effects against AGE formation with $IC_{50}$ values of $0.6{\pm}0.1{\mu}M$ and $0.4{\pm}0.1{\mu}M$, respectively. Furthermore, all isolated dicaffeoylquinic acid derivatives were evaluated for their radical scavenging activities using 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical, and compound 3 exhibited the most potent inhibitory effect in a concentration-dependent manner. This result suggests that the caffeoylquinic acid dimers isolated from A. acerifolia might be beneficial for the prevention of diabetic complications and related diseases.