• Title/Summary/Keyword: Protein solubilization

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Penicillium menonorum: A Novel Fungus to Promote Growth and Nutrient Management in Cucumber Plants

  • Babu, Anam Giridhar;Kim, Sang Woo;Yadav, Dil Raj;Hyum, Umyong;Adhikari, Mahesh;Lee, Youn Su
    • Mycobiology
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    • v.43 no.1
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    • pp.49-56
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    • 2015
  • The present study is the first report on the isolation of Penicillium menonorum from rhizosphere soil in Korea and its identification based on morphological characteristics and internal transcribed spacer gene sequence. The fungal isolate was named KNU-3 and was found to exhibit plant growth-promoting (PGP) activity through indole acetic acid (IAA) and siderophore production, as well as P solubilization. KNU-3 produced 9.7 mg/L IAA and solubilized 408 mg of $Ca_3PO_4/L$, and inoculation with the isolate significantly (p < 0.05) increased the dry biomass of cucumber roots (57%) and shoots (52%). Chlorophyll, starch, protein, and P contents were increased by 16%, 45%, 22%, and 14%, respectively, compared to plants grown in uninoculated soil. The fungus also increased soil dehydrogenase (30%) and acid phosphatase (19%) activities. These results demonstrate that the isolate KNU-3 has potential PGP attributes, and therefore it can be considered as a new fungus to enhance soil fertility and promote plant growth. Moreover, the discovery of PGP ability and traits of this fungus will open new aspects of research and investigations. In this study, plant growth promotion by P. menonorum KNU-3 is reported for the first time in Korea after its original description.

Separation of Immunoglobulin G from Colostrum by Reverse Micelles of Cationic Surfactant (양이온 계면활성제로 형성된 역미셀을 이용한 초유 Immunoglobulin G의 분리)

  • 노경현;임지영
    • Food Science of Animal Resources
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    • v.24 no.1
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    • pp.80-85
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    • 2004
  • This study was carried out to separate immunoglobulin G(IgG) from colostrum using reverse micellar extraction of cationic surfactant and to suggest suitable extraction conditions. The reconstituted colostrum powder was solubilized into a reverse micellar phase containing CDAB(cetyldimethylethyl ammonium bromide) by mixing equal volume of the aqueous and organic phase with constant stirring. The solubilization of proteins from the aqueous to the organic phase was manipulated by pH and ionic strength of the aqueous phase and concentration of surfactant in the organic phase. Based on the SDS-PAGE and densitometry, about more than 90% of initial IgG was remained in the aqueous phase after reverse micellar extraction. Although the aqueous phase contained lactoferrin and bovine serum albumin as minor components, about 93% of the total protein was IgG. The efficient extraction was achieved by the reaction of sodium phosphate buffer(pH 8) containing 50 mM KCl and organic phase containing 100 mM CDAB. The separation of IgG using reverse micellar extraction was simple, highly efficient and easy to be scaled up.

Biochemical and Molecular Characterization of High Population Density Bacteria Isolated from Sunflower

  • Goes, Kelly Campos Guerra Pinheiro De;Fisher, Maria Luisa De Castro;Cattelan, Alexandre Jose;Nogueira, Marco Antonio;Carvalho, Claudio Guilherme Portela De;Oliveira, Andre Luiz Martinez De
    • Journal of Microbiology and Biotechnology
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    • v.22 no.4
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    • pp.437-447
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    • 2012
  • Natural and beneficial associations between plants and bacteria have demonstrated potential commercial application for several agricultural crops. The sunflower has acquired increasing importance in Brazilian agribusiness owing to its agronomic characteristics such as the tolerance to edaphoclimatic variations, resistance to pests and diseases, and adaptation to the implements commonly used for maize and soybean, as well as the versatility of the products and by-products obtained from its cultivation. A study of the cultivable bacteria associated with two sunflower cultivars, using classical microbiological methods, successfully obtained isolates from different plant tissues (roots, stems, florets, and rhizosphere). Out of 57 plant-growth-promoting isolates obtained, 45 were identified at the genus level and phylogenetically positioned based on 16S rRNA gene sequencing: 42 Bacillus (B. subtilis, B. cereus, B. thuringiensis, B. pumilus, B. megaterium, and Bacillus sp.) and 3 Methylobacterium komagatae. Random amplified polymorphic DNA (RAPD) analysis showed a broad diversity among the Bacillus isolates, which clustered into 2 groups with 75% similarity and 13 subgroups with 85% similarity, suggesting that the genetic distance correlated with the source of isolation. The isolates were also analyzed for certain growth-promoting activities. Auxin synthesis was widely distributed among the isolates, with values ranging from 93.34 to 1653.37 ${\mu}M$ auxin per ${\mu}g$ of protein. The phosphate solubilization index ranged from 1.25 to 3.89, and siderophore index varied from 1.15 to 5.25. From a total of 57 isolates, 3 showed an ability to biologically fix atmospheric nitrogen, and 7 showed antagonism against the pathogen Sclerotinia sclerotiorum. The results of biochemical characterization allowed identification of potential candidates for the development of biofertilizers targeted to the sunflower crop.

Improvement of Solubilization and Anaerobic Biodegradability for Sewage Sludge Using Ultrasonic Pre-treatment (하수슬러지의 초음파 전처리를 통한 가용화 및 혐기성 생분해도 향상)

  • Lee, Chae-Young;Park, Seung-Yong
    • Journal of the Korea Organic Resources Recycling Association
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    • v.16 no.3
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    • pp.83-90
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    • 2008
  • The ultrasonic pre-treatment of sewage sludge (SS) was investigated to increase soluble organic material and to improve anaerobic biodegradability. Ultrasonic disintegration of SS increased the amount of soluble chemical oxygen demand (SCOD), protein and carbohydrate concentrations whereas particle size decreased due to the break-up of cell walls. In terms of anaerobic biodegradability, ultrasonic pre-treatment enhanced the anaerobic biodegradation of SS, leading to the methane gas production improvement. Biochemical methane potential (BMP) of SS was 211.3 ml $CH_4/gVS$ whereas BMP after ultrasonic pre-treatment was 294.3 ml $CH_4/gVS$. The improvement in BMP for SS treated with ultrasonic disintegration was as high as 40 %. This result indicated that disintegration of SS was efficient for enhancing anaerobic biodegradability.

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Characterization of the Outer Membrane-Associated 2-Furaldehyde Dehydrogenase from Klebsiella pneumoniae (Klebsiella pneumoniae 균주의 세포외막에서 분리한 2-furaldehyde dehydrogenase의 특성에 관한 연구)

  • 이준우;강사욱;하영칠;한홍의
    • Korean Journal of Microbiology
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    • v.26 no.3
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    • pp.197-206
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    • 1988
  • An outer membrane-associated 2-furaldehyde dehydrogenase, catalyzing the oxidation of 2-furaldehyde to 2-furoic acid from Klebsiella pneumoniae was purified to homogeneity and characterized. The enzyme showed its highly specific dependency on $\beta$-$NAD^{+}$. Enzyme activity was monitored during purification by using substrate 2-furaldehyde and coenzyme $\beta$-$NAD^{+}$ by means of high performance liquid chromatography. The outer membrane was successfully collected by the methods of Percoll density gradient ultracentrifugation and ultracentrifugation after preferential solubilization of the membrane with $Mg^{2+}$ and Triton X-100. The enzyme was purified by the series of procedures including extraction of outer membrane protein with EDTA and lysozume, and fractionation by column chromatography on QAE-Sephades Q-50, and subsequently Sephadex G-100. The enzume showed its optimal activity at $85^{\circ}C$, pH 9.5, and in the presence of 1.5% (vol/vol) Triton X-100. The enzyme exhibited a native molecular size of 88,000 by nondenaturing polyacrylamide gel electrophoresis and had an apparent Km of 4.72mM for 2-furaldehyde.

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Non-Ionic Surfactants Antagonize Toxicity of Potential Phenolic Endocrine-Disrupting Chemicals, Including Triclosan in Caenorhabditis elegans

  • Alfhili, Mohammad A.;Yoon, Dong Suk;Faten, Taki A.;Francis, Jocelyn A.;Cha, Dong Seok;Zhang, Baohong;Pan, Xiaoping;Lee, Myon-Hee
    • Molecules and Cells
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    • v.41 no.12
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    • pp.1052-1060
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    • 2018
  • Triclosan (TCS) is a phenolic antimicrobial chemical used in consumer products and medical devices. Evidence from in vitro and in vivo animal studies has linked TCS to numerous health problems, including allergic, cardiovascular, and neurodegenerative disease. Using Caenorhabditis elegans as a model system, we here show that short-term TCS treatment ($LC_{50}$: ~0.2 mM) significantly induced mortality in a dose-dependent manner. Notably, TCS-induced mortality was dramatically suppressed by co-treatment with non-ionic surfactants (NISs: e.g., Tween 20, Tween 80, NP-40, and Triton X-100), but not with anionic surfactants (e.g., sodium dodecyl sulfate). To identify the range of compounds susceptible to NIS inhibition, other structurally related chemical compounds were also examined. Of the compounds tested, only the toxicity of phenolic compounds (bisphenol A and benzyl 4-hydroxybenzoic acid) was significantly abrogated by NISs. Mechanistic analyses using TCS revealed that NISs appear to interfere with TCS-mediated mortality by micellar solubilization. Once internalized, the TCS-micelle complex is inefficiently exported in worms lacking PMP-3 (encoding an ATP-binding cassette (ABC) transporter) transmembrane protein, resulting in overt toxicity. Since many EDCs and surfactants are extensively used in commercial products, findings from this study provide valuable insights to devise safer pharmaceutical and nutritional preparations.

Complete genome sequence of Variovorax sp. PMC12, a plant growth-promoting bacterium conferring multiple stress resistance in plants (다양한 스트레스에 대한 식물의 내성을 유도하는 식물생육촉진 세균Variovorax sp. PMC12 균주의 유전체 염기서열)

  • Lee, Shin Ae;Kim, Hyeon Su;Kim, Yiseul;Sang, Mee Kyung;Song, Jaekyeong;Weon, Hang-Yeon
    • Korean Journal of Microbiology
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    • v.54 no.4
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    • pp.471-473
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    • 2018
  • Variovorax sp. PMC12 is a rhizobacterium isolated from tomato rhizosphere and enhanced the plant resistance to abiotic and biotic stresses. Here we present the complete genome sequence of strain PMC12. The genome is comprised of two circular chromosomes harboring 5,873,297 bp and 1,141,940 bp, respectively. A total of 6,436 protein-coding genes, 9 rRNAs, 64 tRNAs, 3 ncRNAs, and 80 pseudogenes were identified. We found genes involved in 1-aminocyclopropane-1-carboxylate (ACC) deaminase, antioxidant activity, phosphate solubilization, and biosynthesis of proline and siderophore. Those genes may be related to capability of improving plant resistance to various stresses including salinity, cold temperature, and phytopathogen.

Extraction and characterization of pepsin-soluble collagen from different mantis shrimp species

  • Hiransuchalert, Rachanimuk;Oonwiset, Nakaweerada;Imarom, Yolrawee;Chindudsadeegul, Parinya;Laongmanee, Penchan;Arnupapboon, Sukchai
    • Fisheries and Aquatic Sciences
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    • v.24 no.12
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    • pp.406-414
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    • 2021
  • The objective of this study was to investigate the yield and characteristics of collagen protein extracted from the muscle of four different species of mantis shrimp: Miyakella nepa, Harpiosquilla harpax, Erugosquilla woodmasoni, and Odontodactylus cultrifer. Mantis shrimp muscle was extracted by using a pepsin-solubilization technique, with 0.5 M acetic acid and 5% pepsin enzyme. The highest collagen yield was from M. nepa muscle (0.478 ± 0.06%), which was significantly greater (p < 0.05) than that from H. harpax, O. cultrifer, and E. woodmasoni (0.313 ± 0.03%, 0.123 ± 0.02%, and 0.015 ± 0.00%, respectively). The freeze-dried collagen appeared as thin fibers, and formed an opaque film. The pepsin-soluble collagen (PSC) from four mantis shrimp species was analyzed by gel electrophoresis. The results showed that all species of mantis shrimp contained type I collagen, consisting of β, α1, and α2 subunits with average molecular weights of 250, 145, and 118 kDa, respectively. The study of the solubility of collagen showed that, for NaCl, collagen had the highest relative solubility in 2% NaCl (80.20 ± 4.95%). In contrast, the solubility decreased at higher NaCl concentrations. However, in terms of pH, collagen had the highest relative solubility at pH 3 (91.32 ± 5.14%), and its solubility decreased at higher pH. FT-IR spectroscopy was used to compare the collagen with a model compound. Five wavenumbers in the spectrum for model collagen were identified: Amide A (3,406-3,421 cm-1), amide B (2,916-2,940 cm-1), amide I (1,639-1,640 cm-1), amide II (1,539-1,570 cm-1), and amide III (1,234-1,250 cm-1).

Regulation of Tumor Neceosis Factor-${\alpha}$ Receptors and Signal Transduction Pathways

  • Han, Hyung-Mee
    • Toxicological Research
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    • v.8 no.2
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    • pp.343-357
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    • 1992
  • Tumor necrosis factor-${\alpha}$(TNF), a polypeptide hormone secreted primarily by activated macrophages, was originally identified on the basis of its ability to cause hemorrhagic necrosis and tumor regression in vivo. Subsequently, TNF has been shown to be an important component of the host responses to infection and cancer and may mediate the wasting syndrome known as cachexia. These systemic actions of TNF are reflected in its diverse effects on target cells in vitro. TNF initiates its diverse cellular actions by binding to specific cell surface receptors. Although TNF receptors have been identified on most of animal cells, regulation of these receptors and the mechanisms which transduce TNF receptor binding into cellular responses are not well understood. Therefore, in the present study, the mechanisms how TNF receptors are being regulated and how TNF receptor binding is being transduced into cellular responses were investigated in rat liver plasma membranes (PM) and ME-180 human cervical carcinoma cell lines. $^{125}I$-TNF bound to high ($K_d=1.51{\pm}0.35nM$)affinity receptors in rat liver PM. Solubilization of PM with 1% Triton X-100 increased both high affinity (from $0.33{\pm}0.04\;to\;1.67{\pm}0.05$ pmoles/mg protein) and low affinity (from $1.92{\pm}0.16\;to\;7.57{\pm}0.50$ pmoles/mg protein) TNF binding without affecting the affinities for TNF, suggesting the presence of a large latent pool of TNF receptors. Affinity labeling of receptors whether from PM or solubilized PM resulted in cross-linking of $^{125}I$-TNF into $M_r$ 130 kDa, 90 kDa and 66kDa complexes. Thus, the properties of the latent TNF receptors were similar to those initially accessible to TNF. To determine if exposure of latent receptors is regulated by TNF, $^{125}I$-TNF binding to control and TNF-pretreated membranes were assayed. Specific binding was increased by pretreatment with TNF (P<0.05), demonstrating that hepatic PM contains latent TNF receptors whose exposure is promoted by TNF. Homologous up-regulation of TNF receptors may, in part, be responsible for sustained hepatic responsiveness during chronic exposure to TNF. As a next step, the post-receptor events induced by TNF were examined. Although the signal transduction pathways for TNF have not been delineated clearly, the actions of many other hormones are mediated by the reversible phosphorylation of specific enzymes or target proteins. The present study demonstrated that TNF induces phosphorylation of 28 kDa protein (p28). Two dimensional soidum dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) resolved the 28kDa phosphoprotein into two isoforms having pIs of 6.2 and 6.1. The pIs and relative molecular weight of p28 were consistent with those of a previously characterized mRNA cap binding protein. mRNA cap binding proteins are a class of translation initiation factors that recognize the 7-methylguanosine cap structure found on the 5' end of eukaryotic mRNAs. In vitro, these proteins are defined by their specific elution from affinity columns composed of 7-methylguanosine 5'-triphosphate($m^7$GTP)-Sepharose. Affinity purification of mRNA cap binding proteins from control and TNF treated ME-180 cells proved that TNF rapidly stimulates phosphorylation of an mRNA cap binding protein. Phosphorylation occurred in several cell types that are important in vitro models of TNF action. The mRNA cap binding protein phosphorylated in response to TNF treatment was purifice, sequenced, and identified as the proto-oncogene product eukaryotic initiation factor-4E(eIF-4E). These data show that phosphorylation of a key component of the cellular translational machinery is a common early event in the diverse cellular actions of TNF.

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Extraction of Freeze Dried Young Antler by Water and Protease (물과 단백질 가수분해 효소에 의한 동결건조 녹용의 추출)

  • 안용근
    • The Korean Journal of Food And Nutrition
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    • v.16 no.4
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    • pp.379-387
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    • 2003
  • The freeze dried young antler was extracted by water and proteases. In case of water extraction, the extraction rate was highest when it was reacted in 5% of concentration for 6 hours at 50$^{\circ}C$. The result of HPLC analysis of extract shows that high molecular peak in water extract was transformed into low molecular polk by proteases. The rate of low molecular peak was highest when bacteria protease was used, and its second highest rate was pepsin, but the effect of papain on it was low, The extraction rate of young antler reacted for 5 hours was 33.4%(absorbance 13.25 at 280nm) of bacteria protease, 22.4%(absorbance 10.06) of papain, and 30.2% (absorbance 11.34) of pepsin. The young antler was boiled for 30min and it was reacted by proteases for 5 hours at 50$^{\circ}C$. The extraction rate of it was 47,6%(absorbance 12,54) of bacteria protease, and 26,4%(absorbance 7,48) of papain, and 45.6%(absorbance 7.23) of pepsin, In protein content, water extract was 52,1%, bacteria protease extract was 37.8%, and in amino acid content, water extract was 16.3%, bacteria protease extract was 31.96%, in ash content, water extract was 8.8%, bacteria protease extract was 5.6% by dry base. In mineral content, water extract contains 3.6% of Ca, 8.6% of P, 0.01% of Mg, 1.4 % of Na, 0.02 % of F, and bacteria protease extract contains 2.5% of Ca, 11.8% of P, 0.046 % of Mg, 2.1 % of Na, 0.018 % of F by dry base.