• Title/Summary/Keyword: natural substrates

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Exploring the Nucleophilic N- and S-Glycosylation Capacity of Bacillus licheniformis YjiC Enzyme

  • Bashyal, Puspalata;Thapa, Samir Bahadur;Kim, Tae-Su;Pandey, Ramesh Prasad;Sohng, Jae Kyung
    • Journal of Microbiology and Biotechnology
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    • v.30 no.7
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    • pp.1092-1096
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    • 2020
  • YjiC, a glycosyltransferase from Bacillus licheniformis, is a well-known versatile enzyme for glycosylation of diverse substrates. Although a number of O-glycosylated products have been produced using YjiC, no report has been updated for nucleophilic N-, S-, and C- glycosylation. Here, we report the additional functional capacity of YjiC for nucleophilic N- and S- glycosylation using a broad substrate spectrum including UDP-α-D-glucose, UDP-N-acetyl glucosamine, UDP-N-acetylgalactosamine, UDP-α-D-glucuronic acid, TDP-α-L-rhamnose, TDP-α-D-viosamine, and GDP-α-L-fucose as donor and various amine and thiol groups containing natural products as acceptor substrates. The results revealed YjiC as a promiscuous enzyme for conjugating diverse sugars at amine and thiol functional groups of small molecules applicable for generating glycofunctionalized chemical diversity libraries. The glycosylated products were analyzed using HPLC and LC/MS and compared with previous reports.

Plant Terpene-Induced Expression of Multiple Aromatic Ring Hydroxylation Oxygenase Genes in Rhodococcus sp. Strain T104

  • Kim, Byung-Hyuk;Oh, Eun-Taex;Ahn, Yeong-Hee;Koh, Sung-Cheol
    • Journal of Microbiology
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    • v.41 no.4
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    • pp.349-352
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    • 2003
  • Recent studies have shown that some of the PCB (polychlorinated biphenyl)-degraders are able to effectively degrade PCB in the presence of monoterpenes, which act as inducers for the degradation pathway. Rhodococcus sp. T104, an effective PCB degrader, has been shown to induce the degradation pathway by utilizing limonenes, cymenes, carvones, and pinenes as sole carbon sources which can be found in the natural environment. Moreover, the strain T104 proved to possess three separate oxidation pathways of limonene, biphenyl, and phenol. Of these three, the limonene can also induce the biphenyl degradation pathway. In this work, we report the presence of three distinct genes for aromatic oxygenase, which are putatively involved in the degradation of aromatic substrates including biphenyl, limonene, and phenol, through PCR amplification and denaturing gradient gel electrophoresis (DGGE). The genes were differentially expressed and well induced by limonene, cymene, and plant extract A compared to biphenyl and/or glucose. This indicates that substrate specificity must be taken into account when biodegradation of the target compounds are facilitated by the plant natural substrates.

Induction of PCB degradative pathway by plant terpenoids as growth substrates or inducers

  • Jeong, Gyeong-Ja;Kim, Eung-Bin;So, Jae-Seong;Go, Seong-Cheol
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.489-492
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    • 2000
  • The eventual goal of this study is to elucidate roles of plant terpenoids (e.g., cymene, limonene and others) as natural substrates in the cometabolic biodegradation of PCBs and to develop an effective PCB bioremediation technology. The aim of this study was to examine how plant terpenoids, as natural substrates or inducers would affect the biodegradation of PCB congeners. Various PCB degraders that could grow on biphenyl and several terpenoids were tested for their PCB degradation capabilities. The PCB congener degradation activities were first monitored through resting cell assay technique that could detect degradation products of the substrate. The congener removal was also confirmed by concommitant GC analysis. The PCB degraders, Pseudononas sp. P166 and Caynebacterium sp. T104 were found to grow on both biphenyl and terpenoids ((S)-(-) limonene, p-cymene and ${\alpha}-terpinene$) whereas Arthrobacter B1B could not grow on the terpenoids as a sole carbon source. The strain B1B grown on biphenyl showed a good degradation activity for 4,4'-dichlorobiphenyl (DCBp) while strains P166 and T104 gave about 25% of B1B activity. Induction of degradation by cymene, limonene and terpine was hardly detected by the resting cell assay technique. This appeared to be due to relatively lower induction effect of these terpenoids compared with biphenyl. However, a subsequent GC analysis showed that the congener could be removed up to 30% by the resting cells of T104 grown on the terpenoids. This indicates that terpenoids, widely distributed in nature, could be utilized as both growth and/or inducer substrate for PCB biodegradation.

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Characterization of High Pressure-High Temperature Treated Gem Diamonds (고압고온 처리된 보석용 다이아몬드의 감별 연구)

  • Song, Oh-Sung
    • Journal of the Korean institute of surface engineering
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    • v.39 no.5
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    • pp.229-234
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    • 2006
  • Diamonds have been widely employed as polishing media for precise machining and noble substrates for microelectronics. The recent development of the split sphere press has led to the enhancement of low quality natural diamonds. Synthesized and treated diamonds are sometimes traded deceptively as high quality natural diamonds because it is hard to distinguish among these diamonds with conventional gemological characterization method. Therefore, we need to develop a new identification method that is non-destructive, fast, and inexpensive. We proposed using new methods of UV fluorescence and X-ray Lang topography for checking the local HPHT stress field to distinguish these diamonds from natural ones. We observe unique differences in the local stress field images in treated diamonds using UV fluorescence and Lang topography characterization. Our result implies that our proposed methods may be appropriate for identification of the treated diamonds.

A REVIEW OF THE MICROBIAL DIGESTION OF FEED PARTICLES IN THE RUMEN

  • McAllister, T.A.;Bae, H.D.;Yanke, L.J.;Cheng, K.J.;Ha, J.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.7 no.3
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    • pp.303-316
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    • 1994
  • Microbial digestion of feed in the rumen involves a sequential attack culminating in the formation of fermentation products and microbial cells that can be utilized by the host animal. Most feeds are protected by a cuticular layer which is in effect a microbial barrier that must be penetrated or circumvented for digestion to proceed. Microorganisms gain access to digestible inner plant tissues through damage to the cuticle, or via natural cell openings (e.g., stomata) and commence digestion from within the feed particles. Primary colonizing bacteria adhere to specific substrates, divide to form sister cells and the resultant microcolonies release soluble substrates which attract additional microorganisms to the digestion site. These newly attracted microorganisms associate with primary colonizers to form complex multi-species consortia. Within the consortia, microorganisms combine their metabolic activities to produce the diversity of enzymes required to digest complex substrates (e.g., cellulose, starch, protein) which comprise plant tissues. Feed characteristics that inhibit the microbial processes of penetration, colonization and consortia formation can have a profound effect on the rate and extent of feed digestion in the rumen. Strategies such as feed processing or plant breeding which are aimed at manipulating feed digestion must be based on an understanding of these basic microbial processes and their concerted roles in feed digestion in the rumen.

Purification and characterization of An Extracellular Serine Protease from Bacillus sp. strain KUN-17 (Bacillus sp. KUN-17 균주가 생산하는 균체외 Serine Protease의 정제 및 특성)

  • 황세영
    • Microbiology and Biotechnology Letters
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    • v.23 no.1
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    • pp.53-59
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    • 1995
  • A protease isolated and purified 51 fold from the culture filtrate of a soil bacterium, Bacillus sp. KUN-17, which was appeared to be a monomeric protein with molecular weight of 38, 000 daltons, was suggested to be involved in the serine (-alkaline) protease (E.C 3.4.21.14) since its activity was selectively inhibited by phenylmethylsulfonyl fluoride (PMSF) and required 40$\circ$C and pH 10.5 for optimal condition. The half-life of the enzyme activity was 1 hr at 55$\circ$C, and the activity was maintained even under high concentrations of SDS or urea. The enzyme was indicated to perform random proteolysis from the fact that most of the chromogenic substrates employed were hydrolyzed by the enzyme. The affinity of the enzyme for natural proteins was approximately 10-times higher than ester compounds, and both substrates showed mutual inhibitory effect competitively for the enzyme activity.

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Inhibition of Tumor Invasion and Metastasis by Calcium Spirulan(Ca-SP), a Novel Sulfated Polysaccharide Derived from a Blue-Green Alga Spirulina Platensis

  • Saiki, Ikuo;Murata, Jun;Fujii, Hideki;Kato, Toshimitsu
    • Nutritional Sciences
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    • v.7 no.3
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    • pp.144-150
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    • 2004
  • We have investigated the effect of calcium spirulan(Ca-SP) isolated from a blue-green alga Spirulina platensis, which is a sulfated polysaccharide chelating calcium and mainly composed of rhamnose and fructose, on invasion of both B16- BL6 melanoma cells, Colon 26 carcinoma and HT-1080 fibrosarcoma cells through reconstituted basement membrane (Matrigel). Ca-SP significantly inhibited the invasion of these tumor cells through Matrigel/fibronectin-coated filters in a concentration-dependent manner. Ca-SP also inhibited the haptotactic migration of tumor cells to laminin, but it had no inhibitory effect on tumor cell migration to fibronectin-coated filters. Ca-SP prevented the adhesion of B16-BL6 cells to Matrigel- and laminin-substrates but did not affect the adhesion to fibronectin. The pretreatment of tumor cells with Ca-SP inhibited the adhesion to laminin in a concentration-dependent fashion, while the pretreatment of laminin-substrates did not. Ca-SP had no effect on the production and activation of type IV collagenase in gelatin zymography. In contraset, Ca-SP significantly inhibited degradation of heparan sulfate by purified heparanase. The experimental lung metastasis was significantly reduced by co-injection of B16-BL6 cells with Ca-SP in a dose-dependent manner. Seven intermittent ⅰ.ⅴ. injection of 100$\mu\textrm{g}$ of Ca-SP caused a marked decrease of lung tumor colonization of B16-BL6 cells in a spontaneous lung metastasis model. These results suggest that Ca-SP, a novel sulfated polysaccharide, could reduce the lung colonization of B16-BL6 melanoma cells in experimental metastasis model, by inhibiting the tumor invasion of basement membrane Matrigel, probably through the prevention of the adhesion and migration of tumor cells to laminin-substrate and of the heparanase activity.

A Study on the Physical Properties of Cromwell-dyed Fabrics (자초염색 직물의 물성 연구)

  • 최인려;최정임
    • Journal of the Korea Fashion and Costume Design Association
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    • v.4 no.1
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    • pp.99-110
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    • 2002
  • This study is performed to investigate the effect of the mordant and chitosan on the colorfastness to laundering and physical properties of the mordanted, chitosan treated and natural - dyed cotton, silk and acrylics substrates. Natural dyes are extracted from Gromwell by boiled water. Three different compounds of Al, Cu, Sn and Chitosan are used as mordanting agents. The result of this study is summarized as follows: 1. Color of the fabrics dyed with Gromwell changes redder, bluer and darker after chitosan treated and mordanting. 2. After washing, the color of natural dyes changes more light and gray, Chitosan and Cu mordanting gives better colorfastness in washing than any others. 3. All chitosan treated fabrics improve air permeability. 4. In the chitosan treated fabrics, a half life of the static electricity is shown good result.

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Lipolytic Properties of Candida Cylindracea Lipase Toward Triacylglycerols with Different Fatty Acyl Chains

  • Park, En-Suk;Yang, Chul-Hak;Park, Myung-Un
    • Korean Journal of Microbiology
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    • v.30 no.3
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    • pp.171-186
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    • 1992
  • Lipolytic characteristics of candia cylindracea lipase was studied by various triacylglycerols with different fatty acyl chains as substrate. The substrate was emulsified with gum arabic and the rate of hydrolysis was determined by pH stat method. The effects of gum concentration, pH, temperature, and $Ca^{2+}$ ion on the enzyme activities were examined. The results show that the effect of these factors are markedly depending on the structurla nature of substrates. The triolein was the best substrate among tested. Present study demonstrates that for characterization of lipolytic enzymes, it is critically important to select proper substrate and activator.r.

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Purification of YPTP1 with Immobilized Phosphonomethylphenylalanine-Containing Peptide as an Affinity Ligand

  • Han, Jun-Pil;Kwon, Mi-Yun;Cho, Hyeong-Jin
    • BMB Reports
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    • v.31 no.2
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    • pp.135-138
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    • 1998
  • A previous study on a yeast protein tyrosine phosphatase, YPTP1, using synthetic phosphotyrosine-containing peptides with various sequences as substrates revealed that DADEpYDA exhibits high affinity ($K_m=4{\mu}M$) toward the enzyme. A modified version of this peptide, GDADEpmFDA, immobilized on a resin, was used in this study as an affinity ligand for the purification of YPTP1. Phosphonomethyl-phenylalanine (pmF) was used as a nonhydrolyzable analog of the phosphotyrosine (pY) residue, with properties similar to pY. A protected form of pmF, $Fmoc-pmF(^{t}Bu)_{2}-OH$, was chemically synthesized and introduced during solid-phase peptide sythesis. YPTP1 was onrexpressed in an E. coli strain carrying a plasmid pT7-7-ptpl. Affinity chromatography of the crude lysate afforded PTPI (39 kDa) of about 50% purity.

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