• Title/Summary/Keyword: SEC/MALLS

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Characterization of Exopolysaccharides Produced by Submerged Culture of an Entomopathogenic Fungus Paecilomyces sinclairii by Using a Multi-Angle Laser Light Scattering System

  • KIM SANG-WOO;HWANG HYE-JIN;CHOI JANG-WON;YUN JONG-WON
    • Journal of Microbiology and Biotechnology
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    • v.15 no.3
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    • pp.560-567
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    • 2005
  • Three groups of exopolysaccharides (EPSs) (designated as Fr-I, Fr-II, and Fr-III) were isolated from the culture filtrates of Paecilomyces sinclairii by gel filtration chromatography on Sepharose CL-4B. Their molecular characteristics were examined by multi-angle laser light scattering (MALLS) connected online to a size exclusion chromatography (SEC) and refractive index (RI) detector system. The weight-average molar mass of Fr-I, Fr-II, and Fr-III of EPSs were determined to be $1.540{\times}10^6,\;6.302{\times}10^4$, and $9.389{\times}10^4\;g/mol$, respectively. All three EPSs showed a fairly low polydispersity indice, ranging from 1.008 to 1.059 (nearly mono disperse behavior), and showed different carbohydrates and amino acids compositions; all fractions of EPSs consisted of mainly cystine, valine, and arginine in the protein moiety, and mainly ribose, galactose, and glucose in the carbohydrate moiety. The determination of gyration radii of the EPSs in SEC/MALLS analysis revealed the molecular shape of the Fr-I to be a rod-like structure, whereas the Fr-II and Fr-III had a random-coil structure in an aqueous solution.

Structural and Molecular Characterization of Extracellular Polysaccharides Produced by a New Fungal Strain, Trichoderma erinaceum DG-312

  • JOO JI-HOON;YUN JONG-WON
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1250-1257
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    • 2005
  • Two groups of exopolysaccharides (designated as Fr-I EPS and Fr-II EPS) were isolated from the culture filtrate of new fungal strain Trichoderma erinaceum DG-312 by Sepharose CL-6B chromatography. The structures of the exopolysaccharides were investigated using gas chromatography (GC), Fourier transform-infrared (FT-IR) spectroscopy, GCMS analysis, and NMR. GC analysis indicated that Fr-I EPS was composed of mainly mannose ($78.9\%$) and galactose ($21.1\%$), whereas Fr-II EPS contained mannose ($68.4\%$), galactose ($26.2\%$), and glucose ($5.4\%$). In the anomeric region ($950-700cm_{-1}$) of the FT-IR spectrum, both EPSs exhibited obvious characteristic absorption of $810\;cm_{-1}$, indicating the existence of mannose. The spectra of $\alpha-and\;\beta$-configurations were assigned at 880 and $914\;cm_{-1}$, respectively. The results of GC-MS analyses confirmed that both EPSs were complex heteropolysaccharides with a ($1{\rightarrow}3$)-linked mannan backbone. The C-1 region that appeared in the $^{13}C-NMR$ spectra of these EPSs indicated a typical anomeric carbon signal. The Fr-I EPS showed two anomeric carbon signals at 102.6 and 99.6 ppm, whereas the Fr-II EPS displayed four anomeric carbon signals at 102.5, 99.6, 98.5, and 94.3 ppm. The molecular characteristics of the EPSs were further investigated using a size exclusion chromatography/multi-angle laser light scattering (SEC/MALLS) system. The SEC/MALLS system revealed that the average molar masses of the EPSs were $6.592{\times}10^{4}$ (Fr-I EPS) and $1.920{\times}10^{4}$ (Fr-II EPS) g/mol, and the molecular conformation of both EPSs in aqueous solution was random coils.

Kinetic Studies on Homopolymerization of $\alpha$-Methylstyrene and Sequential Block Copolymerization of Isobutylene with $\alpha$-Methylstyrene by Living/Controlled Cationic Polymerization (리빙/조절 양이온중합에 의한 알파메틸스티렌 호모중합 및 이소부틸렌과의 블록공중합에 대한 반응속도론 연구)

  • Wu, Yibo;Guo, Wenli;Li, Shuxin;Gong, Huiqing
    • Polymer(Korea)
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    • v.32 no.4
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    • pp.366-371
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    • 2008
  • The controlled/living cationic polymerization of $\alpha$-methylstyrene (${\alpha}MeSt$) and sequential block copolymerization of isobutylene (IB) with ${\alpha}MeSt$ were achieved using 2-chloro-2,4,4-trimethylpentane (TMPCl)/titanium tetrachloride ($TiCl_4$)/titanium isopropoxide ($Ti(OiPr)_4$)/2,6-ditert-butylpyridine (DtBP) initiating system in $CH_3Cl$/hexane(50/50 v/v) solvent mixture at $-80^{\circ}C$. The polymerization rate decreased with increasing $[Ti(OiPr)_4]/[TiCl_4]$ ratio in the homopolymerization of ${\alpha}MeSt$. The effects of $[Ti(OiPr)_4]/[TiCl_4]$ ratios and $PIB^+$ molecular weight on the polymerization rate and blocking efficiency were also investigated. Well-defined poly(isobutylene-b-$\alpha$-methylstyrene)s were demonstrated by $^1H$-NMR and triple detection SEC; refractive index (RI), multiangle laser light scattering (MALLS) and ultraviolet (UV) detectors. Blocking efficiencies for the poly(isobutylene-b-$\alpha$-methylstyrene)s of almost 100% were obtained when ${\alpha}MeSt$ was induced by PIB's of $M_n\;{\geq}\;41000$ at $[Ti(OiPr)_4]/[TiCl_4]=1$. Differential scanning calorimetry (DSC) of the block copolymers showed two glass transition temperatures, thereby demonstrating microphase separation.

Enzymatic Production of Amylopectin Cluster Using Cyclodextrin Glucanotransferase (Cyclodextrin Glucanotransferase를 이용한 아밀로펙틴 클러스터의 생산)

  • Lee, Hye-Won;Jeon, Hye-Yeon;Choi, Hyejeong;Shim, Jae-Hoon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.9
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    • pp.1388-1393
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    • 2014
  • To enzymatically prepare amylopectin cluster (APC), cyclodextrin glucanotransferase (CGTase I-5) and its mutant enzyme from alkalophilic Bacillus sp. I-5 were employed, after which the hydrolysis patterns of CGTase wild-type and its mutant enzyme toward amylopectin were investigated using multi-angle laser light scattering. CGTase wild-type dramatically reduced the molecular weight of waxy rice starch at the initial reaction, whereas the mutant enzyme degraded waxy rice starch relatively slowly. Based on the results, the molecular weight of one cluster of amylopectin could be about $10^4{\sim}10^5g/mol$. To determine production of cyclic glucans from amylopectin, matrix-assisted laser desorption ionization time-of-flight mass spectrometry was performed. CGTase I-5 produced various types of cyclic maltooligosaccharides from amylopectin, whereas the mutant enzyme hardly produced any.