• Title/Summary/Keyword: stationary phase

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Factors Influencing Preferential Utilization of RNA Polymerase Containing Sigma-38 in Stationary-Phase Gene Expression in Escherichia coli

  • Kim, Eun-Young;Shin, Min-Sang;Rhee, Joon-Haeng;Hyon E. Choy
    • Journal of Microbiology
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    • v.42 no.2
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    • pp.103-110
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    • 2004
  • In order to understand the molecular basis of selective expression of stationary-phase genes by RNA polymerase containing$\sigma$$\^$38/ (E$\sigma$$\^$38/) in Escherichia coli, we examined transcription from the stationary-phase promoters, katEP, bo1AP, hdeABP, csgBAP, and mcbP, in vivo and in vitro. Although these pro-moters are preferentially recognized in vivo by E$\sigma$$\^$38/, they are transcribed in vitro by both E$\sigma$$\^$38/ and E$\sigma$$\^$70/ containing the major exponential $\sigma$, $\sigma$$\^$70/. In the presence of high concentrations of glutamate salts, how-ever, oldy E$\sigma$$\^$38/ was able to efficiently transcribe from these promoters, which supports the concept that the promoter selectivity of $\sigma$$\^$38/-containing RNA polymerase is observed only under specific reaction con-ditions. The examination of 6S RNA, which is encoded by the ssr1 gene in vivo, showed that it reduced E$\sigma$$\^$70/ activity during the stationary phase, but this reduction of activity did not result in the elevation of E$\sigma$$\^$38/ activity. Thus, the preferential expression of stationary-phase genes by E$\sigma$$\^$38/ is unlikely the con-sequence of selective inhibition of E$\sigma$$\^$70/ by 6S RNA.

A study of in situ immobilization of lipase by using an LC column with aldehyde-silica stationary phase

  • Seo, U-Yong;Hong, Seon-Hui;Lee, Gi-Se
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.633-634
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    • 2001
  • The in situ immobilization of lipase in an LC column with aldehyde-silica stationary phase was optimized. The effects of feed composition, temperature, mobile phase pH and now rate on immobilization efficiency were investigated. Lipase activity after in situ immobilization was compared with free enzyme and batch immobilized enzyme.

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Resolution of Tocainide and Its Analogues on a Doubly Tethered N-CH3 Amide Chiral Stationary Phase Based on (+)-(18-Crown-6)-2,3,11,12-tetracarboxylic Acid

  • Lee, Kyu Jung;Tak, Kyung Mi;Hyun, Myung Ho
    • Bulletin of the Korean Chemical Society
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    • v.34 no.10
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    • pp.2978-2982
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    • 2013
  • A doubly tethered $N-CH_3$ amide chiral stationary phase (CSP 4) based on (+)-(18-crown-6)-2,3,11,12-tetracarboxylic acid was applied to the resolution of an antiarrythmic agent, tocainide, and its analogues and the chromatographic resolution results were compared with those on a singly tethered N-H amide CSP (CSP 1), a singly tethered $N-CH_3$ amide CSP (CSP 2) and a doubly tethered N-H amide CSP (CSP 3) under an identical aqueous mobile phase condition. CSP 4 was found to be generally better than other CSPs in terms of the separation factors (${\alpha}$) and resolutions (RS). The retention times of analytes denoted by the retention factors ($k_1$) on CSP 4 were quite long compared to those on other CSPs because of the improved lipophilicity of CSP 4. The long retention times of analytes on CSP 4 were successfully controlled by the addition of a small amount of ammonium acetate to aqueous mobile phase without hurting the chiral recognition efficiency. The variation of the content and type of organic and acidic modifier in aqueous mobile phase was found not to change the chiral recognition efficiency significantly.

Degradation of Trichloroethylene by a Growth-Arrested Pseudomonas putida

  • Hahm, Dae-Hyun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.3 no.1
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    • pp.11-14
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    • 1998
  • A toluene-oxidizing strain of Pseudomanas mendocina KR1 containing toluene-4-mono-oxygenase (TMO) completely degrades TCE with the addition of toluene as a co-substrate in aerobic condition. In order to construct in situ bioremediation system for TCE degradation without any growth-stimulating nutrients or toxic inducer such as toluene, we used the carbon-starvation promoter of Pseudomonas putida MK1 (Kim, Y. et al., J. bacteriol., 1995). Upon entry into the stationary phase due to the deprivation of nutrients, this promoter is strongly induced without further cell growth. The TMO gene cluster (4.5 kb) was spliced downstream of the carbon starvation promoter of Pseudomonas putida MK1, already cloned in pUC19. TMO under the carbon starvation promoter was not expressed in E. coli cells either in stationary phase or exponential phase. For TMO expression in Pseudomonas strains, tmo and carbon starvation promoter region were recloned into a modified broad-host range vector pMMB67HES which was made from pMMB67HE(8.9 kb) by deletion of tac promoter and lacIq (about 1.5 kb). Indigo was produced by TMO under the carbon starvation promoter in a Pseudomonas strain of post-exponential phase on M9 (0.2% glucose and 1mM indole) or LB. 18% of TCE was degraded in 14 hours after entering the stationary phase at the initial concentration of 6.6 ${\mu}$M in liquid phase.

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Synthesis and Evaluation of Molecularly Imprinted Polymeric Microspheres for Chloramphenicol by Aqueous Suspension Polymerization as a High Performance Liquid Chromatography Stationary Phase

  • Zhang, Yan;Lei, Jiandu
    • Bulletin of the Korean Chemical Society
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    • v.34 no.6
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    • pp.1839-1844
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    • 2013
  • Molecularly imprinted microsphere for chloramphenicol (CAP) with high adsorption capacity and excellent selectivity is prepared by aqueous suspension polymerization, in which chloramphenicol is used as template molecule and ethyl acetate as porogen. The CAP-imprinted microspheres are used as high performance liquid chromatography (HPLC) stationary phase and packed into stainless steel column ($150mm{\times}4.6mm$ i.d.) for selective separation of chloramphenicol. HPLC analysis suggests that chloramphenicol can be distinguished from not only its structural analogs but also other broad-spectrum antibiotic such as erythromycin and tetracycline. In addition, the binding experiments of CAP-imprinted microspheres are carried out in ethanol/water (1:4, V:V), the results indicate that the maximum apparent static binding capacity of molecularly imprinted microspheres is up to 66.64 mg $g^{-1}$ according to scatchard model.

CSP [Chiral Stationary phase] for SMB Chromatography (SMB 크로마토그래피에서 키랄정지상 [Chiral Stationary Phase, CSP]의 사용)

  • Kim Byung-Lip;Kim In-Ho
    • KSBB Journal
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    • v.20 no.3
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    • pp.177-182
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    • 2005
  • Simulated moving bed (SMB) chromatography was used in the petrochemical industry in the 1960s and its use has been widened to separate chiral compounds in the 1990s. Chiral stationary phase (CSP) is the key component in SMB for the separation of the chiral compounds. CSP has been developed for the analytical use in HPLC, however, its development successfully adapted for the preparative use in SMB chromatography. This review covers the present state-art technology of CSP for SMB chromatography in terms of selectivity, stability, and capacity.

Separation of Amino Acid Enantiomers by Gas Chromatography I With Optically Active N-Benzoyl-L-Valine Derivative as Stationary Phase (가스크로마토그라피에 의한 아미노산 광학이성체의 분리 I 광학활성 N-Benzoyl-L-Valine 유도체의 고정상으로의 응용)

  • 박만기;류재하;강종성
    • YAKHAK HOEJI
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    • v.29 no.6
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    • pp.375-379
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    • 1985
  • The gas chromatographic behavior of optically active N-(N-benzoyl-L-valyl)-anilide used as stationary phase is described. N-(N-benzoyl-L-valyl)-anilide has been synthesized with good yield under mild condition via Schotten-Bauman process and coated on the Chromosorb W AW (80-100mesh) for the purpose of enantiomer separation. The behavior of this compound as optically active stationary phase for the separation of the enantiomers of N-TFA-D, L-amino acid isopropyl esters has been examined with respect to the correlation between the separation factors and column temperatures. All amino acid enantiomers examined were eluted within one hour and the elution pattern showed retention times increasing in the order of alanine, valine, leucine, threonine, proline and methionine.

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Propatation and Application of a New HPLC Chiral Stationary Phase Edrived from 2-Amino-1-indanolo

  • Hyeon, Myeong Ho;Jang, Yeong Su;Min, Hye Jeong;Kim, Do Hun;Eom, Bong Ho
    • Bulletin of the Korean Chemical Society
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    • v.22 no.4
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    • pp.401-404
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    • 2001
  • A new chiral stationary phase (CSP) based on (1R,2S)-2-amino-1-indanol, which is expected to be conformationally rigid because of its cyclic nature, was prepared. The new CSP was applied in resolving various $N-acyl-\alpha-arylalkylamines$. The chromatographic resolution results on the new CSP were compared with those on the other CSP based on (1S,2R)-norephedrine, which is believed to be conformationally flexible. Comparison of the chromatographic resolution results on the two CSPs demonstrated that conformationally flexible analytes are resolved better on the conformationally rigid CSP while conformationally rigid analytes are resolved better on the conformationally flexible CSP. From these results it was concluded that conformational rigidity or flexibility of CSPs is the important factor for the chiral recognition.

Separation of Positional Isomers in a Calix[4]arene-methylsiloxane Polymer as Stationary Phase in Capillary GC

  • Seo, Jeong Gi;Kim, In Hwan;Jang, Seung Hyeon;Kim, Byeong Ok;Ryu, Jae Uk;Park, Jeong Hak
    • Bulletin of the Korean Chemical Society
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    • v.22 no.4
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    • pp.409-412
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    • 2001
  • Poly(p-tert-butyltrimethoxymonopropyloxycalix[4]arene-methylsiloxane) (TBCX-MS) has been prepared and used as a stationary phase in isothermal capillary gas chromatographic separation of some positional isomers. Retention factors (k) and separatio n factors $(\alpha)$ for the isomers were measured and compared with those on poly(p-tert-butyl-dimethoxydipropyloxycalix[4]arene-tetramethyldisiloxane) (TBCX-TMDS), poly(dimethoxydipropyloxycalix[4]arenetetramethyl-disiloxane) (CX-TMDS). Most of the isomers investigated are well resolved on TBCX-MS. Retention of all the compounds decreases on the three phases in the order, TBCX-TMDS ${\geq}$ TBCX-MS > CX-TMDS. Similar retention values on TBCX-TMDS and TBCX-MS seem to indicate that retention property of the two phases is not significantly affected by the spatial position of the calixarene moiety.