• Title/Summary/Keyword: 선택적 가수분해

Search Result 95, Processing Time 0.019 seconds

Functional expression of CalB in E.coli (대장균에서의 Candida antarctica lipase B 최적 발현)

  • Kim, Hyun-Sook;Kim, Yong-Hwan
    • KSBB Journal
    • /
    • v.23 no.5
    • /
    • pp.445-448
    • /
    • 2008
  • Candida antarctica lipase B (CalB) is an efficient biocatalyst for many organic synthesis reactions. To make full use of CalB, we need effective expression system. Previously recombinant CalB was successfully expressed in the methylotropic yeast Pichia pastoris. In addition, we succeed in the functional expression of CalB in the Escherichia coli cytoplasm. This CalB expression system in E.coli has many considerable advantages in comparison with other expression systems and enables high-throughput screening of gene libraries as those derived from directed evolution experiments. To optimize E.coli system, we investigate comparing between OrigamiB (DE3) and BL21 (DE3) and observing effect of IPTG amount.

Synthesis of Novel Kojic Acid Derivative and Its Anti-pigmentation Effect (코직산 유도체의 합성과 미백효과)

  • Kim Ki Ho;Kim Ki Soo;Kim Jin Guk;Han Chang Sung;Kim Young Heui;Park Soo Nam
    • Journal of the Society of Cosmetic Scientists of Korea
    • /
    • v.30 no.3 s.47
    • /
    • pp.409-414
    • /
    • 2004
  • Kojic acid is well known for its anti-pigmentation effect with tyrosinase inhibition activity. However, kojic acid is a unstable compound. In order to improve stability, kojic acid derivative, kojic acid $6-O-2',3',4',6'-tetraacetyl-{\beta}-D-glucopy-ranoside\;(KTGP)$, was synthesized with $O-pentaacetyl-{\beta}-D-glucose$ through the regio- and stereo-selective glycosylation of 6-OH group of kojic acid. High yield $(80\%)$ was obtained by the use of Lewis acid and organic base in nonpolar solvent. Hydrolysis of KTGP with the aid of sodium methoxide in methanol afforded kojic acid $6-O-{\beta}-D-glucopyranoside$ (KGP), which was confirmed by $^1H-NMR\;and\;^{13}C-NMR$ KGP is freely soluble in water and soluble in methanol and ethanol. Inhibition activity of KGP for tyrosinase was investigated by measuring the activity of mushroom tyrosinase compared with those of ascorbic acid, kojic acid, and arbutin. The free radical-scavenger activity was determined by the 1,1-diphenyl- 2-picrylhydrazyl (DPPH) assay. In toxicity assay, KGP was much less toxic than kojic acid and arbutin, Therefore, glycosylation of kojic acid may be useful for the development of stable kojic acid derivatives.

Isomalto-oligosaccharide Production from Maltose by Intact Cells of Aureobasidium pullulaans (Aureobasidium pullulaans를 이용한 Maltose로부터 Isomalto-oligosaccharides의 생산)

  • 윤종원;노지선송주연송승구
    • KSBB Journal
    • /
    • v.9 no.2
    • /
    • pp.122-126
    • /
    • 1994
  • A new method for the production of isomalto-oligosaccharides from maltose was investigated using intact cells of Aureobasidium pullulaans which had been known to produce fructo-oligosaccharides. The cells showed transglucosylation activity producing isomalto-oligosaccharides at high concentrations of maltose, while they showed a hydrolytic activity at low concentrations of substrate when cultivated at $25^{\circ}C$. The optimum reaction conditions for the isomalto-oligosaccharide production were as follows: substrate concentration, 500g/l maltose; pH, 4.5; temperature, $65^{\circ}C$; cell dosage, 10 unit per gram substrate. Under optimized conditions, the maximum yield of isomalto-oligosaccharides achieved was around 48% (w/w). At the early period of reaction, panose was selectively produced from maltose, and thereafter isomaltotriose was synthesized by utilizing panose as a substrate when maltose consumption was discontinued.

  • PDF

Syntheses of Pyrrolo[2,1-b]thiazole Carboxanilides and Their Selective Antifungal Activities against Rice Blast and Wheat Leaf Rust (Pyrrolo[2,1-b]thiazole 카르복스아닐라이드 유도체의 합성 및 그들의 벼 도열병균과 밀 붉은녹병균에 대한 선택적인 항균활성)

  • Hahn, Hoh-Gyu;Nam, Kee-Dal;Yang, Bum-Seung;Choe, Gyeong-Ja;Cho, Kwang-Yun;Lee, Seon-Woo
    • The Korean Journal of Pesticide Science
    • /
    • v.9 no.2
    • /
    • pp.122-131
    • /
    • 2005
  • For development of new agrochemical fungicide, syntheses of pyrrolo[2,1-b]thiazole carboxanilide derivatives 9 and antifungal screening against 6 kinds of plant pathogens were carried out. Functionalization of carboxylic acid on C-2 into carboxanilide in the pyrrolo[2,1-b]thiazole resulted in new 20 candidates, in which are ${\alpha},{\beta}$-unsaturated carboxanilide and methyl groups that are in cis relationship. Treatment of acetoxy-1,4-thiazin with magnesium in refluxing methanol gave pyrrolo[2,1-b]thiazole ethyl ester 4 in higher yield than that of the previous report. Hydrolysis of this compound afforded the corresponding acid 5, which reacted with aniline derivatives in the presence of coupling reagent, DIC to give pyrrolo[2,1-b]thiazole carboxanilides 9. As the result of in vivo antifungal assay of 9 against rice blast, rice sheath blight, cucumber gray mold, tomato late blight, wheat leaf rust, and barley powdery mildew, some compounds showed selectively antifungal activities against the rice blast and wheat leaf rust.

Determination of 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THCCOOH) in human urine by solid-phase extraction and GC/MS (고체상 추출과 GC/MS를 이용한 소변 중 대마 대사체 (THCCOOH) 분석)

  • Cheong, Jae Chul;Kim, Jin Young;In, Moon Kyo;Cheong, Won Jo
    • Analytical Science and Technology
    • /
    • v.19 no.5
    • /
    • pp.441-448
    • /
    • 2006
  • 11-nor-9-carboxy-${\Delta}^9$-tetrahydrocannabinol (THCCOOH) is the major metabolite of tetrahydrocannabinol (THC) which is the primary psychoactive component of marijuana. It is also the target analyte for the discrimination marijuana use. A method using solid-phase extraction (SPE) and gas chromatography/mass spectrometry (GC/MS) was developed for the determination of THCCOOH in human urine. Urine samples (3 mL) were extracted by SPE column with a cation exchange cartridge after basic hydrolysis. The eluents were then evaporated, derivatized, and injected into the GC/MS. The limits of detection (LOD) and quantitation (LOQ) were 0.4 and 1.2 ng/mL, respectively. The response was linear with a correlation coefficient of 0.999 within the concentration range of 1.2 (LLE 1.3)~50.0 ng/mL. The precision and accuracy were stable within 1.20% and the recovery was 83.6~90.7%. The recovery of SPE method was lower than that of liquid-liquid extraction (LLE), but there were no apparent differences in LOD, LOQ, precision and accuracy between the two methods. While SPE method is used as a very effective and rapid procedure for sample pretreatment, and clean extracts, LLE method was not suitable for the extraction procedure of THCCOOH in urine. The applicability of the method was proven by analyzing a urine samples from a marijuana abusers.

Characterization of Pretreatment for Barley straw by Alkaline Solutions (염기 용매를 이용한 보릿짚의 전처리 특성)

  • Kim, Kyoung-Seob;Kim, Jun Seok
    • Korean Chemical Engineering Research
    • /
    • v.50 no.1
    • /
    • pp.18-24
    • /
    • 2012
  • Lignocellulose is difficult to hydrolyze due to the presence of lignin and the technology developed for cellulose fermentation to ethanol is not yet economically viable. However, recent advances in the extremely new field of biotechnology for the ethanol production are making it possible to use of Agriculture residual biomass, e.q., Barley straw, because of their several superior aspects as Agriculture residual biomass; low lignin, high contents of carbohydrates. Barley straw consists of 39.78% cellulose (glucose), 22.56% hemicelluloses and 19.27% lignin. Pretreatment of barley straw using NaOH pretreatment solutions concentration with 2%, temperature $85^{\circ}C$ and reaction times 1 hr were investigates. $NH_4OH$ pretreatment condition was solutions concentration with 15%, temperature $60^{\circ}C$, and reaction times 24hr were investigates. Furthermore, enzymatic saccharification using cellulose at $50^{\circ}C$, pH 4.8, 180 rpm for conversion of cellulose contained in barley straw to monomeric sugar. The pretreatment of barley straw using NaOH and $NH_4OH$ can significantly improve enzymatic saccharification of barley straw by extract more lignin and increasing its accessibility to hydrolytic enzymes. The result showed NaOH pretreatment extracted yield of lignin was 24.15%. $NH_4OH$ pretreatment extracted yield of lignin was 29.09%. Shaccharification of barley straw pretreatment by NaOH for 72hr and pH 4.8 result in maximum glucose concentration 15.39g/L (58.40%) and by $NH_4OH$ for 72hr and pH 4.8 result in maximum glucose concentration 16.01g/L (64.78%).

Horizontal Gene Transfer among Bacteria by Transformation in Soil and Aquatic Environments (토양 및 수계환경에서 Transformation에 의한 세균들간의 수평적 유전물질 전이)

  • 이건형
    • Korean Journal of Environmental Biology
    • /
    • v.18 no.2
    • /
    • pp.205-213
    • /
    • 2000
  • Laboratory studies have revealed that naturally transformable bacteria develop competence under in situ conditions. Thus, the occurrence of competent bacteria in the environment can be considered as a certainty The persistence of free DNA in natural habitats is influenced by nucleolytic degradation and protection from degradation by adsorption to minerals. Although DNA seeded into natural environment was hydrolysed at substantial rates, but was still detectable at low levels after even several weeks. Compared to the number of laboratory based studies, only a few data have been published dealing with transformation of bacteria in the field. Recently, the potential transfer of recombinant DNA (rDNA) from deliberately or accidentally released bacteria to indigenous microbes has raised biosafety issues, since the persistence of rDNA becomes independent of the survival of its original host and leads to unpredictable, long-term ecological effects. The aim of the present review is to summarise recent literature on horizontal gene transfer (HGT) by transformation among bacteria in both soil and aquatic habitat and special emphasis is placed on recent reports which have addressed HGT among bacteria in the field. [Transformation, Horizontal gene transfer (HGT), recombinant DNA (rDNA), Genetically modified microorganisms (GMMs), Biosafety]

  • PDF

Microanalytical Study of Sodium Alginate by Differential Pulse Stripping Voltammetry (시차 펄스 벗김전압전류법을 이용한 Sodium Alginate의 미량분석 연구)

  • Kang, Dae-Joon;Yun, Young-Ja
    • Journal of the Korean Chemical Society
    • /
    • v.44 no.2
    • /
    • pp.109-114
    • /
    • 2000
  • In this study, the optimum analytical condition of Differential pulse Voltammetry for the polymeric Sodium Alginate was studied and its reduction funtional groups were confirmed using Cyclic Vol-tammetry and IR spectroscopy. Optimum conditions were as followed; mercury drop size : medium size, accumulation time : 60sec, accumulation potential : -0.20V vs Ag/AgCl, scan rate : 100mV/sec, supporting electrolyte : 0.10M $NaClO_4$(pH 6.8). After polymeric Sodium Alginate was hydrolized at $100^{\circ}C$ in acidic condition, the current peaks of oligomer were compared with current peak of polymeric Sodium Alginate. In this optimum condition, calibration curve of polymer Sodium Alginate showed good linearity from 0.50ppm to 4.0ppm where as oligomeric Sodium Alginate showed good linearity from 0.05ppm to 0.24ppm.

  • PDF

Cloning and Characterization of Zebrafish Microsomal Epoxide Hydrolase Based on Bioinformatics (생물정보학을 이용한 Zebrafish Microsomal Epoxide Hydrolase 클로닝 및 특성연구)

  • Lee Eun-Yeol;Kim Hee-Sook
    • Microbiology and Biotechnology Letters
    • /
    • v.34 no.2
    • /
    • pp.129-135
    • /
    • 2006
  • A gene encoding for a putative microsomal epoxide hydrolase (mEH) of a zebrafish, Danio rerio, was cloned and characterized. The putative mEH protein of D. rerio exhibited sequence similarity with mammalian mEH and some other bacterial EHs. A structural model for the putative mEH was constructed using homology modeling based on the crystallographic templates, 1 qo7 and 1 ehy. The catalytic triad consisting of $Asp^{233}$, $Glu^{413}$, and $His^{440}$ was identified, and the characteristic features such as two tyrosine residues and oxyanion hole were found to be highly conserved. Based on bioinformatic analysis together with EH activity assay, the putative protein was annotated as mEH of D. rerio. Enantiopure styrene oxide with enantiopurity of 99%ee and yield of 33.5% was obtained from racemic styrene oxide by the enantioselective hydrolysis activity of recombinant mEH of D. rerio for 45 min.

Anti-proliferative Effects of the Isothiocyanate Sulforaphane on the Growth of Human Cervical Carcinoma HeLa Cells (Sulforaphane에 의한 HeLa 인체자궁경부함세포의 증식 억제 기전 연구)

  • Park Soung Young;Bae Song-Ja;Choi Yung Hyun
    • Journal of Life Science
    • /
    • v.15 no.3 s.70
    • /
    • pp.397-405
    • /
    • 2005
  • Sulforaphane, an isothiocyanate derived from hydrolysis of glucoraphanin in broccoli and other cruciferous vegetables, was shown to induce phase II detoxification enzymes and inhibit chemically induced mammary tumors in rodents. Recently, sulforaphane is known to induce cell cycle arrest and apoptosis in human cancer cells, however its molecular mechanisms are poorly understood. In the present study, we demonstrated that sulforaphane acted to inhibit proliferation and induce morphological changes of human cervical carcinoma HeLa cells. Treatment of HeLa cells with $10{\mu}M\;or\;15{\mu}M$ sulforaphane resulted in significant G2/M cell cycle arrest as determined by flow cytometry. Moreover, $20{\mu}M$ sulforaphane significantly induced the population of sub-G1 cells (9.83 fold of control). This anti-proliferative effect of sulforaphane was accompanied by a marked inhibition of cyclin A and cyclin-dependent kinase (Cdk)4 protein and concomitant induction of Cdc2, Cdk inhibitor p16 and p21. However, sulforaphane did not affect the levels of cyelooxygenases and telomere-regulatory gene products. Although further studies are needed, the present work suggests that sulforaphane may be a potential chemoprevetive/ chemotherapeutic agent for the treatment of human cancer cells.