• Title/Summary/Keyword: 가수분해반응

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Enzymatic Characterization of a Thermostable 4-α-Glucanotransferase from Thermotoga neapolitana (Thermotoga neapolitana 유래 내열성 4-알파-글루칸전이효소의 효소적 특성)

  • Choi, Kyoung-Hwa;Seo, Ja-Yeong;Kim, Ji-Eun;Cha, Jae-Ho
    • Journal of Life Science
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    • v.21 no.2
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    • pp.221-226
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    • 2011
  • The gene encoding 4-$\alpha$-glucanotransferase (mgtA) from Thermotoga neapolitana was cloned and expressed in Escherichia coli in order to investigate whether this enzyme was capable of producing cycloamylose for industrial applications. MgtA was purified to homogeneity by HiTrap Q HP and Sephacryl S-200 HR column chromatographies. The size of the enzyme as determined by SDS-PAGE was about 52 kDa, which was in good agreement with its deduced molecular mass of 51.9 kDa. The optimal temperature and pH for the activity of the 4-$\alpha$-glucanotransferase was found to be $85^{\circ}C$ and 6.5, respectively. The enzyme hydrolyzed the 1,4-$\alpha$-glucosidic bonds in oligomeric 1,4-$\alpha$-glucans and transferred oligosaccharides (maltotriose being the shortest one) to acceptor maltodextrins. However, the enzymes had no activity against pullulan, glycogen, and other di- or trioligosaccharides with rare types of $\alpha$-bond. MgtA is distinguished from 4-$\alpha$-glucanotransferase from Thermotoga maritima in that it can convert maltotriose into maltooligosaccharides. The treatment of glucoamylase after the reaction of MgtA with maltotriose, maltotetraose, maltopentaose, or maltohexaose as sole substrate revealed that MgtA yielded linear maltooligosaccharides instead of cycloamylose.

Effects of Types of Catalysts and Solvents on the Water Repellency of Coating Films Prepared from MTMS and TMES (MTMS와 TMES로부터 제조된 코팅 도막의 발수성에 미치는 촉매와 용매 종류의 영향)

  • Kim, Dong Gu;Lee, Byung Wha;Song, Ki Chang
    • Korean Chemical Engineering Research
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    • v.57 no.6
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    • pp.749-757
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    • 2019
  • Methyltrimethoxysilane (MTMS) and trimethylethoxysilane (TMES) as starting materials were dissolved in various types of solvents, and hydrolysis with water and polycondensation reaction were carried out using various types of catalysts to prepare non-fluorinated water-repellent coating solutions. The coating solutions were spin-coated on cold-rolled steel sheets, and thermally cured to prepare water-repellent coating films. The effect of types of catalysts and solvents on the water repellency of the resulting coating films was investigated during this process. When hydrochloric acid and nitric acid, which are strong acids, were used as catalysts, the solutions showed a white opaque state due to the aggregation of siloxane polymers. On the other hand, when acetic acid, phosphoric acid, and oxalic acid, which are weak acids, were used, they were in a stable and transparent state without precipitation. As a result, the contact angles of the coated films, prepared from hydrochloric acid and nitric acid, were $58^{\circ}$ and $92^{\circ}$, respectively, showing low water repellency. On the other hand, when acetic acid, phosphoric acid, and oxalic acid were used, the contact angles of the coated films were $101^{\circ}$, $103^{\circ}$ and $116^{\circ}$, respectively, showing high water repellency. In addition, when isopropanol and ethanol were used as solvents, phase separation occurred in the solutions due to the aggregation of siloxane polymers. On the other hand, when methanol, ethyl acetate, and methyl ethyl ketone were used as solvents, the solutions were transparent and showed a stable state without sedimentation.

Changes in Benthic Environments in Polluted Coastal Sediment Using Granulated Coal Ash as a Cover (석탄회 조립물의 피복에 따른 연안 오염퇴적물의 저서환경 변화)

  • Jeong, Ilwon;Kim, Kyunghoi
    • Journal of the Korean Society of Marine Environment & Safety
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    • v.25 no.1
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    • pp.67-73
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    • 2019
  • We carried out basic research to evaluate covering material for improving and managing contaminated benthic environments in coastal areas. Changes in nutrient concentration such as phosphate, hydrogen sulfide of contaminated sediment, and pH, Oxidation Reduction Potential (ORP) were investigated through mesocosm experiments for 6 months by covering contaminated sediment with granulated coal ash. Calcium oxide eluted from the granulated coal ash was confirmed to neutralize acidified sediment by increasing pH through hydrolysis. Also, calcium oxide and silica eluted from the granulated coal ash adsorbed and precipitated with phosphate in the sediment. The concentration of phosphate in the sediment investigated decreased by ca. 84.31 %. Similarly, the concentration of hydrogen sulfide decreased by 133.5 mg/L in one month. The hydrogen sulfide is considered to have reacted with substances such as manganese oxide which were eluted from the granulated coal ash and precipitated. Also, it was concluded that the hydrogen sulfide was reduced since anaerobic conditions in the sediment weakened. According to the results of these mesocosm experiments, granulated coal ash was found to be effective to remediate and manage benthic environments by covering the surface layer of sediment.

Preparation of Hydrophobic Coating Layers Using Organic-Inorganic Hybrid Compounds Through Particle-to-Binder Process (유-무기 하이브리드 화합물과 Particle-Binder 공정을 이용한 소수성 코팅막 제조)

  • Hwang, Seung-Hee;Kim, Hyo-Won;Kim, Juyoung
    • Journal of Adhesion and Interface
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    • v.21 no.4
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    • pp.143-155
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    • 2020
  • Hydrophobic Organic-Inorganic (O-I) hybrid materials prepared by sol-gel process have been widely used at functional coating fields such as coatings for anti-corrosion, anti-icing, self-cleaning, anti-reflection. The key point for fabricating hydrophobic surface is to optimize the surface energy and roughness of the coating films. There are typical processes to control the surface energy and roughness which are 'In situ fabricating', 'Pre-fluorinating/Post-roughening', 'Pre-roughening/ Post-fluorinating'. In this study, particle-binder process was used for in-situ fabrication of hydrophobic coating films. Various O-I hybrid compounds prepared using several kinds of alkoxysilane compounds were used as a binder for silica nanoparticles at particle-binder process. To study effect of fluorine content and weight ratio of particle : binder on the hydrophobicity and surface morphology, Hydrophobic coating films were prepared onto glass substrate at various content of fluorine content of O-I hybrid binder and weight ratio of particle : binder. The coating films prepared using O-I hybrid binder (GPTi-HF10) having 10 wt% of fluorine content showed the highes water contact angle (107.52±1.6°). The coating films prepared at 1:3 weight ratio of GPTi-HF10 : silica nanoparticle exhibited the highest water contact angle (130.84±1.99°).

Structural elucidation of immuno-stimulating polysaccharide, galactomannan isolated from Colocasia esculenta (토란으로부터 분리한 면역활성 다당 galactomannan의 구조적 특성 규명)

  • Lee, Hee-Won;Shin, Kwang-Soon
    • Korean Journal of Food Science and Technology
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    • v.52 no.6
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    • pp.595-603
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    • 2020
  • To elucidate the structure-function relationship of polysaccharides obtained from Colocasia esculenta, the immuno-stimulating polysaccharide, CE-4a was purified to homogeneity from the crude polysaccharide (CE) extracted from the corms of C. esculenta by two subsequent column chromatographies using DEAE-Sepharose FF and Sephadex G-100, and analysis of their immuno-stimulatory activities and structure were conducted. CE-4a showed an increase in anti-complementary activity in a dose-dependent fashion. The molecular mass was estimated to be 182.4 kDa, which mainly consisted of galactose (43.5%) and mannose (18.2%). Methylation analysis indicated that CE-4a comprised at least 10 different glycosyl linkages, such as terminal Galp, 3-linked Galp, and 4-linked Manp, as well as a characteristic linkage, 2,4,6-branched Manp residue. To analyze the fine structure of CE-4a, it was sequentially digested using endo-α-(1→4)-polygalacturonase, exo-α-galactosidase and endo-β-1,4-D-mannanase. These analyses suggested that CE-4a is to be a highly branched galactomannan with a (1→4)-mannan backbone and galactopyranosyl oligosaccharide side chains.

Synthesis and Characterization of Collagen Peptide Based Copolymer from Shaving Scrap (셰이빙 스크랩으로부터 콜라겐 펩타이드계 공중합체 합성과 특성)

  • Park, Min Seok;Shin, Soo Beom;Kim, Ho Soo;Kim, Min Soo;Kim, Ha Sun;Jang, Jae Hyeok;Lee, Jin Kye;Lee, Dong Kuk
    • Applied Chemistry for Engineering
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    • v.33 no.6
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    • pp.581-587
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    • 2022
  • The leather industry generates a large amount of hazardous leather waste of various types every year. Among them, shaving scrap is difficult to recycle because it contains chromium ions. Many studies in recent years have shown that shaving scraps can be processed into various types of valuable products, such as adsorbent, filler, and poultry feed. In this study, collagen peptides were extracted from shaving scraps and structurally modified to be developed as new materials with improved physicochemical properties. First, the chromium ions contained in the shaving scraps were removed using a sodium hydroxide solution, and purified through concentration and low-temperature crystallization. The purified collagen peptide was used to prepare the powder using a spray dryer. The extracted collagen peptides were structurally modified by introducing double bonds by reacting with methacrylic anhydride (MAA), and the product was confirmed by 1H NMR spectroscopy. Next, a copolymer was prepared by redox polymerization of the modified collagen peptide (MCP) and 2-ethylhexyl acrylate (2-EHA). The structure of the copolymer was qualitatively confirmed by FT-IR. In conclusion, this study confirmed that collagen peptides can be extracted from shaving scrap and converted into new eco-friendly materials through certain treatments.

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
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    • v.19 no.5
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    • pp.441-448
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    • 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.

Characteristics of Functional Components of Red Ginseng Concentrate First Extracted at Low Temperature I - Focused on Ginsenoside - (저온에서 1차 추출한 홍삼농축액의 기능성분 특성 I - Ginsenoside 위주로 -)

  • Su Hyun Lee;Keon Shin;Seon Yeung Jo;Young Sig Park
    • Journal of Food Hygiene and Safety
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    • v.38 no.3
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    • pp.176-183
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    • 2023
  • The extraction and filtration of red ginseng with a mixed solvent of water and alcohol-a common processing method-and the production of a concentrate through heat treatment, such as steaming, leads to its hydrolysis or polymerization. Approximately 200 ginsenosides have consequently been detected in small amounts, in addition to the identification of the functions of approximately 30 major ginsenosides. This complicates the identification of the functionality of red ginseng and its efficacy, and has negative effects as a functional food, as the astringent taste becomes stronger with an increase in the number of extractions. The red ginseng concentrate was, therefore, extracted at a low temperature (less than 40 ℃) and processed to eliminate these negative aspects, with a specific focus on the characteristics of the functional components of ginsenosides.

Characterization of an Extracellular Xylanase from Bacillus sp. HY-20, a Bacterium in the Gut of Apis mellifera (꿀벌(Apis mellifera)의 장내 세균인 Bacillus sp. HY-20이 분비하는 Xylanase의 특성)

  • Lee, Lan-Hee;Kim, Do-Young;Han, Mi-Kyoung;Oh, Hyun-Woo;Ham, Su-Jin;Park, Doo-Sang;Bae, Kyung-Sook;Sok, Dai-Eun;Shin, Dong-Ha;Son, Kwang-Hee;Park, Ho-Yong
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.332-338
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    • 2009
  • A xylan-decomposing bacterium, HY-20, was isolated from the gut of a honeybee, Apis mellifera, and identified as Bacillus sp. The extracellular GH11 xylanase (XylP) gene (687-bp) of strain HY-20 encoded a protein of 228 amino acids with a deduced molecular mass of 25,522 Da and a calculated pI of 9.33. The primary structure of XylP was 97% identical to that of B. pumilus xylanase (GenBank accession no.: AY526092) that has not been characterized yet. The recombinant His-tagged enzyme (rXylP) overexpressed in Escherichia coli BL21 harboring pET-28a(+)/xylP was purified to electrophoretic homogeneity by cation exchange and gel permeation chromatographies. The purified enzyme exhibited the highest catalytic activity toward birchwood xylan at pH 6.5 and $50^{\circ}C$ and retained approximately 50% of its original activity when pre-incubated at $55^{\circ}C$ for 15 min. The recombinant enzyme was completely inactivated by $Hg^{2+}$ (1 mM) and N-bromosuccinimide (5 mM), while its activity was slightly stimulated by approximately 10% in the presence of $Mn^{2+}$ (1 mM), $Fe^{2+}$ (1 mM), and sodium azide (5 mM). rXylP was able to efficiently degrade various polymeric xylose-based substrates but PNP-sugar derivatives and glucose-based polymers were not susceptible to the enzyme.

FMDV 2C Protein of Foot-and-mouth Disease Virus Increases Expression of Pro-inflammatory Cytokine TNFα via Endoplasmic Reticulum Stress (구제역바이러스의 FMDV 2C 단백질은 소포체 스트레스를 통해서 염증 유도 사이토카인 TNFα의 발현을 증가시킴)

  • Kang, Hyo Rin;Seong, Mi So;Nah, Jin Ju;Ryoo, Soyoon;Ku, Bok Kyung;Cheong, JaeHun
    • Journal of Life Science
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    • v.30 no.3
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    • pp.285-290
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    • 2020
  • Foot-and-mouth disease virus (FMDV), a member of the genus Aphthovirus in the Picornaviridae family, affects wild and domesticated ruminants and pigs. FMDV causes various clinical symptoms, including severe inflammation in infected tissue. Genome RNA of FMDV shows a positive single-strand chain approximately 8.3 kb long and encodes a single long open reading frame (ORF). The ORF is translated into structural and non-structural proteins by viral proteases. The FMDV 2C protein is one of the non-structural proteins encoded by FMDV and plays a critical role in FMD pathogenesis, including inflammation, apoptosis, and viral replication. In this study, we examined whether FMDV 2C induces intracellular expression of pro-inflammatory cytokine tumor necrosis factor alpha (TNFα). FMDV 2C expression in pig IBRS-2 cells increased mRNA and protein expression of TNFα at the transcriptional level via activation of TNFα promoter. Treatment with 4-phenylbutyric acid, an endoplasmic reticulum (ER) stress reducer, decreased TNFα expression induced by FMDV 2C. Activating transcription factor 4 (ATF4), a transcription factor mediating ER stress response, induced transactivation of TNFα promoter and expression of mRNA and protein of TNFα. However, the dominant negative mutant of ATF4 did not induce FMDV 2C-mediated TNFα expression. The results indicate that FMDV 2C protein increases clinical inflammation via ATF4-mediated TNFα expression and is associated with ER stress induction.