• Journal of the Korean Applied Science and Technology
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• v.37 no.4
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• pp.723-732
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• 2020

The spent RHDS (Residue HydroDeSulfurization) catalyst is deactivated mainly by deposition of various contaminants such as coke, sulfur and vanadium on the surface of catalyst. To eliminate those contaminants, the following remanufacturing process was conducted. The first, heavy oil on the surface of the spent RHDS catalyst was removed by kerosene and dehydrated. The second, the high temperature incineration was carried out to eliminate coke and sulfur components deposited on the surface of spent RHDS catalyst. The third, the excessive quantity of Vanadium deposited on the surface of catalyst was removed by leaching process as follows: ultrasonic agitation was carried out at 50℃, for 10 seconds with 0.5% and 1% oxalic acid solution. The purpose of this process is to find out regenerated RHDS catalyst can be used as SCR catalyst for NOx reduction by controlling the vanadium residual content of the regenerated RHDS catalyst through leaching process. The composition of regenerated RHDS catalyst was analyzed by XRF and the NOx reduction efficiency was also measured by continuous catalytic fixed bed reactor. As the result, regenerated catalyst, with 0.5% oxalic acid, ultrasonic agitation in 10 seconds, showed the most stable NOx reduction efficiency. Also, in comparison with commercial SCR catalyst, the NOx reduction performance of regenerated catalyst was similar to that of commercial SCR catalyst at the temperature 375℃ and higher whereas was lower than commercial SCR catalyst at the temperature range between 200~250℃. Therefore, it was confirmed that the regenerated catalyst as powder form wash coated on the surface of metal corrugated substrate can be used for commercial SCR catalyst.

• Journal of Microbiology and Biotechnology
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• v.31 no.3
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• pp.483-491
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• 2021

Two putative genes, lip29 and est29, encoding lipolytic enzymes from the thermophilic bacterium Geobacillus thermocatenulatus KCTC 3921 were cloned and overexpressed in Escherichia coli. The recombinant Lip29 and Est29 were purified 67.3-fold to homogeneity with specific activity of 2.27 U/mg and recovery of 5.8% and 14.4-fold with specific activity of 0.92 U/mg and recovery of 1.3%, respectively. The molecular mass of each purified enzyme was estimated to be 29 kDa by SDS-PAGE. The alignment analysis of amino acid sequences revealed that both enzymes belonged to GDSL lipase/esterase family including conserved blocks with SGNH catalytic residues which was mainly identified in plants before. While Est29 showed high specificity toward short-chain fatty acids (C4-C8), Lip29 showed strong lipolytic activity to long-chain fatty acids (C12-C16). The optimal activity of Lip29 toward p-nitrophenyl palmitate as a substrate was observed at 50℃ and pH 9.5, respectively, and its activity was maintained more than 24 h at optimal temperatures, indicating that Lip29 was thermostable. Lip29 exhibited high tolerance against detergents and metal ions. The homology modeling and substrate docking revealed that the long-chain substrates showed the greatest binding affinity toward enzyme. Based on the biochemical and insilico analyses, we present for the first time a GDSL-type lipase in the thermophilic bacteria group.

• Journal of Microbiology and Biotechnology
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• v.25 no.5
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• pp.662-671
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• 2015

To enrich the genetic resource of microbial xylanases with high activity and stability under alkaline conditions, a xylanase gene (xynSL4) was cloned from Planococcus sp. SL4, an alkaline xylanase-producing strain isolated from the sediment of soda lake Dabusu. Deduced XynSL4 consists of a putative signal peptide of 29 residues and a catalytic domain (30-380 residues) of glycosyl hydrolase family 10, and shares the highest identity of 77% with a hypothetical protein from Planomicrobium glaciei CHR43. Phylogenetic analysis indicated that deduced XynSL4 is closely related with thermophilic and alkaline xylanases from Geobacillus and Bacillus species. The gene xynSL4 was expressed heterologously in Escherichia coli and the recombinant enzyme showed some superior properties. Purified recombinant XynSL4 (rXynSL4) was highly active and stable over the neutral and alkaline pH range from 6 to 11, with maximum activity at pH 7 and more than 60% activity at pH 11. It had an apparent temperature optimum of 70℃ and retained stable at this temperature in the presence of substrate. rXynSL4 was highly halotolerant, retaining more than 55% activity with 0.25-3.0 M NaCl and was stable at the concentration of NaCl up to 4M. The enzyme activity was significantly enhanced by β-mercaptoethanol and Ca²⁺ but strongly inhibited by heavy-metal ions and SDS. This thermophilic and alkaline- and salt-tolerant enzyme has great potential for basic research and industrial applications.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2004.06a
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• pp.273-277
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• 2004

$\alpha$-L-Arabinofuranosidase ($\alpha$-L-AFase, EC 3.2.1.55) was isolated from hyperthermophilic microorganism, Thermotoga maritima. The open reading frame (ORF) of $\alpha$-L-AFase gene is 1,455 bp long and encodes 484 amino acid residues with a molecular weight of 55,265 Da. The ORF of $\alpha$-L-AFase gene was introduced into the E. coli expression vector, $_p/RSET-B, and overexpressed in E. coli BL21. The purified recombinant $\alpha$-L-AFase showed the highest activity at 10$0^{\circ}C$ and pH 5.5. The purified enzyme appeared to have no metal cofactor requirement. The Km and specific activity values of the recombinant enzyme were 0.99 mM and 1,200 U/mg on p-nitrophenyl-$\alpha$-L-arabinofuranoside. It released only L-arabinose from sugar beet arabinan, sugar beet debranched arabinan and oat spelts arabinoxylan but had no activity onarabinogalactan and gum arabic. This result suggests that L-arabinose could be produced from natural polysaccharides using this enzyme. Mutant enzymes which Glu26, Glu172 and Glu281 residues were replaced to alanine, aspartic acid or glutamine caused Kcat to decrease by a factor of between 10$^3$ and 10$^4$. Glu172 and Glu281 residues of $\alpha$-L-AFase are seemed to be the acid/base and nucleophile in catalytic reaction, respectively, and Glu26 is supposed to playa key role in substrate binding.ng.

• Proceedings of the Korean Vacuum Society Conference
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• 2012.08a
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• pp.375-375
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• 2012

Graphene is a sp2-hybridized carbon sheet with an atomic-level thickness and a wide range of graphene applications has been intensely investigated due to its unique electrical, optical, and mechanical properties. In particular, hybrid graphene structures combined with various nanomaterials have been studied in energy- and sensor-based applications due to the high conductivity, large surface area and enhanced reactivity of the nanostructures. Conventional metal-catalytic growth method, however, makes useful applications difficult since a transfer process, used to separate graphene from the metal substrate, should be required. Recently several papers have been published on direct graphene growth on the two dimensional planar substrates, but it is necessary to explore a direct growth of hierarchical nanostructures for the future graphene applications. In this study, uniform graphene layers were successfully synthesized on highly dense dielectric nanowires (NWs) without any external catalysts. We also demonstrated that the graphene morphology on NWs can be controlled by the growth parameters, such as temperature or partial pressure in chemical vapor deposition (CVD) system. This direct growth method can be readily applied to the fabrication of nanoscale graphene electrode with designed structures because a wide range of nanostructured template is available. In addition, we believe that the direct growth growth approach and morphological control of graphene are promising for the advanced graphene applications such as super capacitors or bio-sensors.

• Clean Technology
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• v.24 no.2
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• pp.127-134
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• 2018

To reduce the environmental pollution by $NO_x$ from ship engine, International maritime organization (IMO) announced Tier III regulation, which is the emmision regulation of ship's exhaust gas in Emission control area (ECA). Selective catalytic reduction (SCR) process is the most commercial $De-NO_x$ system in order to meet the requirement of Tier III regulation. In generally, commercial ceramic honeycomb SCR catalyst has been installed in SCR reactor inside marine vessel engine. However, the ceramic honeycomb SCR catalyst has some serious issues such as low strength and easy destroution at high velocity of exhaust gas from the marine engine. For these reasons, we design to metallic structured catalyst in order to compensate the defects of the ceramic honeycomb catalyst for applying marine SCR system. Especially, metallic structured catalyst has many advantages such as robustness, compactness, lightness, and high thermal conductivity etc. In this study, in order to support catalyst on metal substrate, coating slurry is prepared by changing binder. we successfully fabricate the metallic structured catalyst with strong adhesion by coating, drying, and calcination process. And we carry out the SCR performance and durability such as sonication and dropping test for the prepared samples. The MFC01 shows above 95% of $NO_x$ conversion and much more robust and more stable compared to the commercial honeycomb catalyst. Based on the evaluation of characterization and performance test, we confirm that the proposed metallic structured catalyst in this study has high efficient and durability. Therefore, we suggest that the metallic structured catalyst may be a good alternative as a new type of SCR catalyst for marine SCR system.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.19 no.1
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• pp.18-24
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• 2018

The policy-making and technological development of eco-friendly automobiles designed to increase their supply is ongoing, but the internal combustion engine still accounts for about 95% of the automobiles in use. Also, in order to meet the stricter emission regulations of internal combustion engines based on fossil fuels, the proportion of after-treatments for vehicles and (ocean going) vessels is gradually increasing. Natural gas is a clean fuel that emits few air pollutants and has been used mainly as a fuel for city buses. In the long term, we intend to develop a new NGOC/LNT+NGCO/SCR combined system that simultaneously reduces the toxic gases, $CH_4$ and NOx, emitted from CNG buses. The objective of this study is to investigate the characteristics of $de-CH_4/NOx$ according to the ceramic and metal substrates of the SCR (Selective Catalytic Reduction) catalysts mounted downstream of the combined system. The V and Cu-SCR catalysts did not affect the $CH_4$ oxidation reaction, the two NGOC/SCR catalysts each coated with two layers began to oxidize $CH_4$ at $400^{\circ}C$, and the amount of $CH_4$ emitted was reduced to about 20% of its initial value at about $550^{\circ}C$. The two NGOC/SCR catalysts each coated with two layers showed a negative (-) NOx conversion rate above $350^{\circ}C$. The ceramic-based combined system reached LOT50 at $500^{\circ}C$, which was about 20% higher in terms of the $CH_4$ conversion rate than the metal-based combined system, showing that the combined system of NGOC/LNT+Cu-SCR is a suitable combination.

• Journal of Microbiology and Biotechnology
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• v.24 no.6
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• pp.748-755
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• 2014

In the genome annotation of Escherichia coli MG1655, the orf382 (1,149 bp) is designated as a gene encoding an alcohol dehydrogenase that may be Fe-dependent. In this study, the gene was amplified from the genome by PCR and overexpressed in Escherichia coli BL21(DE3). The recombinant $6{\times}$His-tag protein was then purified and characterized. In an enzymatic assay using different hydroxyl-containing substrates (n-butanol, $\small{L}$-threonine, ethanol, isopropanol, glucose, glycerol, $\small{L}$-serine, lactic acid, citric acid, methanol, or $\small{D}$-threonine), the enzyme showed the highest activity on $\small{L}$-threonine. Characterization of the mutant constructed using gene knockout of the orf382 also implied the function of the enzyme in the metabolism of $\small{L}$-threonine into glycine. Considering the presence of tested substrates in living E. coli cel ls and previous literature, we believed that the suitable nomenclature for the enzyme should be an $\small{L}$-threonine dehydrogenase (LTDH). When using $\small{L}$-threonine as the substrate, the enzyme exhibited the best catalytic performance at $39^{\circ}C$ and pH 9.8 with $NAD^+$ as the cofactor. The determination of the Km values towards $\small{L}$-threonine (Km = $11.29{\mu}M$), ethanol ($222.5{\mu}M$), and n-butanol ($8.02{\mu}M$) also confirmed the enzyme as an LTDH. Furthermore, the LTDH was shown to be an ion-containing protein based on inductively coupled plasma-atomic emission spectrometry with an isoelectronic point of pH 5.4. Moreover, a circular dichroism analysis revealed that the metal ion was structurally and enzymatically essential, as its deprivation remarkably changed the ${\alpha}$-helix percentage (from 12.6% to 6.3%).

• Journal of Microbiology and Biotechnology
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• v.19 no.9
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• pp.987-996
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• 2009

Aspergillus niger GH1 previously isolated and identified by our group as a wild tannase producer was grown under solid-state (SSC) and submerged culture (SmC) conditions to select the enzyme production system. For tannase purification, extracellular tannase was produced under SSC using polyurethane foam as the inert support. Tannase was purified to apparent homogeneity by ultrafiltration, anion-exchange chromatography, and gel filtration that led to a purified enzyme with a specific activity of 238.14 IU/mg protein with a final yield of 0.3% and a purification fold of 46. Three bands were found on the SDS-PAG with molecular masses of 50, 75, and 100 kDa. PI of 3.5 and 7.1% N-glycosylation were noted. Temperature and pH optima were 600e and 6.0 [methyl 3,4,5-trihydroxybenzoate (MTB) as substrate], respectively. Tannase was found with a $K_M$ value of $0.41{\times}10^{-4}M$ and the value of $V_{max}$ was $11.03{\mu}$moL/min at $60^{\circ}C$ for MTB. Effects of several metal salts, solvents, surfactants, and typical enzyme inhibitors on tannase activity were evaluated to establish the novelty of the enzyme. Finally, the tannase from A. niger GH1 was significantly inhibited by PMSF (phenylmethylsulfonyl fluoride), and therefore, it is possible to consider the presence of a serine or cysteine residue in the catalytic site.

• Microbiology and Biotechnology Letters
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• v.16 no.4
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• pp.310-315
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• 1988

Two isoenzymes of chorismate mutase(E.C.5.4.99.5) designated as chorismate mutase I(CM I) and chorismate mutase II(CM II), were detected and partially purified from a sp. of intrasporangium isolated from soil. CM I and CM II had pH optima of pH 6.5 and 8.0, respectively and showed the same temperature optimum of 45$^{\circ}C$. The activation energy of the enzymatic reaction was estimated to be 14.7kcal/ mole with CM I and 10.8kcal/mole with CM II. The affinity of isoenzyme CM I for substrate(Km= 1.35mM) was almost the same level as that of CM II(Km = 1.22mM). Both isoenzymes were stable at pH values ranged from pH 6.5 to 9.0, but rapidly denaturated at temperatures above 45$^{\circ}C$. CM II was activated about 7$^{\circ}C$ of its activity by $Ba^{++}$ or $Mg^{++}$ while CM I was slightly inhibited by the same metal ions. Thiol compounds were found not to be necessary for stability of the two enzymes but Co$^{++}$ and EDTA had a little stabilizing effect on CM II only. p-Chloromercuribenzoate strongly inactivated the activities of both enzymes but the reducing agents such as dithiothreitol and L-cysteine protected them against the pCMB inhibition.