• Microbiology and Biotechnology Letters
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• v.14 no.5
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• pp.385-389
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• 1986

Both hydrogenase and nitrogenase were found to be involved in hydrogen evolution independently in Rhodopseudomonas sp. KCTC 1437. The hydrogen formation in this bacterium was independent on light illumination and presence of N $H_4^{+}$ After establishment of conditions to measure the amount of hydrogen evolved by each of the enzymes in vivo, the several factors affecting on the hydrogen evolution, e.g. presence of gases ( $C_2$ $H_2$, $H_2$, $O_2$ or $N_2$), C/N ratio, were investigated, Hydrogenase was less inhibited than nitrogenase under $O_2$ and was active independent on the presence of $N_2$ or $C_2$ $H_2$ which were the strong inhibitor of nitrogenase. Besides, the hydrogenase activity was increased after incubation with $H_2$. And it was verified that this bacterium consume hydrogen and photoreduce $CO_2$ by hydrogenase. From above results, it is concluded that hydrogenase in Rhodopseudomonas sp. KCTC 1437 can produce hydrogen under more favorable condition that nitrogenase.e.

• 한국신재생에너지학회:학술대회논문집
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• 2010.06a
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• pp.251.1-251.1
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• 2010

바이오디젤은 동식물성 기름과 메탄올의 트랜스에스테르화 반응에 의해 생산되는 지방산메틸에스테르(FAME, fatty acid methyl esters)로서, 트랜스에스테르화 공정에는 KOH, NaOH, $NaOCH_3$등의 균질계 화학촉매를 이용한 방법, 무촉매 공정인 초임계 메탄올 이용 방법, 그리고 효소촉매를 이용한 방법이 있다. 초임계 공정은 에너지 소비와 장치비가 커서 경제성이 떨어지는 것으로 보고되며 화학촉매 공정은 반응 효율이 높다는 장점을 가지고 있지만, 반응 및 정제단계가 복잡하고 정제과정에 폐수를 발생시키는 문제점을 가지고 있다. 고정화 효소를 사용하는 효소 공정은 에너지 비용의 절감, 후 처리 공정의 단순화, 고 순도의 글리세롤을 얻을 수 있는 장점이 있지만, 반응 속도가 느리고 효소 가격이 비싸다는 단점이 있어 현재까지 상업화되지 못하고 있다. 반응속도가 높고 재사용이 가능한 효소 촉매 공정 개발을 위해 본 연구에서는 Candida rugosa, Rizhopus oryzae 2종을 실리카에 동시 고정화하였다. 고정화 Lipase의 제조는 실리카겔을 과산화수소를 이용하여 전처리를 하고 Acetone과 3-APTES의 혼합용액을 첨가한 후 실리카겔과 (silanization)을 진행 하였다. 그리고 glutaraldehyde를 첨가 하여 공유 결합을 형성 한 후에 증류수를 사용하여 실리카겔을 회수하여 lipase(Rizhopus oryzae, Candida rugosa 10% 용액)를 고정화 하였다. 고정화 효소의 효소 활성을 측정한 결과 3000-3500 Unit(${\mu}mol/g{\cdot}min$)으로 측정되었다. 제조된 고정화 효소를 이용하여 Canola Oil을 바이오디젤로 전환하는 실험을 진행하였으며 생성물로부터 고정화 효소를 분리한 후에 상층의 에스테르층을 취하여 수세한 뒤 원심분리하여 FAME 함량을 측정한 결과 83%의 바이오디젤을 얻을 수 있었다. 그리고 효소 촉매 트랜스에스테르화 반응의 Enzyme, Water, Methanol 투입량의 반응 변수들에 대하여 반응표면분석법(Response Surface Methodology)을 적용하여 최적 반응조건을 도출하는 연구를 수행하였다.

• Investigative Magnetic Resonance Imaging
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• v.10 no.2
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• pp.98-107
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• 2006

Magnetization Transfer (MT) imaging generates contrast dependent on the phenomenon of magnetization exchange between free water proton and restricted proton in macromolecules. In biological materials in knee, MT or cross-relaxation is commonly modeled using two spin pools identified by their different T2 relaxation times. Two models for cross-relaxation emphasize the role of proton chemical exchange between protons of water and exchangeable protons on macromolecules, as well as through dipole-dipole interaction between the water and macromolecule protons. The most essential tool in medical image manipulation is the ability to adjust the contrast and intensity. Thus, it is desirable to adjust the contrast and intensity of an image interactively in the real time. The proton density (PD) and T2-weighted SE MR images allow the depiction of knee structures and can demonstrate defects and gross morphologic changes. The PD- and T2-weighted images also show the cartilage internal pathology due to the more intermediate signal of the knee joint in these sequences. Suppression of fat extends the dynamic range of tissue contrast, removes chemical shift artifacts, and decreases motion-related ghost artifacts. Like fat saturation, phase sensitive methods are also based on the difference in precession frequencies of water and fat. In this study, phase sensitive methods look at the phase difference that is accumulated in time as a result of Larmor frequency differences rather than using this difference directly. Although how MT work was given with clinical evidence that leads to quantitative model for MT in tissues, the mathematical formalism used to describe the MT effect applies to explaining to evaluate knee disorder, such as anterior cruciate ligament (ACL) tear and meniscal tear. Calculation of the effect of the effect of the MT saturation is given in the magnetization transfer ratio (MTR) which is a quantitative measure of the relative decrease in signal intensity due to the MT pulse.

• KSBB Journal
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• v.23 no.5
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• pp.425-430
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• 2008

Hydrogen sulfide ($H_2S$) is a by-product of metabolism of amino acids including sulfur and alcoholic fermentation, it is generally thought of in terms of a poisonous gas. Though $H_2S$ can have a negative impact on the perceived quality of fermented drinks due to an undesirable aroma, it plays prominent roles as a neuromodulator in the mammalian brain as well as a smooth muscle relaxant. Nowadays studies on the proteins which produce $H_2S$ are carried out in various fields such as structure, function, and metabolism. Here we propose to develop a simple and rapid $H_2S$ forming assay method, which will lead to speed up preparing the $H_2S$ forming proteins for identification by MALDI-TOF MS analysis. We detected three kinds of proteins which produce $H_2S$ in the crude extract of Saccharomyces cerevisiae. Those proteins were cystathionie $\beta$-synthase, O-acetylserine sulfhydrylase, and cystathionine $\gamma$-lyase.

• Journal of Life Science
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• v.25 no.2
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• pp.121-129
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• 2015

Alginates are found in marine brown seaweeds and in extracellular biofilms secreted by some bacteria. Previously, we reported an oligoalginate lyase from Sphingomonas sp. MJ-3 (MJ3-Oal) that had an exolytic activity and protein sequence homology with endolytic polymannuronate (polyM) lyase in the N-terminal region. In this study, the MJ3-Oal was tested for both exolytic and endolytic activity by homology modeling using the crystal structure of Alg17c from Saccharophagus degradans 2-40T. The tyrosine residue at the $426^{th}$ position, which possibly formed a hydrogen bond with the substrate, was mutated to phenylalanine. The FPLC profiles showed that MJ3-Oal degraded alginate quickly to monomers as a final product through the oligmers, whereas the Tyr426Phe mutant showed only exolytic alginate lyase activity. $^1H$-NMR spectra also showed that MJ3-Oal degraded the endoglycosidic bond of polyM and polyMG (polymannuronate-guluronate) blocks. These results indicate that oligoalginate lyase from Sphingomonas sp. MJ-3 probably catalyzes the degradation of both exo- and endo-glycosidic bonds of alginate.

• Microbiology and Biotechnology Letters
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• v.26 no.4
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• pp.297-302
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• 1998

$\beta$-Xylosidase B was produced by Escherichia coli HB101/pKMG12 carrying the xylB gene of Bacillus stearothermophilus No.236 on its recombinant plasmid. The $\beta$-xylosidase B produced was purified by ammonium sulfate fractionation, DEAE-Sepharose CL-6B, Sephacryl S-200 and Superdex 200 HR gel filtration. The purified enzyme showed the highest activity at pH 6.5 and 5$0^{\circ}C$. But, the enzyme was observed to be very sensitive to the pH and temperature of the reaction mixture. The enzyme was activated about 35% of its original activity in the presence of 1 mM of $Mn^{2+}$ but it was completely inhibited by $Ag^{+}$, $Cu^{2+}$and $Hg^{2+}$ions. In contrast with the $\beta$-xylosidase A, the B enzyme was found to have $\alpha$-arabinofuranosidase activity though the activity was fairly low compared with the $\alpha$-arabinofuranosidase produced from the arfI gene of the same Bacillus stearothermophilus. Therefore, $\beta$-xylosidase B is considered to be more suitable than $\beta$-xylosidase A at least for the biodegradation of arabinoxylan. The $K_{m}$ and V$_{max}$ values of the $\beta$-xylosidase B for o-nitrophenyl-$\alpha$-D-xylopyranoside were 6.43 mM and 1.45 $\mu$mole/min, respectively. Molecular mass of the enzyme was determind to be about 54 kDa by SDS-PAGE and 160 kDa by Superdex 200HR gel filtration, indicating that the functional $\beta$-xylosidase B was composed of three identical subunits.s.

• Korean Chemical Engineering Research
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• v.61 no.4
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• pp.576-583
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• 2023

This study aimed to investigate the impact of enhancing the electrode conductivity and mitigating the production of hydrogen peroxide - a by-product arising from lactate oxidation - on the performance of lactate electrodes. The electrical conductivity of the electrode was improved by modifying the surface of carbon paper with single-walled carbon nanotubes. Catalase was introduced to effectively eliminate the hydrogen peroxide produced during the lactate oxidation reaction. The carbon paper electrode, with simultaneous immobilization of both lactate oxidase and catalase, yielded a current 1.7 times greater than the electrode where only lactate oxidase was immobilized. The electrode in which lactate oxidase and catalase were co-immobilized on the surface of carbon paper modified with single-walled carbon nanotubes, produced a current of 171 µA, which was more than twice as much current as the carbon paper with only lactate oxidase immobilized. The optimized electrode showed a linear response up to lactate concentration of 20 mM, confirming that it can be used as a sensor electrode.

• Transactions of the Korean hydrogen and new energy society
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• v.20 no.2
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• pp.142-150
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• 2009

This paper presents bio-ethanol production from fruit wastes as it possibly alternate fossil fuel in the future. To illustrate the component ratio in exocarps of fruit wastes such as pears, apples, and persimmons, the amount of moisture, lignin, $\alpha$, $\beta$, $\gamma$-cellulose, and ash content were respectively examined by the ingredient analysis. Also, the amount of the glucose obtained from the enzyme hydrolysis using the axocarps was investigated. It was found in our results that the energy efficient process requires different temperature conditions for the saccharification step($50^{\circ}C$ and the fermentation step($30^{\circ}C$ in ethanol synthesis.

• Transactions of the Korean hydrogen and new energy society
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• v.16 no.3
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• pp.219-228
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• 2005

Fermentative strict anaerobic bacterium, Clostricium butyricum NCIB 9576 (Cl. butyricum) and purple sulfur phototrophic bacterium, Thiocapsa roseopersicina NCIB 8347 (T. roseopersicina) were compared on their temperature and oxygen stabilities of cytoplasmic hydrogenases. Cell growth phase and the specific activities of evolution $H_2ase$ were related for both strains, exhibiting the highest cytoplasmic $H_2ase$ activities during the logarithmic growth phases which were 4 and 18 hrs after the incubation for Cl. butyricum and T. roseopersicina, respectively. The optimum temperatures for the growth of Cl. butyricum and T. roseopersicina were 37$^{\circ}C$ and 27$^{\circ}C$, respectively, while those for $H_2$ evolution of cytoplsmic hydrogenases prepared from Cl. butyricum ($C-H_2ase$) and T. roseopersicina ($T-H_2ase$) were 45$^{\circ}C$ and 65$^{\circ}C$, respectively. $T-H_2ase$ was more thermo-stable than $C-H_2ase$. $T-H_2ase$ retained its full activity for 5 hrs at 50$^{\circ}C$ and retained 90% of its original activity for 5 hrs at 60$^{\circ}C$, however, $C-H_2ase$ lost its activity drastically at 50$^{\circ}C$. The optimum pHs for $H_2$ oxidation of $C-H_2ase$ and $T-H_2ase$ were 9.0 and 7.5 respectively. The both enzymes showed maximum $H_2$ evolution activity at pH 7.0. Under the aerobic condition, 80% of $T-H_2ase$ activity was retained for 10 hrs at 30$^{\circ}C$, and 50% of activity was still remained after 6 days at the same experimental conditions. But the $C-H_2ase$ was labile to oxygen and lost its activity immediately after the exposure to air.

• Microbiology and Biotechnology Letters
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• v.34 no.1
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• pp.7-14
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• 2006

Feather, generated in large quantities as a byproduct of commercial poultry processing, is almost pure keratin, which is not easily degradable by common professes. Four strains, SMMJ-2, FL-3, NO-4 and RM-12 were isolated from soil for production of extracellular keratinolytic protease. They were identified as Bacillus sp. based on their morphological and physiological characteristics. They shown high protease activity on 5.0% skim milk agar medium and produced a substrate like mucoid on keratin agar medium. Bacillus sp. SMMJ-2 had a faster production time for producing keratinolytic protease than other strains. This strain did not completely degrade whole chicken feather for five days in basal medium but completely degraded whole chicken feather when supplied with nitrogen source for 40hours in keratinolytic producing medium ($0.7%\;K_{2}HPO_{4},\;0.2%\;KH_{2}PO_{4},\;0.1%$ fructose, 1.2% whole chicken feather, $0.01%\;Na_{2}CO_3$, pH 7.0). When supplied with chicken feather as nitrogen source, keratinolytic protease activity was 89 units/ml/min. When soybean meal was used as nitrogen source, the keratinolytic protease production reached a maximum of 106 units/ml/min after 48 hours under $30^{\circ}C$, 180 agitation. To isolate the keratinolytic protease, the culture filtrate was precipitated with $(NH_4)_{2}SO_4$ and acetone. The recovery rate of keratinolytic protease was about 96% after treatment with 50% acetone. The enzyme was stable in the range of $30{\sim}50^{\circ}C$ and pH $6.0{\sim}12.0$.