• Journal of Microbiology and Biotechnology
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• 제33권4호
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• pp.552-558
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• 2023

Levulinic acid (LA) is a valuable chemical used in fuel additives, fragrances, and polymers. In this study, we proposed possible biosynthetic pathways for LA production from lignin and poly(ethylene terephthalate). We also created a genetically encoded biosensor responsive to LA, which can be used for screening and evolving the LA biosynthesis pathway genes, by employing an LvaR transcriptional regulator of Pseudomonas putida KT2440 to express a fluorescent reporter gene. The LvaR regulator senses LA as a cognate ligand. The LA biosensor was first examined in an Escherichia coli strain and was found to be non-functional. When the host of the LA biosensor was switched from E. coli to P. putida KT2440, the LA biosensor showed a linear correlation between fluorescence intensity and LA concentration in the range of 0.156-10 mM LA. In addition, we determined that 0.156 mM LA was the limit of LA detection in P. putida KT2440 harboring an LA-responsive biosensor. The maximal fluorescence increase was 12.3-fold in the presence of 10 mM LA compared to that in the absence of LA. The individual cell responses to LA concentrations reflected the population-averaged responses, which enabled high-throughput screening of enzymes and metabolic pathways involved in LA biosynthesis and sustainable production of LA in engineered microbes.

• 청정기술
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• 제20권2호
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• pp.108-115
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• 2014

셀룰로우스(cellulose)를 5-히드록시메틸푸르푸랄(5-hydroxymethylfurfural, 5-HMF)로 직접 전환하기 위해 이온성 액체 용매하에서 다양한 금속염화물과 산 촉매를 비교 연구하였다. 사용한 금속염화물은 Sn(II), Zn(II), Al(III), Fe(III), Cu(II), Cr(III)를 포함한 염화물을 비교하였으며 산 촉매는 산성 이온성 액체를 고정화하여 사용하였다. 비교를 위하여 $H_2SO_4$, HCl, Amberlyst-15와 DOWEX50x8을 사용하였다. 제조한 촉매의 산도와 산 밀도 특성은 Hammett Indicator 지시약을 통하여 분석하였다. 5-HMF의 선택도 및 수율은 반응온도, 반응시간과 촉매 비를 통하여 확인하였다. 사용한 촉매들 중에서 5-HMF의 선택도는 $CrCl_3-6H_2O$와 $SiO_2-[ASBI]HSO_4$를 사용하였을 때에 가장 높게 나타났으며, 상용화 고체 산인 Amberlyst-15와 DOWEX50x8에 비하여 활성이 높다는 것을 확인할 수 있었다. 5-HMF의 선택도는 산 촉매의 산도와 반응에 사용된 촉매비에 영향이 있음을 확인할 수 있었으며, 반응 중 재수화 반응이 일어나 레불린산(levulinic acid)이 생성된다는 것을 확인하였다.

• Bulletin of the Korean Chemical Society
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• 제27권6호
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• pp.933-935
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• 2006

• 한국식품조리과학회지
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• 제7권1호
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• pp.27-34
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• 1991

This study attempted to set up reasonable extraction time of Omija that was put in water for the various components to soak out. Changes of free sugars, organic acids, reducing sugar, total acid and tannin in Omija with various extraction times were investigated (together with the analysis of each components in Omija fruit). 1. High Performance Liquid Chromatography showed fructose, glucose, and sucrose to be the major free sugars of the Omija fruit. Free sugars and reducing sugar value in Omija beverage increased gradually in according with the extraction time, and marked 75.6% per total free sugars and 82.1% per total reducing sugar at 12 hours. 2. Gas Chromatography showed lactic acid, oxalic acid, fumaric acid, levulinic acid, succinic acid, malic acid, citric acid and pyroglutamic acid to be the major organic acids of the Omija fruit. Organic acids and total acids value in Omija beverage increased gradually on proportion to extraction time, and marked 97.0% per total organic acids at 9 hours and 79.0% per total acids at 12 hours. 3. Tannin content in Omija beverage was increased when extraction time was longer but it showed a low percentage as compared with the reducing sugar and total acid. Tannin content marked 48.8% per total tannin at 12 hours. 4. Sensory evaluation revealed that !1 hours of extraction produced the best quality products based in taste, flavor, color and over-all acceptability, considering the data, it seems possible to conclude that the optimum of time for extraction of Omija to water is 9 hours.

• 환경위생공학
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• 제14권3호
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• pp.107-115
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• 1999

The effectiveness of resting cells of a photosynthetic bacterium, Rhodospirillum rubrum N-1, was investigated on the production of extracellular ${\delta}-aminolevulinic$ acid(ALA). The ALA generating system required 3hr-incubation in the presence of 10mg of resting cells per ml to obtain the maximal yield of extracellular ALA. and also, under this condition the effect of ALA inducers, i.e., 30mM levulinic acid (LA) and L-glutamic acid($C_5$ pathway precursor) was relatively higher than that of produced extracellular ALA($83{\mu}M$). The volume of system and proper cell density appeared to be important factors for the effective production of extracellular ALA.

• Applied Biological Chemistry
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• 제40권6호
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• pp.556-560
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• 1997

광합성세균을 이용한 고농도 유기폐수인 두부공업폐수의 정화처리와 동시에 두부공업폐수를 생물제초제인 ${\delta}-aminolevulinic$ acid (ALA) 생산용 대체기질로 이용할 목적으로 Rhodobacter capsulatus KK-10을 사용하여 COD 제거율과 ALA 생산능을 검토하였다. 사용된 두부공업폐수의 성상은 pH 4.2, COD/BOD와 BOD/N의 비율은 0.98 및 17.2, BOD : N : P의 함유비율은 10 : 6 : 4, 젖산 함유량 1,080 ppm 등으로 광합성세균의 생육에 적당한 조건이었다. 두부공업폐수에서 본 균주를 4일 동안 배양한 결과 약 94% COD 값의 감소와 1.2 g/l 균체생육을 보였으며 대수기 중기에서 ALA 탈수효소 저해제인 levulinic acid(LA)를 15 mM 첨가한 결과 55 mg/l의 ALA 생산량을 나타내었다. ALA의 전구물질인 succinate 및 glycine과 LA를 각각 15 mM의 농도로 동시 첨가시 최대 114 mg/l의 생산량을 보였다. 상기의 물질들을 각각 15 mM 농도로 연속 첨가한 후 ALA 생성량을 검토한 결과 LA 1회와 glycine과 succinate를 3회에 걸쳐 첨가하였을 때 120 mg/l로 가장 효과적이었으며 COD 제거율은 92%를 나타내었다.

• Journal of Microbiology and Biotechnology
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• 제14권6호
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• pp.1310-1317
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• 2004

The extracellular production of 5-aminolevulinic acid (ALA) by recombinant E. coli BL21 harboring a fusion gene hemA was investigated in a fermenter. For this purpose, the effects of various physiological factors, such as isopropylthio­$\beta$-D-galactopyranoside (IPTG) concentrations and the time of induction, on enzyme activity were studied. Optimum concentrations of glycine and succinic acid were found to be 30 mM and 90 mM, respectively. When the cells were permitted to grow for 2 h prior to the addition of 0.1 mM IPTG, the activity of ALA synthase was higher than when IPTG was initially added. A 36-fold increase in the activity was observed with only 0.1 mM IPTG added. The pH of the medium also influenced the ALA synthase activity with the maximal activity occurring at pH 6.5. In recombinant E. coli extracts, the repeated addition of glycine and D-glucose increased the production of ALA and the inhibited intracellular ALA dehydratase activity, with up to 32 mM ALA being produced in the cultivation.

• Journal of Microbiology and Biotechnology
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• 제24권9호
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• pp.1245-1253
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• 2014

One-step sulfuric acid saccharification of the red alga Pterocladiella capillacea was optimized, and various detoxification methods (neutralization, overliming, and electrodialysis) of the acid hydrolysate were evaluated for fermentation with the thermotolerant yeast Kluyveromyces marxianus. A proximate composition analysis indicated that P. capillacea was rich in carbohydrates. A significant galactose recovery of $81.1{\pm}5%$ was also achieved under the conditions of a 12% (w/v) biomass load, 5% (v/v) sulfuric acid, $121^{\circ}C$, and hydrolysis for 30 min. Among the various detoxification methods, electrodialysis was identified as the most suitable for fermentable sugar recovery and organic acid removal (100% reduction of formic and levulinic acids), even though it failed to reduce the amount of the inhibitor 5-HMF. As a result, K. marxianus fermentation with the electrodialyzed acid hydrolysate of P. capillacea resulted in the best ethanol levels and fermentation efficiency.

• Applied Biological Chemistry
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• 제30권2호
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• pp.147-152
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• 1987

탈진대두와 밀가루 코오지를 산분해간장에 첨가하여 담금한 각 시험구 간장(시험구 A : 탈지대두와 밀가루 배합량이 동일한 대량 담금구, 시험구 B : 배합량이 A구와 같은 소량 담금구, 시험구 C : 탈지대두 6, 밀가루 4의 비율로 담금한 소량 담금구)의 숙성 과정중 각종 성분을 분석한 결과는 다음과 같다. 총질소와 아미노태 질소는 경시적으로 증가하였고, A구, B구, C구의 순으로 높았다. 환원당은 $20{\sim}40$일에 $12.13{\sim}15.76\;%$ 로 가장 높았고 120일에는 $10.75{\sim}12.40\;%$ 였으며 C구가 높았다. ethyl alcohol은 경시적으로 완만하게 증가되어 120일에 $0.45{\sim}1.23\;%$ 로 가장 높았고 B구와 C구가 높았다. 레블리린산은 담금 직후 A구에서 0.40% 였으나 100일에는 0.17% 로 저하되었고, B구와 C구는 당금 직후 $0.54{\sim}0.60\;%$ 였다. 젖산은 담금 직후보다 100일에 다소 증가되었으나 경시적으로 불규칙적인 변화를 보였고 A구가 높은 경향을 보였다. 유리 아미노산은 glutamic acid, lencine, alanine, phenylalanine이 모든 시험구에서 높았으나 histidine, tyrosine, methionine은 낮았다. 총 유리 아미노산 함량은 담금 직후보다 90일에 시험구 모두 증가되었고, B구, C구, A구의 순으로 높았다.

• 펄프종이기술
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• 제45권1호
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• pp.27-34
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• 2013

Quantitative analysis of 5-hydroxymethylfufural (5-HMF) conversion from fructose by dehydration and rearrangement was investigated by $^1H$-NMR spectroscopic method. Fructose was converted to 5-HMF in dimethylsulfoxide (DMSO)-$d^6$ or acidic deuterium hydroxide at controlled reaction temperature and time. With addition of internal standards (biphenyl for DMSO-$d^6$ solvent, and 2,5-dihydroxybenzoic acid for deuterium oxide solvent), conversion from fructose to 5-HMF was analyzed by $^1H$-NMR spectroscopy. Quantitative analysis was run by comparison with peak area integration between of 5-HMF and internal standard. In DMSO solvent, 5-HMF was stable end product but part of 5-HMF was converted to formic and levulinic acid at acidic aqueous medium.