• Journal of Microbiology and Biotechnology
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• v.33 no.4
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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.

• Clean Technology
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• v.20 no.2
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• pp.108-115
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• 2014

Various metal chlorides and acid catalysts in ionic liquid solvent were investigated to directly convert cellulose into 5-hydroxymethylfurfural (5-HMF). Metal chlorides containing Sn(II), Zn(II), Al(III), Fe(III), Cu(II), and Cr(III) were used and acidic ionic liquid immobilized on silica gel as an acid catalyst and commercial acid catalysts (sulfuric acid, chloric acid, Amberlyst-15,DOWEX50x8) were used for comparison studies. The acid strength and amount of acid catalysts were probed with Hammett indicator. The selectivity and yield of 5-HMF were determined with reaction temperature, reaction time and catalyst ratio. A catalyst containing $CrCl_3-6H_2O$ and $SiO_2-[ASBI]HSO_4$ showed the highest selectivity and it was found that this catalyst had higher activity than commercial solid acid catalysts such as Amberlyst-15 and DOWEX50x8. The selectivity of 5-HMF appeared to be mainly dependent on the acid strength and catalyst ratio, it was found that levulinic acid was produced from 5-HMF by rehydration.

• Bulletin of the Korean Chemical Society
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• v.27 no.6
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• pp.933-935
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• 2006

• Korean journal of food and cookery science
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• v.7 no.1
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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.

• Journal of environmental and Sanitary engineering
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• v.14 no.3
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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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• v.40 no.6
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• pp.556-560
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• 1997

The removal efficiency of COD and the production of ${\delta}-aminolevulinic$ acid (ALA) were concurrently investigated for both purifying the soybean curd wastewater of high BOD and utilizing the wastewater as a renewable substrate of ALA production using Rhodobacter capsulatus KK-10. Its wastewater was a favorable media for the growth of photosynthetic bacteria in terms of its environmental characteristics having COD/BOD rate of 0.98, ratio of BOD : N : P=100 : 6 : 4, BOD/N ratio of 17.2, lactic acid of 1,080 ppm. Its COD value wastewater was decreased to 94% and dry cell weight was approached to about 1.2 g/l after cultivation of the photosynthetic bacteria for 4 days. By the addition of 15 mM levulinic acid (LA) into the wastewater at the middle log phase of cell growth, the amount of ALA secreted was 55 mg/l. The ALA production was considerably increased to 114 mg/l under the cultural condition of 15 mM supplementations of glycine and succinate with LA at the same period. Furthermore the maximum ALA production of 120 mg/l and COD removal efficiency of 92% were accomplished in the soybean curd wastewater enriched with one addition of 15 mM LA and three serial additions 15 mM ALA precursors.

• Journal of Microbiology and Biotechnology
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• v.14 no.6
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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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• v.24 no.9
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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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• v.30 no.2
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• pp.147-152
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• 1987

Soy sauce which was prepared with a mixture of defatted soybeans ant wheat flour koji added into acid-hydrolyzed soy sauce was evaluated for the chemical changes in levulinic acid, ethanol and amino acids composition during fermentation, Results showed that the highest reducing sugar content of $12.13{\sim}15.76%$ was found after $20{\sim}40\;days$ fermentation. The contents of total nitrogen, amino nitrogen, and ethanol increased with increase in fermentation time. A high levulinic acid content was fount at the initial fermentation period which was gradually decreased with further fermentation. The content of lactic acid also increased during fermentation. Amino acids such as Glu, Leu, Ala, Phe, Asp, Thr, Ser, Gly, Met, Tyr, Lys, His, Arg and Pro were detected in all tested groups. Higher values were found particularly for Glu, Leu, Ala and Phe and lower values for His, Tyr and Met. The most of amino acids increased in their contents after 120 days of fermentation.

• Journal of Korea Technical Association of The Pulp and Paper Industry
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• v.45 no.1
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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.