Nho-Eul Song

OPTIMAL PRODUCTION OF L-threo-2,3-DIHYDROXYPHENYLSERINE (L-threo-DOPS) ON A LARGE SCALE BY DIASTEREOSELECTIVITY-ENHANCED VARIANT OF L-THREONINE ALDOLASE EXPRESSED IN Escherichia coli

This study examined the efficient production and optimal separation procedures for pure L-threo-3,4-dihydroxyphenylserine (L-threo-DOPS) from a mixture of diastereomers synthesized by whole-cell aldol condensation reaction, harboring diastereoselectivity-enhanced L-threonine aldolase in Escherichia coli JM109. The addition of the reducing agent sodium sulfite was found to stimulate the production of L-threo-DOPS without affecting the diastereoselectivity ratio, especially at the 50 mM concentration. The optimal pH for diastereoselective synthesis was 6.5. The addition of Triton X-100 also strongly affected the synthesis yield, showing the highest conversion yield at a 0.75% concentration; however, the diastereoselectivity of the L-threonine aldolase was not affected. Lowering the temperature to 10°C did not significantly affect the diastereoselectiviy without affecting the synthesis rate. At the optimized conditions, a mixture of L-threo-DOPS and L-erythro-DOPS was synthesized by diastereoselectivity-enhanced L-threonine aldolase from E. coli in a continuous process for 100 hr, yielding an average of 4.0 mg/mL of L-threo-DOPS and 60% diastereoselectivity (de), and was subjected to two steps of ion exchange chromatography. The optimum separation conditions for the resin and solvent were evaluated in which it was found that a two-step process with the ion-exchange resin Dowex 50 W × 8 and activated carbon by washing with 0.5 N acetic acid was sufficient to separate the L-threo-DOPS. By using two-step ion-exchange chromatography, synthesized high-purity L-threo-DOPS of up to 100% was purified with a yield of 71%. The remaining substrates, glycine and 3,4-dihydroxybenzaldehyde, were recovered successfully with a yield of 71.2%. Our results indicate this potential procedure as an economical purification process for the synthesis and purification of important L-threo-DOPS at the pharmaceutical level.

Bacteria isolated from Korean black raspberry vinegar with low biogenic amine production in wine

A high concentration of histamine, one of the biogenic amines (BAs) usually found in fermented foods, can cause undesirable physiological side effects in sensitive humans. The objective of this study is to isolate indigenous Acetobacter strains from naturally fermented Bokbunja vinegar in Korea with reduced histamine production during starter fermentation. Further, we examined its physiological and biochemical properties, including BA synthesis. The obtained strain MBA-77, identified as Acetobacter aceti by 16S rDNA homology and biochemical analysis and named A. aceti MBA-77. A. aceti MBA-77 showed optimal acidity % production at pH 5; the optimal temperature was 25 °C. When we prepared and examined the BAs synthesis spectrum during the fermentation process, Bokbunja wine fermented with Saccharomyces cerevisiae showed that the histamine concentration increased from 2.72 of Bokbunja extract to 5.29 mg/L and cadaverine and dopamine was decreased to 2.6 and 10.12 mg/L, respectively. Bokbunja vinegar prepared by A. aceti MBA-77 as the starter, the histamine concentration of the vinegar preparation step was decreased up to 3.66 mg/L from 5.29 mg/L in the wine preparation step. To our knowledge, this is the first report to demonstrate acetic acid bacteria isolated from Bokbunja seed vinegar with low spectrum BA and would be useful for wellbeing vinegar preparation.

Microbial community, and biochemical and physiological properties of Korean traditional black raspberry (Robus coreanus Miquel) vinegar.

BACKGROUND The aim of this study was to elucidate the changes in microbial community and biochemical and physiological properties of traditional Muju black raspberry (Robus coreanus Miquel) vinegar (TMBV) during fermentation by culture-independent methods. RESULTS During vinegar fermentation, ethanol produced up to 120 g L(-1) until day 35, with continuously increasing yeast concentration to a total of log 7.6 CFU mL(-1) . After day 35, acetic acid bacteria (AAB) concentrations rose to log 5.8 CFU mL(-1) until day 144. Denaturing gradient gel electrophoresis analysis showed that Saccharomyces cerevisiae was detected until day 87 of the fermentation, at which point Acetobacter pasteurianus gradually took over as the dominant species. Total sugar was reduced to 6.6 °Brix and total acidity produced up to 44 g L(-1) . CONCLUSION In this study, we established the physicochemical analysis and growth dynamics of yeast and AAB during alcoholic and acetic acid fermentation of black raspberry by a traditional method. Overall, S. cerevisiae and A. pasteurianus species appeared to dominate the TMBV fermentation. In conclusion, this study demonstrated a suitable fermentation system for TMBV by the static surface method.

Preparation and characterization of oxygen generating (OG) hydrogels using γ-ray irradiation crosslinking

AbstractOxygen generating (OG) hydrogels immobilizing glucose oxidase (GOx, EC 1.1.3.4) and peroxidase (PO) (EC 1.11.1.7) were prepared using a γ-ray irradiation polymerization technique, and the properties were examined. In physiological tests of the OG hydrogels ranging from 15 to 70 kGy, GOx or PO showed higher gel contents at 87% for the poly(vinyl alcohol) (PVA) hydrogel and 90% for the poly(vinylpyrrolidone) (PVP) hydrogel. The optimal irradiation conditions affecting the entrapment ratio for preparing the OG hydrogel was 35 kGy for the PVP hydrogel, and both enzymes (GOx, 95% and PO, 89%). Optimal GOx and PO activity of the prepared OG hydrogel occurred at pH 6.0 and 35 °C, and both enzymes showed stable activity under acidic conditions. The optimal glucose condition was 4 mg, which resulted in a maximum oxygen generating yield by the OG hydrogels. When oxygen concentration was compared, the OG hydrogels showed higher oxygen pressure than that of control hydrogels without the GOx and PO enzymes. Moreover, the OG hydrogel continuously converted potassium iodide to iodine in a glucose dosage and time dependant manner for 15 h, producing approximately 0.1 M iodine in 5 h.