Young-Ran 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.

Effects of indigenous yeasts on physicochemical and microbial properties of Korean soy sauce prepared by low-salt fermentation.

This study deals with understanding the effects of salt reduction on both the physicochemical and microbiological properties of soy sauce fermentation and also the application of indigenous yeast starters to compensate for undesirable changes occurring in salt-reduced processes. Fermentation was tested in situ at a Korean commercial soy sauce processing unit. Salt reduction resulted in higher acidity as well as lower pH and contents of residual sugar and ethanol. Moreover, undesired flavor characteristics, due to a lack of distinctive compounds, was observed. In addition, putrefactive Staphylococcus and Enterococcus spp. were present only during salt-reduced fermentation. To control these adverse effects, a single or mixed culture of two indigenous yeasts, Torulaspora delbrueckii and Pichia guilliermondii, producing high ethanol and 3-methyl-1-butanol, respectively, were tested. Overall, all types of yeast applications inhibited undesirable bacterial growth despite salt reduction. Of the starter cultures tested, the mixed culture resulted in a balance of more complex and richer flavors with an identical flavor profile pattern to that obtained from high salt soy sauce. Hence, this strategy using functional yeast cultures offers a technological option to manufacture salt-reduced soy sauce while preserving its typical sensory characteristics without affecting safety.

Physicochemical and functional characteristics of a novel fermented pepper (Capsiccum annuum L.) leaves-based beverage using lactic acid bacteria

Two lactic acid bacteria (LAB), Lactobacillus homohiochii JBCC 25 and L. homohiochii JBCC 46, were isolated from a naturally fermented vinegar beverage. Fermented pepper leaves beverage (FPLB-3) which indicated good assessment based on a sensory evaluation test was prepared using the mixed strains. The prepared FPLB-3 showed enhanced functional aspects based on γ-aminobutyric acid (23.6 mg%), total polyphenol (2,828.2 gallic acid equivalents μg/g), and flavonoid contents (2,008.8 catechin equivalents μg/g). Moreover, FPLB-3 showed significantly stronger scavenging activities for the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical (84.6%) and superoxide radical (90.8%) and outstanding inhibitory activity against α-glucosidase of 65.9% at 50 mg/mL compared to the raw material. Based on the results, the newly developed FPLB using Wongi-1 pepper leaves and Lactobacillus is expected to be a novel functional food with natural antioxidant and antidiabetic capacities.

Monitoring of Yeast Communities and Volatile Flavor Changes During Traditional Korean Soy Sauce Fermentation

Flavor development in soy sauce is significantly related to the diversity of yeast species. Due to its unique fermentation with meju, the process of making Korean soy sauce gives rise to a specific yeast community and, therefore, flavor profile; however, no detailed analysis of the identifying these structure has been performed. Changes in yeast community structure during Korean soy sauce fermentation were examined using both culture-dependent and culture-independent methods with simultaneous analysis of the changes in volatile compounds by GC-MS analysis. During fermentation, Candida, Pichia, and Rhodotorula sp. were the dominant species, whereas Debaryomyces, Torulaspora, and Zygosaccharomyces sp. were detected only at the early stage. In addition, Cryptococcus, Microbotryum, Tetrapisispora, and Wickerhamomyces were detected as minor strains. Among the 62 compounds identified in this study, alcohols, ketones, and pyrazines were present as the major groups during the initial stages, whereas the abundance of acids with aldehydes increased as the fermentation progressed. Finally, the impacts of 10 different yeast strains found to participate in fermentation on the formation of volatile compounds were evaluated under soy-based conditions. It was revealed that specific species produced different profiles of volatile compounds, some of which were significant flavor contributors, especially volatile alcohols, aldehydes, esters, and ketones.

Quality Characteristics and Antioxidant Activity of Makgeolli Supplemented with Omija Berries (Schizandra chinensis Baillon)

The objective of this study was to evaluate the quality characteristics of the Korean rice wine, makgeolli, supplemented with omija berries (Schizandra chinensis Baillon) during the fermentation. The changes in pH, total acidity and contents of ethanol, amino acid, total soluble solids, reducing sugar, and total sugar after the completion of fermentation were determined. In comparison with control, omija-supplemented makgeolli showed significantly lower pH (3.46), lower contents of alcohol (17.2%), amino acids (1.85 g/L), and total sugar (17.5 g/L), and higher acidity (12.8 g/L). Moreover, supplementation with omija resulted in significantly higher antioxidant capacity of 1,1-diphenyl-2-picrylhydrazyl (DPPH) and hydroxyl radical scavenging activities, superoxide dismutase-like activity and reducing power, and higher levels of total polyphenol and flavonoid. Sensory evaluation revealed no significant difference on the overall acceptance, although better appearance and refreshing taste of omija-supplemented makgeolli were observed. Our results indicate that omija represents an effective natural additive for enhancing the biological activities of makgeolli.

Optimal production of exopolysaccharide by Bacillus licheniformis KS-17 isolated from kimchi

A strain producing exopolysaccharide (EPS) with strong hydroxyradical scavenging activity and antityrosinase activity has been isolated previously. However, the EPS production rate of the strain (7.3 g/L) was relatively low to utilize it at the industrial level. Therefore, in this study, optimal medium and fermentation conditions were examined to enhance EPS production by Bacillus licheniformis KS-17. Maximum EPS production was obtained in medium with 125 g/L sucrose, 30 g/L ammonium sulfate, and 10 mM calcium chloride without a phosphate source at 37°C for 5 days with the initial pH adjusted to 8.5. Under optimal culture conditions, EPS was produced at up to 27.2 (at 5 days) vs. 12.0 g/L (at 7 days under basal conditions), which was 2.3 times greater and in a shorter time than the production yield possible without optimizing conditions.

Molecular cloning, purification, and characterization of a novel thermostable cinnamoyl esterase from Lactobacillus helveticus KCCM 11223

ABSTRACT A gene encoding cinnamoyl esterase (CE), which breaks down chlorogenic acid (ChA) into caffeic and quinic acids, was cloned from Lactobacillus helveticus KCCM 11223. The gene with an open reading frame of 759 nucleotides was expressed in Escherichia coli, which resulted in a 51.6-fold increase in specific activity compared to L. helveticus KCCM 11223. The recombinant CE exists as a monomeric enzyme having a molecular weight of 27.4 kDa. Although the highest activity was observed at pH 7, the enzyme showed stable activity at pH 4.0–10.0. Its optimum temperature was 65°C, and it also possessed a thermophilic activity: the half-life of CE was 24.4 min at 65°C. The half-life of CE was 145.5, 80.5, and 24.4 min at 60, 62, and 65°C, respectively. The Km and Vmax values for ChA were 0.153 mM and 559.6 µM/min, respectively. Moreover, the CE showed the highest substrate specificity with methyl caffeate among other methyl esters of hydroxycinnamic acids such as methyl ferulate, methyl sinapinate, methyl p-coumarate, and methyl caffeate. Ca2+, Cu2+, and Fe2+ significantly reduced the relative activity on ChA up to 70%. This is the first report on a thermostable CE from lactic acid bacteria that can be useful to hydrolyze ChA from plant cell walls.

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.