Sang-Ho Baik

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.

Improved supercapacitor potential and antibacterial activity of bimetallic CNFs–Sn–ZrO2 nanofibers: fabrication and characterization

The objective of our study was to develop a new class of one-dimensional Sn–ZrO2 nanocrystal decorated CNFs. The utilized CNFs–Sn–ZrO2 composite was prepared by a sol–gel electrospinning method using polyacrylonitrile, ZrO(NO3)3·2H2O and SnCl2·6H2O as precursors. The physicochemical properties of the synthesized samples were characterized by X-ray diffraction (XRD), energy dispersive X-ray analysis (EDX), electron probe microanalysis (EPMA) and scanning electron microscopy (SEM). The bimetallic CNFs–Sn–ZrO2 composite possessed higher electrochemical capacitance and better stability than the monometallic and pristine samples as supercapacitor electrode materials. The CNFs–Sn–ZrO2 composite also exhibited admirable antibacterial activity. From the antimicrobial kinetic test results of E. coli; it was established that the composite (CNFs–Sn–ZrO2) possessed enhanced bactericidal activity compared to monometallic ones. The obtained high supercapacitance and bactericidal potential can be attributed due to the synergistic effect of Sn and ZrO2 in the carbon nanofibrous matrix. These results suggest the applicability of the fabricated nanofibers as electrode materials for supercapacitors and as antibacterial agents for decontamination of water.

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.

Lipid-lowering effects of Pediococcus acidilactici M76 isolated from Korean traditional makgeolli in high fat diet-induced obese mice.

The effect of Pediococcus acidilactici M76 (lactic acid bacteria) isolated from makgeolli on mice fed a high fat diet was investigated to clarify the lipid lowering function. C57BL/6J male mice were randomly divided into a normal diet (ND) group, high fat diet (HD) group, HD plus Pediococcus acidilactici DSM 20284 reference strain (PR) group, and HD plus Pediococcus acidilactici M76 strain (PA) groups. The lyophilized PA and PR strain were dissolved in distilled water at a final concentration of 1.25 × 109 cfu/mL and was given orally to animals at a dose of 4 mL/kg body weight for 12 weeks. The PA group had a lower final body weight, adipose tissue weight, and lipid profile than those in the HD group. Additionally, level of ACC, FAS and PPAR-γ, a key lipid synthesis enzyme, was markedly suppressed in the PA compared to those in the HD group. These data suggest that P. acidilactici M76 may exert a lipid-lowering effect in high fat diet- induced obese mice.

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.

Preparation and characterization of fermented dandelion (Taraxacum officinale) beverage using Lactobacillus acidophilus F46 having cinnamoyl esterase activity

The purpose of this study was to develop probiotic functional beverage with enhanced physiochemical properties by utilizing dandelion due to its high nutritional value. Lactic acid bacteria obtained from human feces were screened based on bacterial survival ability and cinnamoyl esterase (CE) activity. Dandelion extract fermented by Lactobacillus acidophilus F-46 maintained approximately 105–106 log CFU/mL over an 8 days period. Antioxidant activity by using DPPH radical scavenging activity of the prepared fermented dandelion beverage (FDB) were significantly increased compared to that of non-fermented dandelion beverage (NFDB). Moreover, CE activity was significantly enhanced during fermentation and showed the approximately 4.3 times increased concentration of caffeic acid up to 9.91 mg/100 mL after 8 days of incubation compared to NFDB. Therefore, it concluded that dandelion can be a good source for preparing a functional beverage and fermentation by LA-F46 enhanced the food functionality with enhanced caffeic acids.

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.

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.

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.