• Journal of Microbiology and Biotechnology
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• 제20권6호
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• pp.1018-1021
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• 2010

Metallic and non-metallic isomerases can be used to produce commercially important monosaccharides. To determine which category of isomerase is more suitable as a template for directed evolution to improve enzymes for galactose isomerization, L-arabinose isomerase from Escherichia coli (ECAI; E.C. 5.3.1.4) and tagatose-6-phosphate isomerase from Staphylococcus aureus (SATI; E.C. 5.3.1.26) were chosen as models of a metallic and non-metallic isomerase, respectively. Random mutations were introduced into the genes encoding ECAI and SATI at the same rate, resulting in the generation of 515 mutants of each isomerase. The isomerization activity of each of the mutants toward a non-natural substrate (galactose) was then measured. With an average mutation rate of 0.2 mutations/kb, 47.5% of the mutated ECAIs showed an increase in activity compared with wild-type ECAI, and the remaining 52.5% showed a decrease in activity. Among the mutated SATIs, 58.6% showed an increase in activity, whereas 41.4% showed a decrease in activity. Mutant clones showing a significant change in relative activity were sequenced and specific increases in activity were measured. The maximum increase in activity achieved by mutation of ECAI was 130%, and that for SATI was 190%. Based on these results, the characteristics of the different isomerases are discussed in terms of their usefulness for directed evolution of non-natural substrate isomerization.

• Applied Biological Chemistry
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• 제49권4호
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• pp.276-280
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• 2006

본 연구에서는 경기(용인, 안성, 광주) 및 강원지역(오대산)에서 재래식 방법으로 제조된 청국장을 수집하여 지역별 발효 균주들의 동정과 특성분석 및 당 발효능을 조사하였다. 수집된 청국장에서 22종류의 균주를 분리하였고, 16S rDNA의 서열을 분석한 결과 이들은 대부분 Bacillus subtilis, B. licheniformis로 동정되었다. 경기지역의 주요 발효 균주는 B. subtilis와 B. licheniformis, 강원 지역은 B. licheniformis로 동정되었으며, 일반적인 생장 특성은 $5{\sim}7$시간의 유도기를 지나 $23{\sim}40$시간 이후에 휴지기로 접어들었다. 휴지기에 접어든 분리 균주들의 최대 생장 총 균수는 $1{\times}10^6\;CFU/ml$과 $5{\times}10^7\;CFU/ml$ 사이에 있었다. 균주들 간 당 발효능의 차이에 있어서 용인 지역은 D-xylose, xylitol, D-tagatose, Methyl-$\alpha$-D-mannopyranoside가, 안성은 D-lactose, D-tagatose, D-xylose, Methyl-$\alpha$-D-mannopyranoside, amygdalin, arbutin, esculin, 2-keto-gluconate가, 광주는 L-rhamnose, inositol, D-mannitol, D-sorbitol, celibiose, gluconate가, 그리고 오대산은 D-lactose에서 좋은 당 발효능을 보였다.

• Food Science and Biotechnology
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• 제17권6호
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• pp.1349-1351
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• 2008

This study examined the effects of temperature, D-psicose concentration, pH, and various amino acids on the Maillard browning reaction of D-psicose and glycine mixture and compared browning color intensity with those of other sugars, such as sucrose, D-glucose, D-fructose, and D-tagatose. When D-psicose (0.1 M) and glycine (0.1 M) mixture was heated at $70-100^{\circ}C$ for 5 hr, the absorbance at 420 nm increased with increasing reaction temperature and time. The Hunter a, b values, and color difference (${\Delta}E$) increased with increasing D-psicose concentration and pH within the range of pH 3-7 except at pH 6, while the L value decreased. The rate of Maillard browning reaction was in order of D-tagatose>D-psicose $\fallingdotseq$ D-fructose>D-glucose>sucrose. The browning color intensity of the D-psicose-basic and non-polar amino acids mixtures was higher than that of the D-psicose-acidic amino acids.

• Food Science and Biotechnology
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• 제17권4호
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• pp.839-841
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• 2008

The mutagenicity after heating of different sugars (glucose, fructose, galactose, and tagatose) on the non-enzymatic browning reaction in different sugars and glycine model system was investigated. The model system containing 0.2 M glycine and 0.2 M of different sugars in 10 mL water was heated at $150{\pm}5^{\circ}C$ for 30 min. After heating, degree of non-browning reaction intensity and mutagenicity using Salmonella typhimurium TA 98 were examined. Heated glycine model systems containing different sugars increased their mutgenicity ranged from 30 to 372 revertant colonies. After heating for 40 min, mutagenicity was achieved with glycine model systems containing 4 different sugars with by 145, 356, 206, and 369 revertants per plate, respectively. The glycine model systems containing fructose or tagatose were significantly (p<0.05) higher mutagenic activity than glycine model systems containing glucose or galactose after 40 min of heating. The linear regression between Maillard reaction intensity and mutagenic activities (slope=32.38, $R^2=0.93$) indicates that mutagenicity could be fully ascribed to Maillard reaction products.

• BMB Reports
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• 제40권5호
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• pp.801-804
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• 2007

The products of reactions catalyzed by Methanococcus. jannaschii (Mj) aldolase using various substrates were identified by gas chromatography (GC). Although Mj aldolase is considered a fuculose-1-phosphate aldolase based on homology searching after gene sequencing, it has not been proven to be a fuculose-1-phosphate aldolase based on its reaction products. Mj aldolase was found to catalyze reactions between glycoaldehyde or D, L-glyceraldehyde and DHAP (dihydroxyacetone phosphate). Before performing GC the ketoses produced were converted into peracetylated alditol derivatives by sequential reactions, i.e., dephosphorylation, $NaBH_4$ reduction, and acetylation. By comparing the GC data of final products with those of standard alditol samples, it was found that the enzymatic reactions with glycoaldehyde, D-glyceraldehyde, and D, L-glyceraldehyde produced D-ribulose-1-phosphate, D-psicose-1-phosphate, and a mixture of D-psicose and L-tagatose-1-phosphate, respectively. These results provide direct evidence that Mj aldolase is a fuculose-1-phosphate aldolase.

• 한국환경과학회지
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• 제30권10호
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• pp.879-889
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• 2021

This study characterizes the volatile aromatic and metabolite components of domestic native Apocynum lancifolium blossom. The accurate characterization of fragrances collected from the blossom was carried out using gas chromatography-mass. A total of 70 chemical components were identified, including ketones of acetophenone (29.22%), phenylethyl alcohol (10.54%), methyl-benzenemethanol (8.43%), benzyl alcohol (7.97%), natural bicyclic sesquiterpene types of caryophyllene (6.08%), gurjunene (6.20%), humulene (1.90%), and ocimene (1.04%). Overall, the content of ketones, alcohols, and terpenes was higher than that of others. The major metabolite components were pentanoic acid, malic acid, fructofuranoside, quinic acid, tagatose, sorbose, galactose, inositol, galactaric acid, glucopyranoside, and octadecenoic acid.

• Natural Product Sciences
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• 제26권3호
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• pp.214-220
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• 2020

Brassica oleracea var. gongylodes (red kohlrabi) is a biennial herbaceous vegetable whose edible bulbotuber-like stem and leaves are consumed globally. Sliced red kohlrabi tubers were extracted using methanol and the concentrated extract was partitioned successively with dichloromethane (CH₂Cl₂), ethyl acetate (EtOAc), n-butanol (n-BuOH) and water (H₂O). Repeated column chromatography of EtOAc fraction through silica, sephadex LH-20 and RP-18 gel led to isolation of eleven compounds of which compound 1 was a new glycosylated indole alkaloid derivative, 1-methoxyindole 3-carboxylic acid 6-O-β-D-glucopyranoside. Others were known compounds namely, β-sitosterol glucoside (4), 5-hydroxymethyl-2-furaldehyde (5), methyl-1-thio-β-D-glucopyranosyl disulfide (6), 5-hydroxy-2-pyridinemethanol (7), (3S,4R)-2-deoxyribonolactone (8), n-butyl-β-D-fructopyranoside (9), uridine (10) and three fructose derivatives, D-tagatose (11), β-D-fructofuranose (12) and β-D-fructopyranose (13). Similarly, isolation from CH₂Cl₂ fraction gave two known indole alkaloids, indole 3-acetonitrile (2) and N-methoxyindole 3-acetonitrile (3). The structure elucidation and identification of these compounds were conducted with the help of ¹³C and ¹H NMR, HMBC, HMQC, EIMS, HR-ESIMS and IR spectroscopic data, and TLC plate spots visualization. Compounds 2, 3, 4, 5, 6, 7, 8 and 9 are noted to occur in kohlrabi for the first time. Different bioactivities of these isolated compounds have been reported in literature.

• Journal of Dairy Science and Biotechnology
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• 제31권1호
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• pp.51-58
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• 2013

Emerging studies suggest that vegetables or fruit juices deemed to be potential alternative base medium for lactic acid bacteria fermentation. Until now, limited studies have been carried out to evaluate such applications. Thus, the objective of present study is that lactic acid bacteria were evaluated for their viability at low pH, growth during storage at low temperature, and $CO_2$ formation. Furthermore, the effects of grapefruit extract with respect to cell viability, sensory ability, and organic acid production were evaluated for these strains. The probiotic properties of the strains, including acid tolerance, bile tolerance, and adhesion to human intestinal epithelial cells (HT-29 cells), prebiotic characteristics, and safety features were examined. All strains survived in MRS medium broth adjusted to pH 3.8, at $10^{\circ}C$ for 6 days, and did not produce $CO_2$ to check post fermentation. The medium of grapefruit extract fermentation by Lactobacillus plantarum CJIH 203 resulted in maximal viable counts, compared with other strains, and the extract subsequently tasted sour due to the presence of lactic acid. Lactobacillus plantarum CJIH203 was highly resistant to artificial gastric juice and intestinal juice, while Lactococcus lactis SJ09 strongly adhered to HT-29 cells. Tagatose showed the greatest ability to enhance the growth of L. plantarum SJ21, relative to the other strains. All strains were verified by safety tests such as hemolysis, gelatin hydration, and urea degradation. Therefore, these strains could be promising candidates for use in reducing excessive post-fermentation and functional products.

• Journal of Dairy Science and Biotechnology
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• 제34권4호
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• pp.231-237
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• 2016

Kefir is a probiotic food. Probiotics have shown to be beneficial to health, and are currently of great interest to the food industry. Hence, this study was carried out to evaluate the lactase activity of kefir-isolated lactic acid bacteria. Three strains, Lactobacillu kefiri DH5 isolated from the kefir grains and Bifidobacterium animalis subsp. Lactis and Bifidobacteria longum 720, commercial probiotic LAB, were fermented in 10% reconstituted nonfat dry milk suspensions and incubated at $37^{\circ}C$ for 48 h, and then analyzed for various saccarides by HPLC. The results showed that changes in the concentrations of lactose and galactose were significantly decreased and increased, respectively (p<0.05), but all 3 probiotic strains tested in this study showed no increase in glucose concentration during 48 h of incubation. Both DH5 and BL720 showed high lactase activities (p<0.05), whereas BLC exhibited the lowest activity. Additionally, all three lactic acid bacteria showed high tagatose, but did not show high xylose and sedoheptulose. Finally, DH5, a kefir-isolated LAB, may have similar characteristics and properties to typical Bifidobacterium spp. and showed higher lactase activity than commercial Bifidobacterium spp.