• Journal of the Society of Cosmetic Scientists of Korea
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• v.41 no.2
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• pp.135-141
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• 2015

Cell senescence can be identified by cellular changes that occur as a result of intrinsic aging and/or diseases. In case of skin cells, aging and cell senescence caused by external factors results in cessation of cell proliferation and cellular malfunction, which, in turn, accelerates skin aging. In this study, inhibition of cell senescence and enhancement of cell function were studied using cordycepin to evaluate the potential for skin anti-aging agent. By comparing with the number of senescence associated with ${\beta}$-galactosidase (SA-${\beta}$-gal) positive cells in young and replicative aged human fibroblasts, it was found that replicative aged cells showed higher expression of ${\beta}$-galactosidase. Treatment of cordycepin - known as an anti-oxidative and anti-inflammatory agent - reduced ${\beta}$-galactosidase expression in senescent cells and enhanced cell survival in serum-free culture condition. Cordycepin also showed superb inhibition of ROS, which is another indicator of cell senescence. The results of this study proved the anti-aging effect of cordycepin on human fibroblasts and also proposed a possibility of its use as an anti-aging cosmetic ingredient.

• Food Science of Animal Resources
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• v.27 no.3
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• pp.329-336
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• 2007

This study was designed to find the optimum conditions for converting isoflavone glycoside to aglycone by ${\beta}-galactosidase$. Three different forms of the enzyme were tested and the optimum enzyme concentration, incubation temperature, pH, and incubation time were determined. Before treatment with enzyme, isoflavone contained 89.4% glycoside including daidzin, glycitin and genistin, and only 10.6% aglycone including daidzein, glycitein and genistein. Among the enzymes tested, the highest rate of isoflavone hydrolysis to aglycone, 35%, was observed when 3 unit/g Fungal Lactase (Amano Enzyme) was used. Higher incubation temperatures resulted in a higher rate of hydrolysis along with a greater loss of isoflavone mass. Therefore, body temperature $(37^{\circ}C)$ may be adequate for isoflavone conversion, with 44.9% hydrolysis and less than 10% loss of mass. As expected, a higher amount of aglycone was produced at pH 7 compared with other pH values. During 5hr of incubation, the conversion of glycoside to aglycone increased dramatically from 0 to 1hr, and plateaued thereafter. In addition, commercial soy-based milk was hydrolyzed more effectively with ${\beta}-galactosidase$ when incubated for 5hr. Based on the above results, the optimum conditions for isoflavone hydrolysis by ${\beta}-galactosidase$ were for 3 hr at $37^{\circ}C$, pH 7 with 3 unit/g Fungal Lactase (Amano Lactase), yielding an average total amount of aglycone ranging from 40 to 47%.

• Biotechnology and Bioprocess Engineering:BBE
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• v.6 no.5
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• pp.337-340
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• 2001

A galactooligosaccharide(GalOS)-producing yeast, OE-20 was selected from forty seven strains of yeast growing in Korean traditional Meju (cooked soybean) and the yeast was tentatively identified as Kluyveromyces maxianus var lactis by its morphology and fermentation profile. A maximum yield of 25.1%(w/w) GalOS, which corresponds to 25.1 g of GalOS per liter, was obtained from the reaction of 100 g per liter of lactose solution at 3$0^{\circ}C$, pH 7.0 for 18 h with an intracellular crude $\beta$-galactosidase. Glucose and galactosidase were found to inhibit GalOS formation. The GalOS that were purified by active carbon and celite 545 column chromatography were supplemented in MRS media and a stimulated growth was observed of some intestinal bacteria. In particular the growth rate of Bifidobacterium infantis in the GalOS containing MRS broth increased up to 12.5% compared to that of the MRS-glucose broth during a 48h incubation period.

• Journal of the Korean Society of Food Science and Nutrition
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• v.14 no.2
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• pp.157-163
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• 1985

Various cell wall polysaccharides and related enzyme activities in hot pepper fruit were determined at different stages of maturity. The uronic acid content of cell walls decreased between immature green and turning stage fruit and then increased by red ripe stage. In contrast, cellulose content of cell walls changed only a little during ripening. Total neutal sugar content of cell wall material decreased 50% and galactose content of the walls decreased about 80% by the turning stage. Polygalacturonase and ${\beta}-galactosidase$ activities, as well as total hemicellulose from isolated cell walls of ripening hot pepper fruit were studied using gel filtration chromatography. Polygalacturonase activity was not detectable but 5 isozymes of ${\beta}-galactosidase$ were resolved. The activities of the enzymes were relatively high and gel filtration showed that they differed in molecular weight. Hemicellulose content decreased during ripening and softening. The molecular weight profiles shifted from high molecular weight to low molecular weight polymers during ripening. The changes in cell walls that may be associated with fruit softening involve the alteration of hemicellulose prior to the degradation of wall-bound uronic acid. It is suggested that the decrease in cell wall galactose involved changes in turnover of new cell wall components.

• Journal of Microbiology and Biotechnology
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• v.1 no.4
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• pp.274-280
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• 1991

Effects of IPTG induction on cell growth and production of ${\beta}$-galactosidase-preS2 fusion protein (${\beta}$gal-preS2) were studied in a defined medium using a recombinant Escherichia coli JM109/pCMHB30. IPTG was added (0.2 mM) to induce the cloned-gene expression in the early-, mid-, and late-log growth phases. The most serious decreases in growth rate and plasmid stability were observed for the induction in the early-log growth phase. The expression level of ${\beta}$gal-preS2 attained by the induction in the mid-log phase was about 0.51 mg fusion protein/mg total cellular protein, which was 2- and 5-fold improvement over the levels obtained with the inductions in the early- and late-log phases. Formation of acidic byproducts including acetate and pyruvate showed different profiles during the fermentation period for each cases of induction; pyruvate was the major byproduct for the induction in the early-log phase while acetate production became more significant for the cases of inductions in the mid- and late-log phases.

• Korean Journal of Food Science and Technology
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• v.25 no.2
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• pp.191-195
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• 1993

The intestinal microflora of humans is an extraordinarily complex mixture of microorganisms, the majority of which are anaerobic bacteria. Amongst them, most prevalent bacteria are Bacteroides, Eubacterium, Peptococcus, Bifidobacteria. We isolated a Bacteroides fragilis strain from a Korean adult and examined various glycosidase activities of this strain. The activities of $N-acetyl-{\beta}-glucosaminidase,\;{\alpha}-fucosidase$, ${\beta}-glucuronidase$, chitobiase and PNPCase were stronger in Bacteroides fragilis Roid 8 than in other intestinal anaerobic bacteria. $N-acetyl-{\beta}-glucosaminidase$ was strongest, followed by ${\alpha}-fucosidase$, ${\beta}-glucuronidase$ and PNPCase. The activities of ${\beta}-galactosidase$, ${\beta}-xylosidase,\;{\alpha}-arabinofuranosidase$ were not present or very low. The activities of ${\alpha}-glucosidase$, ${\beta}-glucosidase$ and ${\alpha}-galactosidase$ were present but at a lower level than in Bifidobacterium. The effect of the carbon sources on the production of $N-acetyl-{\beta}-glucosaminidase$, ${\alpha}-fucosidase$, ${\beta}-glucuronidase$ and PNPCase of Bacteroides fragilis Roid 8 was investigated. :.actose and glucose lowered the production of the varous glycosidase enzymes studied in this work. In addition, we investigated the optimum temperature and pH of each glycosidase from Bacteroides fragilis Roid-8 using crude enzyme preparations.

• Journal of Microbiology and Biotechnology
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• v.27 no.8
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• pp.1392-1400
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• 2017

Galactooligosaccharides (GOSs) are known to be selectively utilized by Bifidobacterium, which can bring about healthy changes of the composition of intestinal microflora. In this study, ${\beta}-GOS$ were synthesized using bifidobacterial ${\beta}-galactosidase$ (G1) purified from recombinant E. coli with a high GOS yield and with high productivity and enhanced bifidogenic activity. The purified recombinant G1 showed maximum production of ${\beta}-GOSs$ at pH 8.5 and $45^{\circ}C$. A matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis of the major peaks of the produced ${\beta}-GOSs$ showed MW of 527 and 689, indicating the synthesis of ${\beta}-GOSs$ at degrees of polymerization (DP) of 3 and DP4, respectively. The trisaccharides were identified as ${\beta}-{\text\tiny{D}}$-galactopyranosyl-($1{\rightarrow}4$)-O-${\beta}-{\text\tiny{D}}$-galactopyranosyl-($1{\rightarrow}4$)-O-${\beta}-{\text\tiny{D}}$-glucopyranose, and the tetrasaccharides were identified as ${\beta}-{\text\tiny{D}}$-galactopyranosyl-($1{\rightarrow}4$)-O-${\beta}-{\text\tiny{D}}$-galactopyranosyl-($1{\rightarrow}4$)-O-${\beta}-{\text\tiny{D}}$-galactopyranosyl-($1{\rightarrow}4$)-O-${\beta}-{\text\tiny{D}}$-glucopyranose. The maximal production yield of GOSs was as high as 25.3% (w/v) using purified recombinant ${\beta}-galactosidase$ and 36% (w/v) of lactose as a substrate at pH 8.5 and $45^{\circ}C$. After 140 min of the reaction under this condition, 268.3 g/l of GOSs was obtained. With regard to the prebiotic effect, all of the tested Bifidobacterium except for B. breve grew well in BHI medium containing ${\beta}-GOS$ as a sole carbon source, whereas lactobacilli and Streptococcus thermophilus scarcely grew in the same medium. Only Bacteroides fragilis, Clostridium ramosum, and Enterobacter cloacae among the 17 pathogens tested grew in BHI medium containing ${\beta}-GOS$ as a sole carbon source; the remaining pathogens did not grow in the same medium. Consequently, the ${\beta}-GOS$ are expected to contribute to the beneficial change of intestinal microbial flora.

• Applied Biological Chemistry
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• v.38 no.6
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• pp.507-511
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• 1995

In an attempt to improve the productivity of ${\beta}-galactosidase$ from Bacillus sp. A1, which was isolated from soil and has remarkably higher transgalactosylation activity than lactose hydrolysis activity, a chemical mutation procedure using N-methyl-N'-nitro-N-nitrosoguanidine followed by selection was conducted. The final selection, designated as Bacillus sp. A4442, turned out to show a substantially increased enzyme productivity. Catabolite repression by glucose and lactose requirement as an inducer for the enzyme biosynthesis, which were shown in the parent strain, was markedly diminished; instead it was found out that galactose acts as another inducer. Because pH of medium, one of the most important factors for cell growth as well as enzyme production, is closely related with the sugar concentration during culture, it was kept in the optimum range of $6.5{\sim}7.5$; for this the initial glucose concentration was adjusted to be 0.5% which was thereafter maintained by the controlled pumping-in of lactose using the pH-stat technique. By doing so, we were able to increase the productivity of ${\beta}-galactosidase$ with high transgalactosylation activity up to $44\;unit/m{\ell}-broth$.

• Microbiology and Biotechnology Letters
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• v.19 no.2
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• pp.128-134
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• 1991

To study the reduced growth and synthesis, proeviously reported, of ${\beta}$-galactosidase of alkalophilic Bacillus sp. YS-309 at the higher lactose concentration of 0.5% (w/v) in the medium, lactose transport and utilization were examined. The results showed that lactose transport was influenced by the addition of four kinds of antibiotics, and tetracycline stimulated most but not valinomycin. PEP-potentials of the cells grown on lactose was estimated lower than the cells on glucose and on galactose. Thus, the transport of lactose was independent of intracellular PEP and phosphorylation reactions, and was thought to be uptaked directly or oxidized in part in the transport process. In the other hand, once lactose was uptaked into the cells, it was hydrolyzed by ${\beta}$-glactosidase to glucose and galactose. The former was metabolized fast but the latter was accumulated. Galactose and lactose were not utilized until glucose was mostly depleted in the medium. The ${\beta}$-galactosidase synthesis decreased in the presence of glucose over 0.2% and galactose over 0.05 to 0.1%, respectively. In conclusion, it was considered for glucose as a repressor and galactose as a inducer for ${\beta}$-galactosidase synthesis even though the mechanisms were not elucidated. Catabolite repression of glucose on the enzyme synthesis was not relieved by the addition of exogeneous cAMP.

• Microbiology and Biotechnology Letters
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• v.11 no.1
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• pp.15-21
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• 1983

This experiment was carried out to optimize the condition for the enzyme production by selected strain in the basal medium, to purify the enzyme and to characterize the purified enzyme. The results obtained were as follows. 1. The optimal conditions for the $\beta$-galactosidase production were initial pH 7.0 and temperature $65^{\circ}C$. 2. Enzyme was induced by the addition of lactose and galactose, and it was intracellular enzyme. 3. The purified enzyme was obtained with the increased level of activity approximately 28.5 folds as compared with crude enzyme and the yield of 15.2% by means of DEAE-Cellulose column chromatography, Sephadex G-150 gel filtration 4. $\beta$-galactosidase from final step of purification showed a sing1e protein band on polyacrylamide gel disc electrophoresis. 5. The optimal temperature and pH of the purified enzyme were $65^{\circ}C$, pH 6.5 for the hydrolysis of lactose.