• 대한화장품학회지
• /
• 제41권2호
• /
• pp.135-141
• /
• 2015

세포 노쇠화(cell senescence)는 나이 듦에 따른 내인성 노화 및 질병들에서 나타날 수 있는 세포의 노화인자 발현, 세포분열 정지 등의 현상으로 일컬어진다. 피부세포의 경우, 노화 및 외부요인으로 인한 세포 노쇠화가 일어나 세포분열의 정지 및 기능 이상이 관찰되며 이는 피부노화를 가속화시키는 요인이 된다. 본 연구에서는, cordycepin을 이용하여 노화된 피부세포의 세포 노쇠화 억제 및 기능 향상을 유도하여 피부노화 개선의 가능성을 제시하였다. 사람에서 유래한 섬유아세포를 이용하여 세포의 ${\beta}$-galactosidase 활성 세포염색 결과, 많은 계대의 세포에서 발현이 높게 나타남을 알 수 있었다. 항산화 및 항염 효과가 알려진 cordycepin을 많은 계대의 세포에 처리하였을 때 ${\beta}$-galactosidase 활성이 확연히 떨어짐을 확인하였고 무혈청 배지 조건에서 많은 계대 세포의 증식 및 생존율을 높이는 결과를 보였으며 세포 노쇠화와 많은 연관성이 대두되고 있는 미토콘드리아의 기능관련 실험을 진행한 결과, 높은 ROS억제능이 나타났다. 본 연구를 통하여 노화된 사람 피부 섬유아세포에서의 cordycepin의 세포 노화 개선능을 알 수 있었으며, 피부 항노화소재로서의 가능성을 확인하였다.

• 한국축산식품학회지
• /
• 제27권3호
• /
• pp.329-336
• /
• 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
• /
• 제6권5호
• /
• pp.337-340
• /
• 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.

• 한국식품영양과학회지
• /
• 제14권2호
• /
• pp.157-163
• /
• 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
• /
• 제1권4호
• /
• pp.274-280
• /
• 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.

• 한국식품과학회지
• /
• 제25권2호
• /
• pp.191-195
• /
• 1993

인체의 대장은 여러 종류의 균들에 의하여 상재균총이 이루어져 있는데 이들중 혐기성 균들이 주종을 이루고 있다. 이들 혐기성 균들 중 가장 많은 수가 Bacteroides이다. 본 연구에서 한국인으로부터 분리된 Bacteroides fragilis Roid 8은 장내의 다른 혐기성 균주들에 비하여 $N-acetyl-{\beta}-glucosaminidase$, ${\alpha}-fucosidase$, ${\beta}glucuronidase$ chitobiase, PNPCase 등의 활성이 높았다. ${\beta}-galactosidase$, ${\beta}-xylosidase$, ${\alpha}-arabinofuranosidae$활성은 없었고 ${\alpha}-glucosidase$, ${\beta}-glucosidase$, ${\alpha}-galactosidae$ 등의 생산은 Bifidobacteria 에 비하여 낮았다. BHI 기본배지에 여러 종류의 탄수화물을 첨가하여 배양한 뒤 생산된 $N-acetyl-{\beta}-glucosaminidae$, ${\alpha}-fucosidase$, ${\beta}-glucuronidase$ chitobiase, PNPCase, ${\beta}-glucosidase$, ${\beta}-glucosidase$ ${\alpha}-galactosidase$ 활성을 조사한 결과 모두 glucose와 lactose 첨가배지에서 이들 효소들의 활성이 낮았다. 조사된 모든 효소들에 대하여 특이적으로 현저히 생산을 증가시키는 당은 없었다. 이들 8개의 효소에 대하여 최적 pH와 최적온도가 조사되었다.

• Journal of Microbiology and Biotechnology
• /
• 제27권8호
• /
• pp.1392-1400
• /
• 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
• /
• 제38권6호
• /
• pp.507-511
• /
• 1995

토양으로부터 분리된 Bacillus sp.A1 균주는 가수분해활성보다는 전이활성이 훨씬 높은 ${\beta}-galactosidase$를 생산하기 때문에 산업적 응용 가능성이 있으나, glucose에 의한 catabolite repression을 보일 뿐만 아니라 lactose를 inducer로 요구한다는 결정적인 단점이 있어 갈락토올리고당의 제조에 직접 사용하기에는 적합하지 않았다. 따라서 galactose 전이효소의 생산성 제고를 꾀하기 위하여 N-methyl-N'-nitro-N-nitrosoguanidine를 이용한 3단계 변이를 시도하여 A4442변이주를 선발하였다. 이 변이주의 효소생산 능력은 괄목할 만큼 향상하였으며(약 20배 내외) catabolite repression과 lactose 요구성이 상당히 해제되었음을 확인하였다. 아울러 갈락토스가 새로운 inducer로 작용하는 것도 관찰하였다. 변이주를 이용한 발효시에 당의 농도와 배양액의 pH는 상호연관되어 효소의 생산에 영향을 끼쳤다. pH stat 기법을 이용하여 배양중 당의 농도를 0.5% 이하로 조절할 때 pH는 $6.5{\sim}7.5$ 범위내로 유지되었으며 효소활성은 $44\;unit/m{\ell}-broth$로 높게 나타났다.

• 한국미생물·생명공학회지
• /
• 제19권2호
• /
• pp.128-134
• /
• 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.

• 한국미생물·생명공학회지
• /
• 제11권1호
• /
• pp.15-21
• /
• 1983

전보에서 분리 동정한 Y-33 균주의 $\beta$-galactosise 효소생산조건을 검토하고 효소를 정제하여 정제효소의 성질을 조사한 결과는 다음과 같다. 1. 효소생산을 위한 최적초발 pH는 7.0이었고 최적온도는 $65^{\circ}C$이었다. 2. 효소는 lactose와 galactose에 의하여 유도되어졌으며 세포내효소이었다. 3. 조효소액을 1차 DEAE-cellulose, 2차 DEAE-cellulose column chromatography 및 Sephadex G-150로 gel filtration하여 정제도가 28.3배, 수율이 15.2%의 정제효소를 얻었다. 4. 정제효소는 polyacrylamide gel 전기 영동에 의하여 순도가 검정되었다. 5. 정제효소의 유당가수분해를 위한 최적작용 온도는 $65^{\circ}C$. pH는 6.5이었다.