• Biomolecules & Therapeutics
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• 제6권4호
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• pp.364-370
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• 1998

Glycosidase inhibitors from urine of Bombyx mori were isolated and their inhibitory activities on glycosidases were evaluated. Six compounds were isolated by using several ion exchange columns, and their chemical structures were identified by the physicochemical and spectral data. Compound IV, V and Ⅵ were identified as 1-deoxynojirimycin, fagomine and 1,4-dideoxy-1,4-imino-D-arabinitol, respectively. Among six compounds isolated,1-deoxynojirimycin(IV) was the most potent inhibitor on $\alpha$-glucosidase and $\beta$-galactosidase of rat intestine, and its inhibitory activities for trehalase and almond $\beta$-glucosidase were relatively weak. Compound V and Ⅵl retained a little inhibitory potency toward $\alpha$-glucosidase and $\beta$-galactosidase. Compound II and III, however, have been found to have no effect on all glycosidases tested in this study.

• 생약학회지
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• 제38권4호
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• pp.358-362
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• 2007

From the EtOAc fraction of the MeOH extract of Ulmus davidiana, (-)-catechin (1), (-)-catechin-7-O-${\beta}$-D-apiofuranoside (2), and (-)-catechin-7-O-${\beta}$-D-xylopyranoside (3) were isolated and characterized on the basis of $^1H-and\;^{13}C-NMR$, and FABMS spectral data. Compounds 1-3 showed more strong reducing power activities than ${\alpha}-tocopherol$, a positive control.

• 미생물학회지
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• 제24권3호
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• pp.251-262
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• 1986

High-molecular-weight ${\beta}-glucosidase$ (EC 3.2.1.21) was purified from the culture filtrate of Trichoderma koningii through a four-step procedure including chromatography on Bio-Gel P-150, DEAE-Sephadex A-50 and SP-Sephadex C-50; and chromatofocusing on Polybuffer exchanger PBE 94. The molecular weight of the enzyme was determined to be about 101,000 by SDS-polyacrylamide gel electrophoreses, and the isoelectric point was estimated to be 4.96 by analytical isoelectric focusing. The temperature optimum for activity was about $55^{\circ}C$, and the pH optimumwas 3.5. The enzyme was considerably thermostable, for no loss of activity was observed when the enzyme was preincubated at $60^{\circ}C$ for 5h. Km values for cellobiose, gentiobiose, sophorose, salicin and $p-nitrophenyl-{\betha}-D-glucoside$ were 99.2, 14.7, 7.09, 3.15 and 0.70 mM, respectively, which indicates that the enzyme has much higher affinity towards $p-nitrophenyl-{\betha}-D-glucoside$ than towards the other substrates, especially cellobiose. Substrate inhibition by $p-nitrophenyl-{\betha}-D-glucoside$ and salicin was observed at the conecntrations exceeding 5mM. Gluconolactone was a powerful inhibitor against the action of the enzyme on $p-nitrophenyl-{\betha}-D-glucoside\;(K_i\;37.9\;{\mu}M)$, wherease glucose was much less effective ($K_i$ 1.95 mM). Inhibition was of the competitive type in each case. Transglucosylation activity was detected shen the readtion products formed from $p-nitrophenyl-{\betha}-D-glucoside$ by the enzyme were analysed using high-performance liquid chromatography.

• 대한화장품학회지
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• 제38권4호
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• pp.327-338
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• 2012

리퀴리티게닌과 이소리퀴리티게닌은 감초의 주요 플라보노이드 성분이다. 이들 플라보노이드는 수용성 감초 추출물과 ${\beta}$-glucosidase를 생성하는 잎새버섯 HB0071 균사체 발효배양을 통하여 생산하였다. 감초추출물 내 리퀴리티게닌과 이소리퀴리티게닌은 잎새버섯 발효배양 동안 현저히 증가하였다. 이 균주의 ${\beta}$-glucosidase의 활성은 배양 96시간을 기준으로 최고 91.5 mU/mL로 확인되었으며, 감초추출발효물로부터 생성된 리퀴리티게닌과 이소리퀴리티게닌의 함량은 HPLC 분석을 통하여 최대 $568.5{\mu}g/mL$과 $89.6{\mu}g/mL$로 확인되었다. 본 연구에서는 감초추출물의 잎새버섯 발효 전 후의 시료가 처리된 각질형성세포를 이용하여 자외선 UVB에 조사로 발현된 염증유발인자(COX-2)와 사이토카인(IL-$1{\beta}$, IL-6) 모두 감초추출발효물(FLEx)에서 농도의존적으로 발현이 억제되는 것을 확인하였다. 결론적으로 리퀴리티게닌과 이소리퀴리티게닌의 함량이 증가된 감초추출발효물은 자외선으로부터 손상된 피부 염증반응을 완화시켜줄 것으로 사료된다.

• 한국미생물·생명공학회지
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• 제25권2호
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• pp.115-120
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• 1997

Penicillium sp.-L4, a causative fungus of rot in citrus fruits, was isolated and its mode of hydrolytic enzyme production was investigated. Carboxymethylcellulase (CMCase), polygalacturonase(PGase), extra- & intra-cellular $\beta$-glucosidase and cellobiase were produced drastically by addition of substrates in minimal media. Production of the hydrolytic enzymes were induced efficiently by cellobiose and cellooligosaccharides which were the products of cellulose hydrolysis, but repressed by addition of mono-saccharide such as glucose, raffinose, galacturonic acid. The relative activity of p-nitrophenyl-$\beta$-D-glucopyranoside(PNPG) hydrolysis was higher than that of cellobiose hydrolysis in extracellular enzymes, and reverse is true in intracellular enzymes. Intact enzyme production of P. sp.-L4 on lemon peel lesion was sequential. $\beta$-Glucosidase and CMCase were produced first and followed by PGase. The enzyme productivities and pH in lesions were coincident with optimal pH of each enzyme activities.

• 생명과학회지
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• 제16권7호
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• pp.1148-1157
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• 2006

초고온성 세균인 Thermotoga maritima로부터 ${\beta}-glucosidase$ 유전자를 클로닝한 후 대장균 숙주에서 발현시켰다. 이 효소는 salicin, arbutin, $_pNPG$과 같은 탄소원의 ${\beta}$-글루코시드 결합을 가수분해하였다. 721개의 아미노산을 암호화하는 2,166 bp의 DNA 염기서열로된 유전자이였다. 다른 ${\beta}-glucosidase$ 효소들과 단백질 유사성을 비교한 결과 glycosyl hydrolase family 3에 속하였으며 MUG-nondenaturing PAGE와 SDS-PAGE에 의해 확인된 단백질의 크기는 약 81 kDa이었다. 효소활성은 pH 7.0, $80^{\circ}C$에서 가장 높은 활성을 나타냈으며 이 효소의 아미노산 서열에 있는 두 개의 아미노산 잔기 (232번 글루탐산과 242번 아스파르트산 잔기)를 알라닌으로 치환시켜 활성이 없어지는 것으로 보아 이 두 잔기가 효소활성에 중요한 역할을 하는 것으로 추정된다.

• Journal of Microbiology and Biotechnology
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• 제22권12호
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• pp.1705-1713
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• 2012

A gene encoding ${\beta}$-glucosidase was cloned from Weissella cibaria 37, an isolate from human feces. Sequence analysis showed that the gene could encode a protein of 415 amino acids in length, and the translated amino acid sequence showed homology (34-31%) with glycosyl hydrolase family 1 ${\beta}$-glucosidases. The gene was overexpressed in E. coli BL21(DE3) using pET26b(+) and a 50 kDa protein was overproduced, which matched well with the calculated size of the enzyme, 49,950.87 Da. Recombinant ${\beta}$-glucosidase was purified by using a his-tag affinity column. The purified ${\beta}$-glucosidase had an optimum pH and a temperature of 5.5 and $45^{\circ}C$, respectively. Among the metal ions (5mM concentration), $Ca^{2+}$ slightly increased the activity (108.2%) whereas $Cu^{2+}$ (46.1%) and $Zn^{2+}$ (56.7%) reduced the activity. Among the enzyme inhibitors (1 mM concentration), SDS was the strongest inhibitor (16.9%), followed by pepstatin A (45.2%). The $K_m$ and $V_{max}$ values of purified enzyme were 4.04 mM and 0.92 ${\mu}mol/min$, respectively, when assayed using pNPG (p-nitrophenyl-${\beta}$-D-glucopyranoside) as the substrate. The enzyme liberated reducing sugars from carboxymethyl cellulose (CMC).

• 한국미생물·생명공학회지
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• 제17권5호
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• pp.529-532
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• 1989

A strain of Streptomyces sp. (YS-221-B) extracellularly produced an inhibitory substance for $\alpha$-D-Glucosidase. The substance was purified 96-fold from culture filtrate by dialysis, heat treatment, adsorption on active carbon, Bio-Gel P-10 and Sephadex G-75 column chromatography with yield of 9.2%. The substance was stable in pH range from 7.0 to 11.0 at 37$^{\circ}C$, and a treatment at 10$0^{\circ}C$ for 20 min diminished only 15% of the original activity. The inhibitor was not inactivated by the treatment of $\alpha$-, $\beta$-amylases, glucoamylases, trypsin and chymotrypsin but inactivated by pyoteases from Streptomyces griseus and Tritirachium album.

• 한국미생물·생명공학회지
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• 제25권2호
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• pp.189-196
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• 1997

A producer of inhibitor against ${\beta}-glucosidase$ of Penicillium sp.-L4 was screened from Actinomycetes, and the isolated strain was identified as Streptomyces sp. The inhibitor produced was very stable against heat, acidic and alkaline conditions, proteolytic and amylolytic enzymes. The inhibotor was purified from culture broth through activated carbon treatment, ultrafiltration, anion and cation exchange, activated carbon columm, acetone precipitation and preparative HPLC. It showed inhibitory activities against a variety of dissacharide hydrolyzing enzymes produced by P.sp.-L4, and the mode of inhibition was competitive. Its structure and molecular formular was elucidated by IR, $^1H\;and\;^{13}C$ NMR and FAB/Mass spectrometry, which was identified as 1-deoxynojirimycin (dNM). dNM showed inhibitory effects on the cell growth and hydrolytic enzyme action of P.sp.-L4 on agar plate and infected lemon peel.

• 한국미생물·생명공학회지
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• 제42권4호
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• pp.347-353
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• 2014

본 연구는 인삼의 주요성분인 ginsenoside Rb1으로부터 보다 높은 생리기능성을 갖는 것으로 알려져 있는 compound K를 생산하기 위하여 Aspergillus usamii KCTC 6954에서 유래된 ${\beta}$-glucosidase를 사용하여 생물전환을 실시하였다. 15일 동안의 배양 중, 효소활 성 측정은 ${\rho}$-nitrophenyl-${\beta}$-glucopyranoside를 기질로 하여 분해 생성되는 ${\rho}$-nitrophenol (${\rho}NP$)을 비색계로 측정함으로써 실시되었다. 그 결과로서, 균주의 성장 속도는 접종 후 6일 후 최대로 나타났으며 이때의 ${\beta}$-glucosidas 활성도는 $175.93{\mu}M\;ml^{-1}min^{-1}$로 나타났다. 또 한 효소 반응의 최적 조건은 pH 6.0 이내에서는 $60^{\circ}C$인 것으로 나타났다. 배양 중 ginsenosides 분석 결과, 배양 9일 후에는 Rb1는 Rd 로 전환되고 15 days 후에는 compound K로 순차적으로 전환되는 것으로 나타났다. 효소반응에 있어서는 Rb1는 1시간 이내에 ginsenoside Rd로 전환되었고 8시간 이후에 최종산물인 compound K가 측정되었다. 본 연구결과로부터 Rb1으로부터 주요 생물학적 전환 경로는 $Rb1{\rightarrow}Rd{\rightarrow}F2{\rightarrow}$compound K로 나타났으며 이는 차후 Rd나 compound K와 같이 강한 생리기능성을 갖지만 자연에 미 량 존재하는 물질의 대량생산에 응용될 수 있을 것으로 기대된다.