• 한국미생물·생명공학회지
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• 제23권6호
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• pp.710-716
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• 1995

$\beta $-1, 4-D-arabinogalactanase isolated from alkalophilic Bacillus sp. HJ-12, approximate Mw 42 kDa, was generally stable in the range of pH 6-10 and below 50$\circ$C and its highest activity was observed at 60$\circ$C with pH 7-9. The isolated $\beta $-1, 4-D-arabinogalactanase specifically hydrolyzed $\beta $-1, 4-galactosyl linkage that is the major structure of soybean arabinogalactan (SAG) but not $\beta $-1, 3-galactosyl linkage of the other polysaccharides. K. was estimated as 0.67 mg/ml by the method of Hanes-Woolf plot. No metals and chemical reagents inhibited the enzyme activity but urea did. The active site of this enzyme assumed to be tryptophan residue. The hydrolysis products from SAG, assayed by gel chromatography, TLC and HPLC, were predominantly galactotetraose (Gal$_{4}$) and triose (Gal$_{3}$) with a small portion. $\beta $-1, 4-D-arabinogalactanase hydrolyzed ONPG as well as SAG, and the degree of hydrolysis of SAG was 15% which is lower than that by the other $\beta $-1, 4-galactanases from different sources. SAG treated with this enzyme resulted in the reduction of specific viscosity up to 70%.

• KSBB Journal
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• 제5권2호
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• pp.141-149
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• 1990

Aspergillus niger유래의 $\beta$-glucosidase를 (1)glutarald-ehyde를 가교제로 한 chitin과 chitosan담체에 covalent linkage (2)Amberite IRA93과 DEAE-cellulose담체에 succinylation시킨 후 흡착, 그리고 (3)alginarte, polyacry-lamide를 각종 가교제를 이용 entrapment등의 방법으로 고정화 하였다. Glutaraldehyde를 가교제로써 chitosan을 활성화시킨후 $\beta$-glucosidase를 고정화 시켰을때 효소활성 회수율이 31.5%로 가장 높았고, 또한 column형 반응기에서의 15일 경과 후 효소활성 유지도도 69%로서 가장 우수하였다. 또한 succinylation시킨 효소를 Amberite IRA93 담체에 흡착시켰을 때 효소활성 회수율은 24.7%였고 효소 활성유지도는 62%였다. 반면에 entra-pment 방법에 의한 $\beta$-glucosidase의 고정화는 효소의 계속적인 용출로 $\beta$-glucosidase의 고정화에는 적합하지 않았다. Chitosan담체에서의 고정화 최적조건을 조사한 결과 가교제인 glutaraldehyde의 최적농도는 0.4%였고, glutaraldehyde의 최적 반응 pH는 4.8이었다. 또한 column형 반응기를 이용하여 cellobiose로부터 glucose로의 전환율을 조사하여 고정화 효소의 효용성을 검토하였다.

• Journal of Plant Biotechnology
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• 제3권3호
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• pp.151-157
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• 2001

In order to breed barley lines with reduced viscosity of wort, a fractured starch mutant of naked barley cultivar, Nubet, was obtained from the M2 seeds mutated by the diethyl sulfate treatment. Seeds of this fractured starch mutant were opaque and the endosperm consists of angular, irregular and fractured starch. The mutant was caused by single recessive mutation and assigned by the symbol fra. The gene was located on chromosome 4, distal in long arm by linkage recombinations using translocation homozygote lethal test set. The linkage value between the fractured starch mutant and 72-4a, 72-4d were 26.0$\pm$4.9, 34.2$\pm$3.1 percent respectively. In addition to the reduced seed size, fewer kernels per spike and higher tillering ability, lower $\beta$-glucan viscosity and higher lysine content of the grain were associated with this mutant. $\beta$-glucan viscosity of the Nubet grains increased from 3 weeks after anthesis to matury and most of the viscose substances appeared to be stored in the middle of the endosperm tissue. Since the mutant grains showed better milling property as compared to Nubet, it can be used as breeding resources to develope new barley cultivars for maltins and milling purpose.

• 임산에너지
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• 제20권1호
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• pp.35-41
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• 2001

리그닌 모델화합물 Ⅰ-Ⅳ의 시료를 315℃에서 열분해 시킨 결과, 리그닌 모델호합물 Ⅰ, Ⅱ의 열분해에서는 guaiacol이 0.47mol, DMP가 0.57mol로, DMAP는 0.12와 0.23mol로 각각 생성되었으며, 리그닌 모델화합물 Ⅲ, Ⅳ의 열분해에서는 guaiacol의 생성이 0.26mol, DMP가 0.30mol로 TMAP는 0.09mol 과 0.15mol이 각각 생성되었다. 리그닌 모델화합물 열분해 메커니즘으로서는 우선 탈수되고, 이어서 β-O-4 결합이 개열되어 guaiacol, DMP, DMAP와 TMAP가 생성되는 것을 알 수 있었다.

• Applied Biological Chemistry
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• 제17권2호
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• pp.125-137
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• 1974

Selenomonas ruminantium 균체(菌體)를 TCA로 가열(加熱) 분해(分解)한 후 chloroform : methanol (1 : 3)로 추출(抽出)한 당지질(糖脂質)을 분리(分離)하고 이 당지질(糖脂質)을 acetone가용부분(可溶部分) spot A화합물(化合物)과 acetone불용부분(不溶部分)에서 ether로 다시 가용부분(可溶部分)을 추출(抽出)한 spot B화합물(化合物)의 두 부분(部分)으로 분리(分離)하고 이 두 화합물(化合物)에 대(對)하여 각각(各各) 그 화학구조(化學構造)를 구명(究明)하며 당지질(糖脂質)의 구조(構造)를 추정(推定)한 바 다음과 같은 결과(結果)를 얻었다. 1. 두 화합물(化合物)의 적외선흡수분석결과(赤外線吸收分析結果) spot A는 amino당(糖)에 O-acyl 및 N-acyl지방산(脂肪酸)이 결합(結合)하였으며 spot B는 amino당(糖)에 O-acyl 및 N-acyl지방산(脂肪酸)이 결합(結合)하고 인(燐)을 함유(含有)하고 있음을 알았다. 2. 두 화합물(化合物)을 GLC에 의(依)하여 지방산조성(脂肪酸組成)을 조사(調査)한 바 spot A.B화합물중(化合物中)에 있는 O-acyl 및 N-acyl 지방산(脂肪酸)은 ${\beta}-OH\;C_{13:0}$지방산(脂肪酸)이 대부분(大部分)이 였는데 저급(低級)의 hydroxy지방산(脂肪酸) ${\beta}-OH\;C_{9:0}$도 특이적(特異的)으로 함유(含有)되여 있음을 알았다. 3. 두 화합물(化合物)을 hydrazine분해(分解)를 시킨 결과(結果)를 paper chromatography로 조사(調査)한 바 spot A화합물(化合物)은 glucosamine 이 2분자(分子) 결합(結合)하여 있는 chitobiose와 같은 Rf 치(値)를 나타냈음으로 2분자(分子)의 glucosamine이 결합(結合)됨을 확인(確認)하고 spot B 화합물(化合物)의 낮은 Rf치(値)는 glucosamine에 인(燐)이 결합(結合)되여 있음을 알았다. 4. spotA화합물(化合物)의 산분해물(酸分解物)을 다시 ninbydrine으로 산화분해(酸化分解) 시키면 arabinose만이 생기는 것으로 보아 glucosamine의 amino기(基)는 $C_2$의 위치(位置)에 결합(結合)하여 있음을 알았다. 5. N-acetyl화(化)한 spot A에 $NaBH_4$를 추리(處理)한 결과(結果) glucosamine의 전량(全量)이 반감(半減)하는 것으로 보아 2분자(分子)의 glucosamine이 결합(結合)되여 있는 것을 알 수 있고 Morgan-Elson반응(反應) 및 $NaIO_4$분해(分解)에 의(依)하여 2개(個)의 glucosamine은 1.6결합(結合)임을 확인(確認)하였다. 6. N-acetyl화(化)한 spot A.B화합물(化合物)에 ${\beta}-N-acetyl$ glucosarninidase를 반응(反應)시킨 결과(結果) spot A화합물(化合物)은 100% N-acetyl glucosamine으로 분해(分解)되고 spot B화합물(化合物)은 분해(分解)되지 않았으므로 spot A화합물(化合物)만이 ${\beta}$결합(結合)을 하고 있음을 알았다. 7. $^{32}P$함유(含有) spot B화합물(化合物)에 phosphodiesterase 및 phosphomonoesterase를 작용(作用)시킨 결과(結果) phosphodiesterase는 반응(反應)치 않고 phosphomonoesterase에 의(依)하여 100% $^{32}P$가 유리(遊離)되는 것으로 보아 glucosamine 2분자(分子)에 한계의 인(燐)이 monoester결합(結合)을 하고 있음을 알 수 있다. 8. spot A화합물(化合物)은 glucosaminiyl ${\beta}-1.6-glucosamine$의 결합(結合)을 하였고 O-acyl 및 N-acyl지방산(脂肪酸)이 결합(結合)되여 있으며 주지방산(主脂肪酸)은 ${\beta}-OH\;C_{13:0}$임을 알았다. 9. spot B화합물(化合物)도 glucosaminyl ${\beta}-1.6-glucosamine$의 결합(結合)을 하고 O-acyl 및 N-acyl지방산(脂肪酸)이 결합(結合)되여 있으며 주지방산(主脂肪酸)은 ${\beta}-OH\;C_{13:0}$이나 인(燐)이 monoester결합(結合)을 하고 있는 것이 spot A화합물(化合物)과 특이(特異)함을 알았다.

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 1977년도 추계학술발표회 및 특별 강연초록
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• pp.197.2-197
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• 1977

Chitin은 N-acetyl glucosamin의 polymer이며 게새우 같은 절속등물의 골격성분을 이룬다. 이의 $\beta-1,$ 4-linkage를 분해하는 enzyme을 생성하는 Streptomyces속 1균주를 선별하여 기초실험을 하였다. 이 효소가 게로부터 조제한 chitin에 작용할 때 그 최적 pH는 7.0, 최적온도는 $50^{\circ}C이며$ pH6.5~8.0 사이에서 안정하다.(중략)

• 미생물학회지
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• 제29권2호
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• pp.85-91
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• 1991

.betha.-1,4-D-Glucan glucanohydrolase(EC 3.2.1.4;F-II-IV) purified from Trichoderma koningii was identified as a glycoprotein containing 9% carbohydrate. Isoelectric point of the enzyme was estimated to be 4.9 and molecular weight was determined to be approximately 58,000. The porducts of p-nitrophenyl-cellobioside ($PNPG_{2}$) catalyzed by the enzyme were p-nitrophenol(PNP) and p-nitrophenyl-glucoside($PNPG_{1}$). The Km value for $PNPG_{2}$ was estimated to be 0.97 mM in case of the holoside lindage and 10.4 mM in case of the aglycon linkage and their kcat values were $1.8*10^{5}$$ min^{-1}$ and $7.5*10^{5}$ $min^{-1}$ respectively. The product of p-nitrophenyl cellotriose($PNPG_{3}$) was only $PNPG_{1}$. The Km value for $PNPG_{3}$ was 69.5 .$\mu$M and kcat was $1*10^{8}$ $min^{-1}$ which implicates that the enzyme have higher affinity and higher hydrolysis rate toward $PNPG_{3}$ than toward $PNPG_{2}$. The enzyme showed its optimal activity at pH 4.0-4.5 and at 60.deg.C. The effect of gluconolactone on the activity toward $PNPG_{2}$ showed competitive inhibition pattern but glucose and cellobiose did not. The enzyme contained a high content of acidic and hydroxylated amino acids in contrast to basic amino acids.

• Journal of Life Science
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• 제9권1호
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• pp.26-34
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• 1999

A thermostable pullulanase has been isolated and purified from Thermus caldophilus GK-24 to a homogeneity by gel-filtration and ion-exchange chromatography. The specific activity of the purified enzyme was 431-fold increase from the crude culture broth with a recovery of 11.4%. The purified enzyme showed $M_{r}$ of 65 kDa on denaturated and natural conditions. The pI of the enzyme was 6.1 and Schiff staining was negative, suggesting that the enzyme is not a glycoprotein. The enzyme was most active at pH 5.5. The activity was maximal at $75^{\cire}C$ and stable up to $95^{\cire}C$ for 30 min at pH 5.5. The enzyme was stable to incubation from pH 3.5 to pH 8.0 at $4^{\cire}C$ for 24hr. The presence of pullulan protected the enzyme from heat inactivation, the extent depending upon the substrate concentration. The activity of the enzyme was simulated by $Mn^{2+}$ ion, }$Ni^{2+}$, $Ca^{2+}$, $Co^{2+}$ ions. The enzyme hydrolyzed the ${\alpha}$-1,6-linkages of amylopectin, glycogens, ${\alpha}$, ${\beta}$-limited dextrin, and pullulan. The enzyme caused the complete hydrolysis of pullulan to maltotriose and the activity was inhibited by $\alpha$, $\beta$, or $\gamma$-cyclodextrins. The $NH_{2}$-terminal amino acid sequence [(Ala-Pro-Gln-(Asp of Tyr)-Asn-Leu-Leu-Xaa-ILe-Gly-Ala(Ser)] was compared with known sequences of various sources and that was compared with known sequences of various sources and that was different from those of bacterial and plant enzymes, suggesting that the enzymes are structurally different.

• Journal of Applied Biological Chemistry
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• 제61권1호
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• pp.101-108
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• 2018

본 연구에서는 국내산 상황버섯의 효소 가수분해 전처리를 통한 ${\beta}-glucan$의 최적 추출조건을 확립하고 그에 따른 활성을 알아보고자 추출 조건에 따른 생이화학적활성을 측정하였다. 효소가수분해 조건을 최적화하기 위해 실시한 반응표면분석법의 결과 0.66%(v/v)의 viscozyme 농도에서 6.08시간 반응하는 것이 최적이라 예측되었으며($R^2=0.9245$), 이에 따라 최적 추출 조건에서 추출한 시료의 ${\beta}-glucan$ 함량은 1.9594 g/100 g으로 측정되었다. 추출 수율(0.76-16.40%)은 EBE가 NEBE에 비해 약 3배 높았다. ${\beta}-glucan$ 순도(11.15-59.05%)로 가장 높았으며, ${\beta}-glucan$ 함량 또한 0.26-3.38 g/100 g으로 EB (3.38 g/100 g)가 가장 높았다. 총당 함량(0.61-1.17 mg/mL)은 NEB, EB가 NEBE, EBE보다 높았으며, EB가 가장 높았다. 구성당 분석 결과, 모든 추출물에서 glucose의 함량이 가장 높았으며, 대조구와 효소 전처리구 모두 정제하면서 그 비율이 증가하였다. 단백질 함량(0.44-11.73 mg/mL)은 NEBE, EBE가 NEB, EB보다 높았으며, EBE가 가장 높았다. FT-IR 분석 결과 $890cm^{-1}$ 부근에서 peak가 확인되었기에 ${\beta}-glycosidic$ linkage를 가지고 있는 것으로 판단하였다. MTT assay를 통해 B6F10과 SK-MEL-5 세포 독성을 측정한 결과 B6F10의 경우 대조구의 세포 생존율을 100%로 하였을 때 세포 생존율이 80% 이상으로 나타나 세포독성을 보이지 않았으나, SK-MEL-5에서는 EBE를 $100{\mu}g/mL$의 농도로 처리하였을 때 세포 생존율이 75%로 나타나 약간의 세포독성을 보였다. Wound healing assay를 통해 암세포 증식 억제활성 측정 결과, 정제한 NEB, EB가 NEBE, EBE보다 활성이 높았으며, 특히 12시간일 때 EB $30{\mu}g/mL$를 처리한 경우 B6F10과 SK-MEL-5 모두에서 가장 높은 활성을 나타내었다.

• Archives of Pharmacal Research
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• 제16권1호
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• pp.36-42
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• 1993

To find biologically active components of the higher fungi of Korea, the carpophores of Auricularia polytricha, a well-known edible mushroom, were extracted with 0.14 M NaCl solution. The extract was successively fractionated by adding ammonium sulfate at various concentrations, and the respective precipitates were separated by centrifugation, then dialyzed and freeze-dried. When a does of 60 mg/kg of each was injected i.p. into ICR mice, the fraction which precipitated at 20% ammonium sulfate showed the highest toxicity, killing seven out of seven mice within two days. The fraction obtained at 40% ammonium sulfate showed the second highest toxicity. The two fractions were named auritoxin I and II after the genus name. However, they Nere shown to have nearly identical composition by physicochemical and 6.8% protein. The polysaccharide moiety was found to have 12.3% $\alpha$-linkage and 87.7% $\beta$-linkage and to be a heteromannoglucan consisting of 45.1% glucose, 435 mannose and 11.0% xylose. The protein moiety contained ten amino adids. The molecular weight of the toxin was $1.5\times10^6$ dalton by Sepharose CL-4B gel filtration. The modian lethal doses of auritoxin in mice were 56.4, 157.2 and 454.6 mg/kg by i.p., s.c. and p.o.administrations, respectively. The signs of intrxication were convulsion during the first 30 minutes after the injection, coma or sleeping within an hour, termor, lacrimation, nasal bleeding congestion, and death in 24 hours. Smong the various organs, the spleen was found to be enlarged remarkably. Human platelet aggregation was inhibited by the addition of auritoxin. The activity of malic dehydrogenase in vitro was inhibited by the toxin.