• 대한약학회:학술대회논문집
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• 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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• pp.356.3-357
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• 2002

Metoprolol (MT) is one kinds of adrenergic beta-blockers. Its (S)-enantiomer is known to be more active than the (R). Recently. the x-ray structure of beta-blocker. (S)-propranolol (a-naphthyl analogue), complexed in a mould fungal cellulase. Cel7A. was reported and the (R)-form did not build any complex. And in our previous study the conformation and stability of MT in carboxymethylated beta-cyclodextrin (BCD) complex was determined by NMR. HPLC, UV and electrophoresis measurement. (omitted)

• Preventive Nutrition and Food Science
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• 제7권3호
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• pp.310-316
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• 2002

Extracellular cyclodextrin glucanotransferase (CGTase) from Paenibacillus sp. JK-12 was purified through sev-eral purification steps consisting of ammonium sulfate precipitation and chromatographies on DEAE-sephadex A-50 and Mono QIM HR5/5. The purified CGTase exhibited a single band on SDS-PAGE and was estimated to be approximately 82 kDa. The isoelectric point of the enzyme was 7.2 as determined by isoelectric focusing. The CGTase from Paenibacillus sp. JK-12 had a transglucosylation activity at the C-2 position of L-ascorbic acid. The optimum pH and temperature for the CGTase activity were 8.0 and 5$0^{\circ}C$, respectively. The enzyme activity was stable from pH 6.0 to 9.() and at temperatures up to 55$^{\circ}C$ at pB 8.0, having 80% residual activity. The activity of the CGTase was strongly resistant to metals such as A $g^{+}$ and $Ba^{2+}$ but slightly inhibited by H $g^{+}$, N $i^{2+}$ and $Mg^{2+}$. The enzymeproduced $\alpha$ -cyclodextrin ($\alpha$-CD) and $\beta$-CD as the main products from starch, but not ${\gamma}$-CD.X>-CD.

• 한국축산식품학회지
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• 제28권2호
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• pp.211-217
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• 2008

The objective of the present study was to develop cholesterol-reduced and phytosterol-supplemented mayonnaise using crosslinked $\beta$-cyclodextrin and examine its physicochemical and sensory attributes during 10 months of storage. The composition of cholesterol-reduced phytosterol-supplemented mayonnaise was similar to the control. The amount of cholesterol removed ranged from 90.67 to 92.47%. The TBA absorbance of the samples showed that the more phytosterol the sample contained, the lower the TBA absorbance value. The viscosity of cholesterol-reduced mayonnaise with 2.0% phytosterol decreased significantly during storage (p<0.05). The color changes of mayonnaise during storage showed a decrease in the L- and b-values, and an increase in the a-value. The experimental mayonnaise maintained emulsion stability, which was significantly lower in 2.0% phytosterol-supplemented mayonnaise. With regard to sensory attributes, most characteristics were similar to the control mayonnaise, however, the addition of phytosterol had a negative effect on stickiness and bitterness. These results indicate that the cholesterol-reduced and phytosterol-supplemented mayonnaise has decreased oxidation and maintains most of its physicochemical and sensory properties during storage.

• Journal of Microbiology
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• 제42권4호
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• pp.319-327
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• 2004

An additional amylase, besides the typical $\alpha-amylase,$ was detected for the first time in the cytoplasm of B. subtilis SUH4-2, an isolate from Korean soil. The corresponding gene (bbmA) encoded a malto­genic amylase (MAase) and its sequence was almost identical to the yvdF gene of B. subtilis 168, whose function was unknown. Southern blot analysis using bbmA as the probe indicated that this gene was ubiquitous among various B. subtilis strains. In an effort to understand the physiological function of the bbmA gene in B. subtilis, the expression pattern of the gene was monitored by measuring the $\beta-galactosidase$ activity produced from the bbmA promoter fused to the amino terminus of the lacZ struc­tural gene, which was then integrated into the amyE locus on the B. subtilis 168 chromosome. The pro­moter was induced during the mid-log phase and fully expressed at the early stationary phase in defined media containing $\beta--cyclodextrin\;(\beta-CD),$ maltose, or starch. On the other hand, it was kept repressed in the presence of glucose, fructose, sucrose, or glycerol, suggesting that catabolite repression might be involved in the expression of the gene. Production of the $\beta-CD$ hydrolyzing activity was impaired by the spo0A mutation in B. subtilis 168, indicating the involvement of an additional regu­latory system exerting control on the promoter. Inactivation of yvdF resulted in a significant decrease of the $\beta-CD$ hydrolyzing activity, if not all. This result implied the presence of an additional enzyme(s) that is capable of hydrolyzing $\beta-CD$ in B. subtilis 168. Based on the results, MAase encoded by bbmA is likely to be involved in maltose and $\beta-CD$ utilization when other sugars, which are readily usable as an energy source, are not available during the stationary phase.

• Molecules and Cells
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• 제23권1호
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• pp.57-63
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• 2007

Leukotrienes (LTs) are produced by several biosynthetic enzymes including cytosolic phospholipase $A_2$ ($cPLA_2$), 5-lipoxygenase (5-LO), and 5-lipoxygenase activating protein (FLAP) in the perinuclear area. In the present study, we showed that pretreatment with methyl-${\beta}$-cyclodextrin (MβCD), a cholesterol-depleting agent, dramatically reduced the synthesis of LTs in response to A23187 in mast cells. A23187-induced LT synthesis was inhibited by pretreatment with M${\beta}$CD, and this effect was reversed when cholesterol was added. In an approach to identifying the $M{\beta}CD$-sensitive protein(s), we observed that FLAP co-localized with flotillin-1, a lipid raft marker protein, in the lipid raft-rich low-density region of sucrose gradients. In addition, electron microscopic analysis revealed that FLAP co-localized with flotillin-1. Together, these results suggest that FLAP is present in cholesterol-rich lipid raft-like domains and that its localization in these domains is critical for LT synthesis.

• 미생물학회지
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• 제53권3호
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• pp.208-215
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• 2017

Streptococcus pyogenes ATCC 700294 유전체로부터 cyclomaltodextrinase (SPCD)로 예상되는 유전자를 발견하였다. SPCD는 총 567개의 아미노산으로 이루어진 66.8 kDa의 효소이며, 기존에 알려진 CDase 계열 효소들과 37% 미만의 아미노산 서열 상동성을 가진다. 본 연구에서는 SPCD 유전자를 클로닝하였으며, 대장균 내에서 카복시 말단에 6개의 histidine 잔기가 결합된 dimer 형태로 발현 및 정제되었다. SPCD는 pH 7.5, $45^{\circ}C$의 반응조건에서 최대의 활성을 나타내었으며, ${\beta}$-cyclodextrin, starch, maltotriose를 기질로 반응하여 maltose를 주산물로 생성하였다. 또한 pullulan을 panose 단위로 분해하며, acarbose를 glucose와 acarviosine-glucose로 가수분해하는 CDase 계열의 효소로 확인되었다. 그러나, SPCD는 다른 효소에 비해 저분자 소당류인 ${\beta}$-cyclodextrin에 대한 활성이 매우 높고, starch 및 pullulan과 같은 고분자 기질에 대해 매우 낮은 활성을 보였다. 또한 maltotriose 분해 활성이 매우 낮은 반면 acarbose에 대해 상대적으로 높은 가수분해 활성을 가지나, 당전이 활성은 매우 낮아 다른 CDase 계열 효소들과 구별된다.

• 한국축산식품학회:학술대회논문집
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• 한국축산식품학회 2005년도 제36차 추계 학술발표대회
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• pp.271-275
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• 2005

본 연구결과, ${\beta}$-CD의 재활용을 위해 adipic acid로 ${\beta}$-CD를 가교시킨 후, 우유의 콜레스테롤 제거 실험에 사용한 결과, 10회 재활용하는 동안 평균 89.90%의 콜레스테롤이 제거되었으며, 이에 따른 가교화 ${\beta}$-CD의 재활용률은 97.30%로 나타났다. 재활용에 따른 가교화 ${\beta}$-CD의 수율은 8회 재활용할 때까지 감소폭이 매우 적었으며, 가교화 ${\beta}$-CD의 구조 역시, 8회 재활용할때까지는 ${\beta}$-CD간의 가교결합이 유지되다가 이후부터는 대부분 powder ${\beta}$-CD 형태로 존재하는 것으로 관찰되었다. 재활용하는 동안 콜레스테롤 제거가 가능한 ${\beta}$-CD를 정량한 결과, 8회 재활용할 때까지 일정한 수치를 유지하는 것으로 나타났다. 위 실험 결과, adipic acid로 가교시킨 ${\beta}$-CD를 이용하여 우유의 콜레스테롤 제거 시, 여러 번 사용하여도 가교화 ${\beta}$-CD의 물리적 성질에 변화가 거의 없이 자체의 특성을 유지하는 것으로 보아, 이를 유가공 산업에 적용 시, 경제적인 효과가 매우 클 것이라고 사료된다.

• 한국해양환경ㆍ에너지학회지
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• 제7권3호
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• pp.146-151
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• 2004

포접력을 지닌 2-hydroxypropyl-β-cyclodextrin(β-HPCD)을 비롯한 cyclodextrin계 화합물은 소수성 유기물질의 용해도를 증가시킴으로써 미생물에 의한 분해를 촉진시키나 그 외의 자세한 기작은 알려져 있지 않다. 본 실험에서는 β-HPCD 유무에 따라 고분자 PAHs 분해력을 지닌 Novosphingobium pentaromtivorans US6-1 균주의 pyrene라 benzo[a]pyrene(B[a]P)의 분해정도, 이때의 biomass 변화 및 dioxygenase활성을 측정함으로써 PAHs 생분해과정에서 β-HPCD의 역할을 규명하고자 하였다. 실험구는 균주와 PAHs, β-HPCD의 존재 유무에 따라 8개 조건으로 준비하였으며 각 실험구의 배양기간에 따른 PAHs 분해도와 생체량의 변화를 측정하였다. Pyrene의 경우 β-HPCD가 존재함에 따라 분해도가 증진되는 것이 확인되었으며, 특히 B[a]P의 분해에는 β-HPCD가 필수요소로 작용하였다. 생체량의 변화는 PAHs의 존재 유무에 영향을 받지 않았고 β-HPCD의 존재 유무에 따라 차이를 나타내었다. 또한 US6-1 균주는 β-HPCD가 포함된 MM2 무기영양배지에서 전배양 할 때에 ZoBell 배지에서 전배양하는 경우에 비해 catechol-1,2-dioxygenase 효소활성이 높은 것으로 나타났으나 그 값은 빈영양상태에서 배양한 세포의 효소환성과 큰 차이를 보이지는 않았다. 이상의 결과로 볼 때 β-HPCD는 PAHs의 이용성을 높여주는 동시에 탄소원으로 이용되어 균주의 생체량 증가에 기여함으로써 PAMs의 분해력을 증진시키지만 dioxygenase 효소활성에는 영향을 미치지 않는 것으로 사료된다.

• 한국식품영양학회지
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• 제13권4호
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• pp.342-347
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• 2000

과요오드산-산화전분으로 보리 $\beta$-amylase(Bio-zyme ML, 일본 아마노제약)를 변형시켜서 인공당단백질을 만들었다. pH 8.0에서 변형한 효소는 비변형효소의 92%, pH 9.7에서 변형한 효소는 42%의 활성이 남았다. 6$0^{\circ}C$에서의 열안정성은 $\alpha$-cyclodextrin ( $\alpha$-CD) 존재 시에 변형하여 $\alpha$-CD존재시 분석한 효소는 10분 뒤에 비활성의 8%가 남은 반면 변형하지 않은 효소는 4.5%밖에 남지 않았다. pH안정성은 변형 시켜서 $\alpha$-CD존재 하에 분석한 효소가 가장 높아서 pH 2~5와 7~12에서 안정성이 매우 증가하였다. HPLC분석 결과 효소는 두 개의 피크를 나타냈으며 변형시킨 것은 당결합으로 분자량이 커져서 유출시간이 약간 빨라졌다.

• Journal of Microbiology and Biotechnology
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• 제9권6호
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• pp.811-819
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• 1999

The ${\beta}-CGTase$ gene of alkalophilic Bacillus firmus var. alkalophilus was cloned into E. coli using $pZErO^{TM}-2$ as a vector. The cloned gene encoded a total of 710 amino acid residues consisting of 674 amino acids of the matured protein and 36 amino acids of the signal peptide, including 20 amino acids from the lacZ gene in the vector. Although the cloned ${\beta}-CGTase$ gene did not contain the promoter and start codons, it was expressed by the lac promoter and lacZ start codon in the $pZErO^{TM}$ vector. A comparison was made with the amino acid sequence and ten other CGTases from Bacillus sp. Also, ten highly conserved regions, which are important amino acid residues in catalysis of CGTase, were identified. The lac promoter used for expression of the ${\beta}-CGTase$ gene was induced constitutively in recombinant E. coli even without IPTG possibly because of a lack of the lacI gene in both host and vector, repressing the lacZ gene in the lac operon. Its expression was catabolically repressed by glucose, however, its repression was reduced by soluble starch, mainly because of the extremely high increase of the cAMP level. ${\beta}-CGTase$ can be overproduced in the recombinant E. coli by maintaining intracellular cAMP levels mostly through the intermittent feeding of glucose during cultivation.