Earticle

생리활성이 우수한 발효홍삼농축액 특유의 쓴맛과 발효과정 중 생성된 신맛 등의 이미를 개선하기 위해 ${\beta}$-, ${\gamma}$-cyclodextrin(CD) 혼합시료를 이용하여 이미를 포접시켜 관능평가와 ginsenoside Rb1 및 총산도를 분석하고 최대 포접효율을 나타내는 ${\beta}$-, ${\gamma}-CD$ 혼합시료의 농도를 반응표면분석 중심합성법으로 계획하여 기호성이 증진된 발효홍삼농축액을 제조하는 최적화 조건을 확인하였다. ${\beta}$-, ${\gamma}-CD$ 혼합시료를 이용한 쓴맛 포접 효과를 확인한 결과 예측된 정상점은 최저점으로 ${\beta}-CD$가 3.74%이고 ${\gamma}-CD$ 20.63%일 때 쓴맛이 2.07로 최소값을 나타내었다. Ginsenoside Rb1 포접 효과를 확인한 결과 예측된 정상점은 최대값으로 분석한 결과 ${\beta}-CD$가 3.47%이고 ${\gamma}-CD$가 19.89%일 때 ginsenoside Rb1이 96.75%로 최대값을 나타내었다. ${\beta}$-, ${\gamma}-CD$ 혼합시료를 이용한 신맛 포접 효과는 ${\beta}-CD$가 9.34%이고 ${\gamma}-CD$ 7.96%일 때 신맛이 5.63으로 최소값을 나타내었다. 젖산의 포접 효과는 ${\beta}-CD$가 16.00%이고 ${\gamma}-CD$ 13.18%일 때 젖산의 포접효율이 67.73%로 최대값을 나타내었다. 실험 결과를 중심으로 최적 조건을 중첩시켜 ${\beta}-CD$와 ${\gamma}-CD$의 함량을 달리하여 발효홍삼농축액의 이미를 줄이기 위해서는 신맛의 포접 정도는 6, 쓴맛 포접 정도는 4, 산의 포접 정도는 65%, ginsenoside 포접 정도는 95%로 하는 조건을 만족하는 영역으로 ${\beta}-CD$와 ${\gamma}-CD$의 첨가량은 ${\beta}-CD$가 10%이고 ${\gamma}-CD$가 13%임을 알 수 있었다.

Fermented red ginseng concentrate is known as a healthy food source, whereas it has off-flavor such as bitterness and sour flavor based on fermentation. ${\beta}$- and ${\gamma}$-cyclodextrin (CD) were used to encapsulate the off-flavor of fermented red ginseng concentrate by using response surface methodology design on ${\beta}$- and ${\gamma}-CD$ combination. The reducing effects were analyzed by sensory evaluation for bitter and sour tastes, ginsenoside Rb1, and total acidity. The optimized mixing ratio of ${\beta}$- and ${\gamma}-CD$ for reducing bitterness was the least expected value of 2.07 at ${\beta}-CD$ 3.74% versus the soluble solid content of fermented red ginseng concentrate and the ${\gamma}-CD$ 20.63% mixture. The encapsulation effects of ginsenoside Rb1 were the most expected value of 96.75% at ${\beta}-CD$ 3.47% and ${\gamma}-CD$ 19.89% mixture. The encapsulation effects of sour taste were the least expected value of 5.63 at ${\beta}-CD$ 9.34% and ${\gamma}-CD$ 9.96% mixture. The encapsulation effects of lactic acid were the most expected value of 67.73% at ${\beta}-CD$ 16.0% and ${\gamma}-CD$ 13.18% mixture. Based on encapsulation and each optimized combination, the most effective entrapping ${\beta}$-and ${\gamma}-CD$ combination ratio was ${\beta}-CD$ 10% and ${\gamma}-CD$ 13%.

선행연구에서 얻은 alkalophilic Bacillus I-5 유래의 CGTase wild-type과 가수분해능이 강화된 mutant 효소를 활용하여 waxy rice starch로부터 아밀로펙틴 클러스터를 제조하였다. SEC-MALLS-RI 분석법으로 CGTase wild-type과 mutant 효소가 처리된 시료의 평균 분자량을 확인한 결과 10분가량의 효소반응으로 두 반응 모두 평균 분자량은 $10^4{\sim}10^5Da$으로 급격히 감소하였음을 확인하였으며, 일정 반응 시간이 경과한 이후에는 더 이상 분자량의 감소가 일어나지 않음으로 미루어 시료가 아밀로펙틴 클러스터 단위로 분해되었으며 그 분자량은 $10^4{\sim}10^5Da$ 정도임을 알 수 있다. 또한 MALDI-TOF/MS 분석을 통하여 CGTase wild-type은 다양한 종류의 cyclic 형태의 maltodextrin을 생성하고 있으며 mutant 효소는 주로 소량의 maltooligosaccharide 들을 생산함을 확인하였다.

To enzymatically prepare amylopectin cluster (APC), cyclodextrin glucanotransferase (CGTase I-5) and its mutant enzyme from alkalophilic Bacillus sp. I-5 were employed, after which the hydrolysis patterns of CGTase wild-type and its mutant enzyme toward amylopectin were investigated using multi-angle laser light scattering. CGTase wild-type dramatically reduced the molecular weight of waxy rice starch at the initial reaction, whereas the mutant enzyme degraded waxy rice starch relatively slowly. Based on the results, the molecular weight of one cluster of amylopectin could be about $10^4{\sim}10^5g/mol$. To determine production of cyclic glucans from amylopectin, matrix-assisted laser desorption ionization time-of-flight mass spectrometry was performed. CGTase I-5 produced various types of cyclic maltooligosaccharides from amylopectin, whereas the mutant enzyme hardly produced any.

김치는 상온에서 산패 및 연부되고 냄새가 강해져서 오래 보존할 수 없으나, 건조하여 환으로 만들면 변질되지 않고, 상온에서 장기간저장 및 유통시킬 수 있고, 냄새를 줄이고 영양분을 고농축할 수 있다. 그러나 건조시 김치의 유산과 향기 성분이 휘발하여 없어진다. 그래서 cyclodextrin(CD)을 안정제로 사용하여 발효 후보다는 발효 전에, $1\%$보다는 $2\%$ 첨가하는 경우가 유산의 휘발 방지 작용이 컸으나 총당, 환원당, 단백질, 아미노산에 대하여서는 영향이 크지 않았다. 김치에 CD를 $1\%$ 가하여 1차로 동결건조한 다음 분쇄하여 분말로 만들고, 분말에 풀을 가해 반죽하여 환으로 성형하고, 2차로 건조기로 말려서 환을 제조하였다. 2차 건조시 동결건조한 김치환의 색과 냄새가 가장 좋았고 그 다음 자연건조, 열풍건조의 순이었다. 유산도 동결건조가 가장 적게 증발되고, 다음 자연건조, 열풍건조의 순이었다. 열풍건조의 경우 $60^{\circ}C$ 이상에서 장기간 건조하면 김치를 삶은 냄새, 탄내, 탄맛이 생겼다. 발효시키기 전의 김치, CD를 가한 발효김치, 김치환 세 가지의 흡광도를 분광광도계로 분석한 결과 약간의 차이 밖에 없었다. 이들 세 무리 시료를 Superose 12컬럼을 사용한 HPLC로 280nm와 210nm에서 검출 분석한 결과 주피크의 위치와 양은 비슷하였으나 다른 피크들은 분포가 달랐고, 펩티드 및 아미노산으로 보이는 다양한 피크가 나타났다. CD를 $1\%$ 가하여 만든 김치환은 8배 농축되고, 총당 $14.4\%$, 환원당 $8.8\%$, 단백질은 $4.8\%$, 아미노산 $2.4\%$, 기타 성분 $74.4\%$, 산도 32.8, pH는 3.5를 나타냈다. 비만인 사람 20명, 변비환자 20명에게 60일 동안 세 번 30개(하루 총량 30g)의 김치환을 섭취시킨 결과 몸무게는 평균 $2.29\%$ 감소하였고, 변비는20명 중7명이 모두 해소되고, 8명이 '효과가 있다'고 답하였다.

In room temperature, Kimchi becomes acidified and a little decayed, scenting a bad smell, and It couldn't be well kept. But if it should be made into a pill, it could be preserved for a long time for marketing, with nutrition highly concentrated as well as with no scent. Therefore, making Kimchi into a pill needs drying. When dried Kimchi, lactic acid and fragrant ingredient will vanish along with volatilization. The cyclodextrin(CD) as a stabilizer shows that the protecting rate of volatility of lactic acid in Kimchi is higher before than that of after fermentation, and it is higher at the addition $2\%\;than\;of\;1\%$ in case of Kimchi with CD. But it doesn't give much effect on total sugar, reducing sugar, protein and amino acid. Evaporation rate of lactic acid is the least in freeze dry, and natural dry, heat dry come next, respectively. In heat dry, if dried at more than $60^{\circ}C$ for a long time, Kimchi exudes boiling and scorched scent, causing bitter taste. The result of HPLC with superose 12 column at 280nm and 210nm shows that place and amount of main peak is almost the same, but the distribution of other peaks are different, with the revelation of various peaks like peptide and amino acid. The Kimchi pill made by the addition of $1\%$ CD shows that concentration is eight times higher than general Kimchi, total sugar is $14.4\%$, reducing sugar is $8.8\%$, protein is $4.8\%$, amino acid is $2.4\%$, and other contents are $74.4\%$, acidity is 32.8, and pH is 3.5 each. The result of letting 20 people with obesity, 20 patients with constipation have 30 pills(total weight 30g) three times a day for 60 days reveals they lost $2.29\%$ in weight on the average, and 7 among 20 were all relieved in constipation, and 8 responded that they experienced its efficacy.

본 연구에서는 caprylic acid와 cyclodextrin과의 complex(CA-CD complex)의 첨가수준이 in vitro 반추위 발효 성상과 메탄 생성에 미치는 영향을 알아보고자 하였다. CA-CD complex 무첨가구를 대조구로 설정 하였으며, 처리구로는 CA 20mg 단독 첨가구(T1), CD 830mg 단독 첨가구(T2), CA-CD complex 425mg 첨가구(T3, CA 함량 10mg), CA-CD complex 850mg 첨가구(T4, CA 함량 20mg), CA-CD complex 1700 mg 첨가구(T5, CA 함량 40mg)이며, 3반복으로 4, 8, 12시간의 배양 실험을 수행하였다. 모든 시험구에서 배양 12시간까지 pH가 지속적으로 감소하였다. 특히, 배양 8시간 후부터 T2, T3, T4, T5 처리구에서 대조구보다 유의적으로 감소하였다(P<0.05). Total VFA 농도는 대조구에 비해 T2, T3, T4, T5구들이 8시간과 12시간에서 유의적(P<0.05)으로 감소하였다. Propionate 농도는 배양 8시간에서만 다른 처리구보다 T4, T5 처리구가 유의적(P<0.05)으로 높게 나타났다. Acetate:Propionate(A/P) 비율은 모든 배양시간에서 처리구간 유의적인 차이가 없었다. 암모니아 농도는 대조구에 비해 T2, T3, T4, T5 처리구에서 배양 4시간부터 대조구보다 암모니아 농도가 유의적(P<0.05)으로 감소하였다. Total gas는 T1구에서 대조구보다 유의적(P< 0.05)인 감소가 있었으나, T2, T3, T4, T5 처리구에서는 대조구에 비하여 12시간에서 유의적(P<0.05)으로 증가하였다. Total gas 발생량에 대한 메탄 비율(%)은 12시간 배양시간에서 대조구는 3.4%이었지만, 메탄 발생량이 가장 적었던 T1구는 2.8%, 메탄 발생량이 가장 많았던 T2구는 5.1%, 그리고 T3, T4, T5구는 평균 2.7%로 T1구와 통계적 차이가 없었다. 따라서 CA-CD complex를 이용하면, 반추위내 발효에 부정적 영향 없이 메탄을 감소시킬 수 있을 것으로 기대된다.

This study was conducted to evaluate the effects of dietary addition of caprylic acid(CA)-cyclodextrin (CD) complex on in vitro fermentation characteristics, total gas and methane production. Experiment was done with six treatment groups; 1) no CA-CD complex(control), 2) CA 20 mg(T1), 3) CD 830 mg(T2), 4) CA-CD complex 425 mg(T3), CA-CD complex 850mg(T4), CA-CD complex 1,700 mg(T5). Ruminal pH, ammonia and total VFA concentrations of T2, T3, T4 and T5 were lower(P<0.05) than those of control and T1 for the 12h incubation. The increase in molar percentage of propionate was observed in T4 and T5 compared with control and T2 for the 8h incubation(P<0.05), however, the ratio of acetate to propionate was unchanged in all treatments. Total gas of T1 was lower than that of control, but T2, T3, T4 and T5 were higher compared with control for 12h incubation(P<0.05). If the methane ratio (as %) to total gas for all treatments was compared, T3, T4 and T5(CA-CD supplemented groups) averaged 2.7% whereas control, T1 and T2 showed 3.4, 2.8 and 5.1%, respectively. Therefore, according to these results, it might be concluded that supplementation of CA-CD complex could reduce methane production without disrupting ruminal fermentation.

피부는 신체의 가장 바깥에 위치하며 외부환경의 영향은 피부노화의 원인이 되기도 한다. 화장품을 이용한 피부의 노화 지연을 위 한 노력에 있어서 화장품 성분의 안정화 기술 방법은 그 기능을 극대화 시키는데 있어서 중요하다. 마이크로캡슐화, 리포좀, 액정공 법 이외에도 사이클로덱스트린(cyclodextrin)의 구조적 특성을 이용하여 물질의 용해성 및 안정성 증진, 서방형(徐放型)성질을 향상 시켜 생체 이용률을 증가시키는 연구가 다양하게 진행되고 있다. 이에 본 총설에서는 cyclodextrin의 구조적 특성을 이용하여 다양 한 유효성분의 화장품 원료로서의 유용성을 높일 수 있는 선행연구들을 고찰하였다. Esculin, hydrogel, coenzyme Q10, genistein 의 cyclodextrin 포접체와 포접하지 않은 상태의 각각의 물질과 비교한 결과 용해도가 증가하였고, 생물학적 안정성 및 피부투과율 이 개선되었다. 또한, β-Cyclodextrin에 아미노산인 시스테인구조 단위를 결합시켜서 만든 cysteinyl β-cyclodextrin은 플라보노 이드인 baicalein에 특이적인 포접 기능을 보여서 복합체를 만들었다. 물과 강한 친화력이 있는 시스테인 부위가 영향을 줌으로써 복합체는 baicalein이 물에 더욱 잘 용해되도록 도와주는 것으로 나타났다. 따라서 cysteinyl 그룹을 첨가한 구조 이외에도 다양한 cyclodextrin 구조의 유도체를 합성하여 Host 구조의 종류를 증가시킴으로써 이미 검증된 유효성분들의 효능을 부가시킬 수 있는 구조 특이적인 Host를 제시하는 연구도 의미가 있을 것으로 생각된다. 또한 추후에 β-cyclodextrin의 포접기능을 이용한 기능성 화 장품으로서의 효과를 입증하는 임상 연구 등이 보강된다면, cyclodextrin은 화장품 산업에서 유효성분의 안정화 소재로서 상용화될 것으로 기대할 수 있다.

Skin is the outermost barrier of the body, making it vulnerable to various external stimuli that can directly influence skin aging. To maximize the impact of cosmetics that delay skin aging, the stabilization of cosmetic ingredients is necessary. In addition to microencapsulation, liposome, and liquid crystal methods to increase the bioavailability of cyclodextrin, various studies have been conducted that utilize its structural characteristics to improve its properties of solubility, stability, and sustained release. In the present study, we reviewed previous investigations demonstrating that cyclodextrin enhanced the usefulness of various active ingredients as cosmetics materials based on its structural characteristics. Cyclodextrin inclusion complexes of esculin, hydrogel, coenzyme Q10, and genistein were compared with each non-inclusion material. The results showed that cyclodextrin increased solubility and improved skin permeability. Additionally, cysteinyl β-cyclodextrin, formed by combining a structural unit of the amino acid cysteine with β-cyclodextrin, showed a complexing function specific to the flavonoid baicalein. The strongly hydrophilic cysteine site of the complex improved the dissolution of baicalein in water. Therefore, further studies should investigate the various structure-specific host complexes with different cyclodextrin structure derivatives besides the cysteinyl group to enhance the already verified efficacy of the effective ingredients. Furthermore, it is expected that cyclodextrin will be commercialized as a stabilizing material for active ingredients in the cosmetic industry if clinical studies can prove the effectiveness of the β-cyclodextrin inclusion function in functional cosmetics.

皮肤位于身体的最外侧，外部环境的影响会导致皮肤老化。在使用化妆品延缓皮肤衰老的努力中，化妆品成 分的稳定化技术对于最大化其功能很重要。除了微囊化，脂质体和液晶方法，还进行了各种研究，以通过改 善材料的溶解性和稳定性来增加生物利用度，并通过利用环糊精的结构性质来改善徐放型性能。在这篇综述 中，我们回顾了以前的研究，以通过使用环糊精的结构性质来提高各种活性成分作为化妆品成分的实用性。将 Esculin，水凝胶，辅酶Q10和染料木黄酮的环糊精包合物与每种非包合物材料进行了比较。结果表明，环糊精 增加溶解度并改善皮肤渗透性。另外，通过将氨基酸半胱氨酸结构单元与β-环糊精结合而制成的半胱氨酸β-环 糊精显示出对黄酮类baicalein特有的复合功能。通过影响与水有很强亲和力的半胱氨酸部位，该复合物已被 证明可以帮助baicalein更好地溶于水。因此，除了半胱氨酰基团的结构外，一项研究提出了一种结构特异性 宿主，该宿主可以通过合成各种环糊精结构衍生物来增加宿主结构的类型，从而增加已经证实的有效成分的功 效，这被认为是有意义的。另外，如果将来进行临床研究以利用β-环糊精的包合功能证明作为功能性化妆品的 效果的临床研究，则可以期望将环糊精商品化为化妆品工业中的活性成分的稳定材料。

Bacillus firmus가 생산하는 cyclodextrin glycosyltran-sferase(CGTase)를 ammonium sulfate 침전, DEAE-Sephadex A-50 및 Sephdex G-100 column chromatography로 분리, 정제하였다. 이 방법으로 수율은 12%, 정제도는 49배인 SDS-PAGE 상에서 단일 band의 효소 단백질을 얻을 수 있었다. 정제된 CGTase는 Phast system의 SDS-polyacrylamide gel 전기영동에 의하여 분자량은 80,000dalton, isoelectric focusing으로 등전점은 9.6으로 추정되는 단백질이었다. 이 효소의 최적 활성 pH와 온도는 8.0, $65^{\circ}C$이였으며, pH와 열안정성은 $pH\;5.5{\sim}9.0$, $50^{\circ}C$이였다. Soluble starch를 기질로 하여 24시간 효소반응한 액의 ${\alpha}-\;:\;{\beta}-\;:\;{\gamma}-cyclodextrin$의 생성비율은 0.01 : 2.90 : 1.00으로 ${\beta}-$ 및 ${\gamma}-cyclodextrin$을 주로 생산하였다.

The cyclodextrin glycosyltransferase(EC 3.2.1.19) from Bacillus firmus was purified by precipitating with ammonium sulfate followed by, DEAE-Sephadex A-50 column chromatography and Sephadex G-100 column chromatography. In this way, we were able to obtain the single band protein on SDS-PAGE with a yield of 12%, whose purity was 49 fold. The purified CGTase was identified as a protein having molecular weight of approximately 80,000 dalton and isoelectric point of 9.6. The optimum pH and temperature for the enzyme activity were 8.0 and $65^{\circ}C$, respectively. The enzyme was stable at between pH 5.5 and 9.0 and up to $50^{\circ}C$. After 24hr of enzyme reaction using soluble starch as substrate, the ratio of ${\alpha}-$, ${\beta}-$ and ${\gamma}-cyclodextrin$ production was 0.01 : 2.90 : 1.00, respectively. And this CGTase pro-duced mainly ${\beta}-$ and ${\gamma}-cyclodextrin$.

Neohesperidin dihydrochalcone(NHDC) is an intense sweetener with lingering aftertaste, which limits NHDC to use as a sweetener in food products. This study was conducted to examine the changes of teste quality of NHDC when using $\beta$-cyclodextrin($\beta$-CD) as a taste modifier. A series of $\beta$-CD(0.01%, 0.03%, 0.1%, 0.176%) was added to NHDC solution (100ppm) and the taste quality was evaluated by magnitude estimation. It was found that $\beta$-CD produced a significant effect in the reduction of aftertaste of NHDC(p<0.01) and sweetness as well.(p<0.001). Linear regression(log mean magnitude estimate versus $\beta$-CD concentration) analysis showed that the intensity of sweetness(n=-0.31) decreased more rapidly than that of aftertaste(m=-0.17). This result suggests the possibility that $\beta$-CD may form inclusion complex with NHDC, so that the hydrophobic portion is encapsulated in the cavity of $\beta$-CD and the carbohydrate moiety is oriented to the outside of the cavity. Or it may be that $\beta$-CD competes with the NHDC molecule for binding to the taste receptor, resulting in reduced perceived intensity.

This study was undertaken to find out the effect of methyl-beta-cyclodextrin (MBCD) on cold shock and membrane cholesterol quantity of sperm during the freezing process in miniature pigs. For this study, semen ejaculated from PWG M-type miniature pig was diluted that freezing solution (with egg yolk group) and m-Modena B (without egg yolk group) treated with 0, 1, 5, 10 and 20 mM MBCD before freezing process. The diluted semen was monitored sperm ability at room temperature, after cooled until 5℃ and after forzen-thawed for cold shock test of spermatozoa. Also, membrane cholesterol of sperm was extracted by folch solution at the same time. sperm ability was assessed for viability and acrosomal status. The membrane cholesterol quantity was measured by thin-layer chromatography (TLC) method. The result, viability and acrosome integrity in semen diluted without egg yolk groups were decreased at all temperature range by increasing of MBCD concentration. In particular, sperm of egg yolk group was showed that significantly higher viability and lower acrosome damage when treated with 5 mM MBCD (p<0.05). The results of TLC experiment, cholesterol amounts were increased with MBCD cocentration in egg yolk, and decreased with MBCD concentration in m-Modena B. In cryopreservation efficiency, there was no significant difference at viability, and acrosomal state of sperm in 5 mM MBCD concentration was significantly lower in acrosome damage than other groups (p<0.05). Therefore, the addition MBCD in egg yolk was protected spermatozoa from cold shock injury. This protective effect of MBCD may be due to addition of sperm membrane cholesterol.

The objective of this study was to evaluate efficiency of methyl-beta-cyclodextrin (MBCD) in the sperm preservation of bull. For this study, the freezing of diluted semen were added with Triladyl containing 20% egg-yolk and/or 0, 1, 5, 10 and 20 mM MBCD before freezing process. Analysis of viability in frozen-thawed sperm was estimated by SYBR14/PI double stain, hypoosmotic swelling test(HOST) and acrosome damage with FITC-PNA, and mitochondria activation with Rhodamin123 by flow-cytometry. The sperm viability was significantly higher in 0 mM and 5 mM concentrations of MBCD than other groups (p<0.05). However, the HOST was significantly lower at 20 mM concentration of MBCD than other concentrations (p<0.05). In addition, acrosome damage and mitochondria activation rates were significantly lower at 20 mM concentration of MBCD than other groups (p<0.05). In conclusion, the viability of sperm was not significantly different among concentrations of MBCD 0, 5 and 10 mM, but MBCD 20 mM was significantly lower than other groups. In addition, as concentrations of MBCD was high, HOST, acrosome damage and mitochondria activation rates had a negative effect in bull sperm.

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.

Recently, surfactants were frequently used in order to desorb the hydrophobic organic compounds (HOCs) from soil and to enhance the bioavailability. Among them, -cyclodextrin ($\beta$-CD) is one of those. This study was performed to investigate the binding characteristics between benzene and $\beta$-CD and to examine the bioavailability of benzene. First, we investigated binding characteristics between benzene and $\beta$-CD in water and water/soil system. Then, we examined the effect of $\beta$-CD on the biodegradation of benzene in water and water/soil system. Experimental results on the binding characteristics showed that $\beta$-CD resulted in an efficient complex formation with benzene. As -CD concentration increased, the benzene concentration complexed with $\beta$-CD rapidly increased to 30-40% initial benzene added, and reached the equilibrium. We also investigated the effect of $\beta$-CD on the desorption of benzene from soil in the water/soil system. As $\beta$-CD concentration increased, benzene concentration desorbed into water increased up to 90%. How-ever, in its application to biodegradation of benzene in water and water/soil system, the biodegradation rate of benzene did not improved in the presence of $\beta$-CD compared with in the absense of $\beta$-CD. This result indicated that $\beta$-CD was more preferentially used as a carbon source than benzene. Therefore, for remediation of benzene contaminated soils, $\beta$-CD can be used as a surfactant to desert benzene from soil, and then ex-situ chemical treatment can be applied for the remediation.

The 2-O-$\alpha$-D-glucopyranosyl L-ascorbic acid (AA-2G) which was enzymatically glucosylated with the cyclodextrin glucanotransferase (CGTase) [EC 2.4.1.19] from Bacillus sp. JK-43 has been reported previously. The presnet experiments examined the optimal conditons for the productio of AA-2G from AA and soluble starch, and characterized the properties of the CGTase from Bacillus sp. JK-43. The reaction mixture for the maximal production of AA-2G was followings; 12% total substrate concentration, 1,400 usits/mL of CGTase and a mixing ratio of 2 : 3(g or AA : g of soluble starch). Under this condition, 1.76mM of AA-2G, which corresponded to 2.53% yield based on AA, was produced after incubation for 24hrs at 45$^{\circ}C$ (pH 5.5). The optimum pH and temperature for the CGTase activity were 6.0 and 45$^{\circ}C$, respectively. The enzyme was stable at pH 5.5 to 9.5, and at temperature up to 5$0^{\circ}C$. The thermostability of the enzyme could be enhanced up to 6$0^{\circ}C$ by the addition of 30mM CaCl2.

A microorganism capable of producing high level of extracellular cyclodextrin glucanotransferase(EC 2.4.1.19 ; CGTase) was isolated from Kimchi. 2-O-$\alpha$-D-glucopyranosyl L-ascorbic acid(AA-2G) was synthesized by transglycosylation reaction of CGTase using starch as a donor and L-ascorbic acid as an acceptor. The isolated strain S-6 was identified as Bacillus sp. S-6. The maximal CGTase production was observed in a medium containing 0.5% soluble starch, 1% yeast extract, 1% NaCO3, 0.1% K2HPO4, and 0.02% MgSO4 with initial pH 8.0. The strain was cultured at 37$^{\circ}C$ for 40 hr with reciprocal shaking. Using the culture supernatant as crude enzyme, the optimal pH and temperature of the CGTase activity of this strain were 7.0 and 4$0^{\circ}C$. In the effects of pH and temperature on the stability of the enzyme, the enzyme was stable in the range of pH 6.0~10.0 and up to 45$^{\circ}C$, respectively.

A strain of alkalophilic Bacillus sp. C-21 has been Isolated from sold. The strain was capable of producing large amount of cyclodextrin glycosyltransferase (CGTase) in the high alkaline pH medium. The preferable medium composition was determined to be as follows : 1.0% soluble starch, 1.0% peptone, 0.5% yeast extract, 0.1% K2HP04, 0.02% MgSO4.7H2O and 1.0% Na2CO3(pH 10.0) The highest enzyme production was observed after 30hours of cultivation at 33$^{\circ}C$. The optimum temperature and pH for the activity of crude enzyme were 6$0^{\circ}C$ and 6.0, respectively. The enzyme was stable between pH 6.0 and 9.6, and up to 55$^{\circ}C$.

본 연구의 주목적은 돼지고기의 콜레스테롤 함량 감소에 관한 사료 내 ${\beta}CD$의 급여 효과를 조사하는 것이었다. 생체중 50 kg의 3원교잡종 거세 수퇘지 12마리를 이용하여서 4처리구 3반복으로 완전임의배치한 후, 시판체중인 110 kg 도달시까지 약 9주 동안 실험사료를 급여하였다. 일일 사료섭취량, 일일 증체량 및 사료효율은 ${\beta}CD$를 첨가하지 않은 대조구와 비교할 때, ${\beta}CD$ 1.5%, 3.0% 및 5.0% 첨가구의 각 처리구간 통계적인 유의차가 나타나지 않았다. 혈액 총지질, 중성지방 및 총콜레스테롤 함량은 대조구와 비교할 때 ${\beta}CD$ 첨가구가 유의적으로 낮아졌으며, 특히 ${\beta}CD$ 5.0% 첨가구에서 각각의 감소율은 21.80%, 55.58% 및 27.69%로서 가장 높게 나타났다(p<0.05). 돼지고기 삼겹살의 콜레스테롤 함량은 대조구와 비교할 때 ${\beta}CD$ 1.5%, 3.0% 및 5.0% 첨가구에서 각각 5.33mg, 12.70mg및 15.23 mg의 유의적인 감소를 나타났으며, 특히 이 값을 콜레스테롤의 감소율로서 나타냈을 때 각각 6.44%, 15.36%, 18.42%로서 유의적인 차이를 나타냈다(p<0.05). 이 결과는 돼지에서 ${\beta}CD$첨가급여가 콜레스테롤 대사를 조절할 수 있는 식이섬유로서 사용될 수 있음을 시사해준다.

The main objective of the present study was to determine the effect of dietary ${\beta}-cyclodextrin\;({\beta}CD)$ on reducing the cholesterol content of pork. Twelve swine of 50 kg body weight were randomly distributed into four groups of three heads each and fed experimental diets for 9 weeks until they reached a market weight of 110 kg. They were assigned to the four experimental diets containing 0% (control), 1.5%, 3.0% or 5.0% pure ${\beta}CD$. Daily feed intake, body weight gain and feed efficiency were not significantly different between any of the four group. The plasma total lipid, triacylglycerol and total cholesterol content of the swine in the three ${\beta}CD$-fed groups were significantly (p<0.05) decreased when compared to those in the control group, and were significantly (p<0.05) reduced by 21.80%,55.58% and 27.69%, respectively, in the swine fed on 5% ${\beta}CD$. The cholesterol content of pork belly (mg/100g) was significantly (p<0.05) decreased by 5.33 mg, 12.70 mg and 15.23 mg in the swine maintained on 1.5%, 3.0% and 5.0% ${\beta}CD$, respectively. The cholesterol content of pork belly, when expressed as the rate of reduced cholesterol, was significantly (p<0.05) decreased by 6.44%, 15.36% and 18.42% in groups of 1.5%, 3.0% and 5.0% ${\beta}CD$, respectively, when compared to that of the control group. These results suggest that dietary ${\beta}CD$ may be classified as dietary fiber which can modulate cholesterol metabolism in swine.