연구논문 : 생물전환 대두 추출물의 생물학적 활성에 관한 연구

나은지 ( Eun Ji Na ),문지선 ( Ji Sun Moon ) 한국미용학회 2015 한국미용학회지 Vol.21 No.1

In this study, the bioconversion of soybean extract was performed using various enzymes, and then the change of biological activity before and after the bioconversion was characterized. Embryo bud of soybean was firstly extracted using hot water and then extracted again using 95% ethanol, which was named as isoflavone rich soybean extract(IRSE), and this was used for the bioconversion experiments. The bioconversion of IRSE was performed using β-glucosidase, Novozym or Accellerase, which are commercial enzymes. The change of composition of the IRSE before and after the bioconversion was determined using HPLC. The major components of IRSE were genistin and daidzin, which are the glucosides, and genistein and daidzein, which are the aglycones. After 24 hours bioconversionprocess, the major part of genistin and daidzin was converted into genistein or daidzein. The anti-oxidant activities of IRSE were determined before and after bioconversion. The bioconversion process did not influence significantly on the anti-oxidant activities of IRSE. On the other hand, various biological activities were also determined before and after the bioconversion using RAW 264.7 cell. The experimental results showed that both the NO generations in RAW 264.7 effectively after the bioconversion. Taken all together, we could conclude that the bioconversion of IRSE may be useful to the preparation for enhanced functional cosmetic raw materials using soybean extract.

한국형 프로바이오틱스에 의한 쓴메밀 내 rutin의 생물전환

Kwon Chang,Kim Jong Won,Park Young Kwang,Kang Seungbeom,Chung Myung Jun,Kim Su Jeong,Lim Sanghyun 한국미생물·생명공학회 2023 한국미생물·생명공학회지 Vol.51 No.1

In this study, bioconversion of rutin to quercetin was confirmed by the fermentation of Korean indigenous probiotics and tartary buckwheat. Based on whole genome sequencing of 17 probiotics species, α-rhamnosidase, related to bioconversion of isoquercetin (quercetin 3-β-D glucoside) from rutin, is identified in the genome of CBT BG7, LC5, LR5, LP3, LA1, and LGA1. β-Glucosidase, related to bioconversion of isoquercetin to quercetin, is identified in the genome of all 17 species. Among the 17 probiotics species, 6 probiotics including CBT BG7, LR5, LP3, LA1, LGA1 and ST3 performed the bioconversion of rutin to quercetin up to 21.5 ± 0.3% at 7 days after fermentation. The fermentation of each probiotics together with enzyme complex Cellulase KN® was conducted to reduce the time of bioconversion. As a result, CBT LA1 which showed the highest yield of bioconversion of 21.5 ± 0.3% when the enzyme complex was not added showed high bioconversion yield of 84.6 ± 0.5% with adding the enzyme complex at 1 day after fermentation. In particular, CBT ST3 (96.2 ± 0.4%), SL6 (90.1 ± 1.4%) and LP3 (90.0 ± 0.4%) showed high yield of bioconversion more than 90%. In addition, such probiotics including high levels in quercetin indicated the inhibitory effects of NO production in LPS-induced RAW264.7 cells. In this study, we confirmed that the fermentation of Korean indigenous probiotics and enzyme complex together with roasted tartary buckwheat increased the content of quercetin and reduced the time of bioconversion of rutin to quercetin which is a bioactive compound related to anti-inflammatory, antioxidants, anti-obesity, and anti-diabetes.

Antioxidant Effects of Ponciri Fructus Bioconversion Extracts as Cosmetic Ingredients

Gyu Bin Kong,Oh Hyeok Kwon 한국피부과학연구원 2021 아시안뷰티화장품학술지 Vol.19 No.3

목적: 본 연구는 지실의 기능성 화장품 및 식품소재로서의 활용 가능성을 알아보기 위하여 에탄올 추출물과 동충하초 균사체 배양 여액으로 생물전환한 지실추출물을 가지고 실험을 진행하여 보았다.방 법: 지실 에탄올 추출물과 동충하초 균사체 배양여액을 가지 고 총폴리페놀, 총플라보노이드 함량측정과 2,2-diphenyi-1-picrylhydrazyl (DPPH)소거능 활성을 통한 지실 추출물의 항산화 효 과 등을 알아보고 에탄올 추출물과 생물전환 추출물의 결과 비교를 통하여 생물전환방법에 의한 유효성분의 함량변화, 항산화능 차 이를 비교하여 보았다. 결과: 이 실험을 수행한 결과 에탄올 추출물의 경우에는 총 페놀함량이 62.5±4.19 GAE mg/g 있는 것으로 나타났고 생물전환 추출물은 82.8±2.55 GAE mg/g으로 약 32.4% 증가되었고, 총 플라보노이드는 에탄올 추출물의 함량이 30.1 ±3.15 mg/g, 생물전환 추출물은39.5±35.1 mg/g으로 약 31.2% 증가된 것으로 나타났다. DPPH 소거능은 에탄올 추출물의 경우 에는 65.5%, 생물전환 추출물은 75.5%인 것으로 나타났다. 결론: 지실 추출물은 항산화 효과가 우수하였으며 생물전환을 하였을 때 에탄올 추출에 의한 것보다 약간 증가된 것으로 나타났으며, 생물전환을 통하여 지실에 들어있는 기능성 물질에 대한 추출률과 생체 이용률을 높인다면 여러 가지 산화스트레스 요인으로 발생할 수 있는 활성산소에 의한 피부 내. 외적인 문제들을 개선할 수 있 는 화장품, 식품 소재로서의 가능성이 높을 것으로 사료된다. Purpose: This study was conducted to investigate the possible use of Ponciri fructus bioconversion extracts as cosmetic ingredients. Method: Total polyphenol, total flavonoid content, and antioxidant effects of Poncirus trifoliata through the ability of diphenylpicrylhydrazyl (DPPH) radical scavenging were used for antioxidant activity determination. The extracts of Poncirus trifoliata were analyzed using an ultraviolet– visible spectrophotometer Results: The total phenolic content was 62.5±4.19 gallic acid equivalent (GAE) mg/g in the case of alcohol extracts and 82.8±2.55 GAE mg/g for other bioconversion extracts. Therefore, total flavonoids content was found to have increased by 32.4%. The total flavonoid content of ethanol extracts was 30.1±3.15 mg/g, whereas that of other bioconversion extracts was 39.5±35.1 mg/g. Therefore, total flavonoids increased by 31.2%. From DPPH radical results, the alcoholic extract increased from 65.5% to 75.5% after bioconversion. Thus, the bioconversion extracts increased by 10% compared with ethanol extracts. Conclusion: Poncirus trifoliata has excellent antioxidant effects, and it showed increased antioxidant activity after the bioconversion, thereby demonstrating its functional bioutility. Therefore, it can be used as an antioxidant material in the cosmetic and food industry.

생물전환에 의한 발효 목초액의 항산화 활성

조영호(Cho, Young-Ho),조재수(Cho, Jae-Soo),이계원(Lee, Gye-Won) 한국산학기술학회 2011 한국산학기술학회논문지 Vol.12 No.10

활성 산소종은 반응성이 매우 크고 지질, 단백질 및 핵산에 산화를 유발하여 잠재적으로 세포에 매우 해 로운 물질이다. 활성 산소종은 또한 인체에 있어서 노화, 발암, 죽상경화증 유발과 같은 해로운 영향을 준다고 알 려졌다. 본 연구에서는 총산도, 초산, pH 및 총 폴리페놀과 총 플라보노이드의 함량 등을 분석하였고, 생물전환된 목초액의 항산화 활성을 조사하기 위하여 DPPH, 초과산화물 음이온, 과산화수소, 산화질소 라디칼 소거 활성을 측정하였다. 생물전환된 목초액의 총산도와 초산의 양은 생물전환 전보다 낮았지만, pH는 오히려 높게 나타났다. 생물전환된 목초액의 총 폴리페놀과 총 플라보노이드의 함량은 각각 11.17 ㎎/㎖ 과 0.42 ㎎/㎖였다. 생물전환된 목초액의 각종 라디칼을 50% 소거하는 농도는 초과산화물 라디칼 소거활성 < DPPH 라디칼 소거활성 < 과산화 수소 라디칼 소거활성 < 산화질소 라디칼 소거활성 순으로 나타났다. 따라서 목초액을 생물전환할 경우 여러 가 지 라디칼에 대한 소거활성을 높여 천연 의약품 및 화장품 소재로 개발 가능성이 있음을 알 수 있다. Reactive oxygen species (ROS) are reactive and potentially harmful to cells, causing oxidation of lipids, proteins, and DNA. In humans, the deleterious effects of ROS have been linked with aging, carcinogenesis, and atherosclerosis. In order to investigate an antioxidant activity of wood vinegar by bioconversion, we preferentially analyzed the total acidity, acetic acid, pH, and contents of total polyphenols and flavonoids, respectively. Also, we evaluated the scavenging abilities on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals, superoxide anion radicals, hydrogen peroxide radicals, and nitric oxide radicals. The total acidity and amount of acetic acid of wood vinegar after bioconversion were lower than those of wood vinegar before bioconversion, but the pH was higher than that of wood vinegar before bioconversion. The contents of total polyphenols and flavonoids of wood vinegar after bioconversion were 11.17 ㎎/㎖ and 0.42 ㎎/㎖, respectively. The SC50 values were in order of superoxide anion radical scavenging activity < DPPH radical scavenging activity < hydrogen peroxide radical scavenging activity < nitric oxide radical scavenging activity. Therefore, these results suggest that wood vinegar by bioconversion can be useful as primary antioxidants for medicines and cosmetics.

Development of a continuous l-lysine bioconversion system for cadaverine production

Kim, J.H.,Seo, H.M.,Sathiyanarayanan, G.,Bhatia, S.K.,Song, H.S.,Kim, J.,Jeon, J.M.,Yoon, J.J.,Kim, Y.G.,Park, K.,Yang, Y.H. Korean Society of Industrial and Engineering Chemi 2017 Journal of industrial and engineering chemistry Vol.46 No.-

<P>Cadaverine, a five carbon diamine (1,5-diaminopentane), plays a role as a building block of polyamides and it can be made by fermentation or direct bioconversion. To improve its production by increasing reusability of immobilized enzyme and avoid separation of enzyme in bioconversion, a continuous l-lysine bioconversion process for cadaverine production has been developed. Various divalent cations, alginate concentrations, cell density with alginate and flow rate of feed were examined to maximize the lysine decarboxylase activity of the whole-cell immobilized beads. Under the selected conditions, 123 h of continuous cadaverine production has been performed and 5.5 L of 819 mM cadaverine were produced with 14 mL reactor resulting in 466.5 g of cadaverine. Cadaverine production was possible with small volume reactor maintaining relatively high concentration of substrate. (C) 2016 The Korean Society of Industrial and Engineering Chemistry. Published by Elsevier B.V. All rights reserved.</P>

Development of a continuous L-lysine bioconversion system for cadaverine production

김정호,서형민,가네산,Shashi Kant Bhatia,송헌석,김준영,전종민,윤정준,김윤곤,박경문,양영훈 한국공업화학회 2017 Journal of Industrial and Engineering Chemistry Vol.46 No.-

Cadaverine, afive carbon diamine (1,5-diaminopentane), plays a role as a building block of polyamidesand it can be made by fermentation or direct bioconversion. To improve its production by increasingreusability of immobilized enzyme and avoid separation of enzyme in bioconversion, a continuousL-lysine bioconversion process for cadaverine production has been developed. Various divalent cations,alginate concentrations, cell density with alginate andflow rate of feed were examined to maximize thelysine decarboxylase activity of the whole-cell immobilized beads. Under the selected conditions,123 h ofcontinuous cadaverine production has been performed and 5.5 L of 819 mM cadaverine were producedwith 14 mL reactor resulting in 466.5 g of cadaverine. Cadaverine production was possible with smallvolume reactor maintaining relatively high concentration of substrate

Feasibility Study on Production of Liquid Fertilizer in a 1 ㎥ Reactor Using Fishmeal Wastewater for Commercialization

Gwon, Byeong-Geun,Kim, Joong-Kyun Korean Society of Environmental Engineers 2012 Environmental Engineering Research Vol.17 No.1

A scaled-up bioconversion of fishmeal wastewater (FMW) into liquid fertilizer was performed five times in a $1m^3$ reactor in order to examine the feasibility of commercialization. The importance of aeration was marked. Analyses indicated that dissolved oxygen (DO) level was closely related to the value of oxidation-reduction potential (ORP) and it was crucial to achieve high-quality liquid fertilizer. When pure oxygen was supplied through four diffusers into the reactor, DO levels and ORP values were maintained over 1.2 mg/L and 0.2 mV, respectively all the time during 52 hr of bioconversion. The pH changed from 6.8 to 5.9. The average removal percentages of chemical oxygen demand ($COD_{Cr}$) and total nitrogen (TN) were 75.0% and 71.6%, respectively. Compared to the result acquired in a 5-L reactor, bioconversion of FMW into liquid fertilizer was achieved in a shorter time under the same removal percentages of $COD_{Cr}$ and TN. The 52-hr culture of inoculated FMW was phytotoxic-free and it possessed comparable fertilizing ability to a liquid fertilizer made from the fish waste in hydroponic culture with amino acid contents of 5.93 g/ 100 g sample. From all the above results, transferring lab-scale data to large-scale production appeared to be successful. As a result, the commercialization of a liquid fertilizer made from FMW was feasible.

Feasibility Study on Production of Liquid Fertilizer in a 1 m3 Reactor Using Fishmeal Wastewater for Commercialization

Byeong Geun Gwon,Joong Kyun Kim 대한환경공학회 2012 Environmental Engineering Research Vol.17 No.1

A scaled-up bioconversion of fishmeal wastewater (FMW) into liquid fertilizer was performed five times in a 1 m3 reactor in order to examine the feasibility of commercialization. The importance of aeration was marked. Analyses indicated that dissolved oxygen (DO) level was closely related to the value of oxidation-reduction potential (ORP) and it was crucial to achieve high-quality liquid fertilizer. When pure oxygen was supplied through four diffusers into the reactor, DO levels and ORP values were maintained over 1.2 mg/L and 0.2 mV, respectively all the time during 52 hr of bioconversion. The pH changed from 6.8 to 5.9. The average removal percentages of chemical oxygen demand (CODCr) and total nitrogen (TN) were 75.0% and 71.6%, respectively. Compared to the result acquired in a 5-L reactor, bioconversion of FMW into liquid fertilizer was achieved in a shorter time under the same removal percentages of CODCr and TN. The 52-hr culture of inoculated FMW was phytotoxic-free and it possessed comparable fertilizing ability to a liquid fertilizer made from the fish waste in hydroponic culture with amino acid contents of 5.93 g/ 100 g sample. From all the above results, transferring labscale data to large-scale production appeared to be successful. As a result, the commercialization of a liquid fertilizer made from FMW was feasible.

D, L-ATC로부터 L-Cysteine으로의 Bioconversion에 관여하는 효소의 특성

류옥희,신철수 한국산업미생물학회 1990 한국미생물·생명공학회지 Vol.18 No.1

D, L-2-aminothiazoline-4-carboxylic acid(D, L-ATC)로부터 L-cysteine으로의 bioconversion에 대한 특성을 살펴보았다. Pseudomonas species의 배양중에 D, L-ATC를 첨가하여 균체내에 그 관여되는 효소를 유도, 생성시키고 균체만을 모은 후 파쇄하여 조효소액으로 조제하였다. 실험결과, D, L-ATC로부터 L-형의 cysteine만이 생성되며, 이 반응에 관여되는 효소는 cofactor로서 Mn이온을 필요로 하며, Mn이온의 첨가에 의해 L-cysteine의 생성량이 수십배 증가되었다. 그러나, 이 효소는 생성물인 L-cysteine에 의해 feedback inhibition을 받았다. 한편, L-cysteine의 분해효소가 조효소액 내에 존재하며 그 효소반응의 저해제없이는 생성된 L-cysteine의 대부분이 분해되었다. 반면, 매우 효과적인 효소저해제인 hydroxylamine의 첨가로 L-cysteine의 분해를 거의 방지할 수 있었다. The bioconversion of D, L-2-aminothiazoline-4-carboxylic acid(D, L-ATC) to L-cysteine was investigated. After the intracellular enzyme of a Pseudomonas species was inducibly formed by addition of D, L-ATC in the middle of culture, the cells were isolated and treated with sonication to prepare the crude enzyme solution. The results indicated that the cysteine was produced only in the form of L-isomer from D, L-ATC and its production could be enhanced several tens times by addition of managanese ions which were required as a cofactor in this enzymatic reaction. Besides, this r-action suffered from the feedback inhibition of L-cysteine. On the other hand, since L-cysteine-decomposing enzyme coexisted in the crude enzyme solution, most of the L-cysteine formed disappeared in the absence of its inhibitor. However, hydroxylamine was found to be a potent inhibitor which could successfully prevent the decomposition of L-cysteine.

열무김치에서 발굴한 Saccharomyces servazzii의 신규 Strain인 Ceb-kc-011의 생육조건 최적화 및 서목태 생물전환에 따른 α-Glucosidase 억제 효능

박찬호(Chan-Ho Park),최예림(Yae Rim Choi),염규진(Kyu-Jin Yum),김민정(Min Jung Kim) 한국식품영양과학회 2020 한국식품영양과학회지 Vol.49 No.3

본 연구는 열무김치로부터 신규 효모를 분리 및 동정하였고 배양조건을 최적화시킨 후 효모를 활용해 서목태를 생물전환함으로써 α-glucosidase 억제 활성을 확인하였다. 열무김치에서 18S rRNA 염기서열 분석으로 선정된 균주는 S. servazzii Ceb-kc-011로 명명하고, 배양조건은 YPD 배지, 25°C, 호기배양으로 최적화되었다. YPD 기본배지에서 질소원을 서목태로 대체했을 때에는 S. servazzii Ceb-kc-011의 생장속도는 YPD 배지에서 배양했을 때보다 적었으나, 서목태 대사산물에 의한 α-glucosidase 억제능은 더 높아진 것으로 나타났다. 질소원을 서목태로만 설정한 상태에서 서목태 함량을 0.2~4.0% 변화를 시켜주면서 α-glucosidase 억제능을 살펴본 결과 서목태 함량이 0.6~1.0%에서 유의적으로 최고의 억제능이 나타났다. 또한 서목태 함량 0.6% 조건에서 생성된 서목태 대사산물의 α-glucosidase 활성에 대한 IC50은 29.5 mg/mL로 나타났다. α-Glucosidase 억제는 항비만, 항당뇨 소재 개발에 있어서 가장 기초적인 시험법이기 때문에, 이러한 결과는 효모로 서목태를 생물전환함으로써 생성된 서목태 대사산물을 항비만, 항당뇨 소재로의 개발 및 응용에 활용할 수 있을 것이라 예상된다. Food bioconversion and fermentation is a process wherein microorganisms convert food materials to degradation products, including organic acids and bioactive substances. Rhynchosia nulubilis, which is used traditionally as a food and herbal medicine in Asia, has also been biotransformed by microorganisms, and fermented R. nulubilis has anti-hyperglycemic, anti-hyperlipidemic, and anti-obesity activity. On the other hand, the microbes involved in the fermentation of R. nulubilis remain unknown. In particular, the role of yeast in inducing the biotransformation of R. nulubilis has yet to be examined. This study assessed the adaptation of yeast isolated from young radish (Yeolmoo) kimchi for inducing bioconversion using R. nulubilis. A novel yeast strain was isolated from young radish kimchi and called Saccharomyces servazzii Ceb-kc-011 by 18S rRNA gene sequence analysis. The results showed that the optimal culture conditions for Ceb-kc-011 were YPD media [yeast 1% (w/v), peptone 2% (w/v), D-glucose 2% (w/v)], 25°C, and a 24-hour incubation time under aerobic conditions. When R. nulubilis was added to the media, the growth of S. servazzii Ceb-kc-011 was indicated by the presence of yeast and peptones. Still, bioactive substances were produced in the absence of yeast and peptones (2% D-glucose and 4% R. nulubilis). Bioconverted R. nulubilis inhibited α- glucosidase activity (41.43±0.25%). In conclusion, the bioconversion of R. nulubilis using a specific microorganism was observed, highlighting its potential for the production of anti-diabetic agents through bioconversion.