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한국응용생명화학회> Journal of Applied Biological Chemistry> 식물공장에서 생산된 새싹인삼의 크기에 따른 진세노사이드 함량 및 항산화 활성 비교

KCI등재SCOUPUS

식물공장에서 생산된 새싹인삼의 크기에 따른 진세노사이드 함량 및 항산화 활성 비교

Comparison of ginsenoside contents and antioxidant activity according to the size of ginseng sprout has produced in a plant factory

황승하 ( Seung Ha Hwang ) , 김수철 ( Su Cheol Kim ) , 성진아 ( Jin A Seong ) , 이희율 ( Hee Yul Lee ) , 조두용 ( Du Yong Cho ) , 김민주 ( Min Ju Kim ) , 정재각 ( Jea Gack Jung ) , 정은혜 ( Eun Hye Jeong ) , 손기호 ( Ki-ho Son ) , 조계만 ( Kye Man Cho )
  • : 한국응용생명화학회
  • : Journal of Applied Biological Chemistry 64권3호
  • : 연속간행물
  • : 2021년 09월
  • : 253-513(261pages)
Journal of Applied Biological Chemistry

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재료 및 방법
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본 연구에서는 식물공장을 통해 생산된 새싹인삼을 크기에 따라 분류하고 이를 지상부와 지하부로 나누어 진세노사이드 함량과 항산화 활성을 비교하였다. 지상부의 경우 총 phenolic 함량은 중간 크기인 M에서 5.16 GAE mg/g로 가장 높았으며 가장 큰 크기인 L에서 2.23 GAE mg/g으로 가장 낮은 함량을 보였다. 지하부 역시 M 크기에서 가장 높은 함량을 보였으나, 큰 차이를 보이지 않았다. 한편, 총 flavonoid 함량 역시 지상부(5.16 RE mg/g) 와 지하부(1.28 RE mg/g) 모두 M 크기에서 높은 함량을 보였다. 지상부의 주요 진세노사이드는 Re(20.33-24.15 mg/g) > Rd (11.36-27.42 mg/g) > Rg1 (4.48-5.54 mg/g) 순 있었고 지하부는 Rb1 (5.09-8.61 mg/g) > Re (4.48- 5.54 mg/g) > Rc (3.11-4.11 mg/g) 순 있었다. M 크기의 경우 Re와 Rd는 각각 지상부에서 24.15 mg/g과 27.42 mg/g 및 지하부에서 5.20 mg/g과 1.43 mg으로 약 4배와 19배 높은 함량을 보였다. 지상부에는 F3 및 Rh1이 검출되었으나, 지하부에서는 검출되지 않았다. DPPH (74.95%)와 ABTS (94.47%), hydroxyl (70.39%) 라디칼 소거 활성 및 FRAP (2.169) 활성은 다른 크기들보다 M 크기에서 가장 높은 활성을 보였다.
In this study, the ginseng sprout has produced through smart farm was classified according to its size and divided into above-ground (AG) and below-ground (BG) parts to compare ginsenoside contents and antioxidant activity. In the case of the AG part, the total phenolic contents were the highest at 5.16 mg/ g in medium (M) size and the lowest at 2.23 mg/g in largest (L) size. The BG part also showed the highest content in the M size, but there was no significant difference. Also, the total flavonoid contents were also high in the M size in both the AG (5.16 mg/ g) and BG (1.28 mg/g) parts. The major ginsenosides in the AG part were Re (20.33-24.15 mg/g) > Rd (11.36-27.42 mg/g) > Rg1 (4.48-5.54 mg/g) and the main ginsenosides in the BG part were Rb1 (5.09-8.61 mg/g) > Re (4.48-5.54 mg/g) > Rc (3.11-4.11 mg/ g) in orders. In the case of M size, Re and Rd were approximately 4- and 19-folds higher at 24.15 mg/g and at 27.42 mg/g in the AG part and 5.20 mg/g and 1.43 mg in the BG part, respectively. In addition, F3 and Rh1 were detected in the AG part, but not in the BG part. 2,2-diphenyl-1-picrylhydrazyl (74.95%), 2,4,6-azino-bis (3-ethylbenzothiazoline-6-sulphnoic acid) diammonium salt (94.47%), and hydroxyl (70.39%) radical scavenging activities and FRAP (2.169) assay were the highest in M size than other sizes.

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간행물정보

  • : 농학분야  > 농화학
  • : KCI등재
  • : SCOPUS
  • : 계간
  • : 1976-0442
  • : 2234-7941
  • : 학술지
  • : 연속간행물
  • : 1999-2021
  • : 1433


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64권3호(2021년 09월) 수록논문
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1고함량 사포나린 함유 보리 어린 순 재배를 위한 식물공장내 인공광 조건 및 사포나린 추출 분석법 최적화

저자 : 오경열 ( Kyeong-yeol Oh ) , 송영훈 ( Yeong Hun Song ) , 이득영 ( Duek-yeong Lee ) , 이태근 ( Tae-geun Lee ) , 김진효 ( Jin-hyo Kim )

발행기관 : 한국응용생명화학회 간행물 : Journal of Applied Biological Chemistry 64권 3호 발행 연도 : 2021 페이지 : pp. 197-399 (203 pages)

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본 연구에서는 보리 어린 순의 기능성 성분인 saponarin의 정량적 추출을 위한 최적의 추출 분석 조건을 methanol, ethanol, acetonitrile 및 water를 대상으로 반응표면분석법을 통해 실시하였다. 이를 통해 용매의 농도, 추출 시간에 따른 saponarin과 색소 추출율을 확인한 결과, 53.7% methanol 수용액에서 3.9 h 동안 진탕추출하는 것이 높은 saponarin 추출율을 유지하면서, 불필요한 색소의 추출을 최소화하는 최적 추출조건으로 확인되었다. 보리 어린 순의 재배에 필요한 인공광의 saponarin 생성 영향 평가를 광 주기, 광량, 광질을 달리한 조건에서 시험한 결과, saponarin 고함유 보리 어린 순의 인공광 최적 재배 조건은 8 h day-1의 광 주기에서 6500K LED와 적색보광을 통한 총 광량 220-320 μmol m-2 s-1 에서 가장 효과적임을 확인 할 수 있었고, 청색광이 saponarin 생합성에 주요 인자로 작용함을 확인할 수 있었다.


Saponarin is a crucial component of barley sprout, and the production and quantitative analysis are issued to date. In this study, the optimal saponarin extraction conditions were presented on the subject of acetonitrile, ethanol, methanol, and water for the quantitative analysis in barley sprout through the extraction efficiency compared with the solvent concentration and extraction time using the reaction surface methodology. The optimal extraction time and solvent condition for saponarin were 3.9 h and 53.7% of aqueous methanol, respectively. In addition, the effect of LED artificial light on the saponarin production in barley sprouts was evaluated by the light cycle, light quantity, and light quality. The optimal cultivation conditions under artificial light for the growth of barley sprout and saponarin production were most effectively achieved on 220-320 μmol m-2 s-1 of the light quantity with 8 h day-1 of a daylight cycle under 6500K LED combined with red light. Furthermore, blue light was evaluated as the main factor in the biosynthesis of saponarin.

KCI등재 SCOPUS

2Molecular characterization and biological changes caused by Agrobacterium-mediated infiltration of PgTRX1

저자 : Seung Hyuk Choi , Ji Won Seo , Jae Geun Lee , Chang Yeon Yu , Eun Soo Seong

발행기관 : 한국응용생명화학회 간행물 : Journal of Applied Biological Chemistry 64권 3호 발행 연도 : 2021 페이지 : pp. 205-415 (211 pages)

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In order to test the functionality of Panax ginseng thioredoxin 1 (PgTRX1) isolated from fermented wild ginseng roots, a transient effect on physiological activity were performed over a short time frame using the Agrobacterium infiltration technique. The PgTRX1 gene isolated from fermented wild ginseng was confirmed to have a size of 579 bp, and the expression of PgTRX1 was the highest in the sample after 6 h of fermentation. As a result of constructing this gene and confirming the infiltration reaction mediated by Agrobacterium in tobacco leaves, it was found that the expression of the NbHSR203j gene was also induced as PgTRX1 expression increased. As a result of measuring the biological activity of the infiltration samples, the total phenol content increased by 35.45±1.84 to 49.01±1.84 μg GAE/mL compared to the control, and the total flavonoid amount of 9.52±0.41 to 9.82±0.25 μg QE/mL was slightly high. From these results, Agrobacterium-mediated PgTRX1 appears to be related to the hypersensitive response induction mechanism of plants and the production of secondary metabolites such as phenolic substances.

KCI등재 SCOPUS

3Characterization of the recombinant cellulase A from Thermotoga maritima

저자 : Chung Ho Kim

발행기관 : 한국응용생명화학회 간행물 : Journal of Applied Biological Chemistry 64권 3호 발행 연도 : 2021 페이지 : pp. 213-428 (216 pages)

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A gene encoding thermostable cellulase A (TmCelA) was isolated from Thermotoga maritima. The open reading frame of TmCelA gene was 774 bp long which predicted to encode 257 amino acid residues with a molecular weight of 29,732 Da. To examine the biochemical properties, the TmCelA was overexpressed in E. coli BL21, and expressed protein was purified. The optimum temperature of recombinant TmCelA was 90-95 ℃, and the optimum pH of recombinant TmCelA was approximately pH 5.0. Recombinant TmCelA was stable at temperature below 90 ℃.

KCI등재 SCOPUS

4Rhodobacter sphaeroides에서 5-aminolevulinic acid 생산에 대한 850 nm 근적외선 발광다이오드 조사 효과

저자 : 모상준 ( Sangjoon Mo )

발행기관 : 한국응용생명화학회 간행물 : Journal of Applied Biological Chemistry 64권 3호 발행 연도 : 2021 페이지 : pp. 217-439 (223 pages)

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광감각제를 이용한 광역학 치료는 필요한 특정 부위에만 빛을 조사하여 치료 효과를 나타내는 부작용이 적은 방법이다. 5-aminolevulinic acid (ALA)는 다양한 생물체에서 합성되는 대표적 광감제로 암진단과 치료를 포함하는 다양한 분야에서 사용되고 있다. 본 연구에서는 다양한 파장의 LED, 유기산 전구체 및 glucose 농도 변화를 통한 Rhodobacter sphaeroides의 최적 성장 조건과 ALA 생산 조건을 확립하기 위한 실험을 진행하였다. 백열등과 동일한 광도 아래에서 Rhodobacter sphaeroides에 850 nm LED 빛을 조사하면 대조군 대비 균주의 성장과 ALA의 생산 농도를 각각 1.5배 및 1.8배 증가시킬 수 있고, 전구체로 pyruvic acid를 첨가한 경우 850 nm 파장의 LED만 조사한 경우 보다 ALA의 생산 농도를 약 2.8배 증가 시켰으며 동일 배양 조건에 40 mM glucose를 첨가하여 배양한 결과 Rhodobacter sphaeroides의 성장은 850 nm 파장의 LED 조사와 pyruvic acid를 첨가한 것에 비해 약 2.9배, ALA의 생산 농도는 약 3.4배 (20 mM) 증가되었다. 건조체 질량당 ALA의 생산은 20 mM과 40 mM glucose에서 대조군 대비 각각 약 1.4배 높은 결과를 나타냈다. 결론적으로 다양한 파장의 LED 중 850 nm 파장의 LED가 Rhodobacter sphaeroides의 성장률 및 ALA의 생산을 최대로 높였으며, 5 mM pyruvic acid와 40 mM glucose의 농도에서 최적의 Rhodobacter sphaeroides 성장과 ALA 생산을 확인하였다.


5-aminolevulinic acid (ALA) is a representative photosensitizer used in numerous fields including cancer diagnosis and treatment. In this study, experiments were conducted to optimize the growth of Rhodobacter sphaeroides and production of ALA through LED irradiation of various wavelengths, addition of organic acid precursors of ALA, and changes in glucose concentration. After 72 h cultivation, the 850 nm wavelength LED irradiated at the same light intensity as the incandescent lamp increased the growth of R. sphaeroides and the production of ALA about 1.5- and 1.8-fold as compared with the control, respectively (p <0.0001 and p <0.0001). As a result of culturing R. sphaeroides by irradiating an LED with a wavelength of 850 nm after adding organic acid to the final concentration of 5 mM in culture medium, the production of ALA was increased about 2.8- fold in medium supplemented with pyruvic acid compared with the control (p <0.0001). In addition, the growth of the strain and the production of ALA were increased about 2.9- and 3.4-fold in medium supplemented with 40 mM glucose compared to the control which added only 5 mM pyruvic acid, respectively (p <0.0001 and p <0.0001). The yield of ALA per cell dry mass was about 1.4 folds higher than that of the control in 20 and 40 mM glucose, respectively (p <0.001). In conclusion, the growth of R. sphaeroides and production of ALA were increased by 850 nm wavelength LED irradiation. It also optimized the growth of R. sphaeroides and production of ALA through organic acid addition and glucose concentration changes.

KCI등재 SCOPUS

5Cytochrome P450 and the glycosyltransferase genes are necessary for product release from epipyrone polyketide synthase in Epicoccum nigrum

저자 : Eun Ha Choi , Si-hyung Park , Hyung-jin Kwon

발행기관 : 한국응용생명화학회 간행물 : Journal of Applied Biological Chemistry 64권 3호 발행 연도 : 2021 페이지 : pp. 225-459 (235 pages)

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The epipyrone (EPN) biosynthetic gene cluster of Epicoccum nigrum is composed of epnC, epnB, and epnA, which encode cytochrome P450 oxidase, glycosyltransferase, and highly reducing polyketide synthase, respectively. Gene inactivation mutants for epnA, epnB, and epnC were previously generated, and it was found that all of them were incapable of producing EPN and any of its related compounds. It was also reported that epnB inactivation abolished epnA transcription, generating ΔepnAB. This study shows that the introduction of native epnC readily restored EPN production in ΔepnC, suggesting that epnC is essential for polyketide release from EpnA and implies that EpnC works during the polyketide chain assembly of EpnA. Introduction of epnC promoter-epnA restored EPN production in ΔepnA. The ΔepnB genotype was prepared by introducing the epnA expression vector into ΔepnAB, and it was found that the resulting recombinant strain did not produce any EPN-related compounds. A canonical epnB inactivation strain was also generated by deleting its 5′-end. At the deletion point, an Aspergllus nidulans gpdA promoter was inserted to ensure the transcription of epnA, which is located downstream of epnB. Examination of the metabolite profile of the resulting ΔepnB mutant via LC-mass spectrometry verified that no EPN-related compound was produced in this strain. This substantiates that C-glycosylation by EpnB is a prerequisite for the release of EpnA-tethered product. In conclusion, it is proposed that cytochrome P450 oxidase and glycosyltransferase work in concert with polyketide synthase to generate EPN without the occurrence of any free intermediates.

KCI등재 SCOPUS

6Protective effects of Camellia sinensis fruit and fruit peels against oxidative DNA damage

저자 : Joung-jwa Ahn , Tae-won Jang , Jae-ho Park

발행기관 : 한국응용생명화학회 간행물 : Journal of Applied Biological Chemistry 64권 3호 발행 연도 : 2021 페이지 : pp. 237-480 (244 pages)

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Camellia sinensis, Green tea, contains phenolic compounds that act to scavenge reactive oxygen species (ROS), such as catechin, epicatechin, etc. In contrast with the tea leaf, the bioactivity of its fruit and the fruit peels remains still unclear. This study focused on the effects of fruit and fruit peels of C. sinensis (FC and PC) against oxidative DNA damage in NIH/3T3 cells. The scavenging effects of FC and PC on ROS were assessed using 1,1-diphenyl-2-picryl hydrazyl or 2,2'-azino-bis(3-ethylbenzothiazoline- 6-sulfonic acid radicals. The measurement of ROS in cellular levels was conducted by DCFDA reagent and the protein expression of γ-H2AX, H2AX, cleaved caspase-3, p53, and, pp53 was analyzed by immunoblotting. The gene expressions of p53 and H2AX were assessed using polymerase chain reaction techniques. The major metabolites of FC and PC were quantitatively measured analyzed and the amounts of phenolic compounds and flavonoids in PC were greater than those in FC. Further, PC suppressed ROS production, which protects the oxidative stress-induced DNA damage through reducing H2AX, p53, and caspase-3 phosphorylation. These results refer that the protective effects of FC and PC are mediated by inhibition of p53 signaling pathways, probably via the bioactivity of phenolic compounds. Thus, FC and PC can serve as a potential antioxidant in DNA damage-associated diseases.

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7UPLC-QTOF-MS분석를 이용한 국내산 더덕 주산지의 표지물질 선정

저자 : 안영민 ( Young Min An ) , 장현재 ( Hyun-jae Jang ) , 김두영 ( Doo-young Kim ) , 백남인 ( Nam-in Baek ) , 오세량 ( Sei-ryang Oh ) , 이대영 ( Dae Young Lee ) , 류형원 ( Hyung Won Ryu )

발행기관 : 한국응용생명화학회 간행물 : Journal of Applied Biological Chemistry 64권 3호 발행 연도 : 2021 페이지 : pp. 245-495 (251 pages)

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더덕(Codonopsis lanceolata)은 주로 한국, 중국 등 동아시아 지역에 재배되고 있으며, 더덕의 뿌리는 기침, 기관지염, 천식, 결핵, 소화 불량의 증상을 치료하기 위한 기능성 식품 및 전통 의학으로 사용되어져 왔다. 보고된 바에 의하면 phenylpropanoids, polyacetylenes, saponins, flavonoids와 같은 다양한 식물 천연물 성분들이 항비만, 항염, 항암, 항산화, 항미생물 활성과 같은 약리학적 작용에 관여한다고 보고되어 있다. MS기반 대사체학 분석을 이용한 주산지의 마커 성분을 선정하는 것은 다른 지역에서 재배된 약용 식물의 안전성뿐만 아니라 화학적 조성과 생물학적 효능의 변화와도 관련이 있기 때문에 부작용 없이 더덕의 유익한 효과만을 보장하는데 중요하다. 본 연구에서는 국내산 더덕의 주산지 특성을 구별하기 위해 UPLC-QTOF-MS를 기반으로 하는 대사체 프로파일링과 다변량 통계분석 기법인 PCA 분석을 수행하여 판별모델을 확립하였다. 그 결과 인제(강원도), 횡성(강원도), 무주(전라북도)의 3개 그룹이 PCA와 loading plot 분석결과 tangshenoside I, lancemaside A, lancemaside G는 더덕 주산지를 구별하기 위한 잠재적 대사체 마커들로 제안하였다.


Codonopsis lanceolata (Deoduk) was grown in East Asia, including Korea, China, Japan, and Russia, and the roots of C. lanceolata have been used as functional foods and traditional medicine to treat symptoms of cough, bronchitis, asthma, tuberculosis, and dyspepsia. The phytochemicals of C. lanceolata have been reported such as phenylpropanoids, polyacetylenes, saponins, and flavonoids that are involved in pharmacological effects such as anti-obesity, anti-inflammation, anti-tumor, antioxidant, and anti-microbial activities. Selecting marker substances of the main producing area by MS-based metabolomics analysis is important to ensure the beneficial effect of C. lanceolata without side-effects because differences in cultivated areas of plants were related not only to the safety of medicinal plants but also to changes in chemical composition and biological efficacy. In our present study, ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry combined with multivariate statistical analysis was applied to recognize the main producing area of C. lanceolata in South Korea. As a result of Principal Component Analysis and loading plot analysis of three groups, Inje (Kangwon-do), Hoengseong (Kangwon-do), and Muju (Jeonlabuk-do), several secondary metabolites of C. lanceolata including tangshenoside I, lancemaside A, and lancemaside G, were suggested as potential marker substances to distinguish the place of main producing area of C. lanceolata.

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8식물공장에서 생산된 새싹인삼의 크기에 따른 진세노사이드 함량 및 항산화 활성 비교

저자 : 황승하 ( Seung Ha Hwang ) , 김수철 ( Su Cheol Kim ) , 성진아 ( Jin A Seong ) , 이희율 ( Hee Yul Lee ) , 조두용 ( Du Yong Cho ) , 김민주 ( Min Ju Kim ) , 정재각 ( Jea Gack Jung ) , 정은혜 ( Eun Hye Jeong ) , 손기호 ( Ki-ho Son ) , 조계만 ( Kye Man Cho )

발행기관 : 한국응용생명화학회 간행물 : Journal of Applied Biological Chemistry 64권 3호 발행 연도 : 2021 페이지 : pp. 253-513 (261 pages)

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본 연구에서는 식물공장을 통해 생산된 새싹인삼을 크기에 따라 분류하고 이를 지상부와 지하부로 나누어 진세노사이드 함량과 항산화 활성을 비교하였다. 지상부의 경우 총 phenolic 함량은 중간 크기인 M에서 5.16 GAE mg/g로 가장 높았으며 가장 큰 크기인 L에서 2.23 GAE mg/g으로 가장 낮은 함량을 보였다. 지하부 역시 M 크기에서 가장 높은 함량을 보였으나, 큰 차이를 보이지 않았다. 한편, 총 flavonoid 함량 역시 지상부(5.16 RE mg/g) 와 지하부(1.28 RE mg/g) 모두 M 크기에서 높은 함량을 보였다. 지상부의 주요 진세노사이드는 Re(20.33-24.15 mg/g) > Rd (11.36-27.42 mg/g) > Rg1 (4.48-5.54 mg/g) 순 있었고 지하부는 Rb1 (5.09-8.61 mg/g) > Re (4.48- 5.54 mg/g) > Rc (3.11-4.11 mg/g) 순 있었다. M 크기의 경우 Re와 Rd는 각각 지상부에서 24.15 mg/g과 27.42 mg/g 및 지하부에서 5.20 mg/g과 1.43 mg으로 약 4배와 19배 높은 함량을 보였다. 지상부에는 F3 및 Rh1이 검출되었으나, 지하부에서는 검출되지 않았다. DPPH (74.95%)와 ABTS (94.47%), hydroxyl (70.39%) 라디칼 소거 활성 및 FRAP (2.169) 활성은 다른 크기들보다 M 크기에서 가장 높은 활성을 보였다.


In this study, the ginseng sprout has produced through smart farm was classified according to its size and divided into above-ground (AG) and below-ground (BG) parts to compare ginsenoside contents and antioxidant activity. In the case of the AG part, the total phenolic contents were the highest at 5.16 mg/ g in medium (M) size and the lowest at 2.23 mg/g in largest (L) size. The BG part also showed the highest content in the M size, but there was no significant difference. Also, the total flavonoid contents were also high in the M size in both the AG (5.16 mg/ g) and BG (1.28 mg/g) parts. The major ginsenosides in the AG part were Re (20.33-24.15 mg/g) > Rd (11.36-27.42 mg/g) > Rg1 (4.48-5.54 mg/g) and the main ginsenosides in the BG part were Rb1 (5.09-8.61 mg/g) > Re (4.48-5.54 mg/g) > Rc (3.11-4.11 mg/ g) in orders. In the case of M size, Re and Rd were approximately 4- and 19-folds higher at 24.15 mg/g and at 27.42 mg/g in the AG part and 5.20 mg/g and 1.43 mg in the BG part, respectively. In addition, F3 and Rh1 were detected in the AG part, but not in the BG part. 2,2-diphenyl-1-picrylhydrazyl (74.95%), 2,4,6-azino-bis (3-ethylbenzothiazoline-6-sulphnoic acid) diammonium salt (94.47%), and hydroxyl (70.39%) radical scavenging activities and FRAP (2.169) assay were the highest in M size than other sizes.

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9The hyaluronan synthesis inhibitor 7-hydroxy-4-methylcoumarin inhibits LPS-induced inflammatory response in RAW 264.7 macrophage cells

저자 : Gwan Bo Kim , Chang-gu Hyun

발행기관 : 한국응용생명화학회 간행물 : Journal of Applied Biological Chemistry 64권 3호 발행 연도 : 2021 페이지 : pp. 263-530 (268 pages)

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7-Hydroxy-4-methylcoumarin (7H-4MC) inhibits hyaluronan production in multiple cell lines and tissue types both in vitro and in vivo. It is a commercially available drug approved for human use, called hymecromone, in European and Asian countries to prevent biliary spasms. Nevertheless, as the pharmacological efficacy of 7H-4MC has not yet been reported in macrophages, this study investigated its anti-inflammatory effects and mechanism of action using lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. LPS-induced RAW 264.7 cells were treated with various concentrations of 7H-4MC (62.5, 125, 250, and 500 μM). The application of 7H-4MC significantly reduced nitric oxide and prostaglandin E2 production without cytotoxic effects. Additionally, 7H-4MC strongly decreased the expression of inducible nitric oxide synthase and cyclooxygenase. Furthermore, 7H-4MC reduced the production of proinflammatory cytokines, such as tumor necrosis factor-α, interleukin (IL)-1β, and IL-6. Finally, 7H-4MC exerted its potent anti-inflammatory actions via the upregulation of IκB-α production, which led to the inhibition of nuclear factor-κB (NF-κB) activity. These results, obtained in macrophage cell lines, suggest that 7H-4MC prevents inflammatory diseases via the NF-κB signaling pathway and that its use could be beneficial for human health. Ultimately, this is the first report describing the anti-inflammatory activity of 7H-4MC in a macrophage cell line.

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10Anti-adipocyte differentiation activity and flavonoid content determination by HPLC/UV analysis of tree sprouts

저자 : Juree Kim , Taewon Jang , Ji Hyun Kim , Hanna Shin , Jaeho Park , Sanghyun Lee

발행기관 : 한국응용생명화학회 간행물 : Journal of Applied Biological Chemistry 64권 3호 발행 연도 : 2021 페이지 : pp. 269-543 (275 pages)

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The in vitro anti-obesity activity of 12 species of tree sprouts in differentiated 3T3-L1 cells and the mechanisms underlying their activity were evaluated. (+)-Catechin and quercetin concentrations in the sprouts were analyzed by HPLC/ UV at 270 and 254 nm, respectively. Euonymus alatus (EAT) and Fraxinus mandschuria (FMS) extracts at doses of 50 and 100 μg/ mL inhibited the accumulation of lipid droplets in differentiated 3T3-L1 cells. Moreover, EAT and FMS downregulated the expression of the CCAAT/enhancer-binding protein-α, adipogenesisrelated proteins peroxisome proliferator-activated receptor-γ, and adipocyte P-2α in differentiated 3T3-L1 cells. Tree sprouts with an abundant flavonoid content exerted the highest anti-obesity activity. Concentrations of total flavonoids were the highest in FMS (24.281 mg/g DW) sprouts. These findings could be used to develop health-promoting functional foods or supplements derived from tree sprouts.

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1고함량 사포나린 함유 보리 어린 순 재배를 위한 식물공장내 인공광 조건 및 사포나린 추출 분석법 최적화

저자 : 오경열 ( Kyeong-yeol Oh ) , 송영훈 ( Yeong Hun Song ) , 이득영 ( Duek-yeong Lee ) , 이태근 ( Tae-geun Lee ) , 김진효 ( Jin-hyo Kim )

발행기관 : 한국응용생명화학회 간행물 : Journal of Applied Biological Chemistry 64권 3호 발행 연도 : 2021 페이지 : pp. 197-399 (203 pages)

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본 연구에서는 보리 어린 순의 기능성 성분인 saponarin의 정량적 추출을 위한 최적의 추출 분석 조건을 methanol, ethanol, acetonitrile 및 water를 대상으로 반응표면분석법을 통해 실시하였다. 이를 통해 용매의 농도, 추출 시간에 따른 saponarin과 색소 추출율을 확인한 결과, 53.7% methanol 수용액에서 3.9 h 동안 진탕추출하는 것이 높은 saponarin 추출율을 유지하면서, 불필요한 색소의 추출을 최소화하는 최적 추출조건으로 확인되었다. 보리 어린 순의 재배에 필요한 인공광의 saponarin 생성 영향 평가를 광 주기, 광량, 광질을 달리한 조건에서 시험한 결과, saponarin 고함유 보리 어린 순의 인공광 최적 재배 조건은 8 h day-1의 광 주기에서 6500K LED와 적색보광을 통한 총 광량 220-320 μmol m-2 s-1 에서 가장 효과적임을 확인 할 수 있었고, 청색광이 saponarin 생합성에 주요 인자로 작용함을 확인할 수 있었다.

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2Molecular characterization and biological changes caused by Agrobacterium-mediated infiltration of PgTRX1

저자 : Seung Hyuk Choi , Ji Won Seo , Jae Geun Lee , Chang Yeon Yu , Eun Soo Seong

발행기관 : 한국응용생명화학회 간행물 : Journal of Applied Biological Chemistry 64권 3호 발행 연도 : 2021 페이지 : pp. 205-415 (211 pages)

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In order to test the functionality of Panax ginseng thioredoxin 1 (PgTRX1) isolated from fermented wild ginseng roots, a transient effect on physiological activity were performed over a short time frame using the Agrobacterium infiltration technique. The PgTRX1 gene isolated from fermented wild ginseng was confirmed to have a size of 579 bp, and the expression of PgTRX1 was the highest in the sample after 6 h of fermentation. As a result of constructing this gene and confirming the infiltration reaction mediated by Agrobacterium in tobacco leaves, it was found that the expression of the NbHSR203j gene was also induced as PgTRX1 expression increased. As a result of measuring the biological activity of the infiltration samples, the total phenol content increased by 35.45±1.84 to 49.01±1.84 μg GAE/mL compared to the control, and the total flavonoid amount of 9.52±0.41 to 9.82±0.25 μg QE/mL was slightly high. From these results, Agrobacterium-mediated PgTRX1 appears to be related to the hypersensitive response induction mechanism of plants and the production of secondary metabolites such as phenolic substances.

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3Characterization of the recombinant cellulase A from Thermotoga maritima

저자 : Chung Ho Kim

발행기관 : 한국응용생명화학회 간행물 : Journal of Applied Biological Chemistry 64권 3호 발행 연도 : 2021 페이지 : pp. 213-428 (216 pages)

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A gene encoding thermostable cellulase A (TmCelA) was isolated from Thermotoga maritima. The open reading frame of TmCelA gene was 774 bp long which predicted to encode 257 amino acid residues with a molecular weight of 29,732 Da. To examine the biochemical properties, the TmCelA was overexpressed in E. coli BL21, and expressed protein was purified. The optimum temperature of recombinant TmCelA was 90-95 ℃, and the optimum pH of recombinant TmCelA was approximately pH 5.0. Recombinant TmCelA was stable at temperature below 90 ℃.

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4Rhodobacter sphaeroides에서 5-aminolevulinic acid 생산에 대한 850 nm 근적외선 발광다이오드 조사 효과

저자 : 모상준 ( Sangjoon Mo )

발행기관 : 한국응용생명화학회 간행물 : Journal of Applied Biological Chemistry 64권 3호 발행 연도 : 2021 페이지 : pp. 217-439 (223 pages)

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광감각제를 이용한 광역학 치료는 필요한 특정 부위에만 빛을 조사하여 치료 효과를 나타내는 부작용이 적은 방법이다. 5-aminolevulinic acid (ALA)는 다양한 생물체에서 합성되는 대표적 광감제로 암진단과 치료를 포함하는 다양한 분야에서 사용되고 있다. 본 연구에서는 다양한 파장의 LED, 유기산 전구체 및 glucose 농도 변화를 통한 Rhodobacter sphaeroides의 최적 성장 조건과 ALA 생산 조건을 확립하기 위한 실험을 진행하였다. 백열등과 동일한 광도 아래에서 Rhodobacter sphaeroides에 850 nm LED 빛을 조사하면 대조군 대비 균주의 성장과 ALA의 생산 농도를 각각 1.5배 및 1.8배 증가시킬 수 있고, 전구체로 pyruvic acid를 첨가한 경우 850 nm 파장의 LED만 조사한 경우 보다 ALA의 생산 농도를 약 2.8배 증가 시켰으며 동일 배양 조건에 40 mM glucose를 첨가하여 배양한 결과 Rhodobacter sphaeroides의 성장은 850 nm 파장의 LED 조사와 pyruvic acid를 첨가한 것에 비해 약 2.9배, ALA의 생산 농도는 약 3.4배 (20 mM) 증가되었다. 건조체 질량당 ALA의 생산은 20 mM과 40 mM glucose에서 대조군 대비 각각 약 1.4배 높은 결과를 나타냈다. 결론적으로 다양한 파장의 LED 중 850 nm 파장의 LED가 Rhodobacter sphaeroides의 성장률 및 ALA의 생산을 최대로 높였으며, 5 mM pyruvic acid와 40 mM glucose의 농도에서 최적의 Rhodobacter sphaeroides 성장과 ALA 생산을 확인하였다.

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5Cytochrome P450 and the glycosyltransferase genes are necessary for product release from epipyrone polyketide synthase in Epicoccum nigrum

저자 : Eun Ha Choi , Si-hyung Park , Hyung-jin Kwon

발행기관 : 한국응용생명화학회 간행물 : Journal of Applied Biological Chemistry 64권 3호 발행 연도 : 2021 페이지 : pp. 225-459 (235 pages)

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The epipyrone (EPN) biosynthetic gene cluster of Epicoccum nigrum is composed of epnC, epnB, and epnA, which encode cytochrome P450 oxidase, glycosyltransferase, and highly reducing polyketide synthase, respectively. Gene inactivation mutants for epnA, epnB, and epnC were previously generated, and it was found that all of them were incapable of producing EPN and any of its related compounds. It was also reported that epnB inactivation abolished epnA transcription, generating ΔepnAB. This study shows that the introduction of native epnC readily restored EPN production in ΔepnC, suggesting that epnC is essential for polyketide release from EpnA and implies that EpnC works during the polyketide chain assembly of EpnA. Introduction of epnC promoter-epnA restored EPN production in ΔepnA. The ΔepnB genotype was prepared by introducing the epnA expression vector into ΔepnAB, and it was found that the resulting recombinant strain did not produce any EPN-related compounds. A canonical epnB inactivation strain was also generated by deleting its 5′-end. At the deletion point, an Aspergllus nidulans gpdA promoter was inserted to ensure the transcription of epnA, which is located downstream of epnB. Examination of the metabolite profile of the resulting ΔepnB mutant via LC-mass spectrometry verified that no EPN-related compound was produced in this strain. This substantiates that C-glycosylation by EpnB is a prerequisite for the release of EpnA-tethered product. In conclusion, it is proposed that cytochrome P450 oxidase and glycosyltransferase work in concert with polyketide synthase to generate EPN without the occurrence of any free intermediates.

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6Protective effects of Camellia sinensis fruit and fruit peels against oxidative DNA damage

저자 : Joung-jwa Ahn , Tae-won Jang , Jae-ho Park

발행기관 : 한국응용생명화학회 간행물 : Journal of Applied Biological Chemistry 64권 3호 발행 연도 : 2021 페이지 : pp. 237-480 (244 pages)

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Camellia sinensis, Green tea, contains phenolic compounds that act to scavenge reactive oxygen species (ROS), such as catechin, epicatechin, etc. In contrast with the tea leaf, the bioactivity of its fruit and the fruit peels remains still unclear. This study focused on the effects of fruit and fruit peels of C. sinensis (FC and PC) against oxidative DNA damage in NIH/3T3 cells. The scavenging effects of FC and PC on ROS were assessed using 1,1-diphenyl-2-picryl hydrazyl or 2,2'-azino-bis(3-ethylbenzothiazoline- 6-sulfonic acid radicals. The measurement of ROS in cellular levels was conducted by DCFDA reagent and the protein expression of γ-H2AX, H2AX, cleaved caspase-3, p53, and, pp53 was analyzed by immunoblotting. The gene expressions of p53 and H2AX were assessed using polymerase chain reaction techniques. The major metabolites of FC and PC were quantitatively measured analyzed and the amounts of phenolic compounds and flavonoids in PC were greater than those in FC. Further, PC suppressed ROS production, which protects the oxidative stress-induced DNA damage through reducing H2AX, p53, and caspase-3 phosphorylation. These results refer that the protective effects of FC and PC are mediated by inhibition of p53 signaling pathways, probably via the bioactivity of phenolic compounds. Thus, FC and PC can serve as a potential antioxidant in DNA damage-associated diseases.

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7UPLC-QTOF-MS분석를 이용한 국내산 더덕 주산지의 표지물질 선정

저자 : 안영민 ( Young Min An ) , 장현재 ( Hyun-jae Jang ) , 김두영 ( Doo-young Kim ) , 백남인 ( Nam-in Baek ) , 오세량 ( Sei-ryang Oh ) , 이대영 ( Dae Young Lee ) , 류형원 ( Hyung Won Ryu )

발행기관 : 한국응용생명화학회 간행물 : Journal of Applied Biological Chemistry 64권 3호 발행 연도 : 2021 페이지 : pp. 245-495 (251 pages)

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더덕(Codonopsis lanceolata)은 주로 한국, 중국 등 동아시아 지역에 재배되고 있으며, 더덕의 뿌리는 기침, 기관지염, 천식, 결핵, 소화 불량의 증상을 치료하기 위한 기능성 식품 및 전통 의학으로 사용되어져 왔다. 보고된 바에 의하면 phenylpropanoids, polyacetylenes, saponins, flavonoids와 같은 다양한 식물 천연물 성분들이 항비만, 항염, 항암, 항산화, 항미생물 활성과 같은 약리학적 작용에 관여한다고 보고되어 있다. MS기반 대사체학 분석을 이용한 주산지의 마커 성분을 선정하는 것은 다른 지역에서 재배된 약용 식물의 안전성뿐만 아니라 화학적 조성과 생물학적 효능의 변화와도 관련이 있기 때문에 부작용 없이 더덕의 유익한 효과만을 보장하는데 중요하다. 본 연구에서는 국내산 더덕의 주산지 특성을 구별하기 위해 UPLC-QTOF-MS를 기반으로 하는 대사체 프로파일링과 다변량 통계분석 기법인 PCA 분석을 수행하여 판별모델을 확립하였다. 그 결과 인제(강원도), 횡성(강원도), 무주(전라북도)의 3개 그룹이 PCA와 loading plot 분석결과 tangshenoside I, lancemaside A, lancemaside G는 더덕 주산지를 구별하기 위한 잠재적 대사체 마커들로 제안하였다.

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8식물공장에서 생산된 새싹인삼의 크기에 따른 진세노사이드 함량 및 항산화 활성 비교

저자 : 황승하 ( Seung Ha Hwang ) , 김수철 ( Su Cheol Kim ) , 성진아 ( Jin A Seong ) , 이희율 ( Hee Yul Lee ) , 조두용 ( Du Yong Cho ) , 김민주 ( Min Ju Kim ) , 정재각 ( Jea Gack Jung ) , 정은혜 ( Eun Hye Jeong ) , 손기호 ( Ki-ho Son ) , 조계만 ( Kye Man Cho )

발행기관 : 한국응용생명화학회 간행물 : Journal of Applied Biological Chemistry 64권 3호 발행 연도 : 2021 페이지 : pp. 253-513 (261 pages)

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본 연구에서는 식물공장을 통해 생산된 새싹인삼을 크기에 따라 분류하고 이를 지상부와 지하부로 나누어 진세노사이드 함량과 항산화 활성을 비교하였다. 지상부의 경우 총 phenolic 함량은 중간 크기인 M에서 5.16 GAE mg/g로 가장 높았으며 가장 큰 크기인 L에서 2.23 GAE mg/g으로 가장 낮은 함량을 보였다. 지하부 역시 M 크기에서 가장 높은 함량을 보였으나, 큰 차이를 보이지 않았다. 한편, 총 flavonoid 함량 역시 지상부(5.16 RE mg/g) 와 지하부(1.28 RE mg/g) 모두 M 크기에서 높은 함량을 보였다. 지상부의 주요 진세노사이드는 Re(20.33-24.15 mg/g) > Rd (11.36-27.42 mg/g) > Rg1 (4.48-5.54 mg/g) 순 있었고 지하부는 Rb1 (5.09-8.61 mg/g) > Re (4.48- 5.54 mg/g) > Rc (3.11-4.11 mg/g) 순 있었다. M 크기의 경우 Re와 Rd는 각각 지상부에서 24.15 mg/g과 27.42 mg/g 및 지하부에서 5.20 mg/g과 1.43 mg으로 약 4배와 19배 높은 함량을 보였다. 지상부에는 F3 및 Rh1이 검출되었으나, 지하부에서는 검출되지 않았다. DPPH (74.95%)와 ABTS (94.47%), hydroxyl (70.39%) 라디칼 소거 활성 및 FRAP (2.169) 활성은 다른 크기들보다 M 크기에서 가장 높은 활성을 보였다.

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9The hyaluronan synthesis inhibitor 7-hydroxy-4-methylcoumarin inhibits LPS-induced inflammatory response in RAW 264.7 macrophage cells

저자 : Gwan Bo Kim , Chang-gu Hyun

발행기관 : 한국응용생명화학회 간행물 : Journal of Applied Biological Chemistry 64권 3호 발행 연도 : 2021 페이지 : pp. 263-530 (268 pages)

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7-Hydroxy-4-methylcoumarin (7H-4MC) inhibits hyaluronan production in multiple cell lines and tissue types both in vitro and in vivo. It is a commercially available drug approved for human use, called hymecromone, in European and Asian countries to prevent biliary spasms. Nevertheless, as the pharmacological efficacy of 7H-4MC has not yet been reported in macrophages, this study investigated its anti-inflammatory effects and mechanism of action using lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. LPS-induced RAW 264.7 cells were treated with various concentrations of 7H-4MC (62.5, 125, 250, and 500 μM). The application of 7H-4MC significantly reduced nitric oxide and prostaglandin E2 production without cytotoxic effects. Additionally, 7H-4MC strongly decreased the expression of inducible nitric oxide synthase and cyclooxygenase. Furthermore, 7H-4MC reduced the production of proinflammatory cytokines, such as tumor necrosis factor-α, interleukin (IL)-1β, and IL-6. Finally, 7H-4MC exerted its potent anti-inflammatory actions via the upregulation of IκB-α production, which led to the inhibition of nuclear factor-κB (NF-κB) activity. These results, obtained in macrophage cell lines, suggest that 7H-4MC prevents inflammatory diseases via the NF-κB signaling pathway and that its use could be beneficial for human health. Ultimately, this is the first report describing the anti-inflammatory activity of 7H-4MC in a macrophage cell line.

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10Anti-adipocyte differentiation activity and flavonoid content determination by HPLC/UV analysis of tree sprouts

저자 : Juree Kim , Taewon Jang , Ji Hyun Kim , Hanna Shin , Jaeho Park , Sanghyun Lee

발행기관 : 한국응용생명화학회 간행물 : Journal of Applied Biological Chemistry 64권 3호 발행 연도 : 2021 페이지 : pp. 269-543 (275 pages)

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The in vitro anti-obesity activity of 12 species of tree sprouts in differentiated 3T3-L1 cells and the mechanisms underlying their activity were evaluated. (+)-Catechin and quercetin concentrations in the sprouts were analyzed by HPLC/ UV at 270 and 254 nm, respectively. Euonymus alatus (EAT) and Fraxinus mandschuria (FMS) extracts at doses of 50 and 100 μg/ mL inhibited the accumulation of lipid droplets in differentiated 3T3-L1 cells. Moreover, EAT and FMS downregulated the expression of the CCAAT/enhancer-binding protein-α, adipogenesisrelated proteins peroxisome proliferator-activated receptor-γ, and adipocyte P-2α in differentiated 3T3-L1 cells. Tree sprouts with an abundant flavonoid content exerted the highest anti-obesity activity. Concentrations of total flavonoids were the highest in FMS (24.281 mg/g DW) sprouts. These findings could be used to develop health-promoting functional foods or supplements derived from tree sprouts.

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