Biotransformation of Panax ginseng extract by rat intestinal microflora: identification and quantification of metabolites using liquid chromatography-tandem mass spectrometry

Dong, Wei-Wei,Zhao, Jinhua,Zhong, Fei-Liang,Zhu, Wen-Jing,Jiang, Jun,Wu, Songquan,Yang, Deok-Chun,Li, Donghao,Quan, Lin-Hu The Korean Society of Ginseng 2017 Journal of Ginseng Research Vol.41 No.4

Background: In general, after Panax ginseng is administered orally, intestinal microbes play a crucial role in its degradation and metabolization process. Studies on the metabolism of P. ginseng by microflora are important for obtaining a better understanding of their biological effects. Methods: In vitro biotransformation of P. ginseng extract by rat intestinal microflora was investigated at $37^{\circ}C$ for 24 h, and the simultaneous determination of the metabolites and metabolic profile of P. ginseng saponins by rat intestinal microflora was achieved using LC-MS/MS. Results: A total of seven ginsenosides were detected in the P. ginseng extract, including ginsenosides Rg1, Re, Rf, Rb1, Rc, Rb2, and Rd. In the transformed P. ginseng samples, considerable amounts of deglycosylated metabolite compound K and Rh1 were detected. In addition, minimal amounts of deglycosylated metabolites (ginsenosides Rg2, F1, F2, Rg3, and protopanaxatriol-type ginsenosides) and untransformed ginsenosides Re, Rg1, and Rd were detected at 24 h. The results indicated that the primary metabolites are compound K and Rh1, and the protopanaxadiol-type ginsenosides were more easily metabolized than protopanaxatriol-type ginsenosides. Conclusion: This is the first report of the identification and quantification of the metabolism and metabolic profile of P. ginseng extract in rat intestinal microflora using LC-MS/MS. The current study provided new insights for studying the metabolism and active metabolites of P. ginseng.

Biotransformation of Panax ginseng extract by rat intestinal microflora

Wei-Wei Dong,Jinhua Zhao,Fei-Liang Zhong,Wen-Jing Zhu,Jun Jiang,Songquan Wu,Deok-Chun Yang,Donghao Li,Lin-Hu Quan 고려인삼학회 2017 Journal of Ginseng Research Vol.41 No.4

Background: In general, after Panax ginseng is administered orally, intestinal microbes play a crucial role in its degradation and metabolization process. Studies on the metabolism of P. ginseng by microflora are important for obtaining a better understanding of their biological effects. Methods: In vitro biotransformation of P. ginseng extract by rat intestinal microflora was investigated at 37C for 24 h, and the simultaneous determination of the metabolites and metabolic profile of P. ginseng saponins by rat intestinal microflora was achieved using LCeMS/MS. Results: A total of seven ginsenosides were detected in the P. ginseng extract, including ginsenosides Rg1, Re, Rf, Rb1, Rc, Rb2, and Rd. In the transformed P. ginseng samples, considerable amounts of deglycosylated metabolite compound K and Rh1 were detected. In addition, minimal amounts of deglycosylated metabolites (ginsenosides Rg2, F1, F2, Rg3, and protopanaxatriol-type ginsenosides) and untransformed ginsenosides Re, Rg1, and Rd were detected at 24 h. The results indicated that the primary metabolites are compound K and Rh1, and the protopanaxadiol-type ginsenosides were more easily metabolized than protopanaxatriol-type ginsenosides. Conclusion: This is the first report of the identification and quantification of the metabolism and metabolic profile of P. ginseng extract in rat intestinal microflora using LCeMS/MS. The current study provided new insights for studying the metabolism and active metabolites of P. ginseng.

Neospora caninum-antigens and Antigen Displaying Baculovirus Using Silkworm Larvae Targeting for Neospora Vaccine

Enoch Y. Park,Takahiro Otsuki,Jinhua Dong,Tatsuya Kato 한국생물공학회 2013 한국생물공학회 학술대회 Vol.2013 No.10

Single-Step Detection of the Influenza Virus Hemagglutinin Using Bacterially-Produced Quenchbodies

Sanghyun CHA,DongHyun LEE,Dayoung YUN,Dohee LEE,Jinhua DONG,Hiroshi UEDA,Hee-Jin JEONG 한국생물공학회 2019 한국생물공학회 학술대회 Vol.2019 No.4

Discovery of the leinamycin family of natural products by mining actinobacterial genomes

Pan, Guohui,Xu, Zhengren,Guo, Zhikai,Hindra,Ma, Ming,Yang, Dong,Zhou, Hao,Gansemans, Yannick,Zhu, Xiangcheng,Huang, Yong,Zhao, Li-Xing,Jiang, Yi,Cheng, Jinhua,Van Nieuwerburgh, Filip,Suh, Joo-Won,Duan National Academy of Sciences 2017 PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF Vol.114 No.52

<P>Nature's ability to generate diverse natural products from simple building blocks has inspired combinatorial biosynthesis. The knowledge-based approach to combinatorial biosynthesis has allowed the production of designer analogs by rational metabolic pathway engineering. While successful, structural alterations are limited, with designer analogs often produced in compromised titers. The discovery-based approach to combinatorial biosynthesis complements the knowledge-based approach by exploring the vast combinatorial biosynthesis repertoire found in Nature. Here we showcase the discovery-based approach to combinatorial biosynthesis by targeting the domain of unknown function and cysteine lyase domain (DUF-SH) didomain, specific for sulfur incorporation from the leinamycin (LNM) biosynthetic machinery, to discover the LNM family of natural products. By mining bacterial genomes from public databases and the actinomycetes strain collection at The Scripps Research Institute, we discovered 49 potential producers that could be grouped into 18 distinct clades based on phylogenetic analysis of the DUF-SH didomains. Further analysis of the representative genomes from each of the clades identified 28 lnm-type gene clusters. Structural diversities encoded by the LNM-type biosynthetic machineries were predicted based on bioinformatics and confirmed by in vitro characterization of selected adenylation proteins and isolation and structural elucidation of the guangnanmycins and weishanmycins. These findings demonstrate the power of the discovery-based approach to combinatorial biosynthesis for natural product discovery and structural diversity and highlight Nature's rich biosynthetic repertoire. Comparative analysis of the LNM-type biosynthetic machineries provides outstanding opportunities to dissect Nature's biosynthetic strategies and apply these findings to combinatorial biosynthesis for natural product discovery and structural diversity.</P>

Amycolatopsis sp. KCTC 29142로부터 유래된 siderochelin A의 다제 내성 균주에 대한 항균활성

이동령,성금화,이성권,홍희전,송재경,양승환,서주원,Lee, Dong-Ryung,Cheng, Jinhua,Lee, Sung-Kwon,Hong, Hee-Jeon,Song, Jaekyeong,Yang, Seung Hwan,Suh, Joo-Won The Microbiological Society of Korea 2016 미생물학회지 Vol.52 No.3

본 연구에서는 신규 Amycolatopsis 균주 KCTC 29142를 분리하여 형태학적 관찰, 계통분석 및 화학분류학적 분석 등 다상 분류분석을 통해 분석하였다. KCTC 29142 균주의 에틸아세테이트추출물은 강한 항균활성을 나타났고, 활성물질은 철 이온 킬레이트 물질인 siderochelin A로 동정되었다. 본 연구에서 분리된 siderochelin A는 다제내성균인 Acinetobacter baumanii, methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Staphylococcus aureus (VRSA), 및 Escherichia coli (E. coli)에 대해 강한 활성을 보였고, 임상에서 분리된 다제내성균에 대한 MIC를 결정하였다. A novel Amycolatopsis strain KCTC 29142 was isolated and characterized based on the polyphasic taxonomic analysis including morphological observation, phylogenetic analysis, physiological and chemotaxonomic characteristics. The ethyl acetate extract of strain KCTC 29142 culture broth showed strong antibacterial activity and the active compound was identified as siderochelin A, a ferrous-ion chelating compound. In this study, siderochelin A showed good activity against multi-drug resistant pathogens, including Acinetobacter baumanii, methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Staphylococcus aureus (VRSA), and Escherichia coli (E. coli). The minimum inhibitory activity against clinical isolates was also determined.

발효 청미래덩굴잎 용매 추출물 및 분획물의 xanthine 및 aldehyde oxidase 저해활성과 항고요산혈증 효과

이상일 ( Sang Il Lee ),이예경 ( Ye Kyung Lee ),김순동 ( Soon Dong Kim ),성금화 ( Jinhua Cheng ),양승환 ( Seung Hwan Yang ),서주원 ( Joo Won Suh ) 한국응용생명화학회(구 한국농화학회) 2014 Journal of Applied Biological Chemistry (J. Appl. Vol.57 No.1

Aspergillus oryzae로 발효시킨 청미래덩굴잎 용매추출분획물의 xanthine oxidase (XO) 및 aldehyde oxidase (AO) 저해활성과 항고요산혈증에 미치는 영향을 조사하였다. 용매별 추출수율(g/kg)은 80% ethanol (EtOH)은 13.56이었으며, n-hexane, dichloromethane (DICM), ethylacetate (EtOAc) 및 n-butanol fraction (BuOH) 은 1.35.3.33 범위였다. Total polyphenol 함량(mg/g-extract)은 EtOAc fraction 478.07.501.26, BuOH fraction 259.49.289.02, DICM fraction 165.03.232.27, EtOH fraction 134.02.196.54이었으며 EtOAc 및 DICM fraction에서는 발효시킨 경우가 비 발효에 비하여 각각 4.85 및 40.74%가 높았으나 그 외 모든fraction에서는 발효시킨 경우가 낮았다. 총 flavonoid 함량은 EtOAc fraction이 여타 fraction에 비하여 높았다. EtOAc 및 BuOH fraction의 TF 함량은 발효시킨 경우가 비 발효에 비하여 각각 10.56% 및 60.17%가 높았으나 여타 fraction들에서는 발효시킨 경우가 낮았다. XO 저해활성은 모든 분획물에서 발효시킨 경우가 비발효에 비하여 현저하게 높았으며 EtOAc 및 BuOH fraction의 경우 발효시킨 경우는 각각 75.02 및 65.59%로 비발효 경우 39.42 및 5.34% 보다 현저하게 높았다. AO 저해활성은 DICM과 EtOAc fraction에서 각각 81.82 및 77.93%로 여타 fraction들에 비하여 높았으며XO의 경우와 마찬가지로 발효시킨 경우가 비발효에 비하여 현저하게 높았다. 고요산혈증 mouse의 혈중 요산(SU) 함량에 미치는 영향을 조사한 결과, 고요산혈증 대조군의 SU 함량은 6.98mg/dL로 정상대조군 (NC)의 3.82 mg/dL에 비하여 1.83배가 높았으나, EtOAc fraction은 투여량에 의존적으로 SU의 함량이 감소하였으며, 발효시킨 경우가 비발효에 비하여 유의적인 감소를 보였다. 이상의 결과, A. oryzae와 청미래덩굴잎의 발효과정을 통해, glycoside형태의 flavonoid를 aglycone화하여 비발효에 비해 높은 XO 및 AO 저해활성과 함께 항고요산혈증 효과를 나타내는 것으로 사료된다. To evaluate the inhibitory effect of xanthine oxidase (XO) and aldehyde oxidase (AO), and antihyperuricemic effect by Aspergillus oryzae fermented Smilax china L. leaf extracts and fractions, we observed extracted yield by each solvent, the content of total polyphenol and total flavonoid (TF), the activities of XO and AO, and serum uric acid level. Extracted yield (g/kg) by 80% ethanol (EtOH) was 13.56, those of n-hexane, dichloromethane (DICM), ethylacetate (EtOAc) and n-butanol fraction (BuOH) were 1.35.3.33. Furthermore, total polyphenol content (mg/gextract) of EtOAc fraction, BuOH fraction, DICM fraction and EtOH fraction is 478.07.501.26, 259.49.289.02, 165.03.232.27, 134.02.196.54, respectively. Those of fermented EtOAc and DICM fraction was 4.85 and 40.74% higher than that of nonfermented fraction, respectively, while the other fermented fractions were lower than those of non-fermented fractions. And total flavonoid content (mg/g-extract) of EtOAc fraction was higher than those of other fractions. Additionally, TF of fermented EtOAc and BuOH fraction is 10.56 and 60.17% higher, than that of fermented fraction, respectively, although those of the other fermented fractions was lower than that of non-fermented fractions. On the other hand, XO inhibitory activities of all fermented fractions was significantly higher than that of all non-fermented fraction, while those of fermented EtOAc (75.02%) and BuOH fraction (65.59%) was markedly higher than that of non-fermented fraction (39.42 and 5.34%), respectively. In addition, AO inhibitory activities of DICM and EtOAc fraction was 81.82 and 77.93% higher, respectively, than those of the other fractions, and those of fermented fractions as with XO were significantly higher than that of non-fermented fractions. Meanwhile, serum uric acid level (SU) of hyperuricemic control mice (HC, 6.98 mg/dL) was 1.83 folds higher than that of normal control (NC, 3.82 mg/dL). Furthermore, SU in the group treated with EtOAc fraction decreased in a dose dependent manner compared with the allopurinol control group, although those of fermented fractions were significantly lower than those of non-fermented fractions. This study suggests that fermented Smilax china L. leaf extracts may regulate the XO and AO inhibitory activities and antihyperuricemic effect due to aglycone components from glycoside form flavonoids by fermentation of A. oryzae.