Preparation of minor ginsenosides C-Mc, C-Y, F2, and C-K from American ginseng PPD-ginsenoside using special ginsenosidase type-I from Aspergillus niger g.848

Chun-Ying Liu,Rui-Xin Zhou,Chang-Kai Sun,Ying-Hua Jin,Hong-Shan Yu,Tian-Yang Zhang,Long-Quan Xu,Feng-Xie Jin 고려인삼학회 2015 Journal of Ginseng Research Vol.39 No.3

Background: Minor ginsenosides, those having low content in ginseng, have higher pharmacological activities. To obtain minor ginsenosides, the biotransformation of American ginseng protopanaxadiol (PPD)-ginsenoside was studied using special ginsenosidase type-I from Aspergillus niger g.848. Methods: DEAE (diethylaminoethyl)-cellulose and polyacrylamide gel electrophoresis were used in enzyme purification, thin-layer chromatography and high performance liquid chromatography (HPLC) were used in enzyme hydrolysis and kinetics; crude enzyme was used in minor ginsenoside preparation from PPD-ginsenoside; the products were separated with silica-gel-column, and recognized by HPLC and NMR (Nuclear Magnetic Resonance). Results: The enzyme molecular weight was 75 kDa; the enzyme firstly hydrolyzed the C-20 position 20- O-b-D-Glc of ginsenoside Rb1, then the C-3 position 3-O-b-D-Glc with the pathway Rb1/Rd/F2/C-K. However, the enzyme firstly hydrolyzed C-3 position 3-O-b-D-Glc of ginsenoside Rb2 and Rc, finally hydrolyzed 20-O-L-Ara with the pathway Rb2/C-O/C-Y/C-K, and Rc/C-Mc1/C-Mc/C-K. According to enzyme kinetics, Km and Vmax of MichaeliseMenten equation, the enzyme reaction velocities on ginsenosides were Rb1 > Rb2 > Rc > Rd. However, the pure enzyme yield was only 3.1%, so crude enzyme was used for minor ginsenoside preparation. When the crude enzyme was reacted in 3% American ginseng PPD-ginsenoside (containing Rb1, Rb2, Rc, and Rd) at 45C and pH 5.0 for 18 h, the main products were minor ginsenosides C-Mc, C-Y, F2, and C-K; average molar yields were 43.7% for CMc from Rc, 42.4% for C-Y from Rb2, and 69.5% for F2 and C-K from Rb1 and Rd. Conclusion: Four monomer minor ginsenosides were successfully produced (at low-cost) from the PPDginsenosides using crude enzyme.

Kinetics of a Cloned Special Ginsenosidase Hydrolyzing 3-O-Glucoside of Multi-Protopanaxadiol-Type Ginsenosides, Named Ginsenosidase Type 3

( Xue Feng Jin ),( Hong Shan Yu ),( Dong Ming Wang ),( Ting Qiang Liu ),( Chun Ying Liu ),( Dong Shan An ),( Wan Taek Im ),( Song Gun Kim ),( Feng Xie Jin ) 한국미생물 · 생명공학회 2012 Journal of microbiology and biotechnology Vol.22 No.3

In this paper, the kinetics of a cloned special glucosidase, named ginsenosidase type III hydrolyzing 3-O-glucoside of multi-protopanaxadiol (PPD)-type ginsenosides, were investigated. The gene (bgpA) encoding this enzyme was cloned from a Terrabacter ginsenosidimutans strain and then expressed in E. coli cells. Ginsenosidase type III was able to hydrolyze 3-O-glucoside of multi-PPD-type ginsenosides. For instance, it was able to hydrolyze the 3- O-β-D-(1→2)-glucopyranosyl of Rb1 to gypenoside XVII, and then to further hydrolyze the 3-O-β-D-glucopyranosyl of gypenoside XVII to gypenoside LXXV. Similarly, the enzyme could hydrolyze the glucopyranosyls linked to the 3-O- position of Rb2, Rc, Rd, Rb3, and Rg3. With a larger enzyme reaction Km value, there was a slower enzyme reaction speed; and the larger the enzyme reaction Vmax value, the faster the enzyme reaction speed was. The Km values from small to large were 3.85 mM for Rc, 4.08 mM for Rb1, 8.85 mM for Rb3, 9.09 mM for Rb2, 9.70 mM for Rg3(S), 11.4 mM for Rd and 12.9 mM for F2; and Vmax value from large to small was 23.2 mM/h for Rc, 16.6 mM/h for Rb1, 14.6 mM/h for Rb3, 14.3 mM/h for Rb2, 1.81mM/h for Rg3(S), 1.40 mM/h for Rd, and 0.41 mM/h for F2. According to the Vmax and Km values of the ginsenosidase type III, the hydrolysis speed of these substrates by the enzyme was Rc>Rb1>Rb3>Rb2>Rg3(S)>Rd>F2 in order.

Preparation of minor ginsenosides C-Mc, C-Y, F2, and C-K from American ginseng PPD-ginsenoside using special ginsenosidase type-I from Aspergillus niger g.848

Liu, Chun-Ying,Zhou, Rui-Xin,Sun, Chang-Kai,Jin, Ying-Hua,Yu, Hong-Shan,Zhang, Tian-Yang,Xu, Long-Quan,Jin, Feng-Xie The Korean Society of Ginseng 2015 Journal of Ginseng Research Vol.39 No.3

Background: Minor ginsenosides, those having low content in ginseng, have higher pharmacological activities. To obtain minor ginsenosides, the biotransformation of American ginseng protopanaxadiol (PPD)-ginsenoside was studied using special ginsenosidase type-I from Aspergillus niger g.848. Methods: DEAE (diethylaminoethyl)-cellulose and polyacrylamide gel electrophoresis were used in enzyme purification, thin-layer chromatography and high performance liquid chromatography (HPLC) were used in enzyme hydrolysis and kinetics; crude enzyme was used in minor ginsenoside preparation from PPD-ginsenoside; the products were separated with silica-gel-column, and recognized by HPLC and NMR (Nuclear Magnetic Resonance). Results: The enzyme molecular weight was 75 kDa; the enzyme firstly hydrolyzed the C-20 position 20-O-${\beta}$-D-Glc of ginsenoside Rb1, then the C-3 position 3-O-${\beta}$-D-Glc with the pathway $Rb1{\rightarrow}Rd{\rightarrow}F2{\rightarrow}C-K$. However, the enzyme firstly hydrolyzed C-3 position 3-O-${\beta}$-D-Glc of ginsenoside Rb2 and Rc, finally hydrolyzed 20-O-L-Ara with the pathway $Rb2{\rightarrow}C-O{\rightarrow}C-Y{\rightarrow}C-K$, and $Rc{\rightarrow}C-Mc1{\rightarrow}C-Mc{\rightarrow}C-K$. According to enzyme kinetics, $K_m$ and $V_{max}$ of Michaelis-Menten equation, the enzyme reaction velocities on ginsenosides were Rb1 > Rb2 > Rc > Rd. However, the pure enzyme yield was only 3.1%, so crude enzyme was used for minor ginsenoside preparation. When the crude enzyme was reacted in 3% American ginseng PPD-ginsenoside (containing Rb1, Rb2, Rc, and Rd) at $45^{\circ}C$ and pH 5.0 for 18 h, the main products were minor ginsenosides C-Mc, C-Y, F2, and C-K; average molar yields were 43.7% for C-Mc from Rc, 42.4% for C-Y from Rb2, and 69.5% for F2 and C-K from Rb1 and Rd. Conclusion: Four monomer minor ginsenosides were successfully produced (at low-cost) from the PPD-ginsenosides using crude enzyme.

Life Science : Discrimination of Phellodendron amurense and P chinense Based on DNA Analysis and the Simultaneous Analysis of Alkaloids

( Jin Ah Ryuk ),( Ming Shan Zheng ),( Mi Young Lee ),( Chang Seob Seo ),( Ying Li ),( Seung Ho Lee ),( Dong Cheul Moon ),( Hye Won Lee ),( Je Hyun Lee ),( Ju Young Park ),( Jong Keun Son ),( Byung Seo 영남대학교 약품개발연구소 2012 영남대학교 약품개발연구소 연구업적집 Vol.22 No.0

Phellodendri Cortex is the bark of the stems of Phellodendron amurense Ruprecht or P. chinense Schneider (Rutaceae), which is orginated from periderm. The internal transcribed spacer sequences of 20 originated plants and identified samples were analyzed. The result showed that the 99% of the base sequences of P. amurense were identical to that of P. chinense, but the differentiation of P. amurenseand P. chinense was difficult. In addition, the ribulose-1, 5-bisphospate carboxylase large subunit (rbcL) intergenic spacer sequences of specific parts produced the same result. However, when the analysis was carried out by using the RAPD (randomly amplification polymorphism DNA) analysis method, which utilizes 48 randomly primers, it allowed us to confirm the polymorphism of P. amurenseand P. chinense in 12 primers. A high-performance liquid chromatographic (HPLC) method was developed and validated for the simultaneous quantitation of berberine, palmatine and jatrorrhizine in a traditional herbal drug, Phellodendri Cortex. The HPLC method was applied successfully to the quantification of three constituents in the extract of twenty Phellodendri Cortex. The results indicated that the established HPLC and RAPD methods are suitable for the quantitative analysis and the quality control multi-simultaneous discrimination in Phellodendri Cortex.

Discrimination of Phellodendron amurense and P. chinense Based on DNA Analysis and the Simultaneous Analysis of Alkaloids

Jin Ah Ryuk,고병섭,Ming Shan Zheng,Mi Young Lee,서창섭,Ying Li,이승호,문동철,Hye Won Lee,이제현,Ju Young Park,손종근 대한약학회 2012 Archives of Pharmacal Research Vol.35 No.6

Phellodendri Cortex is the bark of the stems of Phellodendron amurense Ruprecht or P. chinense Schneider (Rutaceae), which is orginated from periderm. The internal transcribed spacer sequences of 20 originated plants and identified samples were analyzed. The result showed that the 99% of the base sequences of P. amurense were identical to that of P. chinense, but the differentiation of P. amurense and P. chinense was difficult. In addition, the ribulose-1, 5-bisphospate carboxylase large subunit (rbcL) intergenic spacer sequences of specific parts produced the same result. However, when the analysis was carried out by using the RAPD (randomly amplification polymorphism DNA) analysis method, which utilizes 48 randomly primers, it allowed us to confirm the polymorphism of P. amurense and P. chinense in 12 primers. A high-performance liquid chromatographic (HPLC) method was developed and validated for the simultaneous quantitation of berberine, palmatine and jatrorrhizine in a traditional herbal drug, Phellodendri Cortex. The HPLC method was applied successfully to the quantification of three constituents in the extract of twenty Phellodendri Cortex. The results indicated that the established HPLC and RAPD methods are suitable for the quantitative analysis and the quality control multi-simultaneous discrimination in Phellodendri Cortex.

Baicalin attenuates TNBS-induced colitis in rats by modulating the Th17/Treg paradigm

Ying Zou,Shi-Xue Dai,Hong-Gang Chi,Tao Li,Zhi-Wei He,Jian Wang,Cai-Guo Ye,Guo-Liang Huang,Bing Zhao,Wen-Yang Li,Zheng Wan,Jin-Shan Feng,Xue-Bao Zheng 대한약학회 2015 Archives of Pharmacal Research Vol.38 No.10

Baicalin, a flavonoid, has a wide range ofpharmacological properties, including immunomodulation. The objective of this study was to investigate the effect ofbaicalin on the balance of T helper 17 (Th17) and regulatoryT (Treg) cells in a colitis model. The rat colitis modelwas induced by 2,4,6-trinitrobenzene sulfonic acid(TNBS). Baicalin (10 ml/kg, each) or mesalazine (positivecontrol) was then administered orally for 7 days. Inflammatoryand immunological responses were evaluated bypathology, enzyme-linked immunosorbent assay, real-timepolymerase chain reaction, western blot analysis, and flowcytometry. Our study showed that baicalin not only significantlyattenuated TNBS-induced colitis by reducing thedisease activity index as well as macroscopic and microscopicscores, but it also improved the weight loss andshortening of the colon. Baicalin treatment also induced asignificant decrease in the levels of inflammatory mediators,including the myeloperoxidase activity, the levels oftumor necrosis factor a, IL-1b, and Th1-related cytokinesIL-12 and IFN-c. Furthermore, the beneficial effects ofbaicalin seem to be associated with regulation of the Th17and Treg paradigm. We found that administration ofbaicalin significantly downregulated the number of Th17cells and the levels of Th17-related cytokines (IL-17 andIL-6) and retinoic acid receptor-related orphan receptor ct. In contrast, there was an increase in Treg cells numbers,Treg-related cytokines transforming growth factor-b andIL-10, and forkhead box P3. Our results suggest that theanti-inflammatory effect of baicalin may be linked tomodulation of the balance between Th17 and Treg cells inTNBS-induced ulcerative colitis.

Discrimination of Phellodendron amurense and P. chinense Based on DNA Analysis and the Simultaneous Analysis of Alkaloids

Ryuk, Jin Ah,Zheng, Ming Shan,Lee, Mi Young,Seo, Chang Seob,Li, Ying,Lee, Seung Ho,Moon, Dong Cheul,Lee, Hye Won,Lee, Je-Hyun,Park, Ju Young,Son, Jong Keun,Ko, Byoung Seob 대한약학회 2012 Archives of Pharmacal Research Vol.35 No.6

Phellodendri Cortex is the bark of the stems of Phellodendron amurense Ruprecht or P. chinense Schneider (Rutaceae), which is orginated from periderm. The internal transcribed spacer sequences of 20 originated plants and identified samples were analyzed. The result showed that the 99% of the base sequences of P. amurense were identical to that of P. chinense, but the differentiation of P. amurense and P. chinense was difficult. In addition, the ribulose-1, 5-bisphospate carboxylase large subunit (rbcL) intergenic spacer sequences of specific parts produced the same result. However, when the analysis was carried out by using the RAPD (randomly amplification polymorphism DNA) analysis method, which utilizes 48 randomly primers, it allowed us to confirm the polymorphism of P. amurense and P. chinense in 12 primers. A high-performance liquid chromatographic (HPLC) method was developed and validated for the simultaneous quantitation of berberine, palmatine and jatrorrhizine in a traditional herbal drug, Phellodendri Cortex. The HPLC method was applied successfully to the quantification of three constituents in the extract of twenty Phellodendri Cortex. The results indicated that the established HPLC and RAPD methods are suitable for the quantitative analysis and the quality control multi-simultaneous discrimination in Phellodendri Cortex.

Inhibitory effects of piceatannol on human cytomegalovirus (hCMV) in vitro

Wang San-Ying,Zhang Jing,Xu Xiao-Gang,Su Hui-Li,Xing Wen-Min,Zhang Zhong-Shan,Jin Wei-Hua,Dai Ji-Huan,Wang Ya-Zhen,He Xin-Yue,Sun Chuan,Yan Jing,Mao Gen-Xiang 한국미생물학회 2020 The journal of microbiology Vol.58 No.8

Human cytomegalovirus (hCMV) is a ubiquitous herpesvirus, which results in the establishment of a latent infection that persists throughout the life of the host and can be reactivated when the immunity is low. Currently, there is no vaccine for hCMV infection, and the licensed antiviral drugs mainly target the viral enzymes and have obvious adverse reactions. Thus, it is important to search for compounds with antihCMV properties. The present study aimed to investigate the suppressive effects of piceatannol on hCMV Towne strain infection and the putative underlying mechanisms using human diploid fibroblast WI-38 cells. Piceatannol supplementation prevented the lytic changes induced by hCMV infection in WI-38 cells. Furthermore, piceatannol suppressed the expression of hCMV immediate-early (IE) and early (E) proteins as well as the replication of hCMV DNA in a dose-dependent manner. Moreover, hCMV-induced cellular senescence was suppressed by piceatannol, as shown by a decline in the senescence-associated β-galactosidase (SA-β-Gal) activity and decreased production of intracellular reactive oxygen species (ROS). p16INK4a, a major senescence-associated molecule, was dramatically elevated by current hCMV infection that was attenuated by pre-incubation with piceatannol in a dose-dependent manner. These results demonstrated that piceatannol suppressed the hCMV infection via inhibition of the activation of p16INK4a and cellular senescence induced by hCMV. Together, these findings indicate piceatannol as a novel and potent anti-hCMV agent with the potential to be developed as an effective treatment for chronic hCMV infection.

온라인 협업 도구 활용 한국어 수업 사례 : 상해 삼달대학교를 중심으로

배영련(Pei, Ying Lian),오선자(Wu, Shan Zi),최선진(Choi, Seon Jin),한나(Han, Na),오영화(Wu, Ying Hua) 글로벌교육연구학회 2021 글로벌교육연구 Vol.13 No.4

본 연구는 온라인 수업에서의 상호작용에 도움을 주며, 중국과 한국에서 동시에 사용할수 있는 다양한 온라인 협업 도구를 소개하고, 이 도구 네 가지(mentimeter, zoom, padlet, Quizizz)를 사용하여 중국인 대상 한국어 수업을 운영한 사례를 제시하였다. 그리고본 수업 운영에 대한 학습자 만족도를 살펴보기 위하여 2021학년도 3월 학기에 본 수업을수강한 상해 삼달대학교 한국어 학습자 75명을 대상으로 만족도 조사를 시행하였으며, 그결과 전체적인 수업 만족도는 평균 4.09였다. 또한, 온라인 수업에서의 상호작용 향상을위한 방법으로 활용한 온라인 협업 도구에 대한 전체적인 만족도는 4.24였으며, 각 도구의만족도는 mentimeter가 4.63, zoom이 4.15, padlet이 4.09, Quizizz 3.81 순이었다. 이 중에서 협업 도구에 대한 전체적인 만족도와 padlet 만족도가 수업 전체 만족도에 영향을미치는 것으로 분석되었다. 본 연구 결과에 따른 제언은 다음과 같다. 첫째, 온라인 협업도구 활용에 대한 학습자 만족도가 높기 때문에 한국어 수업에 활용할 수 있는 다양한 협업도구 연구를 바탕으로 수업 적용이 필요하며, 각 국 학습자용 교재 편찬이 필요하다고 할수 있다. 본 연구는 다양한 온라인 수업에서의 상호작용에 필요한 다양한 협업 도구를 소개하고 비대면 수업에서의 실제 활용방안을 제시했으며, 이에 대한 학습자들의 의견을 수렴했다는 점에서 의의가 있다. This study was to present a case of an Online Korean class using four online tools such as Mentimeter, Zoom, Padlet, Quizizz for helping the interaction of Korean and Chinese students. To investigate learner satisfaction, 75 Chinese students from Shanda University participated in a survey after the class. The results were as follows: First, the overall class satisfaction was 4.09, especially students were satisfied with using collaboration tools. Second, online tool satisfaction was 4.63 for Mentimeter, 4.15 for Zoom, 4.09 for Padlet, and 3.81 for Quizizz in the order of the program. Based on the results, it was suggested applications of various online tools into class and textbooks for learners. At last, it was significant that the case of online Korean class was presented with many online tools and learners satisfaction in this study.

Vagococcus zengguangii sp. nov., isolated from yak faeces

Ge Yajun,Jin Dong,Lai Xin-He,Yang Jing,Lu Shan,Huang Ying,Zheng Han,Zhang Xiaoyan,Xu Jianguo 한국미생물학회 2021 The journal of microbiology Vol.59 No.1

Two unknown Gram-stain-positive, catalase- and oxidasenegative, non-motile, and coccus-shaped bacteria, designated MN-17T and MN-09, were isolated from yaks faeces (Bos grunniens) in the Qinghai-Tibet Plateau of China. 16S rRNA gene sequence-based comparative analyses revealed that the two strains were grouped within the genus Vagococcus, displaying the highest similarity with Vagococcus xieshaowenii CGMCC 1.16436T (98.6%) and Vagococcus elongatus CCUG 51432T (96.4%). Both strains grew optimally at 37°C and pH 7.0 in the presence of 0.5% (w/v) NaCl. The complete genome of MN-17T comprises 2,085 putative genes with a total of 2,190,262 bp and an average G + C content of 36.7 mol%. The major fatty acids were C16:0 (31.2%), C14:0 (28.5%), and C18:1ω9c (13.0%); the predominant respiratory quinone was MK-7 (68.8%); the peptidoglycan type was A4α(L-Lys-DAsp); and the major polar lipid was diphosphatidylglycerol. Together, these supported the affiliation of strain MN-17T to the genus Vagococcus. In silico DNA-DNA hybridization and the average nucleotide identity values between MN-17T and all recognized species in the genus were 21.6–26.1% and 70.7–83.0%, respectively. MN-17T produced acid from D-cellobiose, D-fructose, glycerol, D-glucose, N-acetyl-glucosamine, gentiobiose, D-mannose, D-maltose, D-ribose, Dsaccharose, salicin, D-trehalose, and D-xylose. These results distinguished MN-17T and MN-09 from closely related species in Vagococcus. Thus, we propose that strains MN-17T and MN-09 represent a novel species in the genus Vagococcus, with the name Vagococcus zengguangii sp. The type strain is MN-17T (= CGMCC 1.16726T = GDMCC 1.1589T = JCM 33478T).