Mucilaginibacter composti sp. nov., with Ginsenoside Converting Activity, Isolated from Compost

CUICHANGHAO,최태은,Hongshan Yu,Fengxie Jin,이성택,김선창,임완택 한국미생물학회 2011 The journal of microbiology Vol.49 No.3

The Gram-negative, strictly aerobic, non-motile, non-spore-forming, rod shaped bacterial strain designated TR6-03^T was isolated from compost, and its taxonomic position was investigated by using a polyphasic approach. Strain TR6-03^T grew at 4-42°C and at pH 6.0-8.0 on R2A and nutrient agar without NaCl supplement. Strain TR6-03^T had β-glucosidase activity, which was responsible for its ability to transform ginsenoside Re (one of the dominant active components of ginseng) to Rg_2. On the basis of 16S rRNA gene sequence similarity, strain TR6-03^T was shown to belong to the family Sphingobacteriaceae and to be related to Mucilaginibacter lappiensis ANJLI2^T (96.3% sequence similarity), M. dorajii FR-f4^T (96.1%),and M. rigui WPCB133^T (94.1%). The G+C content of the genomic DNA was 45.6%. The predominant respiratory quinone was MK-7 and the major fatty acids were summed feature 3 (comprising C_(16:1) ω7c and/or iso-C_(15:0) 2OH), iso-C_(15:0) and iso-C_(17:0) 3OH. DNA and chemotaxonomic data supported the affiliation of strain TR6-03^T to the genus Mucilaginibacter. Strain TR6-03^T could be differentiated genotypically and phenotypically from the recognized species of the genus Mucilaginibacter. The isolate therefore represents a novel species,for which the name Mucilaginibacter composti sp. nov. is proposed, with the type strain TR6-03^T (=KACC 14956^T =KCTC 12642^T =LMG 23497^T).

Production of ginsenoside F1 using commercial enzyme Cellulase KN

Yu Wang,Kang-Duk Choi,Hongshan Yu,Fengxie Jin,임완택 고려인삼학회 2016 Journal of Ginseng Research Vol.40 No.2

Background: Ginsenoside F1, a pharmaceutical component of ginseng, is known to have antiaging, antioxidant, anticancer, and keratinocyte protective effects. However, the usage of ginsenoside F1 is restricted owing to the small amount found in Korean ginseng. Methods: To enhance the production of ginsenoside F1 as a 10 g unit with high specificity, yield, and purity, an enzymatic bioconversion method was developed to adopt the commercial enzyme Cellulase KN from Aspergillus niger with food grade, which has ginsenoside-transforming ability. The proposed optimum reaction conditions of Cellulase KN were pH 5.0 and 50C. Results: Cellulase KN could effectively transform the ginsenosides Re and Rg1 into F1. A scaled-up biotransformation reaction was performed in a 10 L jar fermenter at pH 5.0 and 50C for 48 h with protopanaxatriol-type ginsenoside mixture (at a concentration of 10 mg/mL) from ginseng roots. Finally, 13.0 g of F1 was produced from 50 g of protopanaxatriol-type ginsenoside mixture with 91.5 1.1% chromatographic purity. Conclusion: The results suggest that this enzymatic method could be exploited usefully for the preparation of ginsenoside F1 to be used in cosmetic, functional food, and pharmaceutical industries.

Purification and Characterization of $Ginsenoside-{\beta}-Glucosidase$

Yu Hongshan,Ma Xiaoqun,Guo Yong,Jin Fengxie The Korean Society of Ginseng 1999 Journal of Ginseng Research Vol.23 No.1

In this paper, the saponin enzymatic hydrolysis of ginsenoside Rg3 was studied. The $ginsenoside-{\beta}-glucosidase$ from FFCDL-48 strain mainly hydrolyzed the ginsenoside Rg3 to Rh2, the enzyme from FFCDL-00 strain hydrolyzed Rg3 to the mixture of Rh2 and protopanaxadiol (aglycon). The $ginsenoside-{\beta}-glucosidase$ from FFCDL-48 strain was purified with a column of DEAE-Cellulose to one spot in the SDS polyacrylamide gel electrophoresis. During the purification, the enzyme specific acitvity was increased about 10 times. The purified $ginsenoside-{\beta}-glucosidase$ can hydrolyze the Rg3 to Rh2, but do not hydrolyze the $p-nitrophenyl-{\beta}-glucoside$ which is a substrate of original exocellulase such as ${\beta}-glucosidase$ of cellulose. The molecular weight of $ginsenoside-{\beta}-glucosidase$ was 34,000, the optimal temperature of enzyme reaction was $50^{\circ}C,$ and the optimal pH was 5.0.

Mucilaginibacter pocheonensis sp. nov., with ginsenoside-converting activity, isolated from soil of a ginseng-cultivating field

Zhao, Yan,Lee, Hyung-Gwan,Kim, Soo-Ki,Yu, Hongshan,Jin, Fengxie,Im, Wan-Taek Microbiology Society 2016 International journal of systematic and evolutiona Vol.66 No.-

Production of ginsenoside F1 using commercial enzyme Cellulase KN

Wang, Yu,Choi, Kang-Duk,Yu, Hongshan,Jin, Fengxie,Im, Wan-Taek The Korean Society of Ginseng 2016 Journal of Ginseng Research Vol.40 No.2

Background: Ginsenoside F1, a pharmaceutical component of ginseng, is known to have antiaging, antioxidant, anticancer, and keratinocyte protective effects. However, the usage of ginsenoside F1 is restricted owing to the small amount found in Korean ginseng. Methods: To enhance the production of ginsenoside F1 as a 10 g unit with high specificity, yield, and purity, an enzymatic bioconversion method was developed to adopt the commercial enzyme Cellulase KN from Aspergillus niger with food grade, which has ginsenoside-transforming ability. The proposed optimum reaction conditions of Cellulase KN were pH 5.0 and $50^{\circ}C$. Results: Cellulase KN could effectively transform the ginsenosides Re and Rg1 into F1. A scaled-up biotransformation reaction was performed in a 10 L jar fermenter at pH 5.0 and $50^{\circ}C$ for 48 h with protopanaxatriol-type ginsenoside mixture (at a concentration of 10 mg/mL) from ginseng roots. Finally, 13.0 g of F1 was produced from 50 g of protopanaxatriol-type ginsenoside mixture with $91.5{\pm}1.1%$ chromatographic purity. Conclusion: The results suggest that this enzymatic method could be exploited usefully for the preparation of ginsenoside F1 to be used in cosmetic, functional food, and pharmaceutical industries.

Identification and Characterization of a Novel Terrabacter ginsenosidimutans sp. nov. β-Glucosidase That Transforms Ginsenoside Rb1 into the Rare Gypenosides XVII and LXXV

An, Dong-Shan,Cui, Chang-Hao,Lee, Hyung-Gwan,Wang, Liang,Kim, Sun Chang,Lee, Sung-Taik,Jin, Fengxie,Yu, Hongshan,Chin, Young-Won,Lee, Hyeong-Kyu,Im, Wan-Taek,Kim, Song-Gun American Society for Microbiology 2010 Applied and environmental microbiology Vol.76 No.17

<B>ABSTRACT</B><P>A new β-glucosidase from a novel strain of <I>Terrabacter ginsenosidimutans</I> (Gsoil 3082<SUP>T</SUP>) obtained from the soil of a ginseng farm was characterized, and the gene, <I>bgpA</I> (1,947 bp), was cloned in <I>Escherichia coli</I>. The enzyme catalyzed the conversion of ginsenoside Rb1 {3-<I>O</I>-[β-d-glucopyranosyl-(1-2)-β-d-glucopyranosyl]-20-<I>O</I>-[β-d-glucopyranosyl-(1-6)-β-d-glucopyranosyl]-20(<I>S</I>)-protopanaxadiol} to the more pharmacologically active rare ginsenosides gypenoside XVII {3-<I>O</I>-β-d-glucopyranosyl-20-<I>O</I>-[β-d-glucopyranosyl-(1-6)-β-d-glucopyranosyl]-20(<I>S</I>)-protopanaxadiol}, gypenoside LXXV {20-<I>O</I>-[β-d-glucopyranosyl-(1-6)-β-d-glucopyranosyl]-20(<I>S</I>)-protopanaxadiol}, and C-K [20-<I>O</I>-(β-d-glucopyranosyl)-20(<I>S</I>)-protopanaxadiol]. A BLAST search of the <I>bgpA</I> sequence revealed significant homology to family 3 glycoside hydrolases. Expressed in <I>E. coli</I>, β-glucosidase had apparent <I>Km</I> values of 4.2 ± 0.8 and 0.14 ± 0.05 mM and <I>V</I>max values of 100.6 ± 17.1 and 329 ± 31 μmol·min<SUP>−1</SUP>·mg of protein<SUP>−1</SUP> against <I>p</I>-nitrophenyl-β-d-glucopyranoside and Rb1, respectively. The enzyme catalyzed the hydrolysis of the two glucose moieties attached to the C-3 position of ginsenoside Rb1, and the outer glucose attached to the C-20 position at pH 7.0 and 37°C. These cleavages occurred in a defined order, with the outer glucose of C-3 cleaved first, followed by the inner glucose of C-3, and finally the outer glucose of C-20. These results indicated that BgpA selectively and sequentially converts ginsenoside Rb1 to the rare ginsenosides gypenoside XVII, gypenoside LXXV, and then C-K. Herein is the first report of the cloning and characterization of a novel ginsenoside-transforming β-glucosidase of the glycoside hydrolase family 3.</P>

Icaritin Preparation from Icariin by a Special Epimedium Flavonoid-Glycosidase from Aspergillus sp.y848 Strain

( Zhenghao Wang ),( Chunying Liu ),( Hongshan Yu ),( Bo Wu ),( Baoyu Huai ),( Ziyu Zhuang ),( Changkai Sun ),( Longquan Xu ),( Fengxie Jin ) 한국미생물생명공학회 2022 Journal of microbiology and biotechnology Vol.32 No.4

In this study, to obtain icaritin with high pharmacological activities from icariin, which has a content ratio of over 58% in the total flavonoids of Epimedium herb, a special Epimedium flavonoid-glycosidase was produced, purified and characterized from Aspergillus sp.y848 strain. The optimal enzyme production was gained in a medium containing 5% (w/v) wheat bran extract and 0.7% (w/v) Epimedium leaf powder as the enzyme inducer, and strain culture at 30℃ for 6-7 days. The molecular weight of the enzyme was approximately 73.2 kDa; the optimal pH and temperature were 5.0 and 40°C. The enzyme Km and Vmax values for icariin were 15.63 mM and 55.56 mM/h. Moreover, the enzyme hydrolyzed the 7-O-glucosides of icariin into icariside II, and finally hydrolyzed 3-Orhamnoside of icariside II into icaritin. The enzyme also hydrolyzed 7-O-glucosides of epimedin B to sagittatoside B, and then further hydrolyzed terminal 3-O-xyloside of sagittatoside B to icarisiede II, before finally hydrolyzing 3-O-rhamnoside of icarisiede II into icaritin. The enzyme only hydrolyzed 7-O-glucoside of epimedin A or epimedin C into sagittatoside A or sagittatoside C. It is possible to prepare icaritin from the high-content icariin in Epimedium herb using this enzyme. When 2.5% icariin was reacted at 40℃ for 18-20 h by the low-cost crude enzyme, 5.04 g icaritin with 98% purity was obtained from 10 g icariin. Also, the icaritin molar yield was 92.5%. Our results showed icaritin was successfully produced via cost-effective and relatively simple methods from icariin by crude enzyme. Our results should be very useful for the development of medicines from Epimedium herb.

Dynamic changes of multi-notoginseng stem-leaf ginsenosides in reaction with ginsenosidase type-I

Xiao, Yongkun,Liu, Chunying,Im, Wan-Teak,Chen, Shuang,Zuo, Kangze,Yu, Hongshan,Song, Jianguo,Xu, Longquan,Yi, Tea-Hoo,Jin, Fengxie The Korean Society of Ginseng 2019 Journal of Ginseng Research Vol.43 No.2

Background: Notoginseng stem-leaf (NGL) ginsenosides have not been well used. To improve their utilization, the biotransformation of NGL ginsenosides was studied using ginsenosidase type-I from Aspergillus niger g.848. Methods: NGL ginsenosides were reacted with a crude enzyme in the RAT-5D bioreactor, and the dynamic changes of multi-ginsenosides of NGL were recognized by HPLC. The reaction products were separated using a silica gel column and identified by HPLC and NMR. Results: All the NGL ginsenosides are protopanaxadiol-type ginsenosides; the main ginsenoside contents are 27.1% Rb3, 15.7% C-Mx1, 13.8% Rc, 11.1% Fc, 7.10% Fa, 6.44% C-Mc, 5.08% Rb2, and 4.31% Rb1. In the reaction of NGL ginsenosides with crude enzyme, the main reaction of Rb3 and C-Mx1 occurred through Rb3${\rightarrow}$C-Mx1${\rightarrow}$C-Mx; when reacted for 1 h, Rb3 decreased from 27.1% to 9.82 %, C-Mx1 increased from 15.5% to 32.3%, C-Mx was produced to 6.46%, finally into C-Mx and a small amount of C-K. When reacted for 1.5 h, all the Rb1, Rd, and Gyp17 were completely reacted, and the reaction intermediate F2 was produced to 8.25%, finally into C-K. The main reaction of Rc (13.8%) occurred through Rc${\rightarrow}$C-Mc1${\rightarrow}$C-Mc${\rightarrow}$C-K. The enzyme barely hydrolyzed the terminal xyloside on 3-O- or 20-O-sugar-moiety of the substrate; therefore, 9.43 g C-Mx, 6.85 g C-K, 4.50 g R7, and 4.71 g Fc (hardly separating from the substrate) were obtained from 50 g NGL ginsenosides by the crude enzyme reaction. Conclusion: Four monomer ginsenosides were successfully produced and separated from NGL ginsenosides by the enzyme reaction.

An Efficient and Robust Data Integrity Verification Algorithm Based on Context Sensitive

Feng Xie,Hu Chen 보안공학연구지원센터 2016 International Journal of Security and Its Applicat Vol.10 No.4

There exist two key problems about data aggregation that should be thoroughly explored - algorithm design in networking layer, and algorithm design in application layer. Those two problems should be subtlety tackled in termers of high efficiency and robustness. Therefore, the former one requires the survivability and highly reliable design at networking layer, the latter one usually asks for high efficiency and robustness at application layer. Moreover, the optimization of algorithms is also considered for further enhancement. The integrity check is a key requirement for optimization. The context-aware and cross-layer design is applied in the optimization. A dynamic fragment odd-even parity checking code is proposed, and a context-aware aggregative integrity check code is proposed.

Arabidopsis MiR396 Mediates the Development of Leaves and Flowers in Transgenic Tobacco

Fengxi Yang,Gang Liang,Dongmei Liu,Diqiu Yu 한국식물학회 2009 Journal of Plant Biology Vol.52 No.5

MicroRNAs (miRNAs) are single-stranded, noncoding small RNAs that usually function as posttranscriptional negative regulators by base pairing to target genes. They are pivotal to plant development. MiR396 is conserved among plant species and is predicted to target GRF (growth-regulating factor) genes in Arabidopsis. Here, overexpression of ath-miR396 in tobacco reduced the levels of three NtGRF-like genes containing an miR396 match site. Furthermore, its elevated expression resulted in a small, narrow leaf phenotype similar to that found with the Arabidopsis grf1grf2grf3 triple mutant. We also demonstrated that 35S:MIR396a transgenic plants were defective in the four whorls of floral organs. These results provide a link between the miR396- mediated regulatory pathway of NtGRF-like gene expression and the developmental processes for leaves and flowers in tobacco.