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.

Pyramiding of 2 QTLs, gw8 and gw9, underlying grain weight using nearly isogenic lines (NILs) in rice

Feng-Xue Jin,Xiao-Bo Xie,Shi-Dong Ji,Hong-Guang Ju,Jung-Pil Suh,Hung-Goo Hwang,Sang-Nag Ahn 한국작물학회 2008 한국작물학회 학술발표대회 논문집 Vol.2008 No.10

Grain weight (GW) or grain size is the most important target not only as a major component of grain yield, but also associated with the cooking quality in rice breeding program. In a previous study, two loci for grain weight, gw8 and gw9, have been fine map to about 306.4kb and 37.4kb, respectively, in backcross populations derived from a cross between the Korean japonica cultivar Hwaseongbyeo and Oryza rufipigon (IRGC 105491). Both O.rufipogon alleles increased GW in the Hwaseongbyeo background despite the fact that O. rufipogon was the small-seeded parent. To test the effect of pyramiding the QTLs for grain weight, an F2 population consisted of 226 individuals was developed derived from the cross between the two QTLs (gw8 and gw9), and used MAS to select nearly isgenic lines for two QTLs and a pyramiding line. Molecular genotyping of F2 population revealed the existence of interaction between gw8 and gw9. Moreover, the interaction was also confirmed by phenotypic analysis of four QTL-NILs. The results suggest that epistasis interaction is important genetic basis in determining yield traits in rice.

Pyramiding of 2 QTLs, gw8 and gw9, underlying grain weight using nearly isogenic Lines (NILs) in rice

Feng Xue Jin,Xiao Bo Xie,Shi Dong Ji,Hong Guang Ju,Jung Pil Suh,Hung Goo Hwang,Sang Nag Ahn 한국육종학회 2008 한국육종학회 학술발표회 발표요지 Vol.40 No.1

Detection of epistatic interaction of two QTLs, <i>gw8.1</i> and <i>gw9.1</i>, underlying grain weight using nearly isogenic lines in rice

Jin, Feng-Xue,Ji, Shi-Dong,Xie, Xiao-Bo,Kang, Ju-Won,Ju, Hong-Guang,Ahn, Sang-Nag Japanese Society of Breeding 2011 Breeding science Vol.61 No.1

<P>Grain weight (GW) is one of the most important targets for grain yield in rice breeding. In previous studies, two quantitative trait loci (QTLs) for GW, <I>gw8.1</I> and <I>gw9.1</I>, have been identified using progeny derived from a cross between the <I>japonica</I> cultivar Hwaseong and <I>Oryza rufipogon </I>(IRGC 105491). To test whether these quantitative trait loci (QTLs) have an epistatic interaction, we developed an F<SUB>2</SUB> population by crossing two nearly isogenic lines (NILs) harboring <I>gw8.1</I> and <I>gw9.1</I>. Simple sequence repeat (SSR) markers tightly linked to the QTLs were used to select F<SUB>3</SUB> QTL-NILs from the F<SUB>2</SUB> population. A two-way ANOVA revealed an epistatic interaction between the two QTLs in the F<SUB>2</SUB> population (<I>P</I> = 0.0084). This interaction was confirmed by an analysis of F<SUB>3</SUB> QTL-NILs indicating that both QTLs are involved in the same genetic mechanism controlling GW. The <I>gw8.1</I> QTL was further mapped between two SSR markers, RM23204 and RM23211, which are 110.1 kb apart. To our knowledge, this is the first report using QTL-NILs to reveal an epistatic interaction between QTLs for GW.</P>

Low Doses of Nonylphenol Promote Growth of Colon Cancer Cells through Activation of ERK1/2 via G Protein–Coupled Receptor 30

Ming Xie,Jin-Long Liang,Han-Dong Huang,Mai-Jian Wang,Tao Zhang,Xue-Feng Yang 대한암학회 2019 Cancer Research and Treatment Vol.51 No.4

Purpose Nonylphenol (NP) is an endocrine disruptor found in products such as cleaners, plastics, and detergents. It exerts actions similar to endogenous 17-estradiol (E2) and is reported to influence various cancers. However, its role in colon cancer remains elusive. Materials and Methods Colon cancer cell lines COLO 205 and SW480 were employed in our study. The cells were treated with NP or E2 followed by measurement of apoptosis and proliferation using flow cytometry and MTT assays, respectively. G protein–coupled estrogen receptor 30 (GPR30) expression was visualized using immunofluorescence and Western blot. To investigate the underlying mechanism, the expression levels of GPR30, p-protein kinase A (PKA), c-myc, cyclin D1, and ERK1/2 were analyzed using Western blot. Meanwhile, the GPR30 antagonist G15 was utilized to validate the role of GPR30 in colon cancer progression. Finally, the effect of a GPR30 inhibitor on tumor growth was determined in vivo using tumor xenograft mouse models. Results NP facilitated the proliferation of colon cancer cells and induced apoptosis failure in vitro. Western blot revealed increased GPR30 expression levels in response to NP treatment. Cyclin D1, p-PKA, c-myc, and proliferating cell nuclear antigen, proteins that regulate the cell cycle, were all upregulated by NP, and NP-mediated ERK1/2 activation and subsequent cell proliferation were abrogated by the GPR30 inhibitor G15. Moreover, colon cancer mice that received G15 administration demonstrated impaired tumor growth in vivo. Conclusion Low dose NP promotes the growth of colon tumors through GPR30-mediated activation of ERK1/2 signaling.

Comparison of Gene Expression between Cumulus Oocyte Complexes and Naked Oocytes by Suppression Subtractive Hybridization in Swine

Xiang, Zhi Feng,Zhang, Jin Zhou,Li, Xue Bin,Xie, Hong Bin,Wang, Qing Hua Asian Australasian Association of Animal Productio 2010 Animal Bioscience Vol.23 No.1

In the antral follicle phase, several layers of cumulus cells surround the oocyte and play an important support and regulation role in oocyte development and maturation via intercellular communications and interactions between oocytes and cumulus cells. However, information on stage specific gene expression in swine during the phase is not well understood. To investigate the function of cumulus cells during in vitro maturation of porcine oocytes and gene expression, suppression subtractive hybridization (SSH) was performed to screen genes that were differentially expressed between cumulus-oocyte complexes (COCs) and naked oocytes (NOs). Utilizing mRNAs from in vitro maturation oocytes, a SSH cDNA library from COCs as the tester and NOs as the driver was constructed. The SSH cDNA library was then screened using dot blot analysis. Results showed that a total of 70 clones randomly selected from the library were differentially expressed. Among these, 41 exhibited high homology to known genes and 11 were novel expressed sequences tags (ESTs). Four differentially expressed genes, including bfgf, sprouty 2, egr and btc, were further studied by real time quantitative PCR; results confirmed an increased expression of respective mRNA in COCs compared with NOs, which suggests that these factors may play an important role in oocyte development and maturation.

Study on Strain Energy Transfer and Efficiency in Spatial Micro-forming of Metal

Zhaojie Chen,Jin Xie,Quanpeng He,Dongsheng Ge,Kuo Lu,Chaolun Feng 한국정밀공학회 2024 International Journal of Precision Engineering and Vol.11 No.2

In spatial micro-fabrication on metallic surface, the mechanical machining consumes material shear deformation energy, while the laser machining energy is greatly converted into material melting heat energy. In production, the micron-scale material-removal machining requires the CNC system to long-time tool path interpolation for high energy-consumption. According to dynamics and kinematics of metallic plastic deformation, a strain energy transfer is proposed to deform micro-topographic shapes by differentiated surface stress. The objective is to realize the precision forming of spatial microstructure surface through the strain energy conversion and conservation. First, the energy transfer and strain variations were modelled in relation to die curvature radius, workpiece thickness, initial microstructure angle and depth. Then, the strain energy consumption was investigated in relation to material properties, die movement, and micro dimensions. Finally, it was applied to industrial cold-pressing. It is shown that the strain energy of a single microstructure formation transfers from centre to outer part. The spatial microstructure forming may change from diversified strain stage to uniform strain state with the highest energy efficiency at a critical strain energy, while the surface roughness remains unchanged. Under the strain energy transfer, the microstructure shape changes with increasing energy consumption to a critical value. The metal compressive strength, die curvature radius and workpiece thickness promotes energy consumption, while descending velocity promotes processing efficiency. By controlling the energy conversion, the spatial microstructure sizes may be fabricated with an error of about 1.0% and the energy consumption of about 10 mm3/J. In industrial production, it contributes high energy efficiency without coolant pollutant in contrast to mechanical machining and laser machining. As a result, the strain energy conversion and conservation may be regarded as an evaluation for an eco-friendly micro-fabrication.

Preparation of Chitooligosaccharides from Chitosan using Crude Enzyme of Bacillus cereus D-11

( Xing Ai Gao ),( Yong Feng Zhang ),( Ro Dong Park ),( Xiao Huang ),( Xin Ying Zhao ),( Jiao Xie ),( Rong De Jin ) 한국응용생명화학회 2012 Journal of Applied Biological Chemistry (J. Appl. Vol.55 No.1

In order to enzymatically produce chitooligosaccharide using the crude enzyme preparation from Bacillus cereus D-11, we first studied the optimal reaction conditions. It was found that the optimal temperature for hydrolysis of chitosan was 55oC. The ratio of enzyme/substrate should not be lower than 0.13 U/mg in the reaction mixture. The enzyme activity was stable below 50oC. The products of enzymatic reaction were analyzed by both thin layer chromatography and high performance liquid chromatography. Under the appropriate condition, chitosan was hydrolyzed using the enzyme preparation. The resulting chitooligosaccharides were purified and separated by Dowex (H+) ion exchange chromatography. From 4 g soluble chitosan, 0.95 g (GlcN)2, 1.43 g (GlcN)3, and 1.18 g (GlcN)4 were recovered.

Cloning of the Correct Full Length cDNA of NF-kB-repressing Factor

Yumin Mao,Dai Jianfeng,Jin Feng,Ji Chaoneng,Zheng Zhongshou,Gu Shaohua,Wu Qihan,Wang Liu,Yin Gang,Xie Yi 한국분자세포생물학회 2003 Molecules and cells Vol.16 No.3

Chloroplast genomic comparison of two sister species Allium macranthum and A. fasciculatum provides valuable insights into adaptive evolution

Hao Li,Deng‑Feng Xie,Jun‑Pei Chen,Song‑Dong Zhou,Xing‑Jin He 한국유전학회 2020 Genes & Genomics Vol.42 No.5

Background Allium macranthum and Allium fasciculatum are two sister species and their natural populations are separated by high mountains and deep valleys with exact opposite habitat. The chloroplast genome in angiosperms has showed useful for investigating plant evolution and systematic studies. Objective Comparative analysis of these genomes revealed potential markers and phylogenetic analysis, and discuss the influence of positive selected sites on adaptive evolution. Methods Here, we sequenced the complete chloroplast genomes of these two species and analyzed the repeat sequences components, nucleotide diversity, selection pressure and the phylogeny relationships with related species. Results A typical quadripartite structure was detected with a genome size changed from 152,148 to 152,931 bp. We identified 67 and 79 simple sequence repeats in A. macranthum and A. fasciculatum, in which the mono-nucleotide repeats A/T possess the highest percentage. Three mutational hotspots (rpl32, rps16 and matK) at the SSC and LSC regions were observed, which showed remarkably higher Pi value (> 0.03). Additionally, eight genes (rpoA, atpF, cemA, rps4, ccsA, rpoC2, rpl14 and clpP) exhibited elevated pairwise Ka/Ks ratios in alpine species. Phylogenetic analyses based on the CDS sequences and the whole complete genomes showed same topologies with high support, and A. macranthum was closely clustered with A. fasciculatum within the fourteen Amaryllidaceae species. Conclusion Their coding proteins of these genes often functioned in chloroplast protein synthesis, gene transcription, energy transformation and regulation and photosynthesis. These results provide valuable insights into the alpine species adaptation and evolution.