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.

Bioactivity of essential oil of Litsea cubeba from China and its main compounds against two stored product insects

Kai Yang,Cheng FangWang,Chun Xue You,Zhu Feng Geng,Rui Qi Sun,Shan Shan Guo,Shu Shan Du,Zhi Long Liu,Zhi Wei Deng 한국응용곤충학회 2014 Journal of Asia-Pacific Entomology Vol.17 No.3

During our screening program for agrochemicals from Chinese medicinal herbs and wild plants, the essential oilof Litsea cubeba fruits was found to possess strong contact toxicity against the cigarette beetle Lasiodermaserricorne adults and the booklouse Liposcelis bostrychophila, with LD50 values of 27.33 μg/adult and71.56 μg/cm2, respectively, and also showed strong fumigant toxicity against the two stored product insectswith LC50 values of 22.97 and 0.73 mg/L, respectively. The essential oil obtained by hydrodistillation wasinvestigated by GC MS. The main components of the essential oil were identified to be E-citral (geranial)(27.49%), Z-citral (neral) (23.57%) and D-limonene (18.82%) followed by β-thujene (3.34%), β-pinene (2.85%), α-pinene (2.57%), 6-methyl-5-hepten-2-one (2.40%) and linalool (2.36%). Citral (Z/E-citral), D-limonene, β-pinene,α-pinene and linalool were separated and purified by silica gel column chromatography and preparative thinlayer chromatography, and further identified by means of physicochemical and spectrometric analysis. Citral andlinalool showed strong contact toxicity against L. serricorne and L. bostrychophila (LD50 = 11.76, 12.74 μg/adultand 20.15, 99.97 μg/cm2, respectively) and fumigant toxicity against L. serricorne and L. bostrychophila (16.54,18.04 mg/L air and 0.14, 0.71 mg/L air, respectively). Otherwise, citral, D-limonene and linalool were strongly repellentagainst the cigarette beetle L. serricorne as the essential oil whereas β-pinene and α-pinene exhibited weakerrepellency against the cigarette beetle compared with the positive control, DEET. Moreover, except α-pinene andlinalool, the other three compounds as well as the essential oil exhibited comparable repellency against thebooklouse relative to DEET.

A hardening model considering grain size effect for ion-irradiated polycrystals under nanoindentation

Liu, Kai,Long, Xiangyun,Li, Bochuan,Xiao, Xiazi,Jiang, Chao Korean Nuclear Society 2021 Nuclear Engineering and Technology Vol.53 No.9

In this work, a new hardening model is proposed for the depth-dependent hardness of ion-irradiated polycrystals with obvious grain size effect. Dominant hardening mechanisms are addressed in the model, including the contribution of dislocations, irradiation-induced defects and grain boundaries. Two versions of the hardening model are compared, including the linear and square superposition models. A succinct parameter calibration method is modified to parametrize the models based on experimentally obtained hardness vs. indentation depth curves. It is noticed that both models can well characterize the experimental data of unirradiated polycrystals; whereas, the square superposition model performs better for ion-irradiated materials, therefore, the square superposition model is recommended. In addition, the new model separates the grain size effect from the dislocation hardening contribution, which makes the physical meaning of fitted parameters more rational when compared with existing hardness analysis models.

Molecular Biology and Omics : Direct Evaluation of the Effect of Gene Dosage on Secretion of Protein from Yeast Pichia pastoris by Expressing EGFP

( Hai Long Liu ),( Yu Feng Qin ),( Yuan Kai Huang ),( Yao Sheng Chen ),( Pei Qing Cong ),( Zu Yong He ) 한국미생물 · 생명공학회 2014 Journal of microbiology and biotechnology Vol.24 No.2

Increasing the gene copy number has been commonly used to enhance the protein expression level in the yeast Pichia pastoris. However, this method has been shown to be effective up to a certain gene copy number, and a further increase of gene dosage can result in a decrease of expression level. Evidences indicate the gene dosage effect is product-dependent, which needs to be determined when expressing a new protein. Here, we describe a direct detection of the gene dosage effect on protein secretion through expressing the enhanced green fluorescent protein (EGFP) gene under the direction of the α-factor preprosequence in a panel of yeast clones carrying increasing copies of the EGFP gene (from one to six copies). Directly examined under fluorescence microscopy, we found relatively lower levels of EGFP were secreted into the culture medium at one copy and two copies, substantial improvement of secretion appeared at three copies, plateau happened at four and five copies, and an apparent decrease of secretion happened at six copies. The secretion of EGFP being limiting at four and five copies was due to abundant intracellular accumulation of proteins, observed from the fluorescence image of yeast and confirmed by western blotting, which significantly activated the unfolded protein response indicated by the up-regulation of the BiP (the KAR2 gene product) and the protein disulfide isomerase. This study implies that tagging a reporter like GFP to a specific protein would facilitate a direct and rapid determination of the optimal gene copy number for high-yield expression.

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.

Ce3+ triggers fenton-like processes in neutral solutions for effective catechol degradation

Xing Chen,Xu Liu,Hai-Bo Wang,Kang-Ping Cui,Rohan Weerasooriya,Shi-Long He,Guang-Hong Li,Jun Pan,Kai Zhou 대한환경공학회 2022 Environmental Engineering Research Vol.27 No.1

Classical Fenton and Fenton-like processes destruct organic pollutants in water non-selectively to complete mineralization. However, the usage of classical Fenton or Fenton-like processes is often limited due to the narrow operational pH window, sludge accumulation, inefficient H₂O₂ and efficiency decline. To overcome these constraints, in this study, we used a homogeneous Fe<SUP>3+</SUP>-Ce<SUP>3+</SUP>-H₂O₂ Fenton-like process to degrade catechol at different experimental conditions. At pH 7, almost 97% of 10 mM catechol can be destructed within 60 min while the degradation by Classical Fenton or Fe<SUP>3+</SUP>-H₂O₂ Fenton-like process only 36.2% and 23.7%. The resultant solution after the degradation contains only traces of cerium ions. The sludge created by the process was extensively characterized by FTIR and XPS spectroscopy to elucidate the fate of cerium ions. Electron spin resonance (ESR) data confirmed •OH as the major free radical in Fe<SUP>3+</SUP>-Ce<SUP>3+</SUP>-H₂O₂ process. Our Fenton-like process widens the optimal pH values to neutral condition.

Acetate-assisted Synthesis of Chromium(III) Terephthalate and Its Gas Adsorption Properties

Jing-jing Zhou,Kai-yu Liu,Chun-long Kong,Liang Chen 대한화학회 2013 Bulletin of the Korean Chemical Society Vol.34 No.6

We report a facile synthetic approach of high-quality chromium(III) terephthalate [MIL-101(Cr)] by acetateassisted method in the absence of toxic HF. Results indicate that the morphology and surface area of the MIL- 101(Cr) can be tuned by modifying the molar ratio of acetate/Cr(NO3)3. The Brunauer-Emmett-Teller (BET) surface area of MIL-101(Cr) synthesized at the optimized condition can exceed 3300 m2/g. It is confirmed that acetate could promote the dissolution of di-carboxylic linker and accelerate the nucleation ratio. So the pure and small size of MIL-101(Cr) with clean pores can be obtained. CO2, CH4 and N2 adsorption isotherms of the samples are studied at 298 K and 313 K. Compared with the traditional method, MIL-101(Cr) synthesized by acetate-assisted method possess enhanced CO2 selective adsorption capacity. At 1.0 bar 298 K, it exhibits 47% enhanced CO2 adsorption capacity. This may be attributed to the high surface area together with clean pores of MIL-101(Cr).

Study on critical buckling load calculation method of piles considering passive and active earth pressure

Chen, Yong-Hui,Chen, Long,Xu, Kai,Liu, Lin,Ng, Charles W.W. Techno-Press 2013 Structural Engineering and Mechanics, An Int'l Jou Vol.48 No.3

Different types of long slender pile shall buckle with weak soil and liquefied stratum surrounded. Different from considering single side earth pressure, it was suggested that the lateral earth pressure can be divided into two categories while buckling: the earth pressure that prevent and promotes the lateral movement. Active and passive earth pressure calculation model was proposed supposing earth pressure changed linearly with displacement considering overlying load, shaft resistance, earth pressure at both sides of the pile. Critical buckling load calculation method was proposed based on the principle of minimum potential energy quoting the earth pressure calculation model. The calculation result was contrasted with the field test result of small diameter TC pile (Plastic Tube Cast-in-place pile). The fix form could be fixed-hinged in the actual calculation assuring the accuracy and certain safety factor. The contributions of pile fix form depend on the pile length for the same geological conditions. There exists critical friction value in specific geological conditions that the side friction has larger impact on the critical buckling load while it is less than the value and has less impact with larger value. The buckling load was not simply changed linearly with friction. The buckling load decreases with increased limit active displacement and the load tend to be constant with larger active displacement value; the critical buckling load will be the same for different fix form for the small values.

Acetate-assisted Synthesis of Chromium(III) Terephthalate and Its Gas Adsorption Properties

Zhou, Jing-Jing,Liu, Kai-Yu,Kong, Chun-Long,Chen, Liang Korean Chemical Society 2013 Bulletin of the Korean Chemical Society Vol.34 No.6

We report a facile synthetic approach of high-quality chromium(III) terephthalate [MIL-101(Cr)] by acetate-assisted method in the absence of toxic HF. Results indicate that the morphology and surface area of the MIL-101(Cr) can be tuned by modifying the molar ratio of acetate/$Cr(NO_3)_3$. The Brunauer-Emmett-Teller (BET) surface area of MIL-101(Cr) synthesized at the optimized condition can exceed 3300 $m^2/g$. It is confirmed that acetate could promote the dissolution of di-carboxylic linker and accelerate the nucleation ratio. So the pure and small size of MIL-101(Cr) with clean pores can be obtained. $CO_2$, $CH_4$ and $N_2$ adsorption isotherms of the samples are studied at 298 K and 313 K. Compared with the traditional method, MIL-101(Cr) synthesized by acetate-assisted method possess enhanced $CO_2$ selective adsorption capacity. At 1.0 bar 298 K, it exhibits 47% enhanced $CO_2$ adsorption capacity. This may be attributed to the high surface area together with clean pores of MIL-101(Cr).

RBM24 exacerbates bladder cancer progression by forming a Runx1t1/TCF4/miR-625-5p feedback loop

Yin Yue-Wei,Liu Kai-Long,Lu Bao-Sai,Li Wei,Niu Ya-Lin,Zhao Chen-Ming,Yang Zhan,Guo Ping-Ying,Qi Jin-Chun 생화학분자생물학회 2021 Experimental and molecular medicine Vol.53 No.-

RNA–binding motif protein 24 (RBM24) acts as a multifunctional determinant of cell fate, proliferation, apoptosis, and differentiation during development by regulating premRNA splicing and mRNA stability. It is also implicated in carcinogenesis, but the functions of RBM24 in bladder cancer (BC) remain unclear. In the present study, we revealed that RBM24 was upregulated in BC tissues. Importantly, we found that a higher level of RBM24 was correlated with poor prognosis in BC patients. Overexpression of RBM24 promoted BC cell proliferation, while depletion of RBM24 inhibited BC cell proliferation in vivo and in vitro. Mechanistically, RBM24 positively regulated Runx1t1 expression in BC cells by binding to and enhancing Runx1t1 mRNA stability. Furthermore, Runx1t1 in turn promoted RBM24 expression by interacting with the transcription factor TCF4 and suppressing the transcription of miR-625-5p, which directly targets RBM24 and suppresses RBM24 expression. RBM24-regulated BC cell proliferation was moderated via the Runx1t1/TCF4/miR-625-5p feedback loop. These results indicate that the RBM24/Runx1t1/TCF4/miR-625-5p positive feedback loop participates in BC progression. Disruption of this pathway may be a potential therapeutic strategy for BC treatment.