Production of bioactive ginsenoside Rg3(S) and compound K using recombinant Lactococcus lactis

Lingli Jiang,Soo Jin Lee,Qiu Ping Yuan,임완택,김선창,NAM SOO HAN 고려인삼학회 2018 Journal of Ginseng Research Vol.42 No.4

Background: Ginsenoside Rg3(S) and compound K (C-K) are pharmacologically active components of ginseng that promote human health and improve quality of life. The aim of this study was to produce Rg3(S) and C-K from ginseng extract using recombinant Lactococcus lactis. Methods: L. lactis subsp. cremoris NZ9000 (L. lactis NZ9000), which harbors b-glucosidase genes (BglPm and BglBX10) from Paenibacillus mucilaginosus and Flavobacterium johnsoniae, respectively, was reacted with ginseng extract (protopanaxadiol-type ginsenoside mixture). Results: Crude enzyme activity of BglBX10 values comprised 0.001 unit/mL and 0.003 unit/mL in uninduced and induced preparations, respectively. When whole cells of L. lactis harboring pNZBglBX10 were treated with ginseng extract, after permeabilization of cells by xylene, Rb1 and Rd were converted into Rg3(S) with a conversion yield of 61%. C-K was also produced by sequential reactions of the permeabilized cells harboring each pNZBgl and pNZBglBX10, resulting in a 70% maximum conversion yield. Conclusion: This study demonstrates that the lactic acid bacteria having specific b-glucosidase activity can be used to enhance the health benefits of Panax ginseng in either fermented foods or bioconversion processes.

Metabolic engineering of Corynebacterium glutamicum for increasing the production of L-ornithine by increasing NADPH availability.

Jiang, Ling-Yan,Zhang, Yuan-Yuan,Li, Zhen,Liu, Jian-Zhong Published by Stockton Press on behalf of the Socie 2013 Journal of industrial microbiology & biotechnology Vol.40 No.10

<P>The experiments presented here were based on the conclusions of our previous proteomic analysis. Increasing the availability of glutamate by overexpression of the genes encoding enzymes in the L-ornithine biosynthesis pathway upstream of glutamate and disruption of speE, which encodes spermidine synthase, improved L-ornithine production by Corynebacterium glutamicum. Production of L-ornithine requires 2 moles of NADPH per mole of L-ornithine. Thus, the effect of NADPH availability on L-ornithine production was also investigated. Expression of Clostridium acetobutylicum gapC, which encodes NADP-dependent glyceraldehyde-3-phosphate dehydrogenase, and Bacillus subtilis rocG, which encodes NAD-dependent glutamate dehydrogenase, led to an increase of L-ornithine concentration caused by greater availability of NADPH. Quantitative real-time PCR analysis demonstrates that the increased levels of NADPH resulted from the expression of the gapC or rocG gene rather than that of genes (gnd, icd, and ppnK) involved in NADPH biosynthesis. The resulting strain, C. glutamicum δAPRE::rocG, produced 14.84 g l?1 of L-ornithine. This strategy of overexpression of gapC and rocG will be useful for improving production of target compounds using NADPH as reducing equivalent within their synthetic pathways.</P>

A Novel Crawler Based on Loginning Simulation for Weibo Social Network

Ling Xing,Ling Jiang,Bao Peng 보안공학연구지원센터 2016 International Journal of Grid and Distributed Comp Vol.9 No.1

With the rapid development of Weibo, which is the most popular microblog in china, more and more attention was paid to relative studies about it. With the objective of gathering precise information data from Weibo, which is the groundwork of these researches, a novel high efficient Weibo crawler (WCrawler) based on loginning simulation is designed. The priority evaluation is described to ensure the correlation between entires. MD5 is introduced to check for duplicates of URL crawled. Experiments demonstrate that the novel crawler has an efficiency and integrity of information collecting compared with API crawler. In addition, we present a summary of the data that collected from Weibo social network by WCrawler.

High-sensitivity detection for cantharidin by ratiometric fluorescent sensor based on molecularly imprinted nanoparticles of quantum dots

Jiang Ling,Wenqi Zhang,Zijia Cheng,Yanjun Ding 한국공업화학회 2022 Journal of Industrial and Engineering Chemistry Vol.112 No.-

Cantharidin (CTD), an effective component of Chinese herbal medicine with unique efficacy, caused poisoningor death constantly for incorrect use. Therefore, there is an intense need for a reliable, selective,sensitive and low-costing analytical method for monitoring the concentration of CTD in biological samples. In this work, a ratiometric fluorescent sensor (CdTe@MIPs/CDs@NIPs) was successfully constructedfor selective determination of CTD by simply integrated the non-imprinted polymers coated CDs(CDs@NIPs) with cadmium telluride quantum dots coated with molecular imprinted polymers(CdTe@MIPs). CDs@NIPs were directly used as reference signal without addition of CTD templates andthe as-prepared CdTe@MIPs were response signal. With the CTD addition, the fluorescence intensity ofCdTe@MIPs decreased, while the fluorescence of CDs@NIPs remained approximately unchanged, resultinga detection limit as low as 0.15 nM. By spiked human blood samples, the ratiometric sensor was successfullyapplied to CTD detection with good recoveries of 96.12–107.40% and relative standard deviation(RSD) of 2.87–3.96%. Moreover, after centrifuging and re-eluting, the developed sensor could be effectivelyrecycled and reused five times. Significantly, the proposed platform with low-cost and highsensitivityopens a door towards the practical applications of ratiometric sensor and shows immensepotential in pursuing the sensitive, reliable and low-cost determination of poisons in forensic toxicologyanalysis.

Does HBV Infection Increase Risk of Endometrial Carcinoma?

Jiang, Xue-Feng,Tang, Qiong-Lan,Zou, Yuan,Xu, Ling,Zeng, Hua,Chi, Cong,Jiang, Jing-Ru,Zhang, Bin-Zhong Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.2

Objective: Connections between chronic inflammation and tumor development and progression are now generally accepted. Recent evidence indicates that hepatitis B is associated with several types of cancer, but whether endometrial carcinoma (EC) is included has not been reported. Methods: We analyzed HBV serum marker status in 398 patients with endometrial cancer, comparing them to 788 control women undergoing health examination. Results: The total prevalence of HBsAg tested positive in cancer group was significantly higher than the control group (12.8% vs 6.0%, P=0.001), while positive HBsAb was significantly lower (41.2% vs 68.5%, P=0.001). Hepatitis B carriers in endometrial cancer group were also more frequent than in the control group (9.3% vs 5.5%, P=0.013). Interestingly, in the endometrial cancer group, 147 cases were HBV serum marker negative, which was also higher than in the control group (36.9% vs 15.6%, P=0.001). Conclusion: There may be a correlation between HBV infection and endometrial carcinoma.

Somatic Cell Nuclear Transfer of Oocytes Aspirated from Postovulatory Ovarian Follicles of Superovulated Rabbits

Shang, Jiang-Hua,Xu, Ru-Xiang,Jiang, Xiao-Dan,Zou, Yu-Xi,Qin, Ling-Sha,Cai, Ying-Qian,Yang, Zhi-Jun,Zheng, Xing,Cui, Sheng Asian Australasian Association of Animal Productio 2007 Animal Bioscience Vol.20 No.9

The aim of this study was to evaluate if oocytes, aspirated from postovulatory ovarian follicles of superovulated rabbits 14 h post-hCG administration, could be efficiently used as ooplasm recipients for somatic cell nuclear transfer (SCNT). Within a common SCNT protocol, a comparison between oocytes recovered by direct aspiration (aspirated) from available ovarian follicles and oocytes flushed out from oviducts (flushed) was carried out. The results showed that maturation and enucleation rates of aspirated oocytes were 70.7% and 69.2%, significantly lower than 95.3% (p<0.01) and 83.6% (p<0.05), respectively, from flushed oocytes. However, following enucleation of matured oocytes as ooplasm recipients for SCNT, no difference was recorded in fusion and cleavage rates, as well as blastocyst development from cleaved embryos or hatching of blastocysts between aspirated and flushed groups. Additionally, some matured aspirated and flushed oocytes were also used for immediate parthenogenetic activation and the resulting embryo development was not significantly different. Results from this study show the following: i) the majority of oocytes aspirated from postovulatory ovarian follicles of superovulated rabbits 14 h post-hCG administration are matured and can be used directly as ooplasm recipients for SCNT; ii) the reconstructed embryos derived from these oocytes have similar in vitro developmental ability to those flushed from the oviducts.

Cloning and Characterization of Functional Trehalose-6-Phosphate Synthase Gene in Maize

Wei Jiang,Feng-Ling Fu,Su-Zhi Zhang,Ling Wu,Wan-Chen Li 한국식물학회 2010 Journal of Plant Biology Vol.53 No.2

Trehalose is a non-reducing disaccharide of glucose that functions as a compatible solute in the stabilization of biological structures under heat and desiccation stress in bacteria, fungi, and some “resurrection plants”. In the plant kingdom, trehalose is biosynthesized by trehalose-6-phosphate synthase (TPS) and trehalose-6-phosphate phosphatase (TPP). Over-expression of exogenous and endogenous genes encoding TPS and TPP is reported to be effective for improving abiotic stress tolerance in tobacco, potato, tomato, rice, and Arabidopsis. On the basis of bioinformatics prediction, we cloned a fragment containing an open reading frame of 2,820 bp from maize,which encodes a protein of 939 amino acids. Phylogenetic analysis showed that this gene belongs to the class I subfamily of the TPS gene family. Analysis of conserved domains revealed the presence of a TPS domain and a TPP domain. Yeast complementation with TPS and TPP mutants demonstrated that this protein has the activity of trehalose-6-phosphate synthase. Semi-quantitative RT-PCR and realtime quantitative PCR indicated that the expression of this gene is upregulated in response to both salt and cold stress.

Two Kinesins from Arabidopsis, KatB and KatC, Have a Second Microtubule-binding Site in the Tail Domain

( Shi Ling Jiang ),( Ming Li ),( Tao Xu ),( Dong Tao Ren ),( Guo Qin Liu ) 생화학분자생물학회 2007 BMB Reports Vol.40 No.1

Ginsenoside Rg5 overcomes chemotherapeutic multidrug resistance mediated by ABCB1 transporter: in vitro and in vivo study

Sen-Ling Feng,Hai-Bin Luo,Liang Cai,Jie Zhang,Dan Wang,Ying-Jiang Chen,Huan-Xing Zhan,Zhi-Hong Jiang,Ying Xie 고려인삼학회 2020 Journal of Ginseng Research Vol.44 No.2

Background: Multidrug resistance (MDR) to chemotherapy drugs remains a major challenge in clinicalcancer treatment. Here we investigated whether and how ginsenoside Rg5 overcomes the MDR mediatedby ABCB1 transporter in vitro and in vivo. Methods: Cytotoxicity and colon formation as well as the intracellular accumulation of ABCB1 substrateswere carried out in MDR cancer cells A2780/T and A549/T for evaluating the reversal effects of Rg5. Theexpressions of ABCB1 and Nrf2/AKT pathway were determined by Western blotting. An A549/T cellxenograft model was established to investigate the MDR reversal activity of Rg5 in vivo. Results: Rg5 significantly reversed ABCB1-mediated MDR by increasing the intracellular accumulation ofABCB1 substrates without altering protein expression of ABCB1. Moreover, Rg5 activated ABCB1 ATPaseand reduced verapamil-stimulated ATPase activity, suggesting a high affinity of Rg5 to ABCB1 bindingsite which was further demonstrated by molecular docking analysis. In addition, co-treatment of Rg5 anddocetaxel (TXT) suppressed the expression of Nrf2 and phosphorylation of AKT, indicating that sensitizingeffect of Rg5 associated with AKT/Nrf2 pathway. In nude mice bearing A549/T tumor, Rg5 and TXTtreatment significantly suppressed the growth of drug-resistant tumors without increase in toxicitywhen compared to TXT given alone at same dose. Conclusion: Therefore, combination therapy of Rg5 and chemotherapy drugs is a strategy for the adjuvantchemotherapy, which encourages further pharmacokinetic and clinical studies.

Optimization of Enzymatic Synthesis of L-ascorbyl Palmitate by Solvent Engineering and Statistical Experimental Designs

Xiang-Jun Jiang,Yi Hu,Ling Jiang,Bin Zou,Ping Song,He Huang 한국생물공학회 2013 Biotechnology and Bioprocess Engineering Vol.18 No.2

Statistical experimental designs combined with solvent engineering for optimization of enzymatic synthesis of L-ascorbyl palmitate were developed. First, the composition of the solvent for co-dissolving polar and apolar substrates was determined. The co-solvent mixture of tert-pentanol:DMSO at a ratio of 9:1 (v/v) and the optimal biocatalyst were obtained. Then, the Plackett-Burman design was implemented to screen the variables that significantly influence the conversion. The method of steepest ascent was used to approach the proximity of optimum. After determining the Plackett-Burman and steepest ascent designs, the optimum values were determined by central composite design under response surface methodology. The statistical analysis showed that the optimum reaction conditions (temperature 50oC, enzyme concentration 5.8 g/L,and substrate molar ratio 11:1, stirring rate 160 rpm, amount of molecular sieve 50 g/L, time 18 h) led to the maximum conversion (66.44%) and production concentration (20.63 g/L). A very satisfactory conversion (64.74%) and production concentration (20.13 g/L) could be achieved in short time (6 h).