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- 저자 : Liangliang Shi (검색결과 3 건)
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( Liangliang Wang ),( Jiajun Wang ),( Hao Shi ),( Huaxiang Gu ),( Yu Zhang ),( Xun Li ),( Fei Wang ) 한국미생물 · 생명공학회 2016 Journal of microbiology and biotechnology Vol.26 No.6
Glycerol dehydrogenases (GlyDHs) are essential for glycerol metabolism in vivo, catalyzing its reversible reduction to 1,3-dihydroxypropranone (DHA). The gldA gene encoding a putative GlyDH was cloned from Thermoanaerobacterium thermosaccharolyticum DSM 571 (TtGlyDH) and expressed in Escherichia coli. The presence of Mn(2+) enhanced its enzymatic activity by 79.5%. Three highly conserved residues (Asp(171), His(254), and His(271)) in TtGlyDH were associated with metal ion binding. Based on an investigation of glycerol oxidation and DHA reduction, TtGlyDH showed maximum activity towards glycerol at 60°C and pH 8.0 and towards DHA at 60°C and pH 6.0. DHA reduction was the dominant reaction, with a lower Km(DHA) of 1.08 ± 0.13 mM and Vmax of 0.0053 ± 0.0001 mM/s, compared with glycerol oxidation, with a Km(glycerol) of 30.29 ± 3.42 mM and Vmax of 0.042 ± 0.002 mM/s. TtGlyDH had an apparent activation energy of 312.94 kJ/mol. The recombinant TtGlyDH was thermostable, maintaining 65% of its activity after a 2-h incubation at 60°C. Molecular modeling and site-directed mutagenesis analyses demonstrated that TtGlyDH had an atypical dinucleotide binding motif (GGG motif) and a basic residue Arg(43), both related to dinucleotide binding.
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Lineage conversion of mouse fibroblasts to pancreatic α-cells
Lijian Hui,Liangliang Sun,Beige Jiang,Limei Li,Jin Cen,Xiaotao Chen,Zhaoyun Zhang,Qinghua Wang,Xin Cheng,Yongquan Shi,Lijian Hui 생화학분자생물학회 2017 Experimental and molecular medicine Vol.49 No.-
α-cells, which synthesize glucagon, also support β-cell survival and have the capacity to transdifferentiate into β-cells. However, the role of α-cells in pathological conditions and their putative clinical applications remain elusive due in large part to the lack of mature α-cells. Here, we present a new technique to generate functional α-like cells. α-like cells (iAlpha cells) were generated from mouse fibroblasts by transduction of transcription factors, including Hhex, Foxa3, Gata4, Pdx1 and Pax4, which induce α-cell-specific gene expression and glucagon secretion in response to KCl and Arg stimulation. The cell functions in vivo and in vitro were evaluated. Lineage-specific and functional-related gene expression was tested by realtime PCR, insulin tolerance test (ITT), glucose tolerance test (GTT), Ki67 and glucagon immunohistochemistry analysis were done in iAlpha cells transplanted nude mice. iAlpha cells possess α-cell function in vitro and alter blood glucose levels in vivo. Transplantation of iAlpha cells into nude mice resulted in insulin resistance and increased β-cell proliferation. Taken together, we present a novel strategy to generate functional α-like cells for the purposes of disease modeling and regenerative medicine.
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Zengli Xiao,Wenlong Qin,Liangliang Shi 보안공학연구지원센터 2016 International Journal of Smart Home Vol.10 No.6
In this paper, a highly sensitive and selective method based on the poly (sulfosalicylic acid) modified electrode (PSA /CPE) to detect phenol was established. The morphologies and interface properties of PSA film were characterized by scanning electron microscopy and electrochemical impedance spectroscopy. It was illustrated that the PSA/CPE had an excellent electrocatalytic ability towards the oxidation of phenol. Meanwhile the influence of parameters such as pH and scan rate effect on the analytical performance of the sensor was evaluated. Moreover, the interference from o-nitrophenol can be neglected. By using differential pulse voltammetry (DPV), linear calibration curves were obtained as 5–175 and 220–555 μmol L−1 for phenol. The detection limits are 2.2 μmol L−1 for phenol. With favorable selectivity and sensitivity, the present method has been applied to the determination of phenol in oilfield wastewater.
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