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https://www.readbyqxmd.com/read/27444330/microbial-surface-displaying-formate-dehydrogenase-and-its-application-in-optical-detection-of-formate
#1
Aihua Liu, Ruirui Feng, Bo Liang
In the present work, NAD(+)-dependent formate dehydrogenase (FDH), encoded by fdh gene from Candida boidinii was successfully displayed on Escherichia coli cell surface using ice nucleation protein (INP) from Pseudomonas borealis DL7 as an anchoring protein. Localization of matlose binding protein (MBP)-INP-FDH fusion protein on the E. coli cell surface was characterized by SDS-PAGE and enzymatic activity assay. FDH activity was monitored through the oxidation of formate catalyzed by cell-surface-displayed FDH with its cofactor NAD(+), and the production of NADH can be detected spectrometrically at 340nm...
September 2016: Enzyme and Microbial Technology
https://www.readbyqxmd.com/read/27022027/piln-binding-modulates-the-structure-and-binding-partners-of-the-pseudomonas-aeruginosa-type-iva-pilus-protein-pilm
#2
Matthew McCallum, Stephanie Tammam, Dustin J Little, Howard Robinson, Jason Koo, Megha Shah, Charles Calmettes, Trevor F Moraes, Lori L Burrows, P Lynne Howell
Pseudomonas aeruginosa is an opportunistic bacterial pathogen that expresses type IVa pili. The pilus assembly system, which promotes surface-associated twitching motility and virulence, is composed of inner and outer membrane subcomplexes, connected by an alignment subcomplex composed of PilMNOP. PilM binds to the N terminus of PilN, and we hypothesize that this interaction causes functionally significant structural changes in PilM. To characterize this interaction, we determined the crystal structures of PilM and a PilM chimera where PilM was fused to the first 12 residues of PilN (PilM·PilN(1-12))...
May 20, 2016: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/26073813/amperometric-l-glutamate-biosensor-based-on-bacterial-cell-surface-displayed-glutamate-dehydrogenase
#3
Bo Liang, Shu Zhang, Qiaolin Lang, Jianxia Song, Lihui Han, Aihua Liu
A novel L-glutamate biosensor was fabricated using bacteria surface-displayed glutamate dehydrogenase (Gldh-bacteria). Here the cofactor NADP(+)-specific dependent Gldh was expressed on the surface of Escherichia coli using N-terminal region of ice nucleation protein (INP) as the anchoring motif. The cell fractionation assay and SDS-PAGE analysis indicated that the majority of INP-Gldh fusion proteins were located on the surface of cells. The biosensor was fabricated by successively casting polyethyleneimine (PEI)-dispersed multi-walled carbon nanotubes (MWNTs), Gldh-bacteria and Nafion onto the glassy carbon electrode (Nafion/Gldh-bacteria/PEI-MWNTs/GCE)...
July 16, 2015: Analytica Chimica Acta
https://www.readbyqxmd.com/read/23912858/comparison-of-cytotoxicity-and-expression-of-metal-regulatory-genes-in-zebrafish-danio-rerio-liver-cells-exposed-to-cadmium-sulfate-zinc-sulfate-and-quantum-dots
#4
COMPARATIVE STUDY
Song Tang, Vinay Allagadda, Hicham Chibli, Jay L Nadeau, Gregory D Mayer
Recent advances in the ability to manufacture and manipulate materials at the nanometer scale have led to increased production and use of many types of nanoparticles. Quantum dots (QDs) are small, fluorescent nanoparticles composed of a core of semiconductor material (e.g. cadmium selenide, zinc sulfide) and shells or dopants of other elements. Particle core composition, size, shell, and surface chemistry have all been found to influence toxicity in cells. The aim of this study was to compare the toxicities of ionic cadmium (Cd) and zinc (Zn) and Cd- and Zn-containing QDs in zebrafish liver cells (ZFL)...
October 2013: Metallomics: Integrated Biometal Science
https://www.readbyqxmd.com/read/23873835/antibody-conjugated-plga-nanoparticles-for-targeted-delivery-of-paclitaxel-palmitate-efficacy-and-biofate-in-a-lung-cancer-mouse-model
#5
Nour Karra, Taher Nassar, Alina Nemirovski Ripin, Ouri Schwob, Jürgen Borlak, Simon Benita
Aberrant signaling of the epidermal growth factor receptor (EGFR) is common to a variety of human cancers and is also found to be over-expressed in most cases of non-small cell lung cancer. For the development of a molecularly targeted therapy, cetuximab-conjugated nanoparticles (immunonanoparticles, INPs) are designed and loaded with the lipophilic paclitaxel palmitate (pcpl) prodrug. Oleyl cysteineamide (OCA) is synthesized whereby its amphiphilic nature enables interfacial anchoring and thiol surface functionalization of PLGA NPs, facilitating bioconjugation to cetuximab by thioether bonds...
December 20, 2013: Small
https://www.readbyqxmd.com/read/23858715/-construction-of-cell-surface-display-system-in-lactic-acid-bacteria-by-using-ice-nucleation-protein
#6
Qiuxiang Zhang, Huili Hou, Ying Lu, Wei Chen, Jin Zhong
OBJECTIVE: The gene of ice nucleation protein (INP), an outer membrane of Pseudomonas syringae was tested to display foreign proteins on the surface of Lactococcus lactis. METHODS: Plasmids pHZ101 and pHZ102 were constructed using gfp (Green Fluorescence Protein) as the reporter gene and N-terminal and NC-terminal of inp as the anchoring units. Plasmids pHZ101 and pHZ102 were subsequently transformed into Escherichia coli JM109 and Lactococcus lactis MG1363. RESULTS: Fluorescence microscope shows that green fluorescence was observed in both recombinant E...
April 4, 2013: Wei Sheng Wu Xue Bao, Acta Microbiologica Sinica
https://www.readbyqxmd.com/read/23804101/cortical-gyrification-induced-by-fibroblast-growth-factor-2-in-the-mouse-brain
#7
Brian G Rash, Simone Tomasi, H David Lim, Carol Y Suh, Flora M Vaccarino
Gyrification allows an expanded cortex with greater functionality to fit into a smaller cranium. However, the mechanisms of gyrus formation have been elusive. We show that ventricular injection of FGF2 protein at embryonic day 11.5-before neurogenesis and before the formation of intrahemispheric axonal connections-altered the overall size and shape of the cortex and induced the formation of prominent, bilateral gyri and sulci in the rostrolateral neocortex. We show increased tangential growth of the rostral ventricular zone (VZ) but decreased Wnt3a and Lef1 expression in the cortical hem and adjacent hippocampal promordium and consequent impaired growth of the caudal cortical primordium, including the hippocampus...
June 26, 2013: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
https://www.readbyqxmd.com/read/23764018/production-of-d-hydantoinase-via-surface-display-and-self-cleavage-system
#8
Chia-Chi Lin, Tzu-Tsen Liu, Shu-Chen Kan, Chi-Zong Zang, Chiung-Wen Yeh, Jiun-Yan Wu, Jiann-Hwa Chen, Chwen-Jen Shieh, Yung-Chuan Liu
In this study, the cell surface expression system was the first time used to directly produce extracellular enzyme. In the plasmid construction, the truncated ice nucleation protein (INP) was fused with intein (INT) and target protein, D-hydantoinase (DHTase), to form the INP-INT-DHTase gene. The plasmid containing this gene was transformed into Escherichia coli ER2566 cells. The gene construct enables the expression of INP-INT-DHTase fusion protein, which might anchor on cell membrane surface. The induction conditions were studied and optimal conditions were as follows: E...
November 2013: Journal of Bioscience and Bioengineering
https://www.readbyqxmd.com/read/23454338/microbial-surface-display-of-glucose-dehydrogenase-for-amperometric-glucose-biosensor
#9
Bo Liang, Liang Li, Xiangjiang Tang, Qiaolin Lang, Hongwei Wang, Feng Li, Jianguo Shi, Wei Shen, Ilaria Palchetti, Marco Mascini, Aihua Liu
A genetically engineered Escherichia coli (E. coli) strain displaying glucose dehydrogenase (GDH) with ice-nucleation protein (INP) as the anchoring motif was first constructed. The surface localization and functionality of the fusion protein containing GDH were verified by SDS-PAGE, Western blotting and enzymatic activity assay. The fusion of INP had no effects on the functionality of GDH cofactor binding domain. The activity assay showed that 74.6% of the cell lysate GDH activity was detected in the outer membrane fractions...
July 15, 2013: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/22991498/molecular-characterization-of-an-ice-nucleation-protein-variant-inaq-from-pseudomonas-syringae-and-the-analysis-of-its-transmembrane-transport-activity-in-escherichia-coli
#10
Qianqian Li, Qi Yan, Jinsi Chen, Yan He, Jing Wang, Hongxing Zhang, Ziniu Yu, Lin Li
The ice nucleation protein (INP) of Pseudomonas syringae has gained scientific interest not only because of its pathogenicity of foliar necroses but also for its wide range of potential applications, such as in snow making, frozen food preparation, and surface-display system development. However, studies on the transport activity of INP remain lacking. In the present study, a newly identified INP-gene variant, inaQ, from a P. syringae MB03 strain was cloned. Its structural domains, signal sequences, and the hydrophilicity or hydrophobicity of each domain, were then characterized...
2012: International Journal of Biological Sciences
https://www.readbyqxmd.com/read/21941936/cloning-protein-expression-and-display-of-synthetic-multi-epitope-mycobacterial-antigens-on-salmonella-typhi-ty21a-cell-surface
#11
Mohammed A A Sarhan, Mustaffa Musa, Zainul F Zainuddin
Expressing proteins of interest as fusion to proteins of bacterial envelope is a powerful technique for biotechnological and medical applications. The synthetic gene (VacII) encoding for T-cell epitopes of selected genes of Mycobacterium tuberculosis namely, ESAT6, MTP40, 38 kDa, and MPT64 was fused with N- terminus of Pseudomonas syringae ice nucleation protein (INP) outer membrane protein. The fused genes were cloned into a bacterial expression vector pKK223-3. The recombinant protein was purified by Ni-NAT column...
September 2011: Indian Journal of Experimental Biology
https://www.readbyqxmd.com/read/21732326/cell-surface-display-of-carbonic-anhydrase-on-escherichia-coli-using-ice-nucleation-protein-for-co%C3%A2-sequestration
#12
Li-Hai Fan, Ning Liu, Ming-Rui Yu, Shang-Tian Yang, Huan-Lin Chen
Carbonic anhydrase (CA) has recently gained renewed interests for its potential as a mass-transfer facilitator for CO(2) sequestration. However, the low stability and high price severely limit its applications. In this work, the expression of α-CA from Helicobacter pylori on the outer membrane of Escherichia coli using a surface-anchoring system derived from ice nucleation protein (INP) from Pseudomonas syringae was developed. To find the best surface anchoring motif, full-length INP (114 kDa), truncated INP (INP-NC, 33 kDa), and INP's N-domain with first two subunits (INP-N, 22 kDa) were evaluated...
December 2011: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/21622793/cell-surface-display-of-poliovirus-receptor-on-escherichia-coli-a-novel-method-for-concentrating-viral-particles-in-water
#13
Morteza Abbaszadegan, Absar Alum, Hamed Abbaszadegan, Valerie Stout
The lack of efficient methods for concentrating viruses in water samples leads to underreporting of viral contamination in source water. A novel strategy for viral concentration was developed using the expression of target virus receptors on bacterial cells. Poliovirus type 1, the most studied enterovirus, was used as a surrogate for enteric viruses. The human poliovirus receptor (hPVR) gene was expressed on the surface of Escherichia coli cells by using the ice nucleation protein (INP) gene. The hPVR gene was ligated to the 3' end of the INP gene after the removal of the stop codon...
August 2011: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/20556484/anchorage-of-gfp-fusion-on-the-cell-surface-of-pseudomonas-putida
#14
Yulan Yuan, Chao Yang, Cunjiang Song, Hong Jiang, Ashok Mulchandani, Chuanling Qiao
Here we report the cell surface display of organophosphorus hydrolase (OPH) and green fluorescent protein (GFP) fusion by employing the N- and C-terminal domains of ice nucleation protein (INPNC) as an anchoring motif. An E. coli-Pseudomonas shuttle vector, pNOG33, coding for INPNC-OPH-GFP was constructed for targeting the fusion onto the cell surface of p-nitrophenol (PNP)-degrading P. putida JS444. The surface localization of INPNC-OPH-GFP was verified by cell fractionation, Western blot, proteinase accessibility, and immunofluorescence microscopy...
February 2011: Biodegradation
https://www.readbyqxmd.com/read/19933341/cotranslocation-of-methyl-parathion-hydrolase-to-the-periplasm-and-of-organophosphorus-hydrolase-to-the-cell-surface-of-escherichia-coli-by-the-tat-pathway-and-ice-nucleation-protein-display-system
#15
Chao Yang, Roland Freudl, Chuanling Qiao, Ashok Mulchandani
A genetically engineered Escherichia coli strain coexpressing organophosphorus hydrolase (OPH) and methyl parathion hydrolase (MPH) was constructed for the first time by cotransforming two compatible plasmids. Since these two enzymes have different substrate specificities, the coexpression strain showed a broader substrate range than strains expressing either one of the hydrolases. To reduce the mass transport limitation of organophosphates (OPs) across the cell membrane, MPH and OPH were simultaneously translocated to the periplasm and cell surface of E...
January 2010: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/19332818/characterization-and-recombinant-expression-of-a-divergent-ice-nucleation-protein-from-pseudomonas-borealis
#16
Zhongqin Wu, Lei Qin, Virginia K Walker
Isolates of 'Pseudomonas borealis' were recovered after ice-affinity selection of summer-collected soils. 'P. borealis' DL7 was further characterized and shown to have ice nucleation activity (INA), a property that allows the crystallization of ice at temperatures close to the melting point, effectively preventing the supercooling of water. INA was optimally detected after culturing at temperatures consistent with psychrophilic growth. The sequence encoding the 'P. borealis' ice nucleation protein (INP) was obtained using both PCR and chromosome walking...
April 2009: Microbiology
https://www.readbyqxmd.com/read/19168665/intermediate-neuronal-progenitors-basal-progenitors-produce-pyramidal-projection-neurons-for-all-layers-of-cerebral-cortex
#17
Tom Kowalczyk, Adria Pontious, Chris Englund, Ray A M Daza, Francesco Bedogni, Rebecca Hodge, Alessio Attardo, Chris Bell, Wieland B Huttner, Robert F Hevner
The developing cerebral cortex contains apical and basal types of neurogenic progenitor cells. Here, we investigated the cellular properties and neurogenic output of basal progenitors, also called intermediate neuronal progenitors (INPs). We found that basal mitoses expressing transcription factor Tbr2 (an INP marker) were present throughout corticogenesis, from embryonic day 10.5 through birth. Postnatally, Tbr2(+) progenitors were present in the dentate gyrus, subventricular zone (SVZ), and posterior periventricle (pPV)...
October 2009: Cerebral Cortex
https://www.readbyqxmd.com/read/18689674/the-role-of-atp-signaling-in-the-migration-of-intermediate-neuronal-progenitors-to-the-neocortical-subventricular-zone
#18
Xiuxin Liu, Kazue Hashimoto-Torii, Masaaki Torii, Tarik F Haydar, Pasko Rakic
Most neurons of the cerebral cortex are generated in the germinal zones near the embryonic cerebral ventricle and migrate radially to the overlying cortical plate. Initially, all dividing cells are attached to the surface of the embryonic ventricle (ventricular zone) until a subset of dividing cells (basal or intermediate neuronal progenitors, INPs), recognized by their immunoreactivity to Tbr2, detach from the ventricular surface and migrate a short distance to establish a secondary proliferative compartment (the subventricular zone)...
August 19, 2008: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/18535860/surface-display-of-gfp-by-pseudomonas-syringae-truncated-ice-nucleation-protein-in-attenuated-vibrio-anguillarum-strain
#19
Yuzhou Xu, Qin Liu, Lingyun Zhou, Zhao Yang, Yuanxing Zhang
Microbial cell surface display of foreign proteins has been widely developed for many potential applications in live vaccine construction, whole-cell biocatalysts, and bioadsorption. To investigate the feasibility of displaying heterologous proteins on the surface of attenuated Vibrio anguillarum strain for potential multivalent live vaccine development, different display systems were built upon a truncated ice nucleation protein (INP) from Pseudomonas syringae ICMP3023 whose N- and C-terminal domains were considered to be the putative membrane-anchoring motifs...
November 2008: Marine Biotechnology
https://www.readbyqxmd.com/read/16889366/surface-display-of-organophosphorus-hydrolase-on-saccharomyces-cerevisiae
#20
Katsumi Takayama, Shin-ichiro Suye, Kouichi Kuroda, Mitsuyoshi Ueda, Tetsuya Kitaguchi, Kouta Tsuchiyama, Takeshi Fukuda, Wilfred Chen, Ashok Mulchandani
The gene encoding organophosphorus hydrolase (OPH) from Flavobacterium species was expressed on the cell surface of Saccharomyces cerevisiae MT8-1 using a glycosylphosphatidylinositol (GPI) anchor linked to the C-terminal region of OPH. Immunofluorescence microscopy confirmed the localization of OPH on the cell surface, and fluorescence intensity measurement of cells revealed that 1.4 x 10(4) molecules of OPH per cell were displayed. Seventy percent of OPH whole-cell activity was detected on the cell surface by protease accessibility assay...
July 2006: Biotechnology Progress
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