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https://www.readbyqxmd.com/read/29115133/pursuing-the-promise-of-enzymatic-enhancement-with-nanoparticle-assemblies
#1
James Nicholas Vranish, Mario G Ancona, Scott A Walper, Igor L Medintz
The growing emphasis on green chemistry, renewable resources, synthetic biology, regio-/stereospecific chemical transformations, and nanotechnology for providing new biological products and therapeutics is reinvigorating research into enzymatic catalysis. Although the promise is profound, many complex issues remain to be addressed before this effort will have a significant impact. Prime among these is to combat the degradation of enzymes frequently seen in ex vivo formats following immobilization to stabilize the enzymes for long-term application and to find ways of enhancing their activity...
November 8, 2017: Langmuir: the ACS Journal of Surfaces and Colloids
https://www.readbyqxmd.com/read/29050588/sequential-one-pot-multienzyme-synthesis-of-hyaluronan-and-its-derivative
#2
Shuang Li, Shuaishuai Wang, Xuan Fu, Xian-Wei Liu, Peng George Wang, Junqiang Fang
Hyaluronan (HA) is a linear polysaccharide composed of repeating disaccharide units. It has been well documented to play an array of biological functions in cancer events. Here, we reported a sequential one-pot multienzyme (OPME) strategy for in vitro synthesis of HA and its derivatives. The strategy, which combined in situ sugar nucleotides generation with HA chain polymerization, could convert cheap monosaccharides into HA polymers without consuming exogenous sugar nucleotide donors. HA polymers (number-average molecular weight ranged from 1...
December 15, 2017: Carbohydrate Polymers
https://www.readbyqxmd.com/read/29022695/high-activity-and-convenient-ratio-control-dna-directed-coimmobilization-of-multiple-enzymes-on-multifunctionalized-magnetic-nanoparticles
#3
Ye Yang, Ruiqi Zhang, Bingnan Zhou, Jiayi Song, Ping Su, Yi Yang
The development of new methods for fabricating artificial multienzyme systems has attracted much interest because of the potential applications and the urgent need for multienzyme catalysts. Controlling the enzyme ratio is critical for improving the cooperative enzymatic activity in multienzyme systems. Herein, we introduce a versatile strategy for fabricating a multienzyme system by coimmobilizing horseradish peroxidase (HRP) and glucose oxidase (GOx) on magnetic nanoparticles multifunctionalized with dopamine derivatives through DNA-directed immobilization...
October 12, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28936496/h-pylori-%C3%AE-1-3-4-fucosyltransferase-hp3-4ft-catalyzed-one-pot-multienzyme-opme-synthesis-of-lewis-antigens-and-human-milk-fucosides
#4
Hai Yu, Yanhong Li, Zhigang Wu, Lei Li, Jie Zeng, Chao Zhao, Yijing Wu, Nova Tasnima, Jing Wang, Huaide Liu, Madhusudhan Reddy Gadi, Wanyi Guan, Peng G Wang, Xi Chen
Helicobacter pylori α1-3/4-fucosyltransferase (Hp3/4FT) was expressed in Escherichia coli at a level of 30 mg L(-1) culture and used as a diverse catalyst in a one-pot multienzyme (OPME) system for high-yield production of l-fucose-containing carbohydrates including Lewis antigens such as Lewis a, b, and x, O-sulfated Lewis x, and sialyl Lewis x and human milk fucosides such as 3-fucosyllactose (3-FL), lacto-N-fucopentaose (LNFP) III, and lacto-N-difuco-hexaose (LNDFH) II and III. Noticeably, while difucosylation of tetrasaccharides was readily achieved using an excess amount of donor, the synthesis of LNFP III was achieved by Hp3/4FT-catalyzed selective fucosylation of the N-acetyllactosamine (LacNAc) component in lacto-N-neotetraose (LNnT)...
October 5, 2017: Chemical Communications: Chem Comm
https://www.readbyqxmd.com/read/28921955/characterization-of-site-specific-glycosylation-in-influenza-a-virus-hemagglutinin-produced-by-spodoptera-frugiperda-insect-cell-line
#5
Yan-Jun Liu, Shiaw-Lin Wu, Kerry R Love, William S Hancock
Influenza hemagglutinin is a surface glycoprotein related to virus invasion and host immune system response. Understanding site specific glycosylation of hemagglutinin will increase our knowledge about virus evolution and can improve the design and quality of vaccines. In our study, we used glycoproteomic analysis based on multienzyme digestion followed by LC tandem MS analysis to determine the glycosylation of Influenza hemagglutinin (H1/A/California/04/2009) using the following steps: PNGaseF treatment combined with trypsin or pepsin digestion was used to determine the glycosites and glycan occupancy...
October 17, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28866761/t-protein-is-present-in-large-excess-over-the-other-proteins-of-the-glycine-cleavage-system-in-leaves-of-arabidopsis
#6
Stefan Timm, Jonas Giese, Nadja Engel, Maria Wittmiß, Alexandra Florian, Alisdair R Fernie, Hermann Bauwe
T-protein is present in large excess over the other proteins of the glycine cleavage system in leaves of Arabidopsis and therefore, exerts little control over the photorespiratory pathway. T-protein is the aminomethyltransferase of the glycine cleavage multienzyme system (GCS), also known as the glycine decarboxylase complex, and essential for photorespiration and one-carbon metabolism. Here, we studied what effects varying levels of the GCS T-protein would have on GCS activity, the operation of the photorespiratory pathway, photosynthesis, and plant growth...
September 2, 2017: Planta
https://www.readbyqxmd.com/read/28813159/enhancing-enzyme-activity-and-immobilization-in-nanostructured-inorganic-enzyme-complexes
#7
Xuye Lang, Lingling Zhu, Yingning Gao, Ian Wheeldon
Understanding the chemical and physical interactions at the interface of protein surfaces and inorganic crystals has important implications in the advancement of immobilized enzyme catalysis. Recently, enzyme-inorganic hybrid complexes have been demonstrated as effective materials for enzyme immobilization. The precipitation of phosphate nanocrystals in the presence of enzymes creates enzyme-Cu3(PO4)2·3H2O particles with high surface-to-volume ratios, enhanced activity, and increased stability. Here, we begin to develop a mechanistic understanding of enzyme loading in such complexes...
August 25, 2017: Langmuir: the ACS Journal of Surfaces and Colloids
https://www.readbyqxmd.com/read/28801642/extreme-anoxia-tolerance-in-crucian-carp-and-goldfish-through-neofunctionalization-of-duplicated-genes-creating-a-new-ethanol-producing-pyruvate-decarboxylase-pathway
#8
Cathrine E Fagernes, Kåre-Olav Stensløkken, Åsmund K Røhr, Michael Berenbrink, Stian Ellefsen, Göran E Nilsson
Without oxygen, most vertebrates die within minutes as they cannot meet cellular energy demands with anaerobic metabolism. However, fish of the genus Carassius (crucian carp and goldfish) have evolved a specialized metabolic system that allows them to survive prolonged periods without oxygen by producing ethanol as their metabolic end-product. Here we show that this has been made possible by the evolution of a pyruvate decarboxylase, analogous to that in brewer's yeast and the first described in vertebrates, in addition to a specialized alcohol dehydrogenase...
August 11, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28694846/synthetic-co2-fixation-enzyme-cascades-immobilized-on-self-assembled-nanostructures-that-enhance-co2-o2-selectivity-of-rubisco
#9
Sriram Satagopan, Yuan Sun, Jon R Parquette, F Robert Tabita
BACKGROUND: With increasing concerns over global warming and depletion of fossil-fuel reserves, it is attractive to develop innovative strategies to assimilate CO2, a greenhouse gas, into usable organic carbon. Cell-free systems can be designed to operate as catalytic platforms with enzymes that offer exceptional selectivity and efficiency, without the need to support ancillary reactions of metabolic pathways operating in intact cells. Such systems are yet to be exploited for applications involving CO2 utilization and subsequent conversion to valuable products, including biofuels...
2017: Biotechnology for Biofuels
https://www.readbyqxmd.com/read/28520394/in-vitro-metabolic-engineering-of-amorpha-4-11-diene-biosynthesis-at-enhanced-rate-and-specific-yield-of-production
#10
Xixian Chen, Congqiang Zhang, Ruiyang Zou, Gregory Stephanopoulos, Heng-Phon Too
In vitro metabolic engineering is an alternative approach to cell-based biosynthesis. It offers unprecedented flexibility for the study of biochemical pathways, thus providing useful information for the rational design and assembly of reaction modules. Herein, we took the advantage of in vitro metabolic engineering to initially gain insight into the regulatory network of a reconstituted amorpha-4,11-diene (AD) synthetic pathway. Guided by lin-log approximation, we rapidly identified the hitherto unrecognized inhibition of adenosine triphosphate (ATP) on both farnesyl pyrophosphate synthase (FPPS) and amorpha-4,11-diene synthase (ADS)...
September 15, 2017: ACS Synthetic Biology
https://www.readbyqxmd.com/read/28429326/wax-ester-fermentation-and-its-application-for-biofuel-production
#11
REVIEW
Hiroshi Inui, Takahiro Ishikawa, Masahiro Tamoi
In Euglena cells under anaerobic conditions, paramylon, the storage polysaccharide, is promptly degraded and converted to wax esters. The wax esters synthesized are composed of saturated fatty acids and alcohols with chain lengths of 10-18, and the major constituents are myristic acid and myristyl alcohol. Since the anaerobic cells gain ATP through the conversion of paramylon to wax esters, the phenomenon is named "wax ester fermentation". The wax ester fermentation is quite unique in that the end products, i...
2017: Advances in Experimental Medicine and Biology
https://www.readbyqxmd.com/read/28424264/identification-of-a-multienzyme-complex-for-glucose-metabolism-in-living-cells
#12
Casey L Kohnhorst, Minjoung Kyoung, Miji Jeon, Danielle L Schmitt, Erin L Kennedy, Julio Ramirez, Syrena M Bracey, Bao Tran Luu, Sarah J Russell, Songon An
Sequential metabolic enzymes in glucose metabolism have long been hypothesized to form multienzyme complexes that regulate glucose flux in living cells. However, it has been challenging to directly observe these complexes and their functional roles in living systems. In this work, we have used wide-field and confocal fluorescence microscopy to investigate the spatial organization of metabolic enzymes participating in glucose metabolism in human cells. We provide compelling evidence that human liver-type phosphofructokinase 1 (PFKL), which catalyzes a bottleneck step of glycolysis, forms various sizes of cytoplasmic clusters in human cancer cells, independent of protein expression levels and of the choice of fluorescent tags...
June 2, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28411640/expression-of-cellulolytic-enzyme-as-a-fusion-protein-that-reacts-specifically-with-a-polymeric-scaffold
#13
Priya Katyal, Yongkun Yang, Olga Vinogradova, Yao Lin
The formation of higher-order assemblies of multiple proteins or enzymes is a general mechanism to achieve more sophisticated biological function in biological systems. For example, cellulosomes are large complexes consisting of multiple cellulolytic enzymes that rely on the concerted actions of different enzymes built onto a common protein scaffold to facilitate the breakdown of the polymeric substrate, cellulose. One strategy for mimicking these highly effective nanomachines may involve the use of synthetic scaffolds that can react to and organize multiple engineered enzymes to promote synergistic action between the enzymes on the scaffold...
2017: Methods in Enzymology
https://www.readbyqxmd.com/read/28393943/enhancing-coupled-enzymatic-activity-by-conjugating-one-enzyme-to-a-nanoparticle
#14
James N Vranish, Mario G Ancona, Eunkeu Oh, Kimihiro Susumu, Igor L Medintz
Enzymes have long been a prime research target for the commercial production of commodity and specialty chemicals, design of sensing devices, and the development of therapeutics and new chemical processes. Industrial applications for enzymes can potentially be enhanced by enzyme immobilization which often allows for increased enzyme stability, facile product purification, and minimized substrate diffusion times in multienzymatic cascades, but this is usually at the cost of a significant decrease in catalytic rates...
April 20, 2017: Nanoscale
https://www.readbyqxmd.com/read/28259128/directed-regulation-of-multienzyme-complexes-of-2-oxo-acid-dehydrogenases-using-phosphonate-and-phosphinate-analogs-of-2-oxo-acids
#15
REVIEW
A V Artiukhov, A V Graf, V I Bunik
2-Oxo acid dehydrogenase complexes are important metabolic checkpoints functioning at the intercept of sugar and amino acid degradation. This review presents a short summary of architectural, catalytic, and regulatory principles of the complexes structure and function, based on recent advances in studies of well-characterized family members. Special attention is given to use of synthetic phosphonate and phosphinate analogs of 2-oxo acids as selective and efficient inhibitors of the cognate complexes in biological systems of bacterial, plant, and animal origin...
December 2016: Biochemistry. Biokhimii︠a︡
https://www.readbyqxmd.com/read/28203522/mass-spectral-determination-of-phosphopantetheinylation-specificity-for-carrier-proteins-in-mycobacterium-tuberculosis
#16
James Jung, Ghader Bashiri, Jodie M Johnston, Edward N Baker
Phosphopantetheinyl transferases (PPTases) are key elements in the modular syntheses performed by multienzyme systems such as polyketide synthases. PPTases transfer phosphopantetheine derivatives from Coenzyme A to carrier proteins (CPs), thus orchestrating substrate supply. We describe an efficient mass spectrometry-based protocol for determining CP specificity for a particular PPTase in organisms possessing several candidate PPTases. We show that the CPs MbtL and PpsC, both involved in synthesis of essential metabolites in Mycobacterium tuberculosis, are exclusively activated by the type 2 PPTase PptT and not the type 1 AcpS...
December 2016: FEBS Open Bio
https://www.readbyqxmd.com/read/28187530/structural-and-dynamic-characterization-of-a-freestanding-acyl-carrier-protein-involved-in-the-biosynthesis-of-cyclic-lipopeptide-antibiotics
#17
Subrata Paul, Hiroaki Ishida, Leonard T Nguyen, Zhihong Liu, Hans J Vogel
Friulimicin is a cyclic lipodecapeptide antibiotic that is produced by Actinoplanes friuliensis. Similar to the related lipopeptide drug daptomycin, the peptide skeleton of friulimicin is synthesized by a large multienzyme nonribosomal peptide synthetase (NRPS) system. The LipD protein plays a major role in the acylation reaction of friulimicin. The attachment of the fatty acid group promotes its antibiotic activity. Phylogenetic analysis reveals that LipD is most closely related to other freestanding acyl carrier proteins (ACPs), for which the genes are located near to NRPS gene clusters...
May 2017: Protein Science: a Publication of the Protein Society
https://www.readbyqxmd.com/read/28181062/progress-in-biocatalysis-with-immobilized-viable-whole-cells-systems-development-reaction-engineering-and-applications
#18
REVIEW
Milan Polakovič, Juraj Švitel, Marek Bučko, Jaroslav Filip, Vilém Neděla, Marion B Ansorge-Schumacher, Peter Gemeiner
Viable microbial cells are important biocatalysts in the production of fine chemicals and biofuels, in environmental applications and also in emerging applications such as biosensors or medicine. Their increasing significance is driven mainly by the intensive development of high performance recombinant strains supplying multienzyme cascade reaction pathways, and by advances in preservation of the native state and stability of whole-cell biocatalysts throughout their application. In many cases, the stability and performance of whole-cell biocatalysts can be highly improved by controlled immobilization techniques...
May 2017: Biotechnology Letters
https://www.readbyqxmd.com/read/28159788/revisiting-the-regulation-of-the-primary-scaffoldin-gene-in-clostridium-thermocellum
#19
Lizett Ortiz de Ora, Iván Muñoz-Gutiérrez, Edward A Bayer, Yuval Shoham, Raphael Lamed, Ilya Borovok
Cellulosomes are considered to be one of the most efficient systems for the degradation of plant cell wall polysaccharides. The central cellulosome component comprises a large, noncatalytic protein subunit called scaffoldin. Multiple saccharolytic enzymes are incorporated into the scaffoldins via specific high-affinity cohesin-dockerin interactions. Recently, the regulation of genes encoding certain cellulosomal components by multiple RNA polymerase alternative σ(I) factors has been demonstrated in Clostridium (Ruminiclostridium) thermocellum In the present report, we provide experimental evidence demonstrating that the C...
April 15, 2017: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/28138383/systematic-chemoenzymatic-synthesis-of-o-sulfated-sialyl-lewis-x-antigens
#20
Abhishek Santra, Hai Yu, Nova Tasnima, Musleh M Muthana, Yanhong Li, Jie Zeng, Nicholas J Kenyond, Angelique Y Louie, Xi Chen
O-Sulfated sialyl Lewis x antigens play important roles in nature. However, due to their structural complexity, they are not readily accessible by either chemical or enzymatic synthetic processes. Taking advantage of a bacterial sialyltransferase mutant that can catalyze the transfer of different sialic acid forms from the corresponding sugar nucleotide donors to Lewis x antigens which are fucosylated glycans as well as an efficient one-pot multienzyme (OPME) sialylation system, O-sulfated sialyl Lewis x antigens containing different sialic acid forms and O-sulfation at different locations were systematically synthesized by chemoenzymatic methods...
April 1, 2016: Chemical Science
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