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biosensor enables yeast glucose

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https://www.readbyqxmd.com/read/26060079/electrochemical-glucose-biosensor-based-on-glucose-oxidase-displayed-on-yeast-surface
#1
Hongwei Wang, Qiaolin Lang, Bo Liang, Aihua Liu
The conventional enzyme-based biosensor requires chemical or physical immobilization of purified enzymes on electrode surface, which often results in loss of enzyme activity and/or fractions immobilized over time. It is also costly. A major advantage of yeast surface display is that it enables the direct utilization of whole cell catalysts with eukaryote-produced proteins being displayed on the cell surface, providing an economic alternative to traditional production of purified enzymes. Herein, we describe the details of the display of glucose oxidase (GOx) on yeast cell surface and its application in the development of electrochemical glucose sensor...
2015: Methods in Molecular Biology
https://www.readbyqxmd.com/read/25984720/an-enzyme-coupled-biosensor-enables-s-reticuline-production-in-yeast-from-glucose
#2
William C DeLoache, Zachary N Russ, Lauren Narcross, Andrew M Gonzales, Vincent J J Martin, John E Dueber
Benzylisoquinoline alkaloids (BIAs) are a diverse family of plant-specialized metabolites that include the pharmaceuticals codeine and morphine and their derivatives. Microbial synthesis of BIAs holds promise as an alternative to traditional crop-based manufacturing. Here we demonstrate the production of the key BIA intermediate (S)-reticuline from glucose in Saccharomyces cerevisiae. To aid in this effort, we developed an enzyme-coupled biosensor for the upstream intermediate L-3,4-dihydroxyphenylalanine (L-DOPA)...
July 2015: Nature Chemical Biology
https://www.readbyqxmd.com/read/24613019/ultra-high-throughput-screening-method-for-the-directed-evolution-of-glucose-oxidase
#3
Raluca Ostafe, Radivoje Prodanovic, Jovana Nazor, Rainer Fischer
Glucose oxidase (GOx) is used in many industrial processes that could benefit from improved versions of the enzyme. Some improvements like higher activity under physiological conditions and thermal stability could be useful for GOx applications in biosensors and biofuel cells. Directed evolution is one of the currently available methods to engineer improved GOx variants. Here, we describe an ultra-high-throughput screening system for sorting the best enzyme variants generated by directed evolution that incorporates several methodological refinements: flow cytometry, in vitro compartmentalization, yeast surface display, fluorescent labeling of the expressed enzyme, delivery of glucose substrate to the reaction mixture through the oil phase, and covalent labeling of the cells with fluorescein-tyramide...
March 20, 2014: Chemistry & Biology
https://www.readbyqxmd.com/read/21504769/immobilized-formaldehyde-metabolizing-enzymes-from-hansenula-polymorpha-for-removal-and-control-of-airborne-formaldehyde
#4
Sasi Sigawi, Oleh Smutok, Olha Demkiv, Oksana Zakalska, Galina Gayda, Yeshayahu Nitzan, Marina Nisnevitch, Mykhaylo Gonchar
Formaldehyde (FA)-containing indoor air has a negative effect on human health and should be removed by intensive ventilation or by catalytic conversion to non-toxic products. FA can be oxidized by alcohol oxidase (AOX) taking part in methanol metabolism of methylotrophic yeasts. In the present work, AOX isolated from a Hansenula polymorpha C-105 mutant (gcr1 catX) overproducing this enzyme in glucose medium, was tested for its ability to oxidize airborne FA. A continuous fluidized bed bioreactor (FBBR) was designed to enable an effective bioconversion of airborne FA by AOX or by permeabilized mutant H...
May 20, 2011: Journal of Biotechnology
https://www.readbyqxmd.com/read/20798321/phosphatidic-acid-is-a-ph-biosensor-that-links-membrane-biogenesis-to-metabolism
#5
Barry P Young, John J H Shin, Rick Orij, Jesse T Chao, Shu Chen Li, Xue Li Guan, Anthony Khong, Eric Jan, Markus R Wenk, William A Prinz, Gertien J Smits, Christopher J R Loewen
Recognition of lipids by proteins is important for their targeting and activation in many signaling pathways, but the mechanisms that regulate such interactions are largely unknown. Here, we found that binding of proteins to the ubiquitous signaling lipid phosphatidic acid (PA) depended on intracellular pH and the protonation state of its phosphate headgroup. In yeast, a rapid decrease in intracellular pH in response to glucose starvation regulated binding of PA to a transcription factor, Opi1, that coordinately repressed phospholipid metabolic genes...
August 27, 2010: Science
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