Sökning: WFRF:(Huang Mingtao 1984)
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High-throughput scr...
High-throughput screening for industrial enzyme production hosts by droplet microfluidics
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- Sjöström, Staffan L. (författare)
- KTH,Proteomik och nanobioteknologi,Science for Life Laboratory, SciLifeLab,Kungliga Tekniska Högskolan (KTH),Royal Institute of Technology (KTH)
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- Bai, Yunpeng (författare)
- KTH,Proteomik och nanobioteknologi,Science for Life Laboratory, SciLifeLab,Kungliga Tekniska Högskolan (KTH),Royal Institute of Technology (KTH)
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- Huang, Mingtao, 1984 (författare)
- Chalmers tekniska högskola,Chalmers University of Technology
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- Liu, Zihe, 1984 (författare)
- Chalmers tekniska högskola,Chalmers University of Technology
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- Nielsen, Jens (författare)
- KTH,Proteomik och nanobioteknologi,Science for Life Laboratory, SciLifeLab,Chalmers tekniska högskola,Chalmers University of Technology
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- Jönsson, Håkan N. (författare)
- KTH,Proteomik och nanobioteknologi,Science for Life Laboratory, SciLifeLab,Kungliga Tekniska Högskolan (KTH),Royal Institute of Technology (KTH)
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- Andersson Svahn, Helene (författare)
- KTH,Proteomik och nanobioteknologi,Science for Life Laboratory, SciLifeLab,Kungliga Tekniska Högskolan (KTH),Royal Institute of Technology (KTH)
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(creator_code:org_t)
- 2014
- 2014
- Engelska.
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Ingår i: Lab on a Chip. - : Royal Society of Chemistry (RSC). - 1473-0197 .- 1473-0189. ; 14:4, s. 806-813
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http://dx.doi.org/10...
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https://urn.kb.se/re...
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Abstract
Ämnesord
Stäng
- A high-throughput method for single cell screening by microfluidic droplet sorting is applied to a whole-genome mutated yeast cell library yielding improved production hosts of secreted industrial enzymes. The sorting method is validated by enriching a yeast strain 14 times based on its a-amylase production, close to the theoretical maximum enrichment. Furthermore, a 105 member yeast cell library is screened yielding a clone with a more than 2-fold increase in a-amylase production. The increase in enzyme production results from an improvement of the cellular functions of the production host in contrast to previous droplet-based directed evolution that has focused on improving enzyme protein structure. In the workflow presented, enzyme producing single cells are encapsulated in 20 pL droplets with a fluorogenic reporter substrate. The coupling of a desired phenotype (secreted enzyme concentration) with the genotype (contained in the cell) inside a droplet enables selection of single cells with improved enzyme production capacity by droplet sorting. The platform has a throughput over 300 times higher than that of the current industry standard, an automated microtiter plate screening system. At the same time, reagent consumption for a screening experiment is decreased a million fold, greatly reducing the costs of evolutionary engineering of production strains.
Ämnesord
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
- TEKNIK OCH TEKNOLOGIER -- Industriell bioteknik (hsv//swe)
- ENGINEERING AND TECHNOLOGY -- Industrial Biotechnology (hsv//eng)
- TEKNIK OCH TEKNOLOGIER -- Kemiteknik (hsv//swe)
- ENGINEERING AND TECHNOLOGY -- Chemical Engineering (hsv//eng)
Nyckelord
- Saccharomyces-Cerevisiae
- Directed Evolution
- Microdroplets
- Selection
- Systems
- Assays
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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