Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Bioanalytical advantages of a novel recombinant apyrase enzyme in ATP-based bioluminescence methods
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science. KTH, Centres, Science for Life Laboratory, SciLifeLab.
Show others and affiliations
2018 (English)In: Analytica Chimica Acta, ISSN 0003-2670, E-ISSN 1873-4324, Vol. 1025, p. 118-123Article in journal (Refereed) Published
Abstract [en]

Ultrasensitive measurements of intracellular ATP (intATP) based on the firefly luciferase reactions are frequently used to enumerate bacterial or mammalian cells. During clinical applications, extracellular ATP (extATP) should be depleted in biological samples since it interferes with intATP and affects the quantification of bacteria. The extATP can be eliminated by ATP-degrading enzymes but complete hydrolysis of extATP remains a challenge for today's commercial enzymes. The catalytic efficiency of ATP-degrading enzymes depends on enzyme characteristics, sample composition and the ability to deplete diphosphates, triphosphates and their complexes generated during the reaction. This phenomenon restricts the usage of bioluminescence-based ATP methods in clinical diagnostics. In light of this, we have developed a recombinant Shigella flexneri apyrase (RSFA) enzyme and analysed its ATP depletion potential with five commercial biochemical sources including potato apyrase, acid phosphatase, alkaline phosphatase, hexokinase and glycerol kinase. The RSFA revealed superior activity by completely eliminating the extracellular ATP and ATP-complexes, even in biological samples like urine and serum. Therefore, our results can potentially unwrap the chemical and bio-analytical applications of ATP-based bioluminescence tests to develop highly sensitive point-of-care diagnostics.

Place, publisher, year, edition, pages
Elsevier, 2018. Vol. 1025, p. 118-123
Keywords [en]
ATP-depletion assays, ATP-diphosphohydrolase, Commercial apyrase, Extracellular ATP removal, Rapid biomedical diagnostics, Shigella spp.
National Category
Biocatalysis and Enzyme Technology
Identifiers
URN: urn:nbn:se:kth:diva-229269DOI: 10.1016/j.aca.2018.04.054ISI: 000433083200010PubMedID: 29801599Scopus ID: 2-s2.0-85046638940OAI: oai:DiVA.org:kth-229269DiVA, id: diva2:1212103
Funder
Science for Life Laboratory - a national resource center for high-throughput molecular bioscience
Note

QC 20180601

Available from: 2018-06-01 Created: 2018-06-01 Last updated: 2018-06-13Bibliographically approved

Open Access in DiVA

No full text in DiVA

Other links

Publisher's full textPubMedScopus

Authority records BETA

Zelenin, SergeyRussom, Aman

Search in DiVA

By author/editor
Zelenin, SergeyArdabili, SaharRussom, Aman
By organisation
Protein ScienceScience for Life Laboratory, SciLifeLab
In the same journal
Analytica Chimica Acta
Biocatalysis and Enzyme Technology

Search outside of DiVA

GoogleGoogle Scholar

doi
pubmed
urn-nbn

Altmetric score

doi
pubmed
urn-nbn
Total: 4 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf