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dc.contributor.authorChacha, Wambura E.
dc.contributor.authorTran, Huu-Tuan
dc.contributor.authorScarlett, William R.
dc.contributor.authorHutchison, Justin M.
dc.identifier.citationChacha WE, Tran HT, Scarlett WR, Hutchison JM. Extraction of active, contaminant degrading enzymes from soil. Appl Soil Ecol. 2023 Jul;187:104841. doi: 10.1016/j.apsoil.2023.104841. Epub 2023 Feb 17. PMID: 37786531; PMCID: PMC10544838en_US
dc.description.abstractSoil microorganisms play critical roles in the degradation of micro-and nano-pollutants, and the corresponding proteins and enzymes play roles in pollutant recognition, transportation, and degradation. Our ability to study these pathways from soil samples is often complicated by the complex processes involved in extracting proteins from soil matrices. This study aimed to develop a new protein soil extraction protocol that yielded active, intracellular enzymes from the perchlorate degradation pathway, particularly perchlorate reductase. An indirect method, which focused on first separating the cells from the soil matrix, followed by cell lysis and enzyme extraction, was evaluated. The optimized indirect method achieved a final extraction efficiency of the active enzyme and total protein of 15.7 % and 3.3 %, respectively. The final step of separating enzymes from residual soil components resulted in the highest activity and protein losses of 67.7 % ± 14.8 % and 91.8 % ± 1.8 %, respectively. Five buffers, each at different concentrations (0.01 M, 0.05 M, and 0.1 M), were tested to enhance enzyme extraction efficiency. The best extractant requires careful consideration between the highest activity and the quality of the recovered enzymes. Coextraction of humic substances could be minimized by using 0.1 M as compared to 0.01 M and 0.05 M of sodium pyrophosphate; however, this resulted in less recovered activity compared to lower extractant concentrations.en_US
dc.publisherUS Department of Health and Human Servicesen_US
dc.rightsPMC Copyright notice This is an open access article under the CC BY license (
dc.subjectIntracellular enzymesen_US
dc.subjectPerchlorate reductaseen_US
dc.subjectSoil microorganismsen_US
dc.subjectHumic substancesen_US
dc.titleExtraction of active, contaminant degrading enzymes from soilen_US
kusw.kuauthorHutchison, Justin M.
kusw.kudepartmentCivil, Environmental and Architectural Engineering Departmenten_US
kusw.oaversionScholarly/refereed, publisher versionen_US
kusw.oapolicyThis item meets KU Open Access policy criteria.en_US

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PMC Copyright notice
This is an open access article under the CC BY license (
Except where otherwise noted, this item's license is described as: PMC Copyright notice This is an open access article under the CC BY license (