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000638202 005__ 20250918160252.0
000638202 037__ $$aPUBDB-2025-04020
000638202 041__ $$aEnglish
000638202 1001_ $$0P:(DE-HGF)0$$aSigurdsson, Gudmundur$$b0$$eCorresponding author$$gmale
000638202 245__ $$aOverexpression and Purification of SLPI, a proteaseinhibitor from Streptomyces Lividans, in E. coli$$f2024-01-01 - 2025-03-31
000638202 260__ $$c2025
000638202 300__ $$a1-65
000638202 3367_ $$2DataCite$$aOutput Types/Dissertation
000638202 3367_ $$2ORCID$$aDISSERTATION
000638202 3367_ $$2BibTeX$$aPHDTHESIS
000638202 3367_ $$02$$2EndNote$$aThesis
000638202 3367_ $$0PUB:(DE-HGF)11$$2PUB:(DE-HGF)$$aDissertation / PhD Thesis$$bphd$$mphd$$s1758203691_3299243
000638202 3367_ $$2DRIVER$$adoctoralThesis
000638202 502__ $$aDissertation, University of Iceland, 2025$$bDissertation$$cUniversity of Iceland$$d2025
000638202 520__ $$aAbstractThis project focuses on the overexpression and purification of Streptomyces lividansprotease inhibitor (SLPI) in transformed E. coli to compare the efficiency of two methods ofexpression induction. SLPI belongs to the Streptomyces subtilisin inhibitor (SSI) proteinfamily, naturally found in S. lividans. Those methods are the IPTG method and autoinductionmethod. A cloned pET vector containing genomic DNA for SLPI will be transformed intoLemo21 E. coli cells, overexpressed, and then purified through a five-step process, includingsalt precipitation and column purification. The total amount of protein and SLPI producedusing either method will be compared using several analyses. Each step of the purificationprocess for both methods will be analyzed using a Bradford assay to determine proteinconcentration. SDS-PAGE will be run on each step to verify concentration results for thepresence of a protein of SLPI’s size and its purity. SLPI presence will also be verifiedthrough a serine protease (VPR) inhibitor activity assay in the presence of a color-changingsubstrate sample. The characteristics of the isolated protein at the end of purification will bemeasured using CD to determine the mean residue ellipticity (MRE) and melting point (TM).This was compared to the reference MSc thesis and showed that SLPI was isolated.Additionally, SAXS characterization of SLPI will be used. Overexpression of SLPI usingthe autoinduction method was more efficient than the IPTG method, primarily due tofacilitating a richer and more optimized expression environment for the E. coli culture thanthe manual IPTG induction method.
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000638202 693__ $$0EXP:(DE-H253)P-P12-20150101$$1EXP:(DE-H253)PETRAIII-20150101$$6EXP:(DE-H253)P-P12-20150101$$aPETRA III$$fPETRA Beamline P12$$x0
000638202 8564_ $$uhttps://skemman.is/handle/1946/50297?locale=en
000638202 8564_ $$uhttps://bib-pubdb1.desy.de/record/638202/files/Overexpression%20and%20Purification%20of%20SLPI%20a%20protease%20inhibitor%20from%20Streptomyces%20Lividans%20in%20E%20coli.pdf$$yRestricted
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