Characterization of a GDS(L)-like hydrolase from Pleurotus sapidus with an unusual SGNH motif

dc.contributor.authorFingerhut, Miriam A.
dc.contributor.authorHenrich, Lea
dc.contributor.authorLauber, Christiane
dc.contributor.authorBroel, Niklas
dc.contributor.authorGhezellou, Parviz
dc.contributor.authorKarrer, Dominik
dc.contributor.authorSpengler, Bernhard
dc.contributor.authorLangfelder, Kim
dc.contributor.authorStressler, Timo
dc.contributor.authorZorn, Holger
dc.contributor.authorGand, Martin
dc.date.accessioned2025-03-21T08:01:21Z
dc.date.available2025-03-21T08:01:21Z
dc.date.issued2024
dc.description.abstractThe GDS(L)-like lipase from the Basidiomycota Pleurotus sapidus (PSA_Lip) was heterologously expressed using Trichoderma reesei with an activity of 350 U L−1. The isoelectric point of 5.0 was determined by isoelectric focusing. The novel PSA_Lip showed only 23.8–25.1%, 25.5%, 26.6% and 28.4% identity to the previously characterized GDSL-like enzymes phospholipase, plant lipase, acetylcholinesterase and acetylxylan esterase, from the carbohydrate esterase family 16, respectively. Therefore, the enzyme was purified from the culture supernatant and the catalytic properties and the substrate specificity of the enzyme were investigated using different assays to reveal its potential function. While no phospholipase, acetylcholinesterase and acetylxylan esterase activities were detected, studies on the hydrolysis of ferulic acid methyl ester (~ 8.3%) and feruloylated carbohydrate 5-O-transferuloyl-arabino-furanose (~ 0.8%) showed low conversions of these substrates. By investigating the hydrolytic activity towards p-nitrophenyl-(pNP)-esters with various chain-lengths, the highest activity was determined for medium chain-length pNP-octanoate at 65 °C and a pH value of 8, while almost no activity was detected for pNP-hexanoate. The enzyme is highly stable when stored at pH 10 and 4 °C for at least 7 days. Moreover, using consensus sequence analysis and homology modeling, we could demonstrate that the PSA_Lip does not contain the usual SGNH residues in the actives site, which are usually present in GDS(L)-like enzymes.en
dc.description.sponsorshipDeutsche Forschungsgemeinschaft (DFG); ROR-ID:018mejw64
dc.identifier.urihttps://jlupub.ub.uni-giessen.de/handle/jlupub/20396
dc.identifier.urihttps://doi.org/10.22029/jlupub-19747
dc.language.isoen
dc.rightsNamensnennung 4.0 International
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.subject.ddcddc:540
dc.titleCharacterization of a GDS(L)-like hydrolase from Pleurotus sapidus with an unusual SGNH motif
dc.typearticle
local.affiliationFB 08 - Biologie und Chemie
local.projectINST 162/500–1 FUGG; “Expression, crystallization and protein engineering of lipases from basidiomycetes for a better understanding of their sequence- and structure-guided specificity and their applications in cheese”- 689680 (PN 514527182).
local.source.articlenumber98
local.source.epage14
local.source.journaltitleAMB express
local.source.spage1
local.source.urihttps://doi.org/10.1186/s13568-024-01752-x
local.source.volume14

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