TY - JOUR

T1 - Sesquiterpene Cyclases from the Basidiomycete Cerrena unicolor

AU - Püth, Nils

AU - Ersoy, Franziska

AU - Krings, Ulrich

AU - Berger, Ralf G.

N1 - Funding Information: Funding: The APC was funded by the Open Access fund of the Gottfried Wilhelm Leibniz Universität Hannover.

PY - 2021/11/12

Y1 - 2021/11/12

N2 - Hundreds of terpenoids have been isolated from Basidiomycota, among them are volatile mono-and sesquiterpenes with amazing sensory qualities, representing a promising alternative to essential oils from endangered plant species. Sesquiterpene synthases (STS) appear to be an abundant class of enzymes in these fungi. The basidiomycete Cerrena unicolor, a known sesquiterpene producer, was in silico screened for sesquiterpene cyclases via homology Basic Local Alignment Search Tool searches. Cyclase genes identified were cloned and heterologously expressed in Escherichia coli Bl21 using pCOLD I as the expression vector. Ten cyclases were successfully produced and purified, and their identity was confirmed using amino acid sequencing of tryptic peptides by nano-liquid chromatography-high resolution-electrospray ionization-tandem mass spectrome-try. Gas chromatography/mass spectrometry analysis was applied to characterize these cyclases according to the formation of sesquiterpene hydrocarbons and oxidized terpenoids. Bioinformatic characterization and phylogenetic determination allowed for the classification of these diverse fungal enzymes. A representative single and a multi-product STS, respectively, were further analyzed for their dependency from divalent metal cations as a cofactor for the catalytic activity.

AB - Hundreds of terpenoids have been isolated from Basidiomycota, among them are volatile mono-and sesquiterpenes with amazing sensory qualities, representing a promising alternative to essential oils from endangered plant species. Sesquiterpene synthases (STS) appear to be an abundant class of enzymes in these fungi. The basidiomycete Cerrena unicolor, a known sesquiterpene producer, was in silico screened for sesquiterpene cyclases via homology Basic Local Alignment Search Tool searches. Cyclase genes identified were cloned and heterologously expressed in Escherichia coli Bl21 using pCOLD I as the expression vector. Ten cyclases were successfully produced and purified, and their identity was confirmed using amino acid sequencing of tryptic peptides by nano-liquid chromatography-high resolution-electrospray ionization-tandem mass spectrome-try. Gas chromatography/mass spectrometry analysis was applied to characterize these cyclases according to the formation of sesquiterpene hydrocarbons and oxidized terpenoids. Bioinformatic characterization and phylogenetic determination allowed for the classification of these diverse fungal enzymes. A representative single and a multi-product STS, respectively, were further analyzed for their dependency from divalent metal cations as a cofactor for the catalytic activity.

KW - Cerrena unicolor

KW - Heterolo-gous expression

KW - Metal ion dependency

KW - Phylogenetic analysis

KW - Sesquiterpenes

KW - Terpene cyclases

UR - http://www.scopus.com/inward/record.url?scp=85118893457&partnerID=8YFLogxK

U2 - 10.3390/catal11111361

DO - 10.3390/catal11111361

M3 - Article

VL - 11

JO - CATALYSTS

JF - CATALYSTS

SN - 2073-4344

IS - 11

M1 - 1361

ER -