Identification of a Novel Lipase with AHSMG Pentapeptide in Hypocreales and Glomerellales Filamentous Fungi

Abstract

Lipases are enzymes that hydrolyze triglycerides to fatty acids and glycerol. A typical element in lipases is a conserved motif of five amino acids (the pentapeptide), most commonly G-X-S-X-G. Lipases with the pentapeptide A-X-S-X-G are present in species of Bacillus, Paucimonas lemoignei, and the yeast Trichosporon asahii; they are usually thermotolerant and solvent resistant. Recently, while searching for true lipases in the Trichoderma harzianum genome, one lipase containing the pentapeptide AHSMG was identified. In this study, we cloned from T. harzianum strain B13-1 the lipase ID135964, renamed here as ThaL, which is 97.65% identical with the reference. We found that ThaL is a lid-containing true lipase of cluster III that belongs to a large family comprising highly conserved proteins in filamentous fungi in the orders Hypocreales and Glomerellales, in which predominantly pathogenic fungi are found. ThaL was expressed in conidia, as well as in T. harzianum mycelium, where it was cultured in liquid minimal medium. These results—together with the amino acid composition, absence of a signal peptide, mitochondrial sorting prediction, disordered regions in the protein, and lineage-specific phylogenetic distribution of its homologs—suggest that ThaL is a non-canonical effector. In summary, AHSMG-lipase is a novel lipase family in filamentous fungi, and is probably involved in pathogenicity.

FungiRegEx: A Tool for Pattern Identification in Fungal Proteomic Sequences Using Regular Expressions

“…The data presented in this study are available in JGI Mycocosm Database at https://mycocosm.jgi.doe.gov/Mycgr3/Mycgr3.home.html (accessed on 16 May 2024) for Mycosphaerella graminicola, reference number [26] and https://mycocosm.jgi.doe.gov/SacceM3 836_1/SacceM3836_1.home.html (accessed on 16 May 2024) for Saccharomyces cerevisiae, reference number [17][18][19][20]. These data were derived from the following resources available in the public domain: https://mycocosm.jgi.doe.gov/mycocosm/home (accessed on 16 May 2024).…”

“…This means that FungiRegEx will launch the scrapper instances to retrieve the data in the JGI Mycocosm database from 2000 proteomes. FungiRegEx was validated and tested by searching the sequence AXSXG as a regular expression, where AXSXG is a pentapeptide of a lipase group that brings thermostability and resistance to solvents of an enzyme [18] and that has been little described in fungi. The considerations to validate and test FungiRegEx with this sequence are as follows:…”

N-terminal lid swapping contributes to the substrate specificity and activity of thermophilic lipase TrLipE

“…Based on their thermostability, organic solvent tolerance, and excellent catalytic ability under extreme conditions, lipases have been widely used in the food ( Asmat et al, 2019 ; Cipolatti et al, 2020 ), detergent, leather ( Verma et al, 2021 ), pharmaceutical, and biodiesel industries ( Cavalcante et al, 2020 ; Zhong et al, 2020 ; Almeida et al, 2021 ). Lipases have a highly conserved catalytic triad consisting of nucleophilic serine or cysteine, aspartic or glutamic acid, and histidine ( Anobom et al, 2014 ; Gutiérrez-Domínguez et al, 2022 ; Liu et al, 2022 ). Most lipases display a very similar fold in which mixed β-sheets are surrounded by α-helices and a domain named lid that controls the substrate channel to the active pocket ( Mohd Din et al, 2021 ; Xing et al, 2021 ; Cui et al, 2022 ).…”