Characterization and regulation of salt upregulated cyclophilin from a halotolerant strain of Penicillium oxalicum
Abstract Penicillium species are an industrially important group of fungi. Cyclophilins are ubiquitous proteins and several members of this family exhibit peptidyl-prolyl cis–trans isomerase (PPIase) activity. We had earlier demonstrated that the salt-induced PPIase activity in a halotolerant strain...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2023-10-01
|
| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-44606-5 |
| _version_ | 1797559840246071296 |
|---|---|
| author | Mangaljeet Singh Harpreet Singh Kirandeep Kaur Shubhankar Shubhankar Supreet Singh Amarjeet Kaur Prabhjeet Singh |
| author_facet | Mangaljeet Singh Harpreet Singh Kirandeep Kaur Shubhankar Shubhankar Supreet Singh Amarjeet Kaur Prabhjeet Singh |
| author_sort | Mangaljeet Singh |
| collection | DOAJ |
| description | Abstract Penicillium species are an industrially important group of fungi. Cyclophilins are ubiquitous proteins and several members of this family exhibit peptidyl-prolyl cis–trans isomerase (PPIase) activity. We had earlier demonstrated that the salt-induced PPIase activity in a halotolerant strain of P. oxalicum was associated with enhanced expression of a cyclophilin gene, PoxCYP18. Cloning and characterization of PoxCYP18 revealed that its cDNA consists of 522 bp encoding a protein of 173 amino acid residues, with predicted molecular mass and pI values of 18.91 kDa and 8.87, respectively. The recombinant PoxCYP18 can catalyze cis–trans isomerization of peptidyl-prolyl bond with a catalytic efficiency of 1.46 × 107 M−1 s−1 and is inhibited specifically only by cyclosporin A, with an inhibition constant of 5.04 ± 1.13 nM. PoxCYP18 consists of two cysteine residues at positions − 45 and − 170, and loses its activity under oxidizing conditions. Substitution of these residues alone or together by site-directed mutagenesis revealed that the PPIase activity of PoxCYP18 is regulated through a redox mechanism involving the formation of disulfide linkages. Heterologous expression of PoxCYP18 conferred enhanced tolerance to salt stress in transgenic E. coli cells, implying that this protein imparts protection to cellular processes against salt-induced damage. |
| first_indexed | 2024-03-10T17:51:55Z |
| format | Article |
| id | doaj.art-83fd6027808c4de19504b0c165c26ddf |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-10T17:51:55Z |
| publishDate | 2023-10-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-83fd6027808c4de19504b0c165c26ddf2023-11-20T09:20:36ZengNature PortfolioScientific Reports2045-23222023-10-0113111510.1038/s41598-023-44606-5Characterization and regulation of salt upregulated cyclophilin from a halotolerant strain of Penicillium oxalicumMangaljeet Singh0Harpreet Singh1Kirandeep Kaur2Shubhankar Shubhankar3Supreet Singh4Amarjeet Kaur5Prabhjeet Singh6Department of Biotechnology, Guru Nanak Dev UniversityDepartment of Bioinformatics, Hans Raj Mahila Maha VidyalayaDepartment of Biotechnology, Guru Nanak Dev UniversityDepartment of Biotechnology, Guru Nanak Dev UniversityDepartment of Biotechnology, Guru Nanak Dev UniversityDepartment of Microbiology, Guru Nanak Dev UniversityDepartment of Biotechnology, Guru Nanak Dev UniversityAbstract Penicillium species are an industrially important group of fungi. Cyclophilins are ubiquitous proteins and several members of this family exhibit peptidyl-prolyl cis–trans isomerase (PPIase) activity. We had earlier demonstrated that the salt-induced PPIase activity in a halotolerant strain of P. oxalicum was associated with enhanced expression of a cyclophilin gene, PoxCYP18. Cloning and characterization of PoxCYP18 revealed that its cDNA consists of 522 bp encoding a protein of 173 amino acid residues, with predicted molecular mass and pI values of 18.91 kDa and 8.87, respectively. The recombinant PoxCYP18 can catalyze cis–trans isomerization of peptidyl-prolyl bond with a catalytic efficiency of 1.46 × 107 M−1 s−1 and is inhibited specifically only by cyclosporin A, with an inhibition constant of 5.04 ± 1.13 nM. PoxCYP18 consists of two cysteine residues at positions − 45 and − 170, and loses its activity under oxidizing conditions. Substitution of these residues alone or together by site-directed mutagenesis revealed that the PPIase activity of PoxCYP18 is regulated through a redox mechanism involving the formation of disulfide linkages. Heterologous expression of PoxCYP18 conferred enhanced tolerance to salt stress in transgenic E. coli cells, implying that this protein imparts protection to cellular processes against salt-induced damage.https://doi.org/10.1038/s41598-023-44606-5 |
| spellingShingle | Mangaljeet Singh Harpreet Singh Kirandeep Kaur Shubhankar Shubhankar Supreet Singh Amarjeet Kaur Prabhjeet Singh Characterization and regulation of salt upregulated cyclophilin from a halotolerant strain of Penicillium oxalicum Scientific Reports |
| title | Characterization and regulation of salt upregulated cyclophilin from a halotolerant strain of Penicillium oxalicum |
| title_full | Characterization and regulation of salt upregulated cyclophilin from a halotolerant strain of Penicillium oxalicum |
| title_fullStr | Characterization and regulation of salt upregulated cyclophilin from a halotolerant strain of Penicillium oxalicum |
| title_full_unstemmed | Characterization and regulation of salt upregulated cyclophilin from a halotolerant strain of Penicillium oxalicum |
| title_short | Characterization and regulation of salt upregulated cyclophilin from a halotolerant strain of Penicillium oxalicum |
| title_sort | characterization and regulation of salt upregulated cyclophilin from a halotolerant strain of penicillium oxalicum |
| url | https://doi.org/10.1038/s41598-023-44606-5 |
| work_keys_str_mv | AT mangaljeetsingh characterizationandregulationofsaltupregulatedcyclophilinfromahalotolerantstrainofpenicilliumoxalicum AT harpreetsingh characterizationandregulationofsaltupregulatedcyclophilinfromahalotolerantstrainofpenicilliumoxalicum AT kirandeepkaur characterizationandregulationofsaltupregulatedcyclophilinfromahalotolerantstrainofpenicilliumoxalicum AT shubhankarshubhankar characterizationandregulationofsaltupregulatedcyclophilinfromahalotolerantstrainofpenicilliumoxalicum AT supreetsingh characterizationandregulationofsaltupregulatedcyclophilinfromahalotolerantstrainofpenicilliumoxalicum AT amarjeetkaur characterizationandregulationofsaltupregulatedcyclophilinfromahalotolerantstrainofpenicilliumoxalicum AT prabhjeetsingh characterizationandregulationofsaltupregulatedcyclophilinfromahalotolerantstrainofpenicilliumoxalicum |