Cloning, expression and characterisation of a novel cold-adapted GDSL family esterase from Photobacterium sp. strain J15
The gene encoding for a novel cold-adapted enzyme from family II of bacterial classification (GDSL family) was cloned from the genomic DNA of Photobacterium sp. strain J15 in an Escherichia coli system, yielding a recombinant 36 kDa J15 GDSL esterase which was purified in two steps with a final yiel...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Springer
2016
|
| Online Access: | http://psasir.upm.edu.my/id/eprint/55300/1/Cloning%2C%20expression%20and%20characterization%20of%20a%20novel%20cold-adapted%20GDSL%20family%20esterase%20.pdf |
| _version_ | 1825931191477338112 |
|---|---|
| author | Shakiba, Mehrnoush Hadaddzadeh Mohamad Ali, Mohd Shukuri Raja Abd Rahman, Raja Noor Zaliha Salleh, Abu Bakar Thean, Chor Leow |
| author_facet | Shakiba, Mehrnoush Hadaddzadeh Mohamad Ali, Mohd Shukuri Raja Abd Rahman, Raja Noor Zaliha Salleh, Abu Bakar Thean, Chor Leow |
| author_sort | Shakiba, Mehrnoush Hadaddzadeh |
| collection | UPM |
| description | The gene encoding for a novel cold-adapted enzyme from family II of bacterial classification (GDSL family) was cloned from the genomic DNA of Photobacterium sp. strain J15 in an Escherichia coli system, yielding a recombinant 36 kDa J15 GDSL esterase which was purified in two steps with a final yield and purification of 38.6 and 15.3 respectively. Characterization of the biochemical properties showed the J15 GDSL esterase had maximum activity at 20 °C and pH 8.0, was stable at 10 °C for 3 h and retained 50 % of its activity after a 6 h incubation at 10 °C. The enzyme was activated by Tween-20, -60 and Triton-X100 and inhibited by 1 mM Sodium dodecyl sulphate (SDS), while β-mercaptoethanol and Dithiothreitol (DTT) enhanced activity by 4.3 and 5.4 fold respectively. These results showed the J15 GDSL esterase was a novel cold-adapted enzyme from family II of lipolytic enzymes. A structural model constructed using autotransporter EstA from Pseudomonas aeruginosa as a template revealed the presence of a typical catalytic triad consisting of a serine, aspartate, and histidine which was verified with site directed mutagenesis on active serine. |
| first_indexed | 2024-03-06T09:23:00Z |
| format | Article |
| id | upm.eprints-55300 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-03-06T09:23:00Z |
| publishDate | 2016 |
| publisher | Springer |
| record_format | dspace |
| spelling | upm.eprints-553002017-11-07T02:46:26Z http://psasir.upm.edu.my/id/eprint/55300/ Cloning, expression and characterisation of a novel cold-adapted GDSL family esterase from Photobacterium sp. strain J15 Shakiba, Mehrnoush Hadaddzadeh Mohamad Ali, Mohd Shukuri Raja Abd Rahman, Raja Noor Zaliha Salleh, Abu Bakar Thean, Chor Leow The gene encoding for a novel cold-adapted enzyme from family II of bacterial classification (GDSL family) was cloned from the genomic DNA of Photobacterium sp. strain J15 in an Escherichia coli system, yielding a recombinant 36 kDa J15 GDSL esterase which was purified in two steps with a final yield and purification of 38.6 and 15.3 respectively. Characterization of the biochemical properties showed the J15 GDSL esterase had maximum activity at 20 °C and pH 8.0, was stable at 10 °C for 3 h and retained 50 % of its activity after a 6 h incubation at 10 °C. The enzyme was activated by Tween-20, -60 and Triton-X100 and inhibited by 1 mM Sodium dodecyl sulphate (SDS), while β-mercaptoethanol and Dithiothreitol (DTT) enhanced activity by 4.3 and 5.4 fold respectively. These results showed the J15 GDSL esterase was a novel cold-adapted enzyme from family II of lipolytic enzymes. A structural model constructed using autotransporter EstA from Pseudomonas aeruginosa as a template revealed the presence of a typical catalytic triad consisting of a serine, aspartate, and histidine which was verified with site directed mutagenesis on active serine. Springer 2016-01 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/55300/1/Cloning%2C%20expression%20and%20characterization%20of%20a%20novel%20cold-adapted%20GDSL%20family%20esterase%20.pdf Shakiba, Mehrnoush Hadaddzadeh and Mohamad Ali, Mohd Shukuri and Raja Abd Rahman, Raja Noor Zaliha and Salleh, Abu Bakar and Thean, Chor Leow (2016) Cloning, expression and characterisation of a novel cold-adapted GDSL family esterase from Photobacterium sp. strain J15. Extremophiles, 20 (1). pp. 45-55. ISSN 1431-0651; ESSN: 1433-4909 10.1007/s00792-015-0796-4 |
| spellingShingle | Shakiba, Mehrnoush Hadaddzadeh Mohamad Ali, Mohd Shukuri Raja Abd Rahman, Raja Noor Zaliha Salleh, Abu Bakar Thean, Chor Leow Cloning, expression and characterisation of a novel cold-adapted GDSL family esterase from Photobacterium sp. strain J15 |
| title | Cloning, expression and characterisation of a novel cold-adapted GDSL family esterase from Photobacterium sp. strain J15 |
| title_full | Cloning, expression and characterisation of a novel cold-adapted GDSL family esterase from Photobacterium sp. strain J15 |
| title_fullStr | Cloning, expression and characterisation of a novel cold-adapted GDSL family esterase from Photobacterium sp. strain J15 |
| title_full_unstemmed | Cloning, expression and characterisation of a novel cold-adapted GDSL family esterase from Photobacterium sp. strain J15 |
| title_short | Cloning, expression and characterisation of a novel cold-adapted GDSL family esterase from Photobacterium sp. strain J15 |
| title_sort | cloning expression and characterisation of a novel cold adapted gdsl family esterase from photobacterium sp strain j15 |
| url | http://psasir.upm.edu.my/id/eprint/55300/1/Cloning%2C%20expression%20and%20characterization%20of%20a%20novel%20cold-adapted%20GDSL%20family%20esterase%20.pdf |
| work_keys_str_mv | AT shakibamehrnoushhadaddzadeh cloningexpressionandcharacterisationofanovelcoldadaptedgdslfamilyesterasefromphotobacteriumspstrainj15 AT mohamadalimohdshukuri cloningexpressionandcharacterisationofanovelcoldadaptedgdslfamilyesterasefromphotobacteriumspstrainj15 AT rajaabdrahmanrajanoorzaliha cloningexpressionandcharacterisationofanovelcoldadaptedgdslfamilyesterasefromphotobacteriumspstrainj15 AT sallehabubakar cloningexpressionandcharacterisationofanovelcoldadaptedgdslfamilyesterasefromphotobacteriumspstrainj15 AT theanchorleow cloningexpressionandcharacterisationofanovelcoldadaptedgdslfamilyesterasefromphotobacteriumspstrainj15 |