1st International and 10th National Iranian Conference on Bioinformatics
Home Accepted Papers
A Computational Approach to Modeling a Thermophilic Psychrophilic Protease Fusion Protein for Laundry Detergent
Paper ID : 1337-ICB10
Authors:
Atena Abed *1, Ladan Mafakher2, Hamed Mirzaei3, Elham Rismani4
1Cellular and Molecular Research Center, Shahrekord University of Medical Sciences, Shahrekord, Iran
2Thalassemia & amp; Hemoglobinopathy Research center, Health research institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
34Research Center for Biochemistry and Nutrition in Metabolic Diseases, Institute for Basic Sciences, Kashan University of Medical Sciences, Kashan, Iran
4Molecular Medicine Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran
Abstract:
Abstract
Protease is one of the main enzyme which is used in laundry industry. As the thermal condition in lanundry industry is varied from high to low temperature, having protease enzyme which could activate in broad thermal range has great potential. One of the approach in this field is fusion two proteins with different thermal activity[1]. For doing this in current work by computational method, Thermolysin protease an extracellular metalloendopeptidase from a gram-positive thermophilic bacterium Bacillus thermoproteolyticus which activate in thermal range from 25 to 88°C[2],[3] was joined by a four repeat of G4S as a flexible linker[4] with a psychrophilic protease from a marine bacterium Flavobacterium YS-80 with thermal range from 0°C to 30°C[5],[6] to produce protease enzyme which activate in broad thermal range from 0 to 88° C.
Material and methods:
As the whole structure of thermolysin did not exist in PDB bank, homology modeling by Lomet webserver was performed and Modeller package was applied to construct whole protein fusion structure. The quality of model structure was verified by Procheck[7], Verify3D[8], ERRAT[9], ProSA[10] webserver. The secondary structure analysis was done by PSIPRED[11]. The solubility and physiochemical properties of fusion protein was determined by Protein-sol[12] and Protparam webserver[13]. Molecular Dynamics simulation was perfomed by Gromacs package to assess stability of fusion protein structure in simulated condition.
Result and Discussion:
The quality structure assessment of fusion protein showed that this protein modelled with high quality compared to non-fusion form. Also, secondary structure analysis revealed that no secondary structure changes was observed in fusion form compared to non-fusion form. The physicochemical properties of fusion protein figured out this structure had stable structure and good solubility in water. the Molecular dynamic simulation result discovered that no drastic structure changes was happened during simulation time.
Keywords:
protease, protein engineering, homology modeling, molecular dynamics simulation
Status : Paper Accepted (Poster Presentation)