Auto-activated Thrombin Expression from Pichia pastorisSMD1168 Optimized by Plackett-Burman Design and Response Surface Methodology
Journal Title: IOSR Journal of Pharmacy (IOSRPHR) - Year 2017, Vol 7, Issue 9
Abstract
Abstract: Recombinant thrombin (rThrombin) has been used topically as one component of fibrin sealant as the tissue adhesive. Recombinant prethrombin-2 is used as the precursor to generate rThrombin which is then activated by ecarin. Autoactivated thrombin (autoTrm) can be produced by altering four amino acids (EDGE) in prethrombin-2 thus obviate the use of ecarin and simplify the purification process. In this study, Pichia pastorisSMD1168 was used to producing autoTrm in flask culture. Influencing factors on P. pastorisSMD1168 culture in producing autoTRM were screened by Plackett-Burman (PB) design and the optimization of was carried out by Response Surface Methodology. The medium used for cultivation was BMGH for growth phase and Pichia Trace Minerals (PTM1) supplemented Basal Salt Medium (BSM) for induction phase. The influencing factors (PTM1 concentration, methanol concentration, induction temperature,induction time, and pH) on autoTrmexpression werescreenedby PB design then were optimized by Response Surface Methodology (RSM) with Central Composite Design.It was revealed that PTM1 concentration (%) and pH had significant influence (P < 0.05) on autoTrm expression level based on PB design. The optimum%PTM1 and pH revealed by (RSM) was 2.05% and 4.2, respectively
Authors and Affiliations
Muhammad Fadhlillah, Wulan Pertiwi, Ersanda Hafiz, Sheila Chairunnisa, Eria S. Widyaningsih, Inten Ihsannunisa, Toto Subroto
Keywords
Related Articles
- EP ID EP253443
- DOI -
- Views 89
- Downloads 0
Evalution of Anti Microbial Activity and Phytochemical analysis of different organic solvent of Calotropis gigantea
The leaf and fruits extract of Calotropis gigantea were screened for its anti microbial and phytochemical activities. The solvent used for the leaves and fruits extraction were n-haxane, benzene, acetonic, eth...
Theoretical Spectral investigation of a bioactive Benzenoid Isolated from Mimusops elengi
The bioactive aromatic aldehyde, vanillin (3-methoxy 4-hydroxy benzaldehyde) is isolated from stem bark of the medicinal plant Mimusops elengi for the first time. Elucidation of structure is achieved with the hel...
[b] A comparative evaluation of in vitro growth inhibitory activities of different solvent extracts of some medicinal plants in Northern Ghana against selected human pathogens[/b]
Normative Data of Motor Free Visual Perception Test among Indian Children.
OBJECTIVE. The purpose of this study was to obtain normative data for Motor-Free Visual Perception Test (MVPT) in Chennai children.To find out the performance difference between genders. METHOD. Three Hundred and Thirty...
Protective effect of curcumin and picroliv on chronic alcoholic liver cirrhosis of rat induced with low caloric diet
The present investigation aims at evaluation of the protective efficacy of picroliv and curcumin alone and combination against chronic ethanol exposure induced liver damage. Alcoholic liver cirrhosis was induced in the r...