OPTIMATION AND CHARACTERIZATION OF LPP-OMPA-LC-CUTINASE FUSSION PROTEIN AND IT̉̉S SYNONIMOUS MUTANT IN WHOLE-CELL BIOCATALYST ON ESCHERICHIA COLI EPI300TM FOR POLYETHYLENE TEREPHTHALATE (PET) DEGRADATION
Polyethylene therephthalate (PET) is a synthetic polyester composed of terephthalic acid and ethylene glycol. It is among the most commonly used polymer. The consumption of PET in Europe on 2015 reached 7% of total polymer usage (2,177 million Ton) and increased 10% annually. The majority of PET is...
Saved in:
| Main Author: | |
|---|---|
| Format: | Theses |
| Language: | Indonesia |
| Online Access: | https://digilib.itb.ac.id/gdl/view/24770 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| Institution: | Institut Teknologi Bandung |
| Language: | Indonesia |
| Summary: | Polyethylene therephthalate (PET) is a synthetic polyester composed of terephthalic acid and ethylene glycol. It is among the most commonly used polymer. The consumption of PET in Europe on 2015 reached 7% of total polymer usage (2,177 million Ton) and increased 10% annually. The majority of PET is used in production of plastic packaging, household and domestic product. PET is mostly used for textile fiber (60%) and plastic bottles (30%). Unavailability of efficient degradation technology and high usage demand with short period of time usage cause PET waste to build up. This build up usually end up on landfills and may cause major impact on environment and towards human, therefore proper degradation technology is urgently required. LC-cutinase ( leaf compost cutinase) that was discovered from metagenomic studies is known for its capability to degrade PET. LC-cutinase is a lipolytic/esterolytic enzyme from cutinase (EC 3.1.1.74) degrade PET by hydrolyzing PET. ITB_Indonesia iGEM team (2014) came up with a proposal of creating whole-cell biocatalyst system to degrade PET. It was done by expressing fusion protein of Lpp-OmpA-LC-cutinase on surface display based system. The aim of this research is to Optimize and characterize the previous system. Optimization of the system was conducted by synonimous mutation using OptimumGeneTm for design and synthesized by GenScript®. Characterization of the enzyme activity is than conducted done by measuring absorbance caused by the degradation of p-Nitrophenyl butyrate (pNPB) in 415 nm wave length. Based on this characterization, the whole-cell biocatalyst highest activity was measured to be 55oC and pH of 8. Synonimous mutation also increased the cell growth rate from 626.058 cells/second to 1914.858 cells/second. Mutant LC-cutinase also had higher thermal stability on pH 8. |
|---|