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Cloning, sequencing and expression of the lipase gene from Pseudomonas fragi IFO-12049 in E. coli
FEBS Letters
|December 19, 1988
Abstract:
The lipase gene from Pseudomonas fragi IFO-12049 was isolated using the expression library and the primary structure of lipase deduced from the nucleotide sequence was determined. It is composed of 277 amino acid residues and a protein of Mr 29,966, which was close to the value of the lipase expressed in E. coli.
Insights
Researchers isolated the lipase gene from Pseudomonas fragi IFO-12049. The deduced primary structure revealed a protein of 277 amino acids and a molecular weight of 29,966, similar to E. coli expressed lipase.
Area of Science:
- Biochemistry
- Molecular Biology
- Microbial Genetics
Background:
- Pseudomonas fragi is a bacterium known to produce enzymes.
- Lipases are crucial enzymes with diverse industrial applications.
- Understanding lipase gene structure is key to enzyme engineering.
Purpose of the Study:
- To isolate and characterize the lipase gene from Pseudomonas fragi IFO-12049.
- To determine the primary structure and molecular weight of the lipase.
- To compare the deduced lipase properties with those expressed in other systems.
Main Methods:
- Gene isolation using an expression library.
- Nucleotide sequencing to deduce the primary amino acid sequence.
- Bioinformatic analysis for protein molecular weight determination.
Main Results:
- The lipase gene from Pseudomonas fragi IFO-12049 was successfully isolated.
- The deduced protein consists of 277 amino acid residues.
- The calculated molecular weight (Mr 29,966) closely matched experimental values for E. coli expressed lipase.
Conclusions:
- The primary structure of the Pseudomonas fragi lipase has been elucidated.
- The characterized lipase shows potential for biotechnological applications.
- This study provides a foundation for further investigation and modification of this lipase.