Esterification of Polymeric Carbohydrate Through Congener Cutinase-Like Biocatalyst
| Autor: | Mayilvahanan Aarthy, Numbi Ramudu Kamini, Puhazhendi Puhazhselvan, Niraikulam Ayyadurai, Perisamy Saravanan, Valappil Sisila, Girimanikandan Sakkeeshyaa |
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| Rok vydání: | 2020 |
| Předmět: |
0106 biological sciences
Cutinase Starch Bioengineering 01 natural sciences Applied Microbiology and Biotechnology Biochemistry Pentapeptide repeat Fungal Proteins chemistry.chemical_compound 010608 biotechnology Organic chemistry Lipase Molecular Biology chemistry.chemical_classification Esterification biology 010405 organic chemistry Fatty acid General Medicine 0104 chemical sciences Amino acid Cryptococcus Enzyme chemistry Biocatalysis biology.protein Carboxylic Ester Hydrolases Oleic Acid Biotechnology |
| Zdroj: | Applied Biochemistry and Biotechnology. 193:19-32 |
| ISSN: | 1559-0291 0273-2289 |
| Popis: | Cutinase-like enzymes (CLEs) are bi-functional hydrolases, which share the conserved catalytic site of lipase and consensus pentapeptide sequence of cutinase. Here, we have genetically replaced the canonical amino acids (CAA) by their non-canonical fluorinated surrogates to biosynthesize a novel class of congener biocatalyst for esterification of polymeric carbohydrate with long-chain fatty acid. It is a new enzyme-engineering approach used to manipulate industrially relevant biocatalyst through genetic incorporation of new functionally encoded non-canonical amino acids (NCAA). Global fluorination of CLE improved its catalytic, functional, and structural stability. Molecular docking studies confirmed that the fluorinated CLE (FCLE) had developed a binding affinity towards different fatty acids compared with the parent CLE. Importantly, FCLE could catalyze starch oleate synthesis in 24 h with a degree of substitution of 0.3 ± 0.001. Biophysical and microscopic analysis substantiated the efficient synthesis of the ester by FCLE. Our data represent the first step in the generation of an industrially relevant fluorous multifunctional enzyme for facile synthesis of high fatty acid starch esters. |
| Databáze: | OpenAIRE |
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