Esterification of Polymeric Carbohydrate Through Congener Cutinase-Like Biocatalyst
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Esterification of Polymeric Carbohydrate Through Congener Cutinase-Like Biocatalyst Valappil Sisila 1,2 & Puhazhendi Puhazhselvan 1 & Mayilvahanan Aarthy 1 & Girimanikandan Sakkeeshyaa 3 & Perisamy Saravanan 3 & Numbi Ramudu Kamini 1,2 & Niraikulam Ayyadurai 1,2 Received: 30 March 2020 / Accepted: 12 August 2020/ # Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract
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. Keywords Cutinase-like enzyme . Global incorporation . Fluorination . Non-canonical amino acid . Esterification . Starch
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s12010-02003415-6) contains supplementary material, which is available to authorized users.
* Niraikulam Ayyadurai [email protected]
1
Division of Biochemistry and Biotechnology, Council of Scientific and Industrial Research (CSIR), Central Leather Research Institute (CLRI), Chennai, India
2
Academy of Scientific and Innovative Research, Ghaziabad, India
3
Department of Biotechnology, Rajalakshmi Engineering College, Chennai, India
Applied Biochemistry and Biotechnology
Introduction Synthesis of starch fatty acid esters has gained momentum owing to its application in different industries and production from renewable resources. Esterification of starch with long-chain fatty acid yields an ester with thermoplastic, hydrophobic, and biodegradable properties [1]. These esters are increasingly used as essential commodity chemicals in food, plastic, pharmaceutical, and biomedical sectors [2]. Although different chemical methods have been reported for the synthesis of starch esters, bioesterification using enzymes is a widely adopted process due to its eco-friendly features of operation under mild reaction conditions [3, 4]. In particular, cutinaselike enzymes (CLEs) of the serine hydrolases superfamily have garnered atte
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