Simultaneous improvements in the activity and stability of Candida antarctica lipase B through multiple-site mutagenesis

Yagonia, Camila Flor J.; Park, Hyun June; Hong, So Yeon; Yoo, Young Je

doi:10.1007/s12257-014-0706-0

Cited by 18 publications

(12 citation statements)

References 26 publications

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“…The influence of CalB mutations in helices α5 and α10 on esterase and lipase activity toward short-chain and long-chain para-nitrophenyl (pNP) compounds is well established. Yagonita et al found that V139E and A151D mutations yielded a 5-fold increase in activity against pNP caprylate. Circular permutations that significantly altered the substrate recognition or adsorption interface of CalB yielded multiple variants with increased activity toward pNP butyrate and DiFMU octanoate .…”

Section: Results and Discussionmentioning

confidence: 99%

Self-Assembly Nanostructures of Triglyceride–Water Interfaces Determine Functional Conformations of Candida antarctica Lipase B

Benson

Pleiss

2017

Langmuir

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Candida antarctica lipase B (CalB) acts as a lipase when adsorbed to an acylglyceride interface and as an esterase when exposed to an aqueous environment. The effect of the molecular self-assembly nanostructure of triglyceride-water interfaces on structural conformations of adsorbed CalB and the implications to its catalytic function were studied by molecular dynamics simulations. Systems of CalB adsorbed to interfaces and solvated in water were compared. The two environments induced relative motions of helices α5 and α10 that resulted in open and closed conformations. The open conformation was stabilized by interactions between the polar and nonpolar amino acids of α5 and α10 and the nanostructure of triglyceride aggregates, which self-assembled into crystalline-like patterns of alternating polar and nonpolar lamellae. Thus, the structure of CalB has been adapted by evolution to the geometric constraints imposed by the interface nanostructure for optimized catalytic activity. Helices α5 and α10 have two functions. As mobile elements, they ensure access of bulky substrates to the active site in the open conformation. As a part of the active site pocket, they ensure binding of substrate molecules in a productive orientation near the active site. In water, access to the binding site is limited, and the smaller substrate binding site is beneficial for the binding of small, water-soluble substrates. The CalB crystal structure commonly used for protein engineering studies represents an intermediate state between open and closed, and may thus not be adequate to assess the function of CalB, neither as lipase nor as esterase.

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Section: Results and Discussionmentioning

confidence: 99%

Self-Assembly Nanostructures of Triglyceride–Water Interfaces Determine Functional Conformations of Candida antarctica Lipase B

Benson

Pleiss

2017

Langmuir

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“…Mutations which increased rigidity in the solvent affecting region (A92E and T245S) showed higher methanol stability, but were less catalytically active, than the variants with activity enhancing mutations (V139E and A151D) and vice versa. The combination of both mutations resulted in mutants (V139E, A92E and V139E, T245S) that were more active and stable in presence of methanol (Yagonia et al 2015).…”

Section: Residue Flexibilitymentioning

confidence: 99%

Solvent stable microbial lipases: current understanding and biotechnological applications

Priyanka¹,

Tan²,

Kinsella³

et al. 2018

Biotechnol Lett

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Objective: This review examines on our current understanding of microbial lipase solvent tolerance, with a specific focus on the molecular strategies employed to improve lipase stability in a non-aqueous environment. Results: It provides an overview of known solvent tolerant lipases and of approaches to improving solvent stability such as; enhancing stabilising interactions, modification of residue flexibility and surface charge alteration. It shows that judicious selection of lipase source supplemented by appropriate enzyme stabilisation, can lead to a wide application spectrum for lipases. Conclusion: Organic solvent stable lipases are, and will continue to be, versatile and adaptable biocatalytic workhorses commonly employed for industrial applications in the food, pharmaceutical and green manufacturing industries.

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“…One approach to engineering enzymes is to introduce mutations at their active sites when the structures are available, in particular for the residues to interact with substrates (Yagonia et al, 2015;Lee et al, 2019). In addition, understanding the catalytic mechanism may provide useful insights into how a particular amino acid residue functions in enzymatic reactions.…”

Section: Engineering Of Coenzyme B 12 -Dependent Gdht Improvement In mentioning

confidence: 99%

Recent Progress in the Understanding and Engineering of Coenzyme B12-Dependent Glycerol Dehydratase

Nasır

Ashok

Shim

et al. 2020

Front. Bioeng. Biotechnol.

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Coenzyme B 12-dependent glycerol dehydratase (GDHt) catalyzes the dehydration reaction of glycerol in the presence of adenosylcobalamin to yield 3-hydroxypropanal (3-HPA), which can be converted biologically to versatile platform chemicals such as 1,3-propanediol and 3-hydroxypropionic acid. Owing to the increased demand for biofuels, developing biological processes based on glycerol, which is a byproduct of biodiesel production, has attracted considerable attention recently. In this review, we will provide updates on the current understanding of the catalytic mechanism and structure of coenzyme B 12-dependent GDHt, and then summarize the results of engineering attempts, with perspectives on future directions in its engineering.

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Simultaneous improvements in the activity and stability of Candida antarctica lipase B through multiple-site mutagenesis

Cited by 18 publications

References 26 publications

Self-Assembly Nanostructures of Triglyceride–Water Interfaces Determine Functional Conformations of Candida antarctica Lipase B

Self-Assembly Nanostructures of Triglyceride–Water Interfaces Determine Functional Conformations of Candida antarctica Lipase B

Solvent stable microbial lipases: current understanding and biotechnological applications

Recent Progress in the Understanding and Engineering of Coenzyme B12-Dependent Glycerol Dehydratase

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