Combinatorial Library Based Engineering of <i>Candida antarctica</i> Lipase A for Enantioselective Transacylation of <i>sec</i>‐Alcohols in Organic Solvent

Wikmark, Ylva; Humble, Maria Svedendahl; Bäckvall, Jan‐E.

doi:10.1002/ange.201410675

Cited by 12 publications

(7 citation statements)

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“…It is reported that many lipases show promiscuous behavior, 4 and Candida antarctica lipase B (CALB) is one of the best wellknown highly promiscuous enzyme. CALB's primary activity comprises the hydrolysis of the ester bond in triacylglycerides (Figure 1A), but it is also capable of catalyzing a plethora of different reactions such as the synthesis of esters and amides, transacylation of alcohols, 13 epoxidation reactions, 14 aldol additions, Michael-type additions, 4,15 among many others. 16 As a carboxylic ester hydrolase, CALB performs its activity by a characteristic catalytic triad composed of Ser−His−Asp (Figure 1B).…”

Section: ■ Introductionmentioning

confidence: 99%

Exploring the Origin of Amidase Substrate Promiscuity in CALB by a Computational Approach

et al. 2019

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Enzyme promiscuity attracts the interest of the industrial and academic sectors because of its application in the design of biocatalysts. The amidase activity of Candida antarctica lipase B (CALB) on two different substrates has been studied by theoretical quantum mechanics/molecular mechanics methods, supported by experimental kinetic measurements. The aim of the study is to understand the substrate promiscuity of CALB in this secondary reaction and the origin of its promiscuous catalytic activity. The computational results predict activation free energies in very good agreement with the kinetic data and confirm that the activity of CALB as an amidase, despite depending on the features of the amide substrate, is dictated by the electrostatic effects of the protein. The protein polarizes and activates the substrate as well as stabilizes the transition state, thus enhancing the rate constant. Our results can provide guides for future designs of biocatalysts based on electrostatic arguments.

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Section: ■ Introductionmentioning

confidence: 99%

Exploring the Origin of Amidase Substrate Promiscuity in CALB by a Computational Approach

et al. 2019

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“…36,79 However, one lipase derived from Candida antarctica has been reported to be useful for transacylation of secondary alcohols in organic reagents. 80 Based on this, Wu and colleagues 81 successfully constructed four enzymes capable of catalyzing the preparation of four stereoisomeric 1-phenylethyl 2-phenylpropionates (53a) using a strategy termed "focused rational iterative site-specific mutation" from wild-type C. antarctica lipase B (CALB, Scheme 21). This strategy aims to simplify traditional iterative saturation mutations with fewer but representative amino acids, with the hope of similar good result.…”

Section: Preparation Of 1-phenylethyl 2-phenylpropanoates and Its Der...mentioning

confidence: 99%

Enzyme- and Chemo-enzyme-Catalyzed Stereodivergent Synthesis

Ding

Huang

et al. 2022

Pharmaceutical Fronts

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Multiple stereoisomers can be found when a substance contains chiral carbons in its chemical structure. To obtain the desired stereoisomers, asymmetric synthesis was proposed in the 1970s and developed rapidly at the beginning of this century. Stereodivergent synthesis, an extension of asymmetric synthesis in organic synthesis with the hope to produce all stereoisomers of chiral substances in high conversion and selectivity, enriches the variety of available products and serves as a reference suggestion for the synthesis of their derivatives and other compounds. Since biocatalysis has outstanding advantages of economy, environmental friendliness, high efficiency, and reaction at mild conditions, the biocatalytic reaction is regarded as an efficient strategy to perform stereodivergent synthesis. Thus, in this review, we summarize the stereodivergent synthesis catalyzed by enzymes or chemo-enzymes in cases where a compound contains two or three chiral carbons, i.e., at most four or eight stereoisomers are present. The types of reactions, including reduction of substituent ketones, cyclization reactions, olefin addition, and nonredox transesterification reactions, are also discussed for the understanding of the progress and application of biocatalysis in stereodivergent synthesis.

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“…A microtiter screening not involving spectrophotometric methods was carried out for the improvement of the enantioselective transacylation of sec-alcohols by CAL A in organic solvents. The library obtained was analyzed by chiral gas chromatography (GC-FID) (Wikmark, Svedendahl Humble & Backvall, 2015).…”

Section: Library Screeningmentioning

confidence: 99%

Protein engineering of enzymes involved in lipid modification

Oroz‐Guinea

Zorn

Brundiek³

2018

Lipid Modification by Enzymes and Engineered Microbes

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This chapter provides an overview of the most important protein engineering tools and strategies applied to enzymes involved in lipid modification. Thus, application of several methodologies is discussed, including directed evolution, rational protein design, construction of chimeric enzymes and de novo design, together with the utility of bioinformatic tools along the protein engineering process. Additionally, different approaches for the creation of semi-rational libraries and the potential use of different high-throughput screenings for the detection of improved protein variants are described. The examples provided cover the recent achievements in the utilization of these techniques on a broad variety of enzymes. This comprises not only engineered lipases with altered fatty acid chain length selectivity, fatty acid specificity and improved performance in esterification reactions, but also successfully altered phospholipases, lipoxygenases, P450 monooxygenases, decarboxylating enzymes and fatty acid hydratases.

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Combinatorial Library Based Engineering of Candida antarctica Lipase A for Enantioselective Transacylation of sec‐Alcohols in Organic Solvent

Cited by 12 publications

References 58 publications

Exploring the Origin of Amidase Substrate Promiscuity in CALB by a Computational Approach

Exploring the Origin of Amidase Substrate Promiscuity in CALB by a Computational Approach

Enzyme- and Chemo-enzyme-Catalyzed Stereodivergent Synthesis

Protein engineering of enzymes involved in lipid modification

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