Ferulic acid esterase-III fromAspergillus niger does not exhibit lipase activity

Aliwan, Fraj O; Kroon, Paul A.; Faulds, Craig B.; Pickersgill, Richard W.; Williamson, Gary

doi:10.1002/(sici)1097-0010(19990301)79:3<457::aid-jsfa283>3.0.co;2-g

Cited by 21 publications

(3 citation statements)

References 10 publications

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“…More generally, the considerable degree of lid rigidity displayed by the Esterase variant could be a distinguishing factor between a lipase and an esterase enzyme. Whilst most lipases exhibit lid flexibility, which is necessary to allow lid displacement (interfacial activation) and allow substrate access of long-chain triglycerides (Reis et al, 2009b;Sarda and Desnuelle, 1958), esterases generally do not require displacement of helical regions, where short-chain ester substrates are able to access the active site without conformational rearrangement (Aliwan et al, 1999;McAuley et al, 2004;Yu et al, 2014). This may be why the SMD simulations were unable to sample the lid opening process for the Esterase variant, suggesting that the lid mutations affect both the overall activity of the lipase, as well as its structural dynamics such as overall lid flexibility.…”

Section: Discussionmentioning

confidence: 99%

The effect of mutations in the lid region of Thermomyces lanuginosus lipase on interactions with triglyceride surfaces: A multi-scale simulation study

Willems

Lelimousin

Skjold-Jørgensen

et al. 2018

Chemistry and Physics of Lipids

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Lipases naturally function at the interface formed between amphiphilic molecules and the aqueous environment. Thermomyces lanuginosus lipase (TLL) is a well-characterised lipase, known to exhibit interfacial activation during which a lid region covering the active site becomes displaced upon interaction with an interface. In this study, we investigate the effect the amino acid sequence of the lid region on interfacial binding and lid dynamics of TLL. Three TLL variants were investigated, a wild-type variant, a variant containing an esterase lid region (Esterase), and a Hybrid variant, containing both wild-type lid residues and esterase lid residues. Multiple coarse-grained molecular dynamics simulations revealed that the interfacial binding orientation of TLL was significantly affected by the nature of amino acids in the lid region, and atomistic simulations indicated effects on the structural dynamics of the lid itself. The atomistic simulations, as well as steered molecular dynamics simulations, also indicated that the Esterase lid region was less flexible than the wild-type lid region, whereas the Hybrid variant displayed superior lid flexibility and stability in the open conformation both at the interface, and in aqueous solution. Additional experiments performed to investigate the activity and binding behaviour of the lipase variants indicated a slightly higher specific activity for the Hybrid variant compared to the wild-type variant, correlating the observations of increased lid flexibility. Together, these results are in line with previous experimental studies, highlighting the importance of the nature of the amino acid residues within the functional lid region of lipases, particularly regarding interfacial binding orientation, activation, and structural stability.

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Section: Discussionmentioning

confidence: 99%

The effect of mutations in the lid region of Thermomyces lanuginosus lipase on interactions with triglyceride surfaces: A multi-scale simulation study

Willems

Lelimousin

Skjold-Jørgensen

et al. 2018

Chemistry and Physics of Lipids

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“…Lipases can exist in two conformational states: an active form where the catalytic residues are exposed and an inactive form where a helical lid involving residues 82±96 (TlL numbering; Brzozowski et al, 2000) covers the active site. (Aliwan et al, 1999), it is unlikely that this helix needs to adopt two conformations. Although the overall structures are similar, the active-site regions have to be different in order to accommodate the differences in substrates.…”

Section: Comparison With Lipases and Bacterial Faesmentioning

confidence: 99%

“…FAE-III contains the characteristic lipase serine active-site motif and, given the sequence similarities, it was predicted that FAE-III would have considerable structural homology and a similar catalytic mechanism to the fungal lipases. However, FAE-III has been shown not to possess lipase activity (Aliwan et al, 1999).…”

Section: Introductionmentioning

confidence: 99%

Structure of a feruloyl esterase fromAspergillus niger

McAuley

Svendsen

Patkar

et al. 2004

Acta Crystallogr D Biol Crystallogr

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The crystallographic structure of feruloyl esterase from Aspergillus niger has been determined to a resolution of 1.5 A Ê by molecular replacement. The protein has an /-hydrolase structure with a Ser-His-Asp catalytic triad; the overall fold of the protein is very similar to that of the fungal lipases. The structure of the enzyme±product complex was determined to a resolution of 1.08 A Ê and reveals dual conformations for the serine and histidine residues at the active site.

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Understanding the activation mechanism of Thermomyces lanuginosus lipase using rational design and tryptophan‐induced fluorescence quenching

Skjold-Jørgensen

Vind

Svendsen

et al. 2016

Euro J Lipid Sci & Tech

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The attractiveness of lipases as industrial biocatalysts underlines the importance of understanding their molecular action in detail, helping future efforts to improve performance and applicability. Lipases represent a special class of carboxyl ester hydrolases, which act on long‐chained water insoluble acyl‐glycerols at the water‐lipid interface. This heterogeneous catalysis makes it very difficult to monitor the structural changes taking place in the enzyme during activation in its preferred milieu. In this review, we cover recent developments toward a deeper understanding of the interfacial activation phenomenon of lipases and discuss a selection of biophysical methods suitable for studying lipase activation. For many lipases, such as that from Thermomyces lanuginosus, a structural domain called the “lid” is directly involved in the activation process. This has been shown through alteration of the lid‐residue composition using rational design. The resulting lid‐mutants have highlighted the lid's role in governing enzymatic activity and interfacial activation. Moreover, a recently developed fluorescence‐based technique called the Tryptophan Induced Quenching method has proved to enable the direct measurement of lipase activation in water‐ethanol mixtures, and thus represents an intriguing method to gain further insight into the structural movements taking place at the water‐lipid interface. Practical applications: Lipases are important industrial biocatalysts widely used in industrial applications including detergent, leather, paper, fuel, fine chemicals, baking, cosmetics, and food. Knowledge of structural changes of lipase upon activation and identification of the amino acids involved in lipase activation is crucial for design of new variants with improved or new properties. Rational design of the lid‐region in lipases has an interesting potential for optimization of lipase function and adapting them to different environments. In order to leverage on the design of new lipase variants and their altered function, methods that enable direct measurement of lipase action directly in solution, in real time, are desired. In this context, fluorescence based methods have shown great potential and provides interesting perspectives for probing the delicate movements occurring in lipases upon activation. Interfacial activation of lipases and the lid movements studied by rational design, fluorescence quenching methods, and solvent effect – polarity driven activation.

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Ferulic acid esterase-III fromAspergillus niger does not exhibit lipase activity

Cited by 21 publications

References 10 publications

The effect of mutations in the lid region of Thermomyces lanuginosus lipase on interactions with triglyceride surfaces: A multi-scale simulation study

The effect of mutations in the lid region of Thermomyces lanuginosus lipase on interactions with triglyceride surfaces: A multi-scale simulation study

Structure of a feruloyl esterase fromAspergillus niger

Understanding the activation mechanism of Thermomyces lanuginosus lipase using rational design and tryptophan‐induced fluorescence quenching

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