Bacterial lipoxygenases: Biochemical characteristics, molecular structure and potential applications

Chrisnasari, Ruth; Hennebelle, Marie; Vincken, Jean‐Paul; Berkel, W.J.H. van; Ewing, Tom A.

doi:10.1016/j.biotechadv.2022.108046

Cited by 20 publications

(10 citation statements)

References 128 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The question remains as to the interplay between the emergence of quantum effects and the evolution of thermal networks starting with ancestral enzymes where tunneling is expected to be less efficient and protein scaffolds are more thermostable . Future studies in this laboratory will be aimed at investigating more evolutionarily distant LOXs, including those from prokaryotes that present varying sizes, catalytic properties, and N-terminal structural domains . Resolving the relationships between productive, protein thermal motions and effective wave function overlap for hydrogen tunneling in catalysis is expected to provide guidelines to enhance the design of proteins that exploit nontrivial quantum phenomena for their biological output.…”

Section: Discussionmentioning

confidence: 99%

“…83 Future studies in this laboratory will be aimed at investigating more evolutionarily distant LOXs, including those from prokaryotes that present varying sizes, catalytic properties, and N-terminal structural domains. 84 Resolving the relationships between productive, protein thermal motions and effective wave function overlap for hydrogen tunneling in catalysis is expected to provide guidelines to enhance the design of proteins that exploit nontrivial quantum phenomena for their biological output.…”

Section: Mutation-dependent Tdhdx-ms Identifies Two Distinct Network ...mentioning

confidence: 99%

See 1 more Smart Citation

Identification of the Thermal Activation Network in Human 15-Lipoxygenase-2: Divergence from Plant Orthologs and Its Relationship to Hydrogen Tunneling Activation Barriers

Ohler,

Taylor,

Bledsoe

et al. 2024

ACS Catal.

View full text Add to dashboard Cite

The oxidation of polyunsaturated fatty acids by lipoxygenases (LOXs) is initiated by a C−H cleavage step in which the hydrogen atom is transferred quantum mechanically (i.e., via tunneling). In these reactions, protein thermal motions facilitate the conversion of ground-state enzyme−substrate complexes to tunneling-ready configurations and are thus important for transferring energy from the solvent to the active site for the activation of catalysis. In this report, we employed temperaturedependent hydrogen−deuterium exchange mass spectrometry (TDHDX-MS) to identify catalytically linked, thermally activated peptides in a representative animal LOX, human epithelial 15-LOX-2. TDHDX-MS of wild-type 15-LOX-2 was compared to two active site mutations that retain structural stability but have increased activation energies (E a ) of catalysis. The E a value of one variant, V427L, is implicated to arise from suboptimal substrate positioning by increased active-site side chain rotamer dynamics, as determined by X-ray crystallography and ensemble refinement. The resolved thermal network from the comparative E a s of TDHDX-MS between wild-type and V426A is localized along the front face of the 15-LOX-2 catalytic domain. The network contains a clustering of isoleucine, leucine, and valine side chains within the helical peptides. This thermal network of 15-LOX-2 is different in location, area, and backbone structure compared to a model plant lipoxygenase from soybean that exhibits a low E a value of catalysis compared to the human ortholog. The presented data provide insights into the divergence of thermally activated protein motions in plant and animal LOXs and their relationships to the enthalpic barriers for facilitating hydrogen tunneling.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Mutation-dependent Tdhdx-ms Identifies Two Distinct Network ...mentioning

confidence: 99%

Identification of the Thermal Activation Network in Human 15-Lipoxygenase-2: Divergence from Plant Orthologs and Its Relationship to Hydrogen Tunneling Activation Barriers

Ohler,

Taylor,

Bledsoe

et al. 2024

ACS Catal.

View full text Add to dashboard Cite

show abstract

“…Lipoxygenases (LOs) are a family of enzymes that catalyze the addition of oxygen to polyunsaturated fatty acids (PUFAs), specifically those containing a 1,4-pentadiene structure [ 245 ]. This reaction results in the formation of hydroperoxides, which can then be further metabolized into a variety of bioactive molecules, including leukotrienes, hydroxyeicosatetraenoic acids (HETEs), and fatty acid epoxides [ 246 ]. Los are found in a wide range of organisms, including plants, animals, and fungi, and dysregulation of LO activity can have significant implications in various diseases [ 247 ].…”

Section: Polyphenol-mediated Enzyme Regulationmentioning

confidence: 99%

Antioxidant Metabolism Pathways in Vitamins, Polyphenols, and Selenium: Parallels and Divergences

Andrés,

Pérez de la Lastra,

Juan

et al. 2024

IJMS

View full text Add to dashboard Cite

Free radicals (FRs) are unstable molecules that cause reactive stress (RS), an imbalance between reactive oxygen and nitrogen species in the body and its ability to neutralize them. These species are generated by both internal and external factors and can damage cellular lipids, proteins, and DNA. Antioxidants prevent or slow down the oxidation process by interrupting the transfer of electrons between substances and reactive agents. This is particularly important at the cellular level because oxidation reactions lead to the formation of FR and contribute to various diseases. As we age, RS accumulates and leads to organ dysfunction and age-related disorders. Polyphenols; vitamins A, C, and E; and selenoproteins possess antioxidant properties and may have a role in preventing and treating certain human diseases associated with RS. In this review, we explore the current evidence on the potential benefits of dietary supplementation and investigate the intricate connection between SIRT1, a crucial regulator of aging and longevity; the transcription factor NRF2; and polyphenols, vitamins, and selenium. Finally, we discuss the positive effects of antioxidant molecules, such as reducing RS, and their potential in slowing down several diseases.

show abstract

“…LOX catalyzes the oxygenation of fatty acids through four reaction steps: hydrogen abstraction, radical rearrangement, oxygen insertion, and radical reduction-protonation. The figure is adopted from Chrisnasari et al ( 2022 ) with slight modification …”

Section: Introductionmentioning

confidence: 99%

“…The industrial potential of LOX has led to the exploration of this enzyme from various sources. Among these sources, microbial LOXs have gained increasing attention in recent years for their ability to act on a wide range of PUFAs (Banthiya et al 2016 ; Newie et al 2016 ; An et al 2018 ; Goloshchapova et al 2018 ; Qi et al 2020 ; Chrisnasari et al 2022 ; Kim et al 2022 ) and some of them were optimally active at extreme pH values (Qian et al 2017 ; Goloshchapova et al 2018 ). These advantages stimulate their exploitation as an industrial biocatalyst in a wide range of applications.…”

Section: Introductionmentioning

confidence: 99%

Versatile ferrous oxidation–xylenol orange assay for high-throughput screening of lipoxygenase activity

Chrisnasari,

Ewing,

Hilgers

et al. 2024

Appl Microbiol Biotechnol

Self Cite

View full text Add to dashboard Cite

Lipoxygenases (LOXs) catalyze dioxygenation of polyunsaturated fatty acids (PUFAs) into fatty acid hydroperoxides (FAHPs), which can be further transformed into a number of value-added compounds. LOXs have garnered interest as biocatalysts for various industrial applications. Therefore, a high-throughput LOX activity assay is essential to evaluate their performance under different conditions. This study aimed to enhance the suitability of the ferrous-oxidized xylenol orange (FOX) assay for screening LOX activity across a wide pH range with different PUFAs. The narrow linear detection range of the standard FOX assay restricts its utility in screening LOX activity. To address this, the concentration of perchloric acid in the xylenol orange reagent was adjusted. The modified assay exhibited a fivefold expansion in the linear detection range for hydroperoxides and accommodated samples with pH values ranging from 3 to 10. The assay could quantify various hydroperoxide species, indicating its applicability in assessing LOX substrate preferences. Due to sensitivity to pH, buffer types, and hydroperoxide species, the assay required calibration using the respective standard compound diluted in the same buffer as the measured sample. The use of correction factors is suggested when financial constraints limit the use of FAHP standard compounds in routine LOX substrate preference analysis. FAHP quantification by the modified FOX assay aligned well with results obtained using the commonly used conjugated diene method, while offering a quicker and broader sample pH range assessment. Thus, the modified FOX assay can be used as a reliable high-throughput screening method for determining LOX activity. Key points • Modifying perchloric acid level in FOX reagent expands its linear detection range • The modified FOX assay is applicable for screening LOX activity in a wide pH range • The modified FOX assay effectively assesses substrate specificity of LOX

show abstract

Bacterial lipoxygenases: Biochemical characteristics, molecular structure and potential applications

Cited by 20 publications

References 128 publications

Identification of the Thermal Activation Network in Human 15-Lipoxygenase-2: Divergence from Plant Orthologs and Its Relationship to Hydrogen Tunneling Activation Barriers

Identification of the Thermal Activation Network in Human 15-Lipoxygenase-2: Divergence from Plant Orthologs and Its Relationship to Hydrogen Tunneling Activation Barriers

Antioxidant Metabolism Pathways in Vitamins, Polyphenols, and Selenium: Parallels and Divergences

Versatile ferrous oxidation–xylenol orange assay for high-throughput screening of lipoxygenase activity

Contact Info

Product

Resources

About