Enhanced P450 fatty acid decarboxylase catalysis by glucose oxidase coupling and co-assembly for biofuel generation

Zhang, Libo; Manley, Olivia M.; Ma, Dumei; Yin, Yingwu; Makris, Thomas M.; Wang, Qian

doi:10.1016/j.biortech.2020.123538

Cited by 20 publications

(16 citation statements)

References 45 publications

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“…Not like the DFSM system which assists to activate H 2 O 2 in the H 2 O 2 ‐innert P450BM3 variant, [25] this class of small molecules worked on wild type peroxygenase OleT. The authors proposed that the small molecules lead to more efficiently utilization of H 2 O 2 in the enzymatic reaction, meanwhile released the oxidative stress of H 2 O 2 , which was concluded based on a H 2 O 2 titration assay and a glucose oxidase (GOx) related H 2 O 2 releasing assay [82,75] . The limitations of the system was that the small molecules showed varied effect on different substrates.…”

Section: Small Molecule Auxiliaries To Control P450s’ Activity For Sy...mentioning

confidence: 99%

Harnessing P450 Enzyme for Biotechnology and Synthetic Biology

Zhang

Wang

2021

ChemBioChem

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Cytochrome P450 enzymes (P450s, CYPs) catalyze the oxidative transformation of a wide range of organic substrates. Their functions are crucial to xenobiotic metabolism and steroid transformation in humans and other organisms. The enzymes are promising for synthetic biology applications but limited by several drawbacks including low turnover rates, poor stability, the dependance of expensive cofactors and redox partners, and the narrow substrate scope. To conquer these obstacles, emerging strategies including substrate engineering, usage of decoy and decoy‐based small molecules auxiliaries, designing of artificial enzyme cascades and the incorporation of materials have been explored based on the unique properties of P450s. These strategies can be applied to a wide range of P450s and can be combined with protein engineering to improve the enzymatic activities. This minireview will focus on some recent developments of these strategies which have been used to leverage P450 catalysis. Remaining challenges and future opportunities will also be discussed.

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Section: Small Molecule Auxiliaries To Control P450s’ Activity For Sy...mentioning

confidence: 99%

Harnessing P450 Enzyme for Biotechnology and Synthetic Biology

Zhang

Wang

2021

ChemBioChem

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“…With increasing concentration of H 2 O 2 , OleT SA showed decreased activity likely due to the oxidative toxicity of H 2 O 2 . [20] The enhanced effect of the small molecule was highly related to the H 2 O 2 concentration. For example, when 0.2 mM of H 2 O 2 was used, only around 20 % enhancement was obtained in the presence of 3 (2 mM).…”

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“…[15] While H 2 O 2 has served as a desirable attribute for industrial transformations because of its low cost and easy scalability by comparing to other redox reagents, the H 2 O 2 dependent decarboxylation is highly inefficient compared to most O 2 dependent CYPs, primarily due to the potential for oxidative modification of heme prosthetic group. [16] To overcome this limitation in the OleT catalytic reaction, different strategies including protein engineering, [17] photochemistry [18] and process engineering [19,20] that could generate H 2 O 2 in a controlled manner have been reported.…”

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confidence: 99%

“…It has been well established that UV-Vis spectra of cytochrome P450s are sensitive to the redox state of the heme (Figure 1A), which could be applied to monitor the change of local environment in varied conditions. [16,20,31] Therefore, UV-Vis spectroscopy was employed to study the interaction of these small molecules (1-7) with OleT in the presence and absence of substrates. As shown in Figure 2A, with treating by 5 μM of eicosanoic acid (C20 FA), the spin state of heme iron occurred, while with the addition of 3 (2.5 mM), a 'Type II' spectral change was observed, [35] which suggests a change of the heme environment and possible small molecule-substrate interactions.…”

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confidence: 99%

“…To confirm this hypothesis, we tested a bi-enzymatic method recently developed in our group, which used glucose oxidase (GOx) and glucose to generate H 2 O 2 in situ to afford an optimized OleT catalytic efficiency by minimizing H 2 O 2 toxicity. [20] When this method was used, the small molecule' enhanced effect on the enzyme activity was completely diminished (Figure 4B). This result further supports our hypothesis that the origin of the enhancement of small molecules is from enzyme oxidative step.…”

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Using Small Molecules to Enhance P450 OleT Enzyme Activity in Situ

Zhang

Yin

et al. 2021

Chemistry A European J

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Cytochrome P450 OleT is a fatty acid decarboxylase that catalyzes the production of olefins with biofuel and synthetic applications. However, the relatively sluggish catalytic efficiency of the enzyme limits its applications. Here, we report the application of a novel class of benzene containing small molecules to improve the OleT activity. The UV-Vis spectroscopy study and molecular docking results confirmed the high proximity of the small molecules to the heme group of OleT. Up to 6-fold increase of product yield has been achieved in the small molecule-modulated enzymatic reactions. Our work thus sheds the light to the application of small molecules to increase the OleT catalytic efficiency, which could be potentially used for future olefin productions.

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Microbial P450 repertoire (P450ome) and its application feasibility in pharmaceutical industry, chemical industry, and environmental protection

Wang,

Pan,

Wang

et al. 2023

Biotech & Bioengineering

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Cytochrome P450s (CYPs) are heme‐thiolated enzymes that catalyze the oxidation of C‐H bonds in a regio‐ and stereo‐selective manner. CYPs are widely present in the biological world. With the completion of more biological genome sequencing, the number and types of P450 enzymes have increased rapidly. P450 in microorganisms is easy to clone and express, rich in catalytic types, and strong in substrate adaptability, which has good application potential. Although the number of P450 enzymes found in microorganisms is huge, the function of most of the microorganism P450s has not been studied, and it contains a large number of excellent biocatalysts to be developed. This review is based on the P450 groups in microorganisms. First, it reviews the distribution of P450 groups in different microbial species, and then studies the application of microbial P450 enzymes in the pharmaceutical industry, chemical industry and environmental pollutant treatment in recent years. And focused on the application fields of P450 enzymes of different families to guide the selection of suitable P450s from the huge P450 library. In view of the current shortcomings of microbial P450 in the application process, the final solution is the most likely to assist the application of P450 enzymes in large‐scale, that is, whole cell transformation combined with engineering, fusion P450 combined with immobilization technology.

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Enhanced P450 fatty acid decarboxylase catalysis by glucose oxidase coupling and co-assembly for biofuel generation

Cited by 20 publications

References 45 publications

Harnessing P450 Enzyme for Biotechnology and Synthetic Biology

Harnessing P450 Enzyme for Biotechnology and Synthetic Biology

Using Small Molecules to Enhance P450 OleT Enzyme Activity in Situ

Microbial P450 repertoire (P450ome) and its application feasibility in pharmaceutical industry, chemical industry, and environmental protection

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