In vitro characterization of CYP102G4 from Streptomyces cattleya: A self-sufficient P450 naturally producing indigo

Kim, Joonwon; Lee, Pyung‐Gang; Jung, E.; Kim, Byung‐Gee

doi:10.1016/j.bbapap.2017.08.002

Cited by 25 publications

(16 citation statements)

References 57 publications

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“…CYP102G4 is a self-sufficient microbial cytochrome P450 isolated from Streptomyces cattelya . Similar to other self-sufficient P450s, CYP102G4 directly consumes NAD(P)H as an electron source to activate heme, forming ‘Compound I’ Fe=O (IV) that finally breaks the substrate C-H bond for monooxygenation ( Rittle and Green, 2010 ; Kim et al, 2018 ). In our previous study, the CYP102G4, was characterized to show a naturally high monooxygenation activity for various polyaromatic substrates including benzophenone and flavone owing to its wide cavity in the active site that enables bulky substrates to easily access the activated heme species ( Kim et al, 2018 ).…”

Section: Resultsmentioning

confidence: 99%

“…In brief, a two-component flavin-dependent monooxygenase, Ec HpaBC, was newly cloned from the genomic DNA of Escherichia coli ATCC 8739, and Ma FMO was cloned from the genomic DNA of Methylophaga aminisulfidivorans . The vector encoding CYP102G4 was previously constructed ( Kim et al, 2018 ). Respective E. coli BL21 (DE3) transformants were verified for the soluble expression in SDS-PAGE ( Supplementary Figure S2 ) and were then used as microbial strains for the whole-cell biotransformation.…”

Section: Discussionmentioning

confidence: 99%

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Regioselective One-Pot Synthesis of Hydroxy-(S)-Equols Using Isoflavonoid Reductases and Monooxygenases and Evaluation of the Hydroxyequol Derivatives as Selective Estrogen Receptor Modulators and Antioxidants

Song

Lee

Kim

et al. 2022

Front. Bioeng. Biotechnol.

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Several regiospecific enantiomers of hydroxy-(S)-equol (HE) were enzymatically synthesized from daidzein and genistein using consecutive reduction (four daidzein-to-equol–converting reductases) and oxidation (4-hydroxyphenylacetate 3-monooxygenase, HpaBC). Despite the natural occurrence of several HEs, most of them had not been studied owing to the lack of their preparation methods. Herein, the one-pot synthesis pathway of 6-hydroxyequol (6HE) was developed using HpaBC (EcHpaB) from Escherichia coli and (S)-equol-producing E. coli, previously developed by our group. Based on docking analysis of the substrate or products, a potential active site and several key residues for substrate binding were predicted to interpret the (S)-equol hydroxylation regioselectivity of EcHpaB. Through investigating mutations on the key residues, the T292A variant was verified to display specific mono-ortho-hydroxylation activity at C6 without further 3′-hydroxylation. In the consecutive oxidoreductive bioconversion using T292A, 0.95 mM 6HE could be synthesized from 1 mM daidzein, while 5HE and 3′HE were also prepared from genistein and 3′-hydroxydaidzein (3′HD or 3′-ODI), respectively. In the following efficacy tests, 3′HE and 6HE showed about 30∼200-fold higher EC50 than (S)-equol in both ERα and ERβ, and they did not have significant SERM efficacy except 6HE showing 10% lower β/α ratio response than that of 17β-estradiol. In DPPH radical scavenging assay, 3′HE showed the highest antioxidative activity among the examined isoflavone derivatives: more than 40% higher than the well-known 3′HD. In conclusion, we demonstrated that HEs could be produced efficiently and regioselectively through the one-pot bioconversion platform and evaluated estrogenic and antioxidative activities of each HE regio-isomer for the first time.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Regioselective One-Pot Synthesis of Hydroxy-(S)-Equols Using Isoflavonoid Reductases and Monooxygenases and Evaluation of the Hydroxyequol Derivatives as Selective Estrogen Receptor Modulators and Antioxidants

Song

Lee

Kim

et al. 2022

Front. Bioeng. Biotechnol.

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“…Subsequent oxidation of indoxyl leads to the generation of indoxyl radicals, and two molecules of indoxyl radicals are converted to a well‐known blue‐colored indigo dye‐a symmetric dimer‐through a self‐dimerization reaction 53–57. According to recent studies, several monooxygenases such as flavin monooxygenase (FMO), cytochrome P450 monooxygenase (CYP), toluene dioxygenase (TDO), and naphthalene dioxygenase (NDO) were involved in indole C3‐hydroxylation and bio‐indigo formation from L‐tryptophan 58–64.…”

Section: Deamination Of Aromatic Amino Acids and Cosmetic And Pharmaceutical Applicationsmentioning

confidence: 99%

Nitrogen‐Neutral Amino Acids Refinery: Deamination of Amino Acids for Bio‐Alcohol and Ammonia Production

Choi

2021

ChemBioEng Reviews

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Amino acids are a valuable bioresource that contains both the acid form of carbon and the amine group of nitrogen. Amino acids not only constitute proteins and the building blocks for living organisms but also provide a key intermediate for controlling the nitrogen balance in cells. The production of amino acids using microorganisms has, along with the development of biotechnology, created a large market for biochemicals. In parallel, there have been various biochemical production methods using pulmonary proteins or direct amino acids as biomass through enzymatic bioconversion or whole‐cell biotransformation. The deamination of amino acids facilitated the applications of 2‐keto and acrylic acids, in particular, for biofuels of bio alcohol, pharmaceuticals, cosmetics, as building blocks for polymers, and other versatile industrial chemicals. In this review, we report on the application of amino acids derived from biochemicals initiated by deamination reactions using deaminase, ammonia‐lyase, and oxidase enzymes for biotechnology applications.

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“…The improvement was brought about by merely adding one more mutation. Moreover, in the last two decades, more microbial CYPs have been reported for their capacity of indigo formation (Brixius-Anderko et al 2017;Fiorentini et al 2018;Kim et al 2018) which may also represent good starting points for creating an optimized CYP for indigo production. The feature that conversion of indole into indigo can be seen by eye makes it attractive to apply directed evolution protocols.…”

Section: Heme-containing Oxygenasesmentioning

confidence: 99%

An overview of microbial indigo-forming enzymes

Fabara

Fraaije

2019

Appl Microbiol Biotechnol

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Indigo is one of the oldest textile dyes and was originally prepared from plant material. Nowadays, indigo is chemically synthesized at a large scale to satisfy the demand for dyeing jeans. The current indigo production processes are based on fossil feedstocks; therefore, it is highly attractive to develop a more sustainable and environmentally friendly biotechnological process for the production of this popular dye. In the past decades, a number of natural and engineered enzymes have been identified that can be used for the synthesis of indigo. This mini-review provides an overview of the various microbial enzymes which are able to produce indigo and discusses the advantages and disadvantages of each biocatalytic system.

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In vitro characterization of CYP102G4 from Streptomyces cattleya: A self-sufficient P450 naturally producing indigo

Cited by 25 publications

References 57 publications

Regioselective One-Pot Synthesis of Hydroxy-(S)-Equols Using Isoflavonoid Reductases and Monooxygenases and Evaluation of the Hydroxyequol Derivatives as Selective Estrogen Receptor Modulators and Antioxidants

Regioselective One-Pot Synthesis of Hydroxy-(S)-Equols Using Isoflavonoid Reductases and Monooxygenases and Evaluation of the Hydroxyequol Derivatives as Selective Estrogen Receptor Modulators and Antioxidants

Nitrogen‐Neutral Amino Acids Refinery: Deamination of Amino Acids for Bio‐Alcohol and Ammonia Production

An overview of microbial indigo-forming enzymes

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