Characterization of a New Cyclohexylamine Oxidase From Acinetobacter sp. YT-02

Zhou, Hui; Han, Zhenggang; Fang, Ti; Chen, Yuanyuan; Ning, Shang-bo; Gan, Ya-Ting; Ding, Yan

doi:10.3389/fmicb.2018.02848

Cited by 4 publications

(2 citation statements)

References 29 publications

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“…32 Amino acid residues T198, L199, M226, and F351 (CHAO IH-35A numbering) were proposed as gating residues separating these two cavities. 32,33 Interestingly, our ternary structure contains two 2a molecules, with one buried deep inside and the other located close to the surface (Figure S8), therefore strongly supporting the experimentally proposed presence of two cavities. According to the sequence alignment (Figure 2), seven of the nine residues mentioned above were found to be highly conserved in CHAO CCH12-C2 , meaning either the same amino acid or amino acids with very similar chemical properties are present.…”

Section: ■ Introductionmentioning

confidence: 99%

Asymmetric Synthesis of a Key Dextromethorphan Intermediate and Its Analogues Enabled by a New Cyclohexylamine Oxidase: Enzyme Discovery, Reaction Development, and Mechanistic Insight

Huang

Wang

et al. 2020

J. Org. Chem.

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Metrics & MoreArticle Recommendations * sı Supporting Information ABSTRACT: (S)-1-(4-Methoxybenzyl)-1,2,3,4,5,6,7,8-octahydroisoquinoline [(S)-1-(4-methoxybenzyl)-OHIQ, (S)-1a] is a key synthetic intermediate in the industrial production of dextromethorphan, one of the most widely used over-the-counter antitussives. We report here that a new cyclohexylamine oxidase discovered by genome mining, named CHAO CCH12-C2 , was able to completely deracemize 100 mM 1a under Turner's deracemization conditions to afford (S)-1a in 80% isolated yield and 99% ee at a semipreparative scale (0.4 mmol). When this biocatalytic reaction was scaled up to a gram scale (5.8 mmol), without reaction optimization (S)-1a was still isolated in 67% yield and 96% ee. The relatively higher k cat determined for CHAO CCH12-C2 was rationalized as one major factor rendering this enzyme capable of oxidizing 1a effectively at elevated substrate concentrations. Protein sequence alignment, analysis of our co-crystal structure of CHAO CCH12-C2 complexed with the product 1-(4-methoxybenzyl)-3,4,5,6,7,8-hexahydroisoquinoline [1-(4-methoxybenzyl)-HHIQ, 2a], and the structure-guided mutagenesis study together indicated L295 is one of the critical residues for this efficient enzymatic oxidation process and supported the presence of two cavities as well as a catalytically important "aromatic cage" formed by F342, Y433, and FAD. The synthetic applicability of CHAO CCH12-C2 was further underscored by the stereoselective synthesis of various enantioenriched 1-benzyl-OHIQ derivatives of potential pharmaceutical importance at a semipreparative scale.

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Section: ■ Introductionmentioning

confidence: 99%

Asymmetric Synthesis of a Key Dextromethorphan Intermediate and Its Analogues Enabled by a New Cyclohexylamine Oxidase: Enzyme Discovery, Reaction Development, and Mechanistic Insight

Huang

Wang

et al. 2020

J. Org. Chem.

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“…to degrade hydrocarbons, including n-alkanes [99][100][101], aromatic compounds [25, [102][103][104], and diesel fuel [23,105]. The latest information appeared on the participation of representatives of this genus in the decomposition of pesticides [106][107][108] and insecticides [109,110]. It should be noted that the ability to decompose xenobiotics was established by researchers during the cultivation of Acinetobacter genus bacteria on media containing toxic compounds as growing media (Table 6).…”

Section: Ability Of Acinetobacter Bacteria To Phosphates Solubilization and Xenobiotics Degradationmentioning

confidence: 99%

Biotechnological Potential of the Acinetobacter Genus Bacteria

Пирог¹,

Lutsai²,

Muchnyk³

2021

Mikrobiol. Z.

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Until recently, there were rare scientific reports on the biotechnological potential of non-pathogenic bacteria of the Acinetobacter genus. Although the first reports about the practically valuable properties of these bacteria date back to the 70s and 80s of the twentieth century and concerned the synthesis of the emulsan bioemulsifier. In the last decade, interest in representatives of the Acinetobacter genus as objects of biotechnology has significantly increased. The review presents current literature data on the synthesis by bacteria of this genus of high-molecular emulsifiers, low-molecular biosurfactants of glyco- and aminolipid nature, enzymes (lipase, agarase, chondroitinase), phytohormones, as well as their ability to solubilize phosphates and decompose various xenobiotics (aliphatic and aromatic hydrocarbons, pesticides, insecticides). Prospects for practical application of Acinetobacter bacteria and the metabolites synthesized by them in environmental technologies, agriculture, various industries and medicine are discussed. The data of own experimental studies on the synthesis and biological activity (antimicrobial, anti-adhesive, ability to destroy biofilms) of biosurfactants synthesized by A. calcoaceticus IMV B-7241 strain and their role in the degradation of oil pollutants, including complex ones with heavy metals, are presented. The ability of A. calcoaceticus IMV B-7241 to the simultaneous synthesis of phytohormones (auxins, cytokinins, gibberellins) and biosurfactants with antimicrobial activity against phytopathogenic bacteria allows us to consider this strain as promising for practical use in crop production to increase crop yields.

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X-ray crystal structure of Vibrio alkaline phosphatase with the non-competitive inhibitor cyclohexylamine

Ásgeirsson

Markússon

Hlynsdóttir

et al. 2020

Biochemistry and Biophysics Reports

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Background Para -nitrophenyl phosphate, the common substrate for alkaline phosphatase (AP), is available as a cyclohexylamine salt. Here, we report that cyclohexylamine is a non-competitive inhibitor of APs. Methods Cyclohexylamine inhibited four different APs. Co-crystallization with the cold-active Vibrio AP (VAP) was performed and the structure solved. Results Inhibition of VAP fitted a non-competitive kinetic model (K m unchanged, V max reduced) with IC 50 45.3 mM at the pH optimum 9.8, not sensitive to 0.5 M NaCl, and IC 50 27.9 mM at pH 8.0, where the addition of 0.5 M NaCl altered the inhibition to the level observed at pH 9.8. APs from E. coli and calf intestines were less sensitive to cyclohexylamine, whereas an Antarctic bacterial AP was similar to VAP in this respect. X-ray crystallography at 2.3 Å showed two binding sites, one in the active site channel and another at the surface close to dimer interface. Antarctic bacterial AP and VAP have Trp274 in common in their active-sites, that takes part in binding cyclohexylamine. VAP variants W274A, W274K, and W274H gave IC 50 values of 179 mM, 188 mM and 187 mM, respectively, at pH 9.8. Conclusions The binding of cyclohexylamine in locations at the dimeric interface and/or in the active site of APs may delay product release or reduce the rate of catalytic step(s) involving conformational changes and intersubunit communications. General significance Cyclohexylamine is a common chemical in industries and used as a counterion in substrates for alkaline phosphatase, a clinically important and common enzyme in the biosphere.

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Characterization of a New Cyclohexylamine Oxidase From Acinetobacter sp. YT-02

Cited by 4 publications

References 29 publications

Asymmetric Synthesis of a Key Dextromethorphan Intermediate and Its Analogues Enabled by a New Cyclohexylamine Oxidase: Enzyme Discovery, Reaction Development, and Mechanistic Insight

Asymmetric Synthesis of a Key Dextromethorphan Intermediate and Its Analogues Enabled by a New Cyclohexylamine Oxidase: Enzyme Discovery, Reaction Development, and Mechanistic Insight

Biotechnological Potential of the Acinetobacter Genus Bacteria

X-ray crystal structure of Vibrio alkaline phosphatase with the non-competitive inhibitor cyclohexylamine

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