Biodegradation of Cyclohexylamine by
            <i>Brevibacterium oxydans</i>
            IH-35A

Iwaki, Hiroaki; Shimizu, Minako; Tokuyama, Tai; Hasegawa, Yoshie

doi:10.1128/aem.65.5.2232-2234.1999

Cited by 37 publications

(11 citation statements)

References 15 publications

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“…A genome mining approach was employed for discovery of more efficient CHAO. Using the amino acid sequence of CHAO IH‑35A as a template, , a BLAST search was performed, and four candidate enzymes were selected, with the level of sequence identity between them and the template being 50–78% (Table S1). The corresponding codon-optimized genes were synthesized and cloned into the pET-28a vector.…”

Section: Resultsmentioning

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: Resultsmentioning

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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“…The low endogenous metabolism of Brevibacterium is attributed to their ability to survive nutrient starvation and plays a role in the slow growth rate (Forquin‐Gomez & Weimer, 2014). The genus Brevibacterium is capable of metabolizing many different carbon and nitrogen sources for growth and shows diverse metabolic properties that could be exploited in new applications such as bioremediation and biodegradation of aromatic compounds and polymers (Iwaki et al, 1999; Rodriguez et al, 2011; van Afferden et al, 1990). Plastics including PVC mix plasticized with di‐iso‐octyl sebacate Z (Booth et al, 1968), polyethylene (Hadad et al, 2005) and Linear Low‐Density Polyethylene (Dwicania et al, 2019) were also reported to biodegrade by the strains in this genus.…”

Section: Discussionmentioning

confidence: 99%

Biodegradation of polystyrene by three bacterial strains isolated from the gut of Superworms (Zophobas atratus larvae)

Arunrattiyakorn

Ponprateep

Kaennonsang

et al. 2022

Journal of Applied Microbiology

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Aims: To isolate polystyrene-degrading bacteria from the gut of superworms and investigate their ability to degrade polystyrene (PS). Methods and Results:Three PS-degrading bacteria identified as Pseudomonas sp.EDB1, Bacillus sp. EDA4 and Brevibacterium sp. EDX were successfully isolated from the gut of superworms (Zophobas atratus Larvae) that ingest PS. Incubating PS with each strain for 30-day led to the formation of biofilm on the PS film. Scanning electron microscopy (SEM) revealed considerable damage (in terms of pits formation) on the surface of the PS films. FTIR analysis suggested the incorporation of carbonyl group into the carbon backbone of PS. Decreasing of WCA of microbial-treated PS film confirmed a chemical change from hydrophobicity to hydrophilicity on the PS surface. Based on these results, we conclude that all isolates had the ability to degrade PS.Conclusions: Brevibacterium sp. EDX (GenBank MZ32399) was isolated as the most efficient PS-degrading strain based on the most changing in both PS surface morphology (SEM and WCA analyses) and chemical modification (FTIR analysis) in its PS degradation process. Significance and Impact of the Study:This was the first study to describe the PS degradation by Brevibacterium sp. EDX, and thus provided for its development in the plastic remediation process.

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“…In higher organisms, they play an important role in polyamine metabolism, whereas in lower eukaryotes and bacteria amine oxidases are used to provide a source of ammonium ( Mirza et al, 2013 ). CHAO is a member of flavin-containing amine oxidases and is highly specific for catalyzing oxidative deamination of CHAM, forming the oxidation product cyclohexanone ( Tokieda et al, 1977 ; Iwaki et al, 1999a , b ; Mirza et al, 2013 ). The enzyme has great potential in eliminating CHAM pollution ( Iwaki et al, 1999b ; Yan et al, 2017 ) and synthesis of chiral amines in pharmaceutical industry ( Li et al, 2014 ; Li et al, 2016 ).…”

Section: Discussionmentioning

confidence: 99%

“…To date, limited studies showed that bacteria utilized CHAM as the only source of carbon and nitrogen. These bacteria included Brevibacterium oxydans IH-35A ( Iwaki et al, 1999a ). Pseudomonas plecoglossicida NyZ12 ( Shen et al, 2008 ; Yan et al, 2017 ), and Acinetobacter sp.…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2018

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Cyclohexylamine (CHAM) is widely used in various industries, but it is harmful to human beings and the environment. Acinetobacter sp. YT-02 can degrade CHAM via cyclohexanone as an intermediate. In this study, the cyclohexylamine oxidase (CHAO) gene from Acinetobacter sp. YT-02 was cloned. Amino acid sequence alignment indicated that the cyclohexylamine oxidase (CHAOYT–02) was 48% identical to its homolog from Brevibacterium oxydans IH-35A (CHAOIH–35). The enzyme was expressed in Escherichia coli BL21 (DE3), and purified to apparent homogeneity by Ni-affinity chromatography. The purified enzyme was proposed to be a dimer of molecular mass of approximately 91 kDa. The enzyme exhibited its maximum activity at 50°C and at pH 7.0. The enzyme was thermolabile as demonstrated by loss of important percentage of its maximal activity after 30 min incubation at 50°C. Metal ions Mg2+, Co2+, and K+ had certain inhibitory effect on the enzyme activity. The kinetic parameters Km and Vmax were 0.25 ± 0.02 mM and 4.3 ± 0.083 μM min−1, respectively. The biochemical properties, substrate specificities, and three-dimensional structures of CHAOYT–02 and CHAOIH–35 were compared. Our results are helpful to elucidate the mechanism of microbial degradation of CHAM in the strain YT-02. In addition, CHAOYT–02, as a potential biocatalyst, is promising in controlling CHAM pollution and deracemization of chiral amines.

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Biodegradation of Cyclohexylamine by Brevibacterium oxydans IH-35A

Cited by 37 publications

References 15 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

Biodegradation of polystyrene by three bacterial strains isolated from the gut of Superworms (Zophobas atratus larvae)

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

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