A novel cold-adapted nitroreductase from Psychrobactersp. ANT206: Heterologous expression, characterization and nitrobenzene reduction capacity

Wang, Yifan; Hou, Yanhua; Wang, Yatong; Zheng, Lu; Wang, Quanfu

doi:10.1016/j.enzmictec.2019.109434

Cited by 17 publications

(6 citation statements)

References 45 publications

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“…In most cases, the hydroxylamino derivative was the final product of the NB reduction possibly because the second two‐electron reaction is more rapid than the first two‐electron transfer (Race et al ., 2005). A large body of literature revealed that enzyme‐catalysed conversion of nitrobenzene to aniline depends on nitroreductase and mutase (Roldan et al ., 2008), but the final product of nitrobenzene reduction catalysed by novel cold‐adapted nitroreductase was aniline (Wang et al ., 2019). Two‐electron midpoint potential of NR may explain why hydroxylaminobenzene, and not aniline, is the end product of the enzyme‐catalysed reduction of nitrobenzene (Koder and Miller, 1998).…”

Section: Resultsmentioning

confidence: 99%

Cu(II) nonspecifically binding chromate reductase NfoR promotes Cr(VI) reduction

Han

Zheng

Zhou

et al. 2020

Environmental Microbiology

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Summary Cu(II)‐enhanced microbial Cr(VI) reduction is common in the environment, yet its mechanism is unknown. The specific activity of chromate reductase, NfoR, from Staphylococcus aureus sp. LZ‐01 was augmented 1.5‐fold by Cu(II). Isothermal titration calorimetry and spectral data show that Cu(II) binds to NfoR nonspecifically. Further, Cu(II) stimulates the nitrobenzene reduction of NfoR, indicating that Cu(II) promotes electron transfer. The crystal structure of NfoR in complex with CuSO4 (1.46 Å) was determined. The overall structure of NfoR‐Cu(II) complex is a dimer that covalently binds with FMN and Cu(II)‐binding pocket is located at the interface of the NfoR dimer. Structural superposition revealed that NfoR resembles the structure of class II chromate reductase. Site‐directed mutagenesis revealed that Leu46 and Phe123 were involved in NADH binding, whereas Trp70 and Ser45 were the key residues for nitrobenzene binding. Furthermore, His100 and Asp171 were preferential affinity sites for Cu(II) and that Cys163 is an active site for FMN binding. Attenuation reductase activity in C163S can be partially restored to 54% wild type by increasing Cu(II) concentration. Partial restoration indicates dual‐channel electron transfer of NfoR via Cu(II) and FMN. We propose a catalytic mechanism for Cu(II)‐enhanced NfoR activity in which Cu(I) is formed transiently. Together, the current results provide an insight on Cu (II)‐induced enhancement and benefit of Cr(VI) bioremediation.

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Section: Resultsmentioning

confidence: 99%

Cu(II) nonspecifically binding chromate reductase NfoR promotes Cr(VI) reduction

Han

Zheng

Zhou

et al. 2020

Environmental Microbiology

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“…Likewise, a similar feature was found in the cold‐adapted enzymes, including the RNase R from Antarctic sea‐ice bacterium Psychrobacter sp. [24] and the esterase from the psychrophilic yeast Glaciozyma antarctica [25]. Moreover, the flexibility of PsSHMT could be increased due to the lower content of Pro [25].…”

Section: Discussionmentioning

confidence: 99%

Molecular cloning, characterization, and homology modeling of serine hydroxymethyltransferase from psychrophilic bacterium Psychrobacter sp.

Wang

Hou

et al. 2022

J Basic Microbiol

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Serine hydroxymethyltransferase (SHMT) plays a significant role in the synthesis of L-serine, purine, and thymidylate, which could be extensively applied in the treatment of cancers and the development of antibiotics. In this study, cloned from Psychrobacter sp. ANT206, a novel cold-adapted SHMT gene (psshmt, 1257 bp) encoding a protein of 418 amino acids was expressed in Escherichia coli. The homology modeling result revealed that PsSHMT owned

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“…(engineered to co-express Cpn60 and Cpn10 chaperons from Oleispira antarctica) compared to a standard E. coli BL21(DE3). Likewise, other enzymes from Psychrobacter isolates, including nitroreductase (Wang et al, 2019), lipase (Xuezheng et al, 2010), esterase (Novototskaya-Vlasova et al, 2012), have been successfully produced in heterologous E. coli cells.…”

Section: Discussionmentioning

confidence: 99%

Discovery and Characterization of a New Cold-Active Protease From an Extremophilic Bacterium via Comparative Genome Analysis and in vitro Expression

et al. 2020

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Following a screening of Antarctic glacier forefield-bacteria for novel cold-active enzymes, a psychrophilic strain Psychrobacter sp. 94-6PB was selected for further characterization of enzymatic activities. The strain produced lipases and proteases in the temperature range of 4-18 • C. The coding sequence of an extracellular serine-protease was then identified via comparative analysis across Psychrobacter sp. genomes, PCRamplified in our strain 94-6PB and expressed in the heterologous host E. coli. The purified enzyme (80 kDa) resulted to be a cold-active alkaline protease, performing best at temperatures of 20-30 • C and pH 7-9. It was stable in presence of common inhibitors [β-mercaptoethanol (β-ME), dithiothreitol (DTT), urea, phenylmethylsulfonyl fluoride (PMSF) and ethylenediaminetetraacetic acid (EDTA)] and compatible with detergents and surfactants (Tween 20, Tween 80, hydrogen peroxide and Triton X-100). Because of these properties, the P94-6PB protease may be suitable for use in a new generation of laundry products for cold washing. Furthermore, we assessed the microdiversity of this enzyme in Psychrobacter organisms from different cold habitats and found several gene clusters that correlated with specific ecological niches. We then discussed the role of habitat specialization in shaping the biodiversity of proteins and enzymes and anticipate far-reaching implications for the search of novel variants of biotechnological products.

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A novel cold-adapted nitroreductase from Psychrobactersp. ANT206: Heterologous expression, characterization and nitrobenzene reduction capacity

Cited by 17 publications

References 45 publications

Cu(II) nonspecifically binding chromate reductase NfoR promotes Cr(VI) reduction

Cu(II) nonspecifically binding chromate reductase NfoR promotes Cr(VI) reduction

Molecular cloning, characterization, and homology modeling of serine hydroxymethyltransferase from psychrophilic bacterium Psychrobacter sp.

Discovery and Characterization of a New Cold-Active Protease From an Extremophilic Bacterium via Comparative Genome Analysis and in vitro Expression

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