| Title |
Characterization and Soluble Expression of d-Hydantoinase from Pseudomonas fluorescens for the Synthesis of d-Amino Acids
|
|---|---|
| Published in |
Applied Biochemistry and Biotechnology, January 2016
|
| DOI | 10.1007/s12010-015-1975-6 |
| Pubmed ID | |
| Authors |
Guo-Chao Xu, Lei Li, Rui-Zhi Han, Jin-Jun Dong, Ye Ni |
| Abstract |
An active D-hydantoinase from Pseudomonas fluorescens was heterogeneously overexpressed in Escherichia coli BL21(DE3) and designated as D-PfHYD. Sequence and consensus analysis suggests that D-PfHYD belongs to the dihydropyrimidinase/hydantoinase family and possesses catalytic residues for metal ion and hydantoin binding. D-PfHYD was purified to homogeneity by nickel affinity chromatography for characterization. D-PfHYD is a homotetramer with molecular weight of 215 kDa and specific activity of 20.9 U mg(-1). D-PfHYD showed the highest activity at pH 9.0 and 60 °C. Metal ions such as Mn(2+), Fe(2+), and Fe(3+) could activate D-PfHYD with 20 % improvement. Substrate specificity analysis revealed that purified D-PfHYD preferred aliphatic to aromatic 5'-monosubstituted hydantoins. Among various strategies tested, chaperone GroES-GroEL was efficient in improving the soluble expression of D-PfHYD. Employing 1.0 g L(-1) recombinant E. coli BL21(DE3)-pET28-hyd/pGRO7 dry cells, 100 mM isobutyl hydantoin was converted into D-isoleucine with 98.7 % enantiomeric excess (ee), isolation yield of 78.3 %, and substrate to biocatalyst ratio of 15.6. Our results suggest that recombinant D-PfHYD could be potentially applied in the synthesis of D-amino acids. |
X Demographics
The data shown below were collected from the profile of 1 X user who shared this research output. Click here to find out more about how the information was compiled.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 1 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 1 | 100% |
Mendeley readers
The data shown below were compiled from readership statistics for 19 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 19 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 4 | 21% |
| Student > Ph. D. Student | 3 | 16% |
| Student > Master | 3 | 16% |
| Student > Bachelor | 2 | 11% |
| Other | 1 | 5% |
| Other | 0 | 0% |
| Unknown | 6 | 32% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 5 | 26% |
| Agricultural and Biological Sciences | 4 | 21% |
| Engineering | 2 | 11% |
| Immunology and Microbiology | 1 | 5% |
| Medicine and Dentistry | 1 | 5% |
| Other | 1 | 5% |
| Unknown | 5 | 26% |
Attention Score in Context
This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 29 January 2016.
All research outputs
#15,355,821
of 22,842,950 outputs
Outputs from Applied Biochemistry and Biotechnology
#1,554
of 2,508 outputs
Outputs of similar age
#233,198
of 396,721 outputs
Outputs of similar age from Applied Biochemistry and Biotechnology
#9
of 37 outputs
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So far Altmetric has tracked 2,508 research outputs from this source. They receive a mean Attention Score of 3.2. This one is in the 31st percentile – i.e., 31% of its peers scored the same or lower than it.
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We're also able to compare this research output to 37 others from the same source and published within six weeks on either side of this one. This one has gotten more attention than average, scoring higher than 59% of its contemporaries.