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Characterization of ML-005, a Novel Metaproteomics-Derived Esterase

Overview of attention for article published in Frontiers in Microbiology, August 2018
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Title
Characterization of ML-005, a Novel Metaproteomics-Derived Esterase
Published in
Frontiers in Microbiology, August 2018
DOI 10.3389/fmicb.2018.01925
Pubmed ID
Authors

Premankur Sukul, Natalie Lupilov, Lars I. Leichert

Abstract

A novel gene encoding for a lipolytic enzyme, designated ML-005, was recently identified using a functional metaproteomics approach. We heterologously expressed this protein in Escherichia coli and biochemically characterized it. ML-005 exhibited lipolytic activity toward short-chained substrates with the preferred substrate being p-nitrophenyl-butyrate, suggesting that ML-005 is an esterase. According to homology analysis and site-directed mutagenesis, the catalytic triad of the enzyme was identified as Ser-99, Asp-164, and His-191. Its optimal pH was determined to be at pH 8. Optimal activity was observed at 45°C. It also exhibited temperature, pH and salt tolerance. Residual relative activity after incubating at 50-60°C for 360 min was above 80% of its initial activity. It showed tolerance over a broad range of pH (5-12) and retained most of its initial activity. Furthermore, incubating ML-005 in 1 - 5M NaCl solution had negligible effect on its activity. DTT, EDTA, and ß-mercaptoethanol had no significant effect on ML-005's activity. However, addition of PMSF led to almost complete inactivation consistent with ML-005 being a serine hydrolase. ML-005 remains stable in the presence of a range of metal ions, but addition of Cu2+ significantly reduces its relative activity. Organic solvents have an inhibitory effect on ML-005, but it retained 21% of activity in 10% methanol. SDS had the most pronounced inhibitory effect on ML-005 among all detergents tested and completely inactivated it. Furthermore, the Vmax of ML-005 was determined to be 59.8 μM/min along with a Km of 137.9 μM. The kcat of ML-005 is 26 s-1 and kcat/Km is 1.88 × 105 M-1 s-1.

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Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 22 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 22 100%

Demographic breakdown

Readers by professional status Count As %
Student > Master 5 23%
Student > Ph. D. Student 4 18%
Student > Postgraduate 3 14%
Researcher 1 5%
Student > Bachelor 1 5%
Other 0 0%
Unknown 8 36%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 4 18%
Agricultural and Biological Sciences 4 18%
Immunology and Microbiology 2 9%
Pharmacology, Toxicology and Pharmaceutical Science 1 5%
Arts and Humanities 1 5%
Other 2 9%
Unknown 8 36%
Attention Score in Context

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 14 September 2018.
All research outputs
#17,990,045
of 23,103,436 outputs
Outputs from Frontiers in Microbiology
#17,518
of 25,285 outputs
Outputs of similar age
#239,835
of 334,084 outputs
Outputs of similar age from Frontiers in Microbiology
#502
of 717 outputs
Altmetric has tracked 23,103,436 research outputs across all sources so far. This one is in the 19th percentile – i.e., 19% of other outputs scored the same or lower than it.
So far Altmetric has tracked 25,285 research outputs from this source. They typically receive a little more attention than average, with a mean Attention Score of 6.4. This one is in the 22nd percentile – i.e., 22% of its peers scored the same or lower than it.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 334,084 tracked outputs that were published within six weeks on either side of this one in any source. This one is in the 23rd percentile – i.e., 23% of its contemporaries scored the same or lower than it.
We're also able to compare this research output to 717 others from the same source and published within six weeks on either side of this one. This one is in the 22nd percentile – i.e., 22% of its contemporaries scored the same or lower than it.