This web resource is supported by a Research Resource from the National Institute of General Medical Sciences (R24GM141196-01).
The tools are available without charge or license to both academic and commercial users.
Reorganization of UniProtKB
With the current 2026_02 release, the UniProtKB database is reorganized to include an
expanded number of Reference Proteomes to better capture biodiversity.
This includes the removal of proteins from taxonomically unclassified organisms,
i.e., those without a binomial species name (genus and species). The total number of
accessions in UniProtKB has been reduced from 253,635,358 in the “legacy” 2025_03
release to 149,810,139 in the current 2026_02 release.
We are providing the option to select either the “legacy” 2025_03 database or the
current UniProtKB database (now 2026_02) when generating SSNs. You can select the
database in the “Database” accordion on the pages for the EFI-EST options, the EFI-GNT
tool, and the Taxonomy Tool. We suggest that you compare the SSNs, GNNs, and GNDs
generated from both databases as you explore the information you are seeking.
Update of EFI Tools
The EFI Tools are being upgraded on 10/21/26. While the updated site is functionally
identical to the old site, jobs run prior to that date will only be available
through the results link in the finalization email (not the Previous Jobs tab).
User accounts created on the old site will no longer be available, and users
are encouraged to create new accounts when the upgraded site becomes available.
Rémi Zallot, Nils Oberg, and John A. Gerlt, The EFI Web Resource for Genomic Enzymology Tools: Leveraging Protein, Genome, and Metagenome Databases to Discover Novel Enzymes and Metabolic Pathways. Biochemistry 2019 58 (41), 4169-4182. https://doi.org/10.1021/acs.biochem.9b00735
Nils Oberg, Rémi Zallot, and John A. Gerlt, EFI-EST, EFI-GNT, and EFI-CGFP: Enzyme Function Initiative (EFI) Web Resource for Genomic Enzymology Tools. J Mol Biol 2023. https://doi.org/10.1016/j.jmb.2023.168018
The SSN submited has been edited so that the markers and their abundances in the
selected metagenomes are included as node attributes.
File
Size
SSN with quantify results (ZIP)
CGFP Family and Marker Data
The CD-HIT ShortBRED families by cluster file contains mappings of ShortBRED
families to SSN cluster number as well as a color that is assigned to each unique
ShortBRED family. The ShortBRED marker data file lists the markers
that were identified. Finally, the Description of selected metagenomes file
provides available metadata associated with the selected metagenomes.
File
Size
CD-HIT ShortBRED families by cluster
ShortBRED marker data
Description of selected metagenomes
<1 MB
The default is for ShortBRED to report the abundance of metagenome hits for
CD-HIT families using the "median method." The numbers of metagenome hits
identified by all of the markers for a CD-HIT consensus sequence are arranged
in increasing numerical order; the value for the median marker is used as the
abundance. This method assumes that the distribution of hits across the markers
for CD-HIT consensus sequence is uniform (expected if the metagenome sequencing
is "deep," i.e., multiple coverage). For seed sequences with an even number of
markers, the average of the two "middle" markers is used as the abundance.
Files detailing the abundance information are available for download.
Raw Abundance Data
Raw results for the individual proteins in the SSN (Protein abundance data (median))
as well as summarized by SSN cluster (Cluster abundance data (median))
are provided. Units are in reads per kilobase of sequence per million sample reads (RPKM).
File
Size
Protein abundance data (median)
Cluster abundance data (median)
Average Genome Size-Normalized Abundance Data
Data are provided using Average Genome Size (AGS) normalization for
individual proteins in the SSN
as well as summarized by SSN cluster.
Units are have been converted from RPKM to counts per microbial genome, using AGS estimated by MicrobeCensus.
File
Size
Average genome size (AGS) normalized protein abundance data (median)
Average genome size (AGS) normalized cluster abundance data (median)
In the mean method for reporting abundances, the average value the abundances
identified by the markers for each CD-HIT consensus sequence marker is used to
report abundance. This method reports the presence of "any" hit for a marker
for a seed sequence. An asymmetric distribution of hits a seed sequence with
multiple markers is expected for "false positives," so the mean method should
be used with caution.
Files detailing the abundance information are available for download.
Raw Abundance Data
Raw results for the individual proteins in the SSN (Protein abundance data (mean))
as well as summarized by SSN cluster (Cluster abundance data (mean))
are provided. Units are in reads per kilobase of sequence per million sample reads (RPKM).
File
Size
Protein abundance data (mean)
Cluster abundance data (mean)
Average Genome Size-Normalized Abundance Data
Data are provided using Average Genome Size (AGS) normalization for
individual proteins in the SSN
as well as summarized by SSN cluster.
Units are have been converted from RPKM to counts per microbial genome, using AGS estimated by MicrobeCensus.
File
Size
Average genome size (AGS) normalized protein abundance data (mean)
Average genome size (AGS) normalized cluster abundance data (mean)
Heatmaps representing the quantification of sequences from SSN clusters per
metagenome are available.
The y-axis lists the SSN cluster numbers for which metagenome hits were
identified; the x-axis lists the metagenome datasets selected on the Identify
Results page. A color scale is located on the right that displays the AGS
normalized abundance of the number of gene copies for the "hit" per microbial
genome in the metagenome sample.
The metagenomes are grouped according to body site so that trends/consensus
across the six body sites can be easily discerned. The default heat map is
calculated using the median method to report abundances.
This heatmap presents information for SSN cluster/metagenome hit pairs.
This heatmap presents information for SSN singleton/metagenome hit pairs instead of SSN cluster/metagenome hit pairs.
This heatmap combines the information obtained for SSN cluster and singleton/metagenome hit pairs.
Tools for downloading and manipulating the heat map can be accessed by hovering and
clicking above and to the right of the plot.
Several filters are available for manipulating the heatmap.
Show specific clusters: input individual cluster numbers separated by
commas and/or a range of cluster numbers. Only these input clusters are displayed
in the heatmap.
Abundance to display: hide any data values that are outside of the minimum
and/or maximum. These hidden values appear as a zero value cell (i.e. the lowest
color range).
Use mean:
display the heatmap using the mean method for reporting abundances instead of
the defaut median method.
Display hits only: show a black and white heatmap showing presence/absence
of "hits" (which makes it easier to see low abundance hits).
Body Sites: checkboxes are provided for each body site in the heatmap;
selecting one or more of these checkboxes will show data for those body sites only.