Alignments for a candidate for puuA in Escherichia coli BW25113

GapMind for catabolism of small carbon sources

Alignments for a candidate for puuA in Escherichia coli BW25113

Align Gamma-glutamyl-putrescine synthetase (EC 6.3.1.11) (characterized)
to candidate 17910 b3870 glutamine synthetase (NCBI)

Query= reanno::BFirm:BPHYT_RS23160
         (444 letters)



>FitnessBrowser__Keio:17910
          Length = 469

 Score =  134 bits (337), Expect = 6e-36
 Identities = 123/398 (30%), Positives = 175/398 (43%), Gaps = 38/398 (9%)

Query: 62  GTLTGVTDPDMVCVPDASTIRMIPWAVDPTAQVIHDCVHFDGTPVAIS--PRRVLRRVLE 119
           G   G+ + DMV +PDAST  + P+  D T  +  D +   GT       PR + +R  +
Sbjct: 56  GGWKGINESDMVLMPDASTAVIDPFFADSTLIIRCDILE-PGTLQGYDRDPRSIAKRAED 114

Query: 120 LYKAKGWKPVI--APELEFYLVD-----------------MNKDPDLPLQPPIGRTG-RP 159
             ++ G    +   PE EF+L D                 +    +   Q   G  G RP
Sbjct: 115 YLRSTGIADTVLFGPEPEFFLFDDIRFGSSISGSHVAIDDIEGAWNSSTQYEGGNKGHRP 174

Query: 160 ETGRQAYSIEAVNEFDPLFEDIYEYCEVQELEVDTLIHEVGAA-QMEINFMHGDPLKLAD 218
                 + +  V+    +  ++    E   L V+   HEV  A Q E+        K AD
Sbjct: 175 AVKGGYFPVPPVDSAQDIRSEMCLVMEQMGLVVEAHHHEVATAGQNEVATRFNTMTKKAD 234

Query: 219 SVFLFKRTVREAALRHKMYATFMAKPMEGEPGSAMHMHQSLVDEETGHNLFTGPDGKPTS 278
            + ++K  V   A R    ATFM KPM G+ GS MH H SL   + G NLF G      S
Sbjct: 235 EIQIYKYVVHNVAHRFGKTATFMPKPMFGDNGSGMHCHMSL--SKNGVNLFAGDKYAGLS 292

Query: 279 -LFTSYIAGLQKYTPALMPIFAPYINSYRRLSRFMAAPINVAWGYDNRTVGFRIP-HSGP 336
                YI G+ K+  A+  +  P  NSY+RL     AP+ +A+   NR+   RIP  S P
Sbjct: 293 EQALYYIGGVIKHAKAINALANPTTNSYKRLVPGYEAPVMLAYSARNRSASIRIPVVSSP 352

Query: 337 AARRIENRIPGVDCNPYLAIAATLAAGYLGMTQKLEATEPLLSDGYELP-------YQLP 389
            ARRIE R P    NPYL  AA L AG  G+  K+   E +  + Y+LP        Q+ 
Sbjct: 353 KARRIEVRFPDPAANPYLCFAALLMAGLDGIKNKIHPGEAMDKNLYDLPPEEAKEIPQVA 412

Query: 390 RNLEEGLTLMGACEPIAE---VLGEKFVKAYLALKETE 424
            +LEE L  +       +   V  ++ + AY+AL+  E
Sbjct: 413 GSLEEALNELDLDREFLKAGGVFTDEAIDAYIALRREE 450


Lambda     K      H
   0.321    0.138    0.418 

Gapped
Lambda     K      H
   0.267   0.0410    0.140 


Matrix: BLOSUM62
Gap Penalties: Existence: 11, Extension: 1
Number of Sequences: 1
Number of Hits to DB: 480
Number of extensions: 40
Number of successful extensions: 4
Number of sequences better than 1.0e-02: 1
Number of HSP's gapped: 1
Number of HSP's successfully gapped: 1
Length of query: 444
Length of database: 469
Length adjustment: 33
Effective length of query: 411
Effective length of database: 436
Effective search space:   179196
Effective search space used:   179196
Neighboring words threshold: 11
Window for multiple hits: 40
X1: 16 ( 7.4 bits)
X2: 38 (14.6 bits)
X3: 64 (24.7 bits)
S1: 41 (21.8 bits)
S2: 51 (24.3 bits)

This GapMind analysis is from Sep 17 2021. The underlying query database was built on Sep 17 2021.

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About GapMind

Each pathway is defined by a set of rules based on individual steps or genes. Candidates for each step are identified by using ublast (a fast alternative to protein BLAST) against a database of manually-curated proteins (most of which are experimentally characterized) or by using HMMer with enzyme models (usually from TIGRFam). Ublast hits may be split across two different proteins.

A candidate for a step is "high confidence" if either:

ublast finds a hit to a characterized protein at above 40% identity and 80% coverage, and bits >= other bits+10.
- (Hits to curated proteins without experimental data as to their function are never considered high confidence.)
HMMer finds a hit with 80% coverage of the model, and either other identity < 40 or other coverage < 0.75.

where "other" refers to the best ublast hit to a sequence that is not annotated as performing this step (and is not "ignored").

Otherwise, a candidate is "medium confidence" if either:

ublast finds a hit at above 40% identity and 70% coverage (ignoring otherBits).
ublast finds a hit at above 30% identity and 80% coverage, and bits >= other bits.
HMMer finds a hit (regardless of coverage or other bits).

Other blast hits with at least 50% coverage are "low confidence."

Steps with no high- or medium-confidence candidates may be considered "gaps." For the typical bacterium that can make all 20 amino acids, there are 1-2 gaps in amino acid biosynthesis pathways. For diverse bacteria and archaea that can utilize a carbon source, there is a complete high-confidence catabolic pathway (including a transporter) just 38% of the time, and there is a complete medium-confidence pathway 63% of the time. Gaps may be due to:

our ignorance of proteins' functions,
omissions in the gene models,
frame-shift errors in the genome sequence, or
the organism lacks the pathway.

GapMind relies on the predicted proteins in the genome and does not search the six-frame translation. In most cases, you can search the six-frame translation by clicking on links to Curated BLAST for each step definition (in the per-step page).

For more information, see:

the paper from 2019 on GapMind for amino acid biosynthesis
the paper from 2022 on GapMind for carbon sources
the source code
instructions for running GapMind on your computer

If you notice any errors or omissions in the step descriptions, or any questionable results, please let us know

by Morgan Price, Arkin group, Lawrence Berkeley National Laboratory

GapMind for catabolism of small carbon sources