Alignments for a candidate for ech in Escherichia coli BW25113

GapMind for catabolism of small carbon sources

Alignments for a candidate for ech in Escherichia coli BW25113

Align Probable enoyl-CoA hydratase; EC 4.2.1.17 (uncharacterized)
to candidate 15516 b1394 enoyl-CoA hydratase (NCBI)

Query= curated2:P24162
         (257 letters)



>FitnessBrowser__Keio:15516
          Length = 262

 Score =  213 bits (541), Expect = 4e-60
 Identities = 114/256 (44%), Positives = 157/256 (61%), Gaps = 10/256 (3%)

Query: 10  ISEGLAVITLDRPEVMNALNAAMRHELTAALHRARGE--ARAIVLTGSGRAFCSGQDLGD 67
           + +G+  +TL+RPE +N+ N  M  +L   L +   +   R ++LTG+GR FC+GQDL D
Sbjct: 9   VEKGVMTLTLNRPERLNSFNDEMHAQLAECLKQVERDDTIRCLLLTGAGRGFCAGQDLND 68

Query: 68  ------GAAEGLNLETVLREEYEPLLQAIYSCPLPVLAAVNGAAAGAGANLALAADVVIA 121
                 G A  L +   +   Y PL++ +   P PV+ AVNG AAGAGA LAL  D+VIA
Sbjct: 69  RNVDPTGPAPDLGMS--VERFYNPLVRRLAKLPKPVICAVNGVAAGAGATLALGGDIVIA 126

Query: 122 AQSAAFMQAFTRIGLMPDAGGTWWLPRQVGMARAMGMALFAEKIGAEEAARMGLIWEAVP 181
           A+SA F+ AF+++GL+PD GGTW LPR  G ARAMG+AL   ++ AE+A   G+IW+ V 
Sbjct: 127 ARSAKFVMAFSKLGLIPDCGGTWLLPRVAGRARAMGLALLGNQLSAEQAHEWGMIWQVVD 186

Query: 182 DVDFEHHWRARAAHLARGPSAAFAAVKKAFHAGLSNPLPAQLALEARLQGELGQSADFRE 241
           D       +  A HLA  P+     +K+A ++  +N L  QL LE   Q   G+SAD+RE
Sbjct: 187 DETLADTAQQLARHLATQPTFGLGLIKQAINSAETNTLDTQLDLERDYQRLAGRSADYRE 246

Query: 242 GVQAFLEKRPPHFTGR 257
           GV AFL KR P FTG+
Sbjct: 247 GVSAFLAKRSPQFTGK 262


Lambda     K      H
   0.321    0.133    0.397 

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: 157
Number of extensions: 6
Number of successful extensions: 3
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: 257
Length of database: 262
Length adjustment: 24
Effective length of query: 233
Effective length of database: 238
Effective search space:    55454
Effective search space used:    55454
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 (21.9 bits)
S2: 47 (22.7 bits)

Align candidate 15516 b1394 (enoyl-CoA hydratase (NCBI))
to HMM TIGR02280 (paaB: phenylacetate degradation probable enoyl-CoA hydratase PaaB (EC 4.2.1.17))

# hmmsearch :: search profile(s) against a sequence database
# HMMER 3.3.1 (Jul 2020); http://hmmer.org/
# Copyright (C) 2020 Howard Hughes Medical Institute.
# Freely distributed under the BSD open source license.
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
# query HMM file:                  ../tmp/path.carbon/TIGR02280.hmm
# target sequence database:        /tmp/gapView.25888.genome.faa
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

Query:       TIGR02280  [M=256]
Accession:   TIGR02280
Description: PaaB1: phenylacetate degradation probable enoyl-CoA hydratase PaaB
Scores for complete sequences (score includes all domains):
   --- full sequence ---   --- best 1 domain ---    -#dom-
    E-value  score  bias    E-value  score  bias    exp  N  Sequence                       Description
    ------- ------ -----    ------- ------ -----   ---- --  --------                       -----------
   1.2e-149  482.9   0.7   1.3e-149  482.7   0.7    1.0  1  lcl|FitnessBrowser__Keio:15516  b1394 enoyl-CoA hydratase (NCBI)


Domain annotation for each sequence (and alignments):
>> lcl|FitnessBrowser__Keio:15516  b1394 enoyl-CoA hydratase (NCBI)
   #    score  bias  c-Evalue  i-Evalue hmmfrom  hmm to    alifrom  ali to    envfrom  env to     acc
 ---   ------ ----- --------- --------- ------- -------    ------- -------    ------- -------    ----
   1 !  482.7   0.7  1.3e-149  1.3e-149       1     256 []       5     262 .]       5     262 .] 1.00

  Alignments for each domain:
  == domain 1  score: 482.7 bits;  conditional E-value: 1.3e-149
                       TIGR02280   1 illelekgvlrltlnrpdklnsfteemhaelaealerverdd.vrallltGaGrGfcaGqdlsernvtkg.aapdlGet 77 
                                     il+++ekgv++ltlnrp++lnsf++emha+lae+l++verdd +r+llltGaGrGfcaGqdl++rnv+++ +apdlG++
  lcl|FitnessBrowser__Keio:15516   5 ILSHVEKGVMTLTLNRPERLNSFNDEMHAQLAECLKQVERDDtIRCLLLTGAGRGFCAGQDLNDRNVDPTgPAPDLGMS 83 
                                     89****************************************9**************************999******* PP

                       TIGR02280  78 vekfynplvrrlaalpkpvvvavnGvaaGaGanlalagdivlaaesakfiqafaklGlipdsGGtwllprlvGrarakg 156
                                     ve+fynplvrrla+lpkpv++avnGvaaGaGa+lal+gdiv+aa+sakf++af+klGlipd+GGtwllpr++Grara+g
  lcl|FitnessBrowser__Keio:15516  84 VERFYNPLVRRLAKLPKPVICAVNGVAAGAGATLALGGDIVIAARSAKFVMAFSKLGLIPDCGGTWLLPRVAGRARAMG 162
                                     ******************************************************************************* PP

                       TIGR02280 157 lallgekldaetaaewGliwqvvddealadevqalaahlakqptrglalikralqaaetnsldtqldlerdlqrelGrs 235
                                     lallg++l+ae+a+ewG+iwqvvdde+lad++q+la+hla+qpt+gl+lik+a+++aetn+ldtqldlerd+qr +Grs
  lcl|FitnessBrowser__Keio:15516 163 LALLGNQLSAEQAHEWGMIWQVVDDETLADTAQQLARHLATQPTFGLGLIKQAINSAETNTLDTQLDLERDYQRLAGRS 241
                                     ******************************************************************************* PP

                       TIGR02280 236 adyaeGvaafldkrepefkGk 256
                                     ady+eGv+afl+kr+p+f+Gk
  lcl|FitnessBrowser__Keio:15516 242 ADYREGVSAFLAKRSPQFTGK 262
                                     ********************7 PP



Internal pipeline statistics summary:
-------------------------------------
Query model(s):                            1  (256 nodes)
Target sequences:                          1  (262 residues searched)
Passed MSV filter:                         1  (1); expected 0.0 (0.02)
Passed bias filter:                        1  (1); expected 0.0 (0.02)
Passed Vit filter:                         1  (1); expected 0.0 (0.001)
Passed Fwd filter:                         1  (1); expected 0.0 (1e-05)
Initial search space (Z):                  1  [actual number of targets]
Domain search space  (domZ):               1  [number of targets reported over threshold]
# CPU time: 0.01u 0.01s 00:00:00.02 Elapsed: 00:00:00.01
# Mc/sec: 5.79
//
[ok]

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