Alignments for a candidate for pcaF in Pseudomonas fluorescens FW300-N2C3

GapMind for catabolism of small carbon sources

Alignments for a candidate for pcaF in Pseudomonas fluorescens FW300-N2C3

Align subunit of β-ketoadipyl CoA thiolase (EC 2.3.1.174; EC 2.3.1.16) (characterized)
to candidate AO356_04930 AO356_04930 beta-ketoadipyl CoA thiolase

Query= metacyc::MONOMER-3207
         (400 letters)



>FitnessBrowser__pseudo5_N2C3_1:AO356_04930
          Length = 400

 Score =  715 bits (1846), Expect = 0.0
 Identities = 359/400 (89%), Positives = 385/400 (96%)

Query: 1   MRDVFICDAIRTPIGRFGGALAGVRADDLAAVPLKALIEPNPAVQWDQVDEVFFGCANQA 60
           MRDV+ICDAIRTPIGRFGG L+ VRADDLAAVP+KAL+E NP+V W+ VDEVF GCANQA
Sbjct: 1   MRDVYICDAIRTPIGRFGGGLSAVRADDLAAVPIKALMERNPSVDWNAVDEVFLGCANQA 60

Query: 61  GEDNRNVARMALLLAGLPESIPGVTLNRLCASGMDAIGTAFRAIASGEMELAIAGGVESM 120
           GEDNRNVARMALLLAGLP+SIPGVTLNRLCASGMDAIGTAFRAIASGEMELAIAGGVESM
Sbjct: 61  GEDNRNVARMALLLAGLPQSIPGVTLNRLCASGMDAIGTAFRAIASGEMELAIAGGVESM 120

Query: 121 SRAPFVMGKAESGYSRNMKLEDTTIGWRFINPLMKSQYGVDSMPETADNVADDYQVSRAD 180
           SRAPFVMGKA++ +SRNMKLEDTTIGWRFINPLMK+QYGVD+MP+TADNVADDY VSRAD
Sbjct: 121 SRAPFVMGKADTAFSRNMKLEDTTIGWRFINPLMKAQYGVDAMPQTADNVADDYAVSRAD 180

Query: 181 QDAFALRSQQKAAAAQAAGFFAEEIVPVRIAHKKGETIVERDEHLRPETTLEALTKLKPV 240
           QDAFALRSQQ+ AAAQAAGFFAEEIVPVRIAHKKGET+VE+DEH R +TTLE L+KLKPV
Sbjct: 181 QDAFALRSQQRTAAAQAAGFFAEEIVPVRIAHKKGETVVEQDEHPRADTTLETLSKLKPV 240

Query: 241 NGPDKTVTAGNASGVNDGAAALILASAEAVKKHGLTPRARVLGMASGGVAPRVMGIGPVP 300
           NGPDKTVTAGNASGVNDGAAALILASAEAVKKHGLTPR +VLGMAS GVAPRVMGIGPVP
Sbjct: 241 NGPDKTVTAGNASGVNDGAAALILASAEAVKKHGLTPRGKVLGMASAGVAPRVMGIGPVP 300

Query: 301 AVRKLTERLGVAVSDFDVIELNEAFASQGLAVLRELGVADDAPQVNPNGGAIALGHPLGM 360
           AVRKLTERLG+AV+DFDVIELNEAFASQGLAVLR+LG+ADDAPQVNPNGGAIALGHPLGM
Sbjct: 301 AVRKLTERLGLAVADFDVIELNEAFASQGLAVLRDLGLADDAPQVNPNGGAIALGHPLGM 360

Query: 361 SGARLVLTALHQLEKSGGRKGLATMCVGVGQGLALAIERV 400
           SGARLV+TALH LEK+GG+KGLATMCVGVGQGLALAIERV
Sbjct: 361 SGARLVMTALHHLEKTGGKKGLATMCVGVGQGLALAIERV 400


Lambda     K      H
   0.318    0.134    0.383 

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: 668
Number of extensions: 15
Number of successful extensions: 1
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: 400
Length of database: 400
Length adjustment: 31
Effective length of query: 369
Effective length of database: 369
Effective search space:   136161
Effective search space used:   136161
Neighboring words threshold: 11
Window for multiple hits: 40
X1: 16 ( 7.3 bits)
X2: 38 (14.6 bits)
X3: 64 (24.7 bits)
S1: 41 (21.7 bits)
S2: 50 (23.9 bits)

Align candidate AO356_04930 AO356_04930 (beta-ketoadipyl CoA thiolase)
to HMM TIGR02430 (pcaF: 3-oxoadipyl-CoA thiolase (EC 2.3.1.174))

# 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/TIGR02430.hmm
# target sequence database:        /tmp/gapView.26792.genome.faa
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

Query:       TIGR02430  [M=400]
Accession:   TIGR02430
Description: pcaF: 3-oxoadipyl-CoA thiolase
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
    ------- ------ -----    ------- ------ -----   ---- --  --------                                       -----------
     3e-227  740.2  10.3   3.4e-227  740.0  10.3    1.0  1  lcl|FitnessBrowser__pseudo5_N2C3_1:AO356_04930  AO356_04930 beta-ketoadipyl CoA 


Domain annotation for each sequence (and alignments):
>> lcl|FitnessBrowser__pseudo5_N2C3_1:AO356_04930  AO356_04930 beta-ketoadipyl CoA thiolase
   #    score  bias  c-Evalue  i-Evalue hmmfrom  hmm to    alifrom  ali to    envfrom  env to     acc
 ---   ------ ----- --------- --------- ------- -------    ------- -------    ------- -------    ----
   1 !  740.0  10.3  3.4e-227  3.4e-227       1     400 []       2     400 .]       2     400 .] 1.00

  Alignments for each domain:
  == domain 1  score: 740.0 bits;  conditional E-value: 3.4e-227
                                       TIGR02430   1 kevyivdairtpiGrygGslssvraddlaavplkallarnpsldaaaiddvilGcanqaGedn 63 
                                                     ++vyi+dairtpiGr+gG+ls+vraddlaavp+kal++rnps+d++a+d+v+lGcanqaGedn
  lcl|FitnessBrowser__pseudo5_N2C3_1:AO356_04930   2 RDVYICDAIRTPIGRFGGGLSAVRADDLAAVPIKALMERNPSVDWNAVDEVFLGCANQAGEDN 64 
                                                     689************************************************************ PP

                                       TIGR02430  64 rnvarmaallaGlpvsvpgttvnrlcgsgldalglaaraikaGeadlviaGGvesmsrapfvl 126
                                                     rnvarma+llaGlp+s+pg+t+nrlc+sg+da+g+a+rai++Ge +l iaGGvesmsrapfv+
  lcl|FitnessBrowser__pseudo5_N2C3_1:AO356_04930  65 RNVARMALLLAGLPQSIPGVTLNRLCASGMDAIGTAFRAIASGEMELAIAGGVESMSRAPFVM 127
                                                     *************************************************************** PP

                                       TIGR02430 127 GkadsafsrsakledttiGwrfvnpklkalyGvdsmpetaenvaeefgvsredqdafalrsqq 189
                                                     Gkad+afsr+ kledttiGwrf+np +ka+yGvd+mp+ta+nva++++vsr+dqdafalrsqq
  lcl|FitnessBrowser__pseudo5_N2C3_1:AO356_04930 128 GKADTAFSRNMKLEDTTIGWRFINPLMKAQYGVDAMPQTADNVADDYAVSRADQDAFALRSQQ 190
                                                     *************************************************************** PP

                                       TIGR02430 190 rtaaaqakGffaeeivpveikqkkGeetvvdkdehlraettlealaklkavvredgtvtaGna 252
                                                     rtaaaqa+Gffaeeivpv+i++kkG etvv++deh+ra+ttle+l+klk+v+ +d+tvtaGna
  lcl|FitnessBrowser__pseudo5_N2C3_1:AO356_04930 191 RTAAAQAAGFFAEEIVPVRIAHKKG-ETVVEQDEHPRADTTLETLSKLKPVNGPDKTVTAGNA 252
                                                     *************************.899********************************** PP

                                       TIGR02430 253 sGvndGaaalllaseeavkrhgltprarilaaasaGveprvmGlgpvpavkkllaraglsled 315
                                                     sGvndGaaal+las+eavk+hgltpr+++l++asaGv+prvmG+gpvpav+kl +r+gl+++d
  lcl|FitnessBrowser__pseudo5_N2C3_1:AO356_04930 253 SGVNDGAAALILASAEAVKKHGLTPRGKVLGMASAGVAPRVMGIGPVPAVRKLTERLGLAVAD 315
                                                     *************************************************************** PP

                                       TIGR02430 316 ldvielneafaaqalavlrelgladddarvnpnGGaialGhplGasGarlvltalkqleksgg 378
                                                     +dvielneafa+q+lavlr+lgladd+ +vnpnGGaialGhplG+sGarlv+tal++lek+gg
  lcl|FitnessBrowser__pseudo5_N2C3_1:AO356_04930 316 FDVIELNEAFASQGLAVLRDLGLADDAPQVNPNGGAIALGHPLGMSGARLVMTALHHLEKTGG 378
                                                     *************************************************************** PP

                                       TIGR02430 379 ryalatlciGvGqGialvierv 400
                                                     + +lat+c+GvGqG+al+ierv
  lcl|FitnessBrowser__pseudo5_N2C3_1:AO356_04930 379 KKGLATMCVGVGQGLALAIERV 400
                                                     *********************8 PP



Internal pipeline statistics summary:
-------------------------------------
Query model(s):                            1  (400 nodes)
Target sequences:                          1  (400 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: 0.02u 0.01s 00:00:00.03 Elapsed: 00:00:00.02
# Mc/sec: 6.91
//
[ok]

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

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Protein sequences (fasta format)
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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