Alignments for a candidate for dapF in Pseudomonas putida KT2440

GapMind for Amino acid biosynthesis

Alignments for a candidate for dapF in Pseudomonas putida KT2440

Align Diaminopimelate epimerase; DAP epimerase; PLP-independent amino acid racemase; EC 5.1.1.7 (characterized)
to candidate PP_5228 PP_5228 diaminopimelate epimerase

Query= SwissProt::P44859
         (274 letters)



>FitnessBrowser__Putida:PP_5228
          Length = 287

 Score =  328 bits (841), Expect = 8e-95
 Identities = 160/274 (58%), Positives = 204/274 (74%)

Query: 1   MQFSKMHGLGNDFVVVDGVTQNVFFTPETIRRLANRHCGIGFDQLLIVEAPYDPELDFHY 60
           ++F+KMHGLGNDF+V+D V+Q+    P+  ++  +RH G+GFDQLLIVEAP +PE+DF Y
Sbjct: 14  LRFTKMHGLGNDFMVLDLVSQHAHIQPKHAKQWGDRHTGVGFDQLLIVEAPNNPEVDFRY 73

Query: 61  RIFNADGSEVSQCGNGARCFARFVTLKGLTNKKDISVSTQKGNMVLTVKDDNQIRVNMGE 120
           RIFNADGSEV QCGNGARCFARFV  K LT KK I V T+ G +VL V++D Q+ V+MG 
Sbjct: 74  RIFNADGSEVEQCGNGARCFARFVLDKRLTAKKRIRVETKSGIIVLDVQNDGQVSVDMGP 133

Query: 121 PIWEPAKIPFTANKFEKNYILRTDIQTVLCGAVSMGNPHCVVQVDDIQTANVEQLGPLLE 180
           P + PA+IPF A+    NY L  D Q     AVSMGNPH V++VDD+QTA V +LGP +E
Sbjct: 134 PRFIPAEIPFVADAQALNYPLEVDGQLHSIAAVSMGNPHAVLRVDDVQTAPVHELGPKIE 193

Query: 181 SHERFPERVNAGFMQIINKEHIKLRVYERGAGETQACGSGACAAVAVGIMQGLLNNNVQV 240
           +H RFP+RVNAGF+Q+I++    LRV+ERGAGETQACG+GACAA    I QG +++ V +
Sbjct: 194 NHPRFPQRVNAGFIQVIDRHRANLRVWERGAGETQACGTGACAAAVAAISQGWMDSPVSL 253

Query: 241 DLPGGSLMIEWNGVGHPLYMTGEATHIYDGFITL 274
           DLPGG L IEW G G P+ MTG A  +Y+G + L
Sbjct: 254 DLPGGRLHIEWAGPGKPVMMTGPAVRVYEGQVRL 287


Lambda     K      H
   0.321    0.139    0.423 

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: 270
Number of extensions: 5
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: 274
Length of database: 287
Length adjustment: 26
Effective length of query: 248
Effective length of database: 261
Effective search space:    64728
Effective search space used:    64728
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 PP_5228 PP_5228 (diaminopimelate epimerase)
to HMM TIGR00652 (dapF: diaminopimelate epimerase (EC 5.1.1.7))

# 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.aa/TIGR00652.hmm
# target sequence database:        /tmp/gapView.6747.genome.faa
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

Query:       TIGR00652  [M=270]
Accession:   TIGR00652
Description: DapF: diaminopimelate epimerase
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-97  312.6   0.0    1.3e-97  312.4   0.0    1.0  1  lcl|FitnessBrowser__Putida:PP_5228  PP_5228 diaminopimelate epimeras


Domain annotation for each sequence (and alignments):
>> lcl|FitnessBrowser__Putida:PP_5228  PP_5228 diaminopimelate epimerase
   #    score  bias  c-Evalue  i-Evalue hmmfrom  hmm to    alifrom  ali to    envfrom  env to     acc
 ---   ------ ----- --------- --------- ------- -------    ------- -------    ------- -------    ----
   1 !  312.4   0.0   1.3e-97   1.3e-97       2     268 ..      15     285 ..      14     287 .] 0.95

  Alignments for each domain:
  == domain 1  score: 312.4 bits;  conditional E-value: 1.3e-97
                           TIGR00652   2 eFlkmhGlgNdFvlvdevdeelvkeeaelvrkvcdrhtgvgaDgvllvep.sseeadvklrifNsDGSeaemCGN 75 
                                         +F+kmhGlgNdF+++d v ++   + ++ +++  drhtgvg+D++l+ve+ +++e d+++rifN+DGSe+e+CGN
  lcl|FitnessBrowser__Putida:PP_5228  15 RFTKMHGLGNDFMVLDLVSQHAHIQ-PKHAKQWGDRHTGVGFDQLLIVEApNNPEVDFRYRIFNADGSEVEQCGN 88 
                                         8******************888888.9*********************987999********************* PP

                           TIGR00652  76 giRcfakfvyekglkekkelsvetlaglikveveeenkkvkvdmgepkfkkeeipltvekeeekeellalev... 147
                                         g+Rcfa+fv +k l+ kk+++vet++g+i++ v++++ +v+vdmg p+f ++eip++++ ++ ++ l+       
  lcl|FitnessBrowser__Putida:PP_5228  89 GARCFARFVLDKRLTAKKRIRVETKSGIIVLDVQNDG-QVSVDMGPPRFIPAEIPFVADAQALNYPLEVDGQlhs 162
                                         *************************************.*******************988888665544333444 PP

                           TIGR00652 148 l.vvdvGnPHlvvfvedvekldleelgklleaheefpegvNvefvevkkedeiklrvyERGageTlaCGtGavAs 221
                                         + +v++GnPH+v+ v+dv++++++elg+++e+h++fp++vN  f++v ++++  lrv+ERGageT+aCGtGa+A+
  lcl|FitnessBrowser__Putida:PP_5228 163 IaAVSMGNPHAVLRVDDVQTAPVHELGPKIENHPRFPQRVNAGFIQVIDRHRANLRVWERGAGETQACGTGACAA 237
                                         36************************************************************************* PP

                           TIGR00652 222 avvalklgktkkkvtvhleggeLeievkedg.kvyltGpavlvlegel 268
                                         av+a+ +g +++ v   l+gg+L+ie+   g  v++tGpav+v+eg++
  lcl|FitnessBrowser__Putida:PP_5228 238 AVAAISQGWMDSPVSLDLPGGRLHIEWAGPGkPVMMTGPAVRVYEGQV 285
                                         *******************************99*************97 PP



Internal pipeline statistics summary:
-------------------------------------
Query model(s):                            1  (270 nodes)
Target sequences:                          1  (287 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 0.00s 00:00:00.01 Elapsed: 00:00:00.00
# Mc/sec: 8.06
//
[ok]

This GapMind analysis is from Apr 09 2024. The underlying query database was built on Apr 09 2024.

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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
changes to Amino acid biosynthesis since the publication.

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 Amino acid biosynthesis