Alignments for a candidate for astE in Pseudomonas putida KT2440

GapMind for catabolism of small carbon sources

Alignments for a candidate for astE in Pseudomonas putida KT2440

Align Succinylglutamate desuccinylase (EC 3.5.1.96) (characterized)
to candidate PP_4475 PP_4475 succinylglutamate desuccinylase

Query= reanno::pseudo13_GW456_L13:PfGW456L13_1977
         (336 letters)



>FitnessBrowser__Putida:PP_4475
          Length = 335

 Score =  509 bits (1312), Expect = e-149
 Identities = 257/335 (76%), Positives = 292/335 (87%), Gaps = 1/335 (0%)

Query: 1   MLALGKLLELTLAGREPAEKTQLTVEGVRMRWLSEGALEVRPPEARDNGLDLLLSAGIHG 60
           MLALGKLLELTL   EPAEKTQ+T +G R+RWL EGALEVRP E+ D GLDLLLSAGIHG
Sbjct: 1   MLALGKLLELTLTDHEPAEKTQVTPKGARLRWLGEGALEVRPAESDDCGLDLLLSAGIHG 60

Query: 61  NETAPIELLDRLLHDIARGDLKPRARILFLFGNPEAIRKGERFVEQDVNRLFNGRHEQSS 120
           NETAPIELL+RLLH +A G ++PRAR+LFLFGNP AIRKGERF+EQD+NRLFNGRHE SS
Sbjct: 61  NETAPIELLERLLHGVANGKIRPRARVLFLFGNPAAIRKGERFIEQDINRLFNGRHELSS 120

Query: 121 GCEALRACELERLAASFFSLPDRQRLHYDLHTAIRGSKIEQFALYPWKEDRQHSRQELAR 180
           G EALRA ELE+ A  FFS P R RLHYDLHTAIRGSKIEQFALYP+KE R+HSR+ELAR
Sbjct: 121 GFEALRAAELEQFARVFFSKPGRNRLHYDLHTAIRGSKIEQFALYPYKEGRKHSRRELAR 180

Query: 181 LRAAGMEAVLLQNKPSIVFSSYTYDKLGAEAFTLELGKARPFGQNDGVNVSLLEKRLKQI 240
           L AAGMEAVLLQ+K SI FS++TY++L AEAFTLELGKARPFGQN+ VN+  LE+RL +I
Sbjct: 181 LAAAGMEAVLLQSKSSITFSAFTYEQLEAEAFTLELGKARPFGQNEQVNLDKLEERLIRI 240

Query: 241 IEGTEPELTEDALDGLQLYSVAREIIKHSDSFRLNLPADIENFSELEVGYLLAEDIANTR 300
           IE TEPE  E +LDGLQL+SV+REIIKHSDSF L+LPADIENFSEL  GYLLAED+A  R
Sbjct: 241 IEATEPE-DESSLDGLQLFSVSREIIKHSDSFHLHLPADIENFSELSKGYLLAEDLAEMR 299

Query: 301 WIIEEEGARIIFPNPRVKNGLRAGILVVPTTDENL 335
           W++EEEGARIIFPNP+VKNGLRAGIL+VP + + L
Sbjct: 300 WVVEEEGARIIFPNPKVKNGLRAGILIVPDSGQRL 334


Lambda     K      H
   0.319    0.138    0.394 

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: 435
Number of extensions: 18
Number of successful extensions: 2
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: 336
Length of database: 335
Length adjustment: 28
Effective length of query: 308
Effective length of database: 307
Effective search space:    94556
Effective search space used:    94556
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 (21.7 bits)
S2: 49 (23.5 bits)

Align candidate PP_4475 PP_4475 (succinylglutamate desuccinylase)
to HMM TIGR03242 (astE: succinylglutamate desuccinylase (EC 3.5.1.96))

# 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/TIGR03242.hmm
# target sequence database:        /tmp/gapView.7137.genome.faa
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

Query:       TIGR03242  [M=319]
Accession:   TIGR03242
Description: arg_catab_astE: succinylglutamate desuccinylase
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
    ------- ------ -----    ------- ------ -----   ---- --  --------                           -----------
   6.2e-135  435.5   0.0   7.1e-135  435.4   0.0    1.0  1  lcl|FitnessBrowser__Putida:PP_4475  PP_4475 succinylglutamate desucc


Domain annotation for each sequence (and alignments):
>> lcl|FitnessBrowser__Putida:PP_4475  PP_4475 succinylglutamate desuccinylase
   #    score  bias  c-Evalue  i-Evalue hmmfrom  hmm to    alifrom  ali to    envfrom  env to     acc
 ---   ------ ----- --------- --------- ------- -------    ------- -------    ------- -------    ----
   1 !  435.4   0.0  7.1e-135  7.1e-135       1     319 []       6     327 ..       6     327 .. 0.97

  Alignments for each domain:
  == domain 1  score: 435.4 bits;  conditional E-value: 7.1e-135
                           TIGR03242   1 dflaltlekkep.evtqgeaknvklrwldeGvlelePe..aeaekslvisaGihGnetaPielleqllsdiaagk 72 
                                         ++l+ltl+++ep e+tq++ k+++lrwl+eG+le++P+  +++  +l++saGihGnetaPielle+ll+ +a+gk
  lcl|FitnessBrowser__Putida:PP_4475   6 KLLELTLTDHEPaEKTQVTPKGARLRWLGEGALEVRPAesDDCGLDLLLSAGIHGNETAPIELLERLLHGVANGK 80 
                                         589********98999999******************9888999******************************* PP

                           TIGR03242  73 lqlkvrlLlilGnpaalrkgkRyleedlnRlfgGryqeleeskeklRaeeLeqvveaffeagkasearyhyDlht 147
                                         ++ ++r+L+++Gnpaa+rkg+R++e+d+nRlf+Gr+ el++++e+lRa+eLeq++++ff++++++  r+hyDlht
  lcl|FitnessBrowser__Putida:PP_4475  81 IRPRARVLFLFGNPAAIRKGERFIEQDINRLFNGRH-ELSSGFEALRAAELEQFARVFFSKPGRN--RLHYDLHT 152
                                         ************************************.9**************************9..8******* PP

                           TIGR03242 148 aiRasklekfallPyq.ekpfdkellewlaaadldavllhkekggtfshfssekleaeactlelGkarPfGendl 221
                                         aiR+sk+e+fal+Py+ ++++++++l++laaa+++avll++++++tfs+f++e+leaea+tlelGkarPfG+n++
  lcl|FitnessBrowser__Putida:PP_4475 153 AIRGSKIEQFALYPYKeGRKHSRRELARLAAAGMEAVLLQSKSSITFSAFTYEQLEAEAFTLELGKARPFGQNEQ 227
                                         ****************99********************************************************* PP

                           TIGR03242 222 sqfqaitealralisdeaiparkke..elklfevvesilkksdsfelhvaedasnftefakGtllaedkde.ryr 293
                                         +++++++e+l+++i ++++p+++++   l+lf+v ++i+k+sdsf+lh+++d++nf+e++kG+llaed +e r++
  lcl|FitnessBrowser__Putida:PP_4475 228 VNLDKLEERLIRII-EATEPEDESSldGLQLFSVSREIIKHSDSFHLHLPADIENFSELSKGYLLAEDLAEmRWV 301
                                         **************.7788887776679****************************************9988*** PP

                           TIGR03242 294 veeeeerilfPnakvanGlRaglllv 319
                                         veee++ri+fPn+kv+nGlRag+l+v
  lcl|FitnessBrowser__Putida:PP_4475 302 VEEEGARIIFPNPKVKNGLRAGILIV 327
                                         ***********************996 PP



Internal pipeline statistics summary:
-------------------------------------
Query model(s):                            1  (319 nodes)
Target sequences:                          1  (335 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: 00:00:00.01
# Mc/sec: 9.00
//
[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