Alignments for a candidate for leuD in Azospirillum brasilense Sp245

GapMind for Amino acid biosynthesis

Alignments for a candidate for leuD in Azospirillum brasilense Sp245

Align 3-isopropylmalate dehydratase subunit LeuD (EC 4.2.1.33) (characterized)
to candidate AZOBR_RS27810 AZOBR_RS27810 isopropylmalate isomerase

Query= ecocyc::LEUD-MONOMER
         (201 letters)



>FitnessBrowser__azobra:AZOBR_RS27810
          Length = 208

 Score =  182 bits (461), Expect = 5e-51
 Identities = 99/199 (49%), Positives = 131/199 (65%), Gaps = 3/199 (1%)

Query: 1   MAEKFIKHTGLVVPLDAANVDTDAIIPKQFLQKVTRTGFGAHLFNDWRFLDEKGQQPNPD 60
           M E   + TG+  PL  ANVDTDAIIPK  L  + R+G GA LF++WRF D+ G++  PD
Sbjct: 1   MPEPVNRITGIAAPLPRANVDTDAIIPKAHLLTIHRSGLGAGLFSEWRF-DDDGRE-RPD 58

Query: 61  FVLNFPQYQGASILLARENFGCGSSREHAPWALTDYGFKVVIAPSFADIFYGNSFNNQLL 120
           FVLN   ++ A ILLA ENFGCGSSREHA W+L D+G + VIAPSFA IF+ N   N L 
Sbjct: 59  FVLNQAPWREAKILLAGENFGCGSSREHAVWSLMDFGIRCVIAPSFASIFHENCQKNGLA 118

Query: 121 PVKLSDAEVDELFALVKANPGIHFDVDLEAQEVKAGE-KTYRFTIDAFRRHCMMNGLDSI 179
            V L +A +  L A  +A P     VD++A  V A + + + FT++A RR  ++ GLD I
Sbjct: 119 AVTLDEAALAHLVACAQATPAATTTVDIKASRVTAPDGREFPFTMEAARRQALLEGLDEI 178

Query: 180 GLTLQHDDAIAAYEAKQPA 198
           G +L+HD A+AA+EA+  A
Sbjct: 179 GASLRHDAAMAAFEARDRA 197


Lambda     K      H
   0.322    0.138    0.421 

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: 144
Number of extensions: 5
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: 201
Length of database: 208
Length adjustment: 21
Effective length of query: 180
Effective length of database: 187
Effective search space:    33660
Effective search space used:    33660
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: 45 (21.9 bits)

Align candidate AZOBR_RS27810 AZOBR_RS27810 (isopropylmalate isomerase)
to HMM TIGR00171 (leuD: 3-isopropylmalate dehydratase, small subunit (EC 4.2.1.33))

# 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/TIGR00171.hmm
# target sequence database:        /tmp/gapView.9927.genome.faa
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

Query:       TIGR00171  [M=188]
Accession:   TIGR00171
Description: leuD: 3-isopropylmalate dehydratase, small subunit
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-68  216.7   0.0    1.3e-68  216.6   0.0    1.0  1  lcl|FitnessBrowser__azobra:AZOBR_RS27810  AZOBR_RS27810 isopropylmalate is


Domain annotation for each sequence (and alignments):
>> lcl|FitnessBrowser__azobra:AZOBR_RS27810  AZOBR_RS27810 isopropylmalate isomerase
   #    score  bias  c-Evalue  i-Evalue hmmfrom  hmm to    alifrom  ali to    envfrom  env to     acc
 ---   ------ ----- --------- --------- ------- -------    ------- -------    ------- -------    ----
   1 !  216.6   0.0   1.3e-68   1.3e-68       3     186 ..       4     186 ..       1     188 [. 0.97

  Alignments for each domain:
  == domain 1  score: 216.6 bits;  conditional E-value: 1.3e-68
                                 TIGR00171   3 efkkltGlvvpldkanvdtdaiipkqflkkikrtGfgkhlfyewryldekGkepnpefvlnvpqyqgas 71 
                                               ++ + tG+++pl +anvdtdaiipk  l +i+r G+g+ lf ewr+ d++G  + p+fvln+ ++++a+
  lcl|FitnessBrowser__azobra:AZOBR_RS27810   4 PVNRITGIAAPLPRANVDTDAIIPKAHLLTIHRSGLGAGLFSEWRF-DDDG-RERPDFVLNQAPWREAK 70 
                                               57789*****************************************.9999.678************** PP

                                 TIGR00171  72 illarenfGcGssrehapwalkdyGfkviiapsfadifynnsfkngllpirlseeeveellalvk.nkg 139
                                               illa+enfGcGssreha w+l d+G++ +iapsfa if++n+ kngl  + l+e+ + +l+a  +  ++
  lcl|FitnessBrowser__azobra:AZOBR_RS27810  71 ILLAGENFGCGSSREHAVWSLMDFGIRCVIAPSFASIFHENCQKNGLAAVTLDEAALAHLVACAQaTPA 139
                                               **************************************************************9997899 PP

                                 TIGR00171 140 lkltvdleaqkvkdsegkvysfeidefrkhcllnGldeigltlqked 186
                                                  tvd++a +v++ +g+   f++++ r++ ll+Gldeig +l+++ 
  lcl|FitnessBrowser__azobra:AZOBR_RS27810 140 ATTTVDIKASRVTAPDGREFPFTMEAARRQALLEGLDEIGASLRHDA 186
                                               9*******************************************986 PP



Internal pipeline statistics summary:
-------------------------------------
Query model(s):                            1  (188 nodes)
Target sequences:                          1  (208 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: 6.64
//
[ok]

This GapMind analysis is from Aug 03 2021. The underlying query database was built on Aug 03 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, or view the source code, or see 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