Alignments for a candidate for maiA in Pseudomonas fluorescens FW300-N1B4

GapMind for catabolism of small carbon sources

Alignments for a candidate for maiA in Pseudomonas fluorescens FW300-N1B4

Align Maleylacetoacetate isomerase (EC 5.2.1.2) (characterized)
to candidate Pf1N1B4_1023 Maleylacetoacetate isomerase (EC 5.2.1.2) @ Glutathione S-transferase, zeta (EC 2.5.1.18)

Query= reanno::pseudo3_N2E3:AO353_15560
         (211 letters)



>FitnessBrowser__pseudo1_N1B4:Pf1N1B4_1023
          Length = 211

 Score =  392 bits (1008), Expect = e-114
 Identities = 190/211 (90%), Positives = 198/211 (93%)

Query: 1   MELFTYYRSTSSFRVRIALALKGLEYQALPINLIAPQGGEHQQPAYLHINPQGRVPALRT 60
           MEL+TYYRSTSS+RVRIALALKGL+YQALPINLIAPQGGEH+QP YL INPQGRVPALRT
Sbjct: 1   MELYTYYRSTSSYRVRIALALKGLDYQALPINLIAPQGGEHRQPTYLGINPQGRVPALRT 60

Query: 61  DEGELLIQSPAIIEYLEERYPQVPLLSKDLARRAHERGVAALIGCDIHPLHNVSVLNQLR 120
           DEGELLIQSPAIIEYLEERYPQVPLLSKDL  RAHERGVAA+IGCD+HPLHNVSVLNQLR
Sbjct: 61  DEGELLIQSPAIIEYLEERYPQVPLLSKDLVARAHERGVAAVIGCDVHPLHNVSVLNQLR 120

Query: 121 QLGHDEPQVVHWIGHWITQGLAAVEHLIGDEGYCFGNAPGLADVYLIPQLYAAERFNISL 180
           QLGHDEPQVV WIGHWI+QGLA VE LIGD GYCFG+ PGLADVYLIPQLYAAERFNISL
Sbjct: 121 QLGHDEPQVVEWIGHWISQGLATVEQLIGDTGYCFGDQPGLADVYLIPQLYAAERFNISL 180

Query: 181 EAYPRIRRVAALAVQHPAFIKAHPANQPDTP 211
           EAYPRIRRVAALA  HPAF  AHPANQPDTP
Sbjct: 181 EAYPRIRRVAALAATHPAFFLAHPANQPDTP 211


Lambda     K      H
   0.322    0.139    0.427 

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: 317
Number of extensions: 9
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: 211
Length of database: 211
Length adjustment: 21
Effective length of query: 190
Effective length of database: 190
Effective search space:    36100
Effective search space used:    36100
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 Pf1N1B4_1023 (Maleylacetoacetate isomerase (EC 5.2.1.2) @ Glutathione S-transferase, zeta (EC 2.5.1.18))
to HMM TIGR01262 (maiA: maleylacetoacetate isomerase (EC 5.2.1.2))

# 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/TIGR01262.hmm
# target sequence database:        /tmp/gapView.6306.genome.faa
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

Query:       TIGR01262  [M=211]
Accession:   TIGR01262
Description: maiA: maleylacetoacetate isomerase
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
    ------- ------ -----    ------- ------ -----   ---- --  --------                                      -----------
    9.9e-86  272.7   0.0    1.1e-85  272.6   0.0    1.0  1  lcl|FitnessBrowser__pseudo1_N1B4:Pf1N1B4_1023  Maleylacetoacetate isomerase (EC


Domain annotation for each sequence (and alignments):
>> lcl|FitnessBrowser__pseudo1_N1B4:Pf1N1B4_1023  Maleylacetoacetate isomerase (EC 5.2.1.2) @ Glutathione S-transferase,
   #    score  bias  c-Evalue  i-Evalue hmmfrom  hmm to    alifrom  ali to    envfrom  env to     acc
 ---   ------ ----- --------- --------- ------- -------    ------- -------    ------- -------    ----
   1 !  272.6   0.0   1.1e-85   1.1e-85       2     210 ..       3     210 ..       2     211 .] 0.98

  Alignments for each domain:
  == domain 1  score: 272.6 bits;  conditional E-value: 1.1e-85
                                      TIGR01262   2 lYsyfrSsasyRvRiaLaLkgidyesvpvnLlkd..GeqkkeefkalNPqelvPtLkidegevl 63 
                                                    lY+y+rS++syRvRiaLaLkg+dy++ p+nL++   Ge++++ +  +NPq++vP+L++dege l
  lcl|FitnessBrowser__pseudo1_N1B4:Pf1N1B4_1023   3 LYTYYRSTSSYRVRIALALKGLDYQALPINLIAPqgGEHRQPTYLGINPQGRVPALRTDEGELL 66 
                                                    9******************************986679*************************** PP

                                      TIGR01262  64 tqSlAiieyLeetypepaLlpkdpakrarvralalliacdihPlqNlrvlqlleeklgvdeeek 127
                                                     qS AiieyLee+yp+++Ll+kd  +ra++r +a++i+cd+hPl+N++vl++l+ +lg+de + 
  lcl|FitnessBrowser__pseudo1_N1B4:Pf1N1B4_1023  67 IQSPAIIEYLEERYPQVPLLSKDLVARAHERGVAAVIGCDVHPLHNVSVLNQLR-QLGHDEPQV 129
                                                    ******************************************************.779****** PP

                                      TIGR01262 128 kewlkhwiekGlaalEellkekagafcvGdevtladvcLvpqvynAerfevdlaqyPtlkriee 191
                                                     ew+ hwi++Gla++E+l+ + +g +c+Gd+++ladv+L+pq+y+Aerf+++l++yP+++r+++
  lcl|FitnessBrowser__pseudo1_N1B4:Pf1N1B4_1023 130 VEWIGHWISQGLATVEQLIGD-TG-YCFGDQPGLADVYLIPQLYAAERFNISLEAYPRIRRVAA 191
                                                    *******************98.56.*************************************** PP

                                      TIGR01262 192 alaelpafqeahpenqpdt 210
                                                     +a++paf  ahp+nqpdt
  lcl|FitnessBrowser__pseudo1_N1B4:Pf1N1B4_1023 192 LAATHPAFFLAHPANQPDT 210
                                                    ******************8 PP



Internal pipeline statistics summary:
-------------------------------------
Query model(s):                            1  (211 nodes)
Target sequences:                          1  (211 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: 6.60
//
[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