Alignments for a candidate for PAH in Pseudomonas putida KT2440

GapMind for catabolism of small carbon sources

Alignments for a candidate for PAH in Pseudomonas putida KT2440

Align Phenylalanine 4-monooxygenase (EC 1.14.16.1) (characterized)
to candidate PP_4490 PP_4490 phenylalanine-4-hydroxylase

Query= reanno::pseudo13_GW456_L13:PfGW456L13_4394
         (261 letters)



>FitnessBrowser__Putida:PP_4490
          Length = 262

 Score =  493 bits (1268), Expect = e-144
 Identities = 233/261 (89%), Positives = 249/261 (95%)

Query: 1   MKQTQYVAREPDAQGFIDYPAEEHAVWNTLITRQLKVIEGRACQEYLDGIEKLGLPHDRI 60
           MKQTQYVAREPDA GFIDYP +EHAVWNTLITRQLKVIEGRACQEYLDGI++L LPHDRI
Sbjct: 1   MKQTQYVAREPDAHGFIDYPQQEHAVWNTLITRQLKVIEGRACQEYLDGIDQLKLPHDRI 60

Query: 61  PQLGEINKVLAETTGWQVARVPALIPFQTFFELLASKQFPVATFIRTREELDYLQEPDIF 120
           PQLGEINKVL  TTGWQVARVPALIPFQTFFELLASK+FPVATFIRT EELDYLQEPDIF
Sbjct: 61  PQLGEINKVLGATTGWQVARVPALIPFQTFFELLASKRFPVATFIRTPEELDYLQEPDIF 120

Query: 121 HEIFGHCPLLTNPWFAEFTHTYGKLGLKASKEERVYLARLYWMTIEFGLLDTPQGQRIYG 180
           HEIFGHCPLLTNPWFAEFTHTYGKLGL A+KE+RVYLARLYWMTIEFGL++T QG++IYG
Sbjct: 121 HEIFGHCPLLTNPWFAEFTHTYGKLGLAATKEQRVYLARLYWMTIEFGLMETAQGRKIYG 180

Query: 181 GGILSSPKETVYSLSNEPEHQAFDPLECMRTPYRIDILQPLYFVLPNLKRLFDLAHEDIM 240
           GGILSSPKETVYSLS+EPEHQAFDP+E MRTPYRIDILQP+YFVLPN+KRLFDLAHEDIM
Sbjct: 181 GGILSSPKETVYSLSDEPEHQAFDPIEAMRTPYRIDILQPVYFVLPNMKRLFDLAHEDIM 240

Query: 241 GMVKQAMQMGLHTPKFPPKAA 261
           GMV +AMQ+GLH PKFPPK A
Sbjct: 241 GMVHKAMQLGLHAPKFPPKVA 261


Lambda     K      H
   0.322    0.140    0.435 

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: 385
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: 261
Length of database: 262
Length adjustment: 25
Effective length of query: 236
Effective length of database: 237
Effective search space:    55932
Effective search space used:    55932
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_4490 PP_4490 (phenylalanine-4-hydroxylase)
to HMM TIGR01267 (phhA: phenylalanine-4-hydroxylase (EC 1.14.16.1))

# 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/TIGR01267.hmm
# target sequence database:        /tmp/gapView.28353.genome.faa
# - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

Query:       TIGR01267  [M=248]
Accession:   TIGR01267
Description: Phe4hydrox_mono: phenylalanine-4-hydroxylase
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
    ------- ------ -----    ------- ------ -----   ---- --  --------                           -----------
   1.5e-128  413.3   0.0   1.7e-128  413.1   0.0    1.0  1  lcl|FitnessBrowser__Putida:PP_4490  PP_4490 phenylalanine-4-hydroxyl


Domain annotation for each sequence (and alignments):
>> lcl|FitnessBrowser__Putida:PP_4490  PP_4490 phenylalanine-4-hydroxylase
   #    score  bias  c-Evalue  i-Evalue hmmfrom  hmm to    alifrom  ali to    envfrom  env to     acc
 ---   ------ ----- --------- --------- ------- -------    ------- -------    ------- -------    ----
   1 !  413.1   0.0  1.7e-128  1.7e-128       3     247 ..      11     254 ..       9     255 .. 0.99

  Alignments for each domain:
  == domain 1  score: 413.1 bits;  conditional E-value: 1.7e-128
                           TIGR01267   3 tvaqdldryseeehavwatlidrqlkllegracdeyldGveklgldadripdleevneklraltGwkivavpgli 77 
                                         ++a++++ y+++ehavw+tli+rqlk++egrac+eyldG+++l+l++drip+l e+n++l a+tGw++++vp+li
  lcl|FitnessBrowser__Putida:PP_4490  11 PDAHGFIDYPQQEHAVWNTLITRQLKVIEGRACQEYLDGIDQLKLPHDRIPQLGEINKVLGATTGWQVARVPALI 85 
                                         78999********************************************************************** PP

                           TIGR01267  78 padvffehlanrrfpvttflrtpeeldylqepdvfhdlfGhvpllsnpvfadfleayGkkgvkakalgaallarl 152
                                         p+++ffe+la++rfpv+tf+rtpeeldylqepd+fh++fGh+pll+np+fa+f+++yGk+g+ a++++++ larl
  lcl|FitnessBrowser__Putida:PP_4490  86 PFQTFFELLASKRFPVATFIRTPEELDYLQEPDIFHEIFGHCPLLTNPWFAEFTHTYGKLGLAATKEQRVYLARL 160
                                         *************************************************************************** PP

                           TIGR01267 153 ywytvefGlvetaaglriyGaGilsskkelvyaleskeplrvafdllevmrtryridklqkayfvlpslkrlfda 227
                                         yw+t+efGl+eta+g +iyG+Gilss+ke+vy+  s+ep+++afd++e+mrt+yrid+lq++yfvlp++krlfd+
  lcl|FitnessBrowser__Putida:PP_4490 161 YWMTIEFGLMETAQGRKIYGGGILSSPKETVYS-LSDEPEHQAFDPIEAMRTPYRIDILQPVYFVLPNMKRLFDL 234
                                         *********************************.8**************************************** PP

                           TIGR01267 228 aqedfealvaeakdlkaldp 247
                                         a+ed++ +v++a++l++++p
  lcl|FitnessBrowser__Putida:PP_4490 235 AHEDIMGMVHKAMQLGLHAP 254
                                         ****************9997 PP



Internal pipeline statistics summary:
-------------------------------------
Query model(s):                            1  (248 nodes)
Target sequences:                          1  (262 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: 6.50
//
[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