Alignments for a candidate for pimF in Pseudomonas fluorescens FW300-N2E2

GapMind for catabolism of small carbon sources

Alignments for a candidate for pimF in Pseudomonas fluorescens FW300-N2E2

Align 6-carboxyhex-2-enoyl-CoA hydratase (characterized)
to candidate Pf6N2E2_1850 Enoyl-CoA hydratase [isoleucine degradation] (EC 4.2.1.17) / 3-hydroxyacyl-CoA dehydrogenase (EC 1.1.1.35) / 3-hydroxybutyryl-CoA epimerase (EC 5.1.2.3)

Query= metacyc::MONOMER-20678
         (699 letters)



>FitnessBrowser__pseudo6_N2E2:Pf6N2E2_1850
          Length = 715

 Score =  289 bits (740), Expect = 3e-82
 Identities = 224/712 (31%), Positives = 338/712 (47%), Gaps = 46/712 (6%)

Query: 11  DQVAIVTVDSP-PVNALSAAVRRGILENVNAAVADPAVQAIVLVCAGRTFIAGADITEFG 69
           D +  VT+D P PVNA++   R  +   VN    +  +  +V+  A  TF AG D+ E  
Sbjct: 11  DGIVTVTMDMPGPVNAMNQLYRETMAATVNRLEQENGLTGVVIASAKSTFFAGGDLNELT 70

Query: 70  --KPPQPPAL--------NDVIAALENSPKPTIAAIHGTALGGGLEVALGCHFRVAVKE- 118
             KP              ND +  LE  P P +AAI+G ALGGG E+AL C++RV +   
Sbjct: 71  DFKPGDEADFQAKIELTKND-LRRLERLPVPVVAAINGAALGGGFEIALACNYRVLLNSP 129

Query: 119 -AKLGLPEVKLGLLPGAGGTQRLPRAVGPELAVQMIVGGSPIGAAEALKHGLVEEVVEN- 176
            A +GLPEV LGLLPG GG  RL   +G E A+ +++ G  +  AEAL  GLV E++E  
Sbjct: 130 AAVVGLPEVSLGLLPGGGGIVRLVHHLGLERALPLLLEGKRMKPAEALALGLVNELIEQA 189

Query: 177 --LVAGAVAFAKKVLAEKR-PLRRLRDDDSKLAAAKADR----SIFTNAVAAMTKKARGL 229
             LV  A A+ K  + +    L+       K+    A +     +   A A   K  RGL
Sbjct: 190 DALVPAAKAWIKAQVGDDGVSLQPWDHKGHKIPGGTAQQPHVAQMIAGAQAMTAKNTRGL 249

Query: 230 -EAPFACADAIGAAIDLPFEEGLKKEREGFMKLVVSDQSK-AQRYAFFAEREAAKVDGVP 287
             AP         A  L F+  L  E  G   L  S Q+K      FF   E       P
Sbjct: 250 LPAPERILAVAVEATLLDFDAALLVETRGLTHLAASVQAKNIINTMFFQMNEVNGGSSRP 309

Query: 288 DGTKPRPVSRVAIIGAGTMGGGIAMSFANAGIPVTLIETGEEQLKRGLGIMQKNWEATAA 347
            G     VS++ IIGAG MG GIA   A  GI V L++   E  +RG   ++K    T +
Sbjct: 310 TGFDKSKVSKLGIIGAGMMGNGIAHVSAKVGIEVMLLDVSLEAAERGKANVEKLLSKTVS 369

Query: 348 RGGLPPDAPAKRMALITGLVGLENVKDADLIIEAVFETMAVKKEVFTAVDAHAKPGAVLA 407
           +G L      + +ALI   V   +++  D ++EAVFE++ +K +V    +A     AV  
Sbjct: 370 QGRLSEAKKDEILALIKPTVDYADLQGVDFVVEAVFESVDLKGKVTQQAEAQLPESAVFG 429

Query: 408 SNTSYLSIDEIAATTKRPQDVLGMHFFSPANVMKLCEIVRGAKTAPDALLTAVSIAKKIA 467
           +NTS L I  +A  +KRP + +G+HFFSP   M L EI+ G +T+  AL  A   A++I 
Sbjct: 430 TNTSTLPISLLANASKRPANFIGIHFFSPVERMPLVEIICGEQTSDAALAKAFDFARQIG 489

Query: 468 KVPVVVGVCDGFVGNRMLAARSKQSEKLLFEGALPQQVDAVVTKFGMPMGPFAMGDLAGL 527
           K  +VV    GF  +R   +   +  KLL EG  P  +D +  + GMP+GP  + D   L
Sbjct: 490 KTAIVVNDSLGFFTSRTFGSYFDEGCKLLQEGVDPLLIDNLGKQIGMPVGPLTVLDEVSL 549

Query: 528 DIGWR---SRKDRGIKSEIAD----------ALCEAGRFGQKTGKGYYKYEQGSRAPMPD 574
           ++  +   ++K+ G+ + + D           + E  R G+  G G+Y Y +G    +  
Sbjct: 550 ELMRKVNETQKEMGVFATVFDNSHSDAVGNILIKEYNRPGRHYGGGFYDYPEGGEKTIWP 609

Query: 575 PEVETLINDTLAKLGLKRRDITDEEILERMVYPMINEGARILEEKIAARPSDIDVVWLYG 634
              E  +   +         +  E+I ER+++  + E  + L+E +    +D +V  + G
Sbjct: 610 GLYELFVRPEVV--------LPVEDIKERLLFRQVIEAVKCLQEGVLRSVADGNVGSILG 661

Query: 635 YGWPIYRGGPMHYADSVGLKHIAERLSAYAKATNDPSLEPAPLLARLAAEGK 686
            G P + GG + + +  G++   +R    A+ T      P  LL   AA+G+
Sbjct: 662 IGAPTWTGGFLQFVNGYGVERFVQRARELAQ-TYGERFTPPVLLLEKAAKGE 712


Lambda     K      H
   0.317    0.134    0.383 

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: 1016
Number of extensions: 51
Number of successful extensions: 5
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: 699
Length of database: 715
Length adjustment: 39
Effective length of query: 660
Effective length of database: 676
Effective search space:   446160
Effective search space used:   446160
Neighboring words threshold: 11
Window for multiple hits: 40
X1: 16 ( 7.3 bits)
X2: 38 (14.6 bits)
X3: 64 (24.7 bits)
S1: 41 (21.7 bits)
S2: 55 (25.8 bits)

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