L-leucine catabolism in Klebsiella michiganensis M5al

GapMind for catabolism of small carbon sources

L-leucine catabolism in Klebsiella michiganensis M5al

Best path

livF, livG, livJ, livH, livM, ilvE, ofo, liuA, liuB, liuD, liuC, liuE, atoA, atoD, atoB

Also see fitness data for the top candidates

Rules

Overview: Leucine degradation in GapMind is based on MetaCyc pathway L-leucine degradation I, via branched alpha-keto acid dehydrogenase (link). Other pathways for are not included here because they are not linked to sequence (link) or do not result in carbon incorporation.

all: leucine-transport, ilvE, BKD, liuA, liuB, liuD, liuC, liuE and acetoacetate-degradation

Comment: After transamination to 4-methyl-2-oxopentanoate by ilvE, kbd oxidatively decarboxylates it to isovaleryl-CoA (also known as 3-methylbutanoyl-CoA), liuA oxidizes it to 3-methylcrotonyl-CoA, liuBD carboxylates it to 3-methylglutaconyl-CoA, liuC hydrates it to hydroxymethylglutaryl-CoA, and liuE hydrolyzes it to acetoacetate and acetyl-CoA.

acetoacetate-degradation: acetoacetate-activation and atoB

Comment: The acetoacetate is activated to acetoacetyl-CoA, and cleaved by acetyl-CoA acetyltransferase, giving two acetyl-CoA.

acetoacetate-activation:

atoA and atoD
or aacS
Comment: acetyl-CoA:acetoacetyl-CoA transferase (sometimes given EC 2.8.3.9 or EC 2.8.3.8) or succinyl-CoA:acetoacetyl-CoA transferase (EC 2.8.3.5, also known as 3-oxoacid CoA-transferase) can activate acetoacetate. These have an A and B subunit. Alternatively, an ATP-dependent ligase (aacS) can activate acetoacetate (EC 6.2.1.16).

BKD:

bkdA, bkdB, bkdC and lpd
or vorA, vorB and vorC
or ofoA and ofoB
or ofo
Comment: These decarboxylating dehydrogenases act on 4-methyl-2-oxopentanoate, 3-methyl-2-oxobutanoate (2-oxoisovalerate) and (S)-3-methyl-2-oxopentanoate and are known as the branched-chain alpha-ketoacid dehydrogenases. They can pass electrons to NAD (EC 1.2.1.25) or to ferredoxin (EC 1.2.7.7). The NAD-dependent enzyme is the sum of three activities: EC 1.2.4.4 (the 4-methyl-2-oxopentanoate dehydrogenase, with transfer to the lipopoyllysine residue of 2.3.1.168) which is itself heteromeric, with alpha and beta subunits; EC 2.3.1.168 (dihydrolipoyllysine-residue (3-methylbutanoyl)transferase); and EC 1.8.1.4 (dihydrolipoyl dehydrogenase, transferring electrons to NAD). The well-characterized ferredoxin-dependent enzymes have 3 subunits (vorABC) or 2 subunits (ofoAB). Genetic data identified a fused ferredoxin-dependent enzyme with just 1 subunit (ofo).

leucine-transport:

livF, livG, livJ, livH and livM
or natA, natB, natC, natD and natE
or aapJ, aapQ, aapM and aapP
or leuT
or brnQ
or bcaP
or Bap2
or AAP1
Comment: Transporters were identified using query: transporter:leucine:L-leucine

39 steps (25 with candidates)

Or see definitions of steps

Step	Description	Best candidate	2nd candidate
livF	L-leucine ABC transporter, ATPase component 1 (LivF/BraG)	BWI76_RS26330	BWI76_RS07265
livG	L-leucine ABC transporter, ATPase component 2 (LivG/BraF)	BWI76_RS26335	BWI76_RS07270
livJ	L-leucine ABC transporter, substrate-binding component (LivJ/LivK/BraC/BraC3)	BWI76_RS26365	BWI76_RS26350
livH	L-leucine ABC transporter, permease component 1 (LivH/BraD)	BWI76_RS26345	BWI76_RS07280
livM	L-leucine ABC transporter, permease component 2 (LivM/BraE)	BWI76_RS26340	BWI76_RS07275
ilvE	L-leucine transaminase	BWI76_RS24235	BWI76_RS01925
ofo	branched-chain alpha-ketoacid:ferredoxin oxidoreductase, fused
liuA	isovaleryl-CoA dehydrogenase	BWI76_RS02590
liuB	3-methylcrotonyl-CoA carboxylase, alpha (biotin-containing) subunit	BWI76_RS13985	BWI76_RS25540
liuD	3-methylcrotonyl-CoA carboxylase, beta subunit
liuC	3-methylglutaconyl-CoA hydratase	BWI76_RS13115	BWI76_RS20455
liuE	hydroxymethylglutaryl-CoA lyase
atoA	acetoacetyl-CoA transferase, A subunit	BWI76_RS16145
atoD	acetoacetyl-CoA transferase, B subunit	BWI76_RS16140
atoB	acetyl-CoA C-acetyltransferase	BWI76_RS23445	BWI76_RS01360
Alternative steps:
aacS	acetoacetyl-CoA synthetase	BWI76_RS17800	BWI76_RS23695
AAP1	L-leucine permease AAP1
aapJ	ABC transporter for amino acids (Asp/Asn/Glu/Pro/Leu), substrate-binding component AapJ
aapM	ABC transporter for amino acids (Asp/Asn/Glu/Pro/Leu), permease component 2 (AapM)	BWI76_RS18355	BWI76_RS07580
aapP	ABC transporter for amino acids (Asp/Asn/Glu/Pro/Leu), ATPase component AapP	BWI76_RS08095	BWI76_RS08965
aapQ	ABC transporter for amino acids (Asp/Asn/Glu/Pro/Leu), permease component 1 (AapQ)
Bap2	L-leucine permease Bap2	BWI76_RS19685	BWI76_RS07360
bcaP	L-leucine uptake transporter BcaP
bkdA	branched-chain alpha-ketoacid dehydrogenase, E1 component alpha subunit	BWI76_RS14160
bkdB	branched-chain alpha-ketoacid dehydrogenase, E1 component beta subunit	BWI76_RS14155
bkdC	branched-chain alpha-ketoacid dehydrogenase, E2 component	BWI76_RS04885	BWI76_RS14150
brnQ	L-leucine:Na+ symporter BrnQ/BraB	BWI76_RS06240
leuT	L-leucine:Na+ symporter LeuT
lpd	branched-chain alpha-ketoacid dehydrogenase, E3 component	BWI76_RS04890	BWI76_RS14145
natA	L-leucine ABC transporter, ATPase component 1 (NatA)	BWI76_RS26335	BWI76_RS05980
natB	L-leucine ABC transporter, substrate-binding component NatB
natC	L-leucine ABC transporter, permease component 1 (NatC)	BWI76_RS05985
natD	L-leucine ABC transporter, permease component 2 (NatD)	BWI76_RS05990	BWI76_RS26345
natE	L-leucine ABC transporter, ATPase component 2 (NatE)	BWI76_RS26330	BWI76_RS05975
ofoA	branched-chain alpha-ketoacid:ferredoxin oxidoreductase, alpha subunit OfoA
ofoB	branched-chain alpha-ketoacid:ferredoxin oxidoreductase, beta subunit OfoB
vorA	branched-chain alpha-ketoacid:ferredoxin oxidoreductase, alpha subunit VorA
vorB	branched-chain alpha-ketoacid:ferredoxin oxidoreductase, beta subunit VorB
vorC	branched-chain alpha-ketoacid:ferredoxin oxidoreductase, gamma subunit VorC

Confidence: high confidence medium confidence low confidence
transporter – transporters and PTS systems are shaded because predicting their specificity is particularly challenging.

This GapMind analysis is from Sep 17 2021. The underlying query database was built on Sep 17 2021.

Downloads

Candidates (tab-delimited)
Steps (tab-delimited)
Rules (tab-delimited)
Protein sequences (fasta format)
Organisms (tab-delimited)
SQLite3 databases

Related tools

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