REDAM_ASPOR
ID REDAM_ASPOR Reviewed; 295 AA.
AC Q2TW47;
DT 25-OCT-2017, integrated into UniProtKB/Swiss-Prot.
DT 24-JAN-2006, sequence version 1.
DT 03-AUG-2022, entry version 86.
DE RecName: Full=NADPH-dependent reductive aminase {ECO:0000303|Ref.2};
DE Short=RedAm {ECO:0000303|Ref.2};
DE EC=1.1.1.- {ECO:0000269|Ref.2};
DE Flags: Precursor;
GN ORFNames=AO090010000708;
OS Aspergillus oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Aspergillus;
OC Aspergillus subgen. Circumdati.
OX NCBI_TaxID=510516;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 42149 / RIB 40;
RX PubMed=16372010; DOI=10.1038/nature04300;
RA Machida M., Asai K., Sano M., Tanaka T., Kumagai T., Terai G., Kusumoto K.,
RA Arima T., Akita O., Kashiwagi Y., Abe K., Gomi K., Horiuchi H.,
RA Kitamoto K., Kobayashi T., Takeuchi M., Denning D.W., Galagan J.E.,
RA Nierman W.C., Yu J., Archer D.B., Bennett J.W., Bhatnagar D.,
RA Cleveland T.E., Fedorova N.D., Gotoh O., Horikawa H., Hosoyama A.,
RA Ichinomiya M., Igarashi R., Iwashita K., Juvvadi P.R., Kato M., Kato Y.,
RA Kin T., Kokubun A., Maeda H., Maeyama N., Maruyama J., Nagasaki H.,
RA Nakajima T., Oda K., Okada K., Paulsen I., Sakamoto K., Sawano T.,
RA Takahashi M., Takase K., Terabayashi Y., Wortman J.R., Yamada O.,
RA Yamagata Y., Anazawa H., Hata Y., Koide Y., Komori T., Koyama Y.,
RA Minetoki T., Suharnan S., Tanaka A., Isono K., Kuhara S., Ogasawara N.,
RA Kikuchi H.;
RT "Genome sequencing and analysis of Aspergillus oryzae.";
RL Nature 438:1157-1161(2005).
RN [2]
RP X-RAY CRYSTALLOGRAPHY (1.82 ANGSTROMS) IN COMPLEX WITH SUBSTRATES,
RP COFACTOR, SUBUNIT, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, ACTIVITY REGULATION, AND MUTAGENESIS OF ASP-169; TYR-177;
RP TRP-210 AND GLN-240.
RX DOI=10.1038/nchem.2782;
RA Aleku G.A., France S.P., Man H., Mangas-Sanchez J., Montgomery S.L.,
RA Sharma M., Leipold F., Hussain S., Grogan G., Turner N.J.;
RT "A reductive aminase from Aspergillus oryzae.";
RL Nat. Chem. 0:0-0(2017).
CC -!- FUNCTION: NADPH-dependent reductive aminase that catalyzes the
CC reductive coupling of a broad set of carbonyl compounds with a variety
CC of primary and secondary amines (Ref.2). Possesses remarkably high
CC activity for the reductive amination of ketones and amines, often with
CC high stereoselectivity and in some cases with ketone:amine ratios as
CC low as 1:1 (Ref.2). The cofactor NADPH, the carbonyl compound and the
CC amine are added to the enzyme in that sequence, followed by the release
CC of product, NADP(+) being released at last (Ref.2). RedAm is also able
CC to act in the reverse, oxidative direction and exhibits activity in the
CC dehydrogenation of amines to yield imines (Ref.2). The highest activity
CC is found for 1-methyl-tetrahydroquinoline and acyclic amines are also
CC found to be transformed (Ref.2). {ECO:0000269|Ref.2}.
CC -!- COFACTOR:
CC Name=NADPH; Xref=ChEBI:CHEBI:57783; Evidence={ECO:0000269|Ref.2};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.12 mM for NADPH (with cyclohexanone and methylamine as saturated
CC substrates) {ECO:0000269|Ref.2};
CC KM=2.13 mM for cyclohexanone (with NADPH and methylamine as saturated
CC substrates) {ECO:0000269|Ref.2};
CC KM=8.23 mM for methylamine (with cyclohexanone and NADPH as saturated
CC substrates) {ECO:0000269|Ref.2};
CC KM=1.90 mM for cyclohexanone (with NADPH and propargylamine as
CC saturated substrates) {ECO:0000269|Ref.2};
CC KM=2.31 mM for cyclohexanone (with NADPH and allylamine as saturated
CC substrates) {ECO:0000269|Ref.2};
CC KM=2.06 mM for cyclohexanone (with NADPH and pyrrolidine as saturated
CC substrates) {ECO:0000269|Ref.2};
CC KM=1.90 mM for cyclohexanone (with NADPH and benzylamine as saturated
CC substrates) {ECO:0000269|Ref.2};
CC KM=2.15 mM for cyclohexanone (with NADPH and ammonia as saturated
CC substrates) {ECO:0000269|Ref.2};
CC KM=0.04 mM for indanone (with NADPH and propargylamine as saturated
CC substrates) {ECO:0000269|Ref.2};
CC KM=0.94 mM for 4-phenyl-2-butanone (with NADPH and propargylamine as
CC saturated substrates) {ECO:0000269|Ref.2};
CC KM=0.09 mM for acetophenone (with NADPH and propargylamine as
CC saturated substrates) {ECO:0000269|Ref.2};
CC KM=12.20 mM for 2-allyl cyclohexanone (with NADPH and propargylamine
CC as saturated substrates) {ECO:0000269|Ref.2};
CC KM=1.92 mM for 1-tetralone (with NADPH and propargylamine as
CC saturated substrates) {ECO:0000269|Ref.2};
CC KM=5.40 mM for propargylamine (with cyclohexanone and NADPH as
CC saturated substrates) {ECO:0000269|Ref.2};
CC KM=15.33 mM for isopropylamine (with cyclohexanone and NADPH as
CC saturated substrates) {ECO:0000269|Ref.2};
CC KM=25.12 mM for pyrrolidine (with cyclohexanone and NADPH as
CC saturated substrates) {ECO:0000269|Ref.2};
CC KM=0.08 mM for NADP(+) (with N-methylcyclohexylamine as saturated
CC substrate) {ECO:0000269|Ref.2};
CC KM=11.61 mM for N-methylcyclohexylamine (with NADP(+) as saturated
CC substrate) {ECO:0000269|Ref.2};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|Ref.2}.
CC -!- SIMILARITY: Belongs to the HIBADH-related family. {ECO:0000305}.
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DR EMBL; AP007175; BAE66526.1; -; Genomic_DNA.
DR RefSeq; XP_001827659.1; XM_001827607.1.
DR PDB; 5G6R; X-ray; 1.82 A; A/B=1-295.
DR PDB; 5G6S; X-ray; 2.35 A; A/B/C/D/E/F/G/H=1-295.
DR PDBsum; 5G6R; -.
DR PDBsum; 5G6S; -.
DR AlphaFoldDB; Q2TW47; -.
DR SMR; Q2TW47; -.
DR EnsemblFungi; BAE66526; BAE66526; AO090010000708.
DR GeneID; 5999793; -.
DR KEGG; aor:AO090010000708; -.
DR VEuPathDB; FungiDB:AO090010000708; -.
DR HOGENOM; CLU_035117_2_1_1; -.
DR OMA; MYGMFAG; -.
DR Proteomes; UP000006564; Chromosome 8.
DR GO; GO:0050661; F:NADP binding; IEA:InterPro.
DR GO; GO:0016491; F:oxidoreductase activity; IEA:UniProtKB-KW.
DR Gene3D; 1.10.1040.10; -; 1.
DR InterPro; IPR013328; 6PGD_dom2.
DR InterPro; IPR006115; 6PGDH_NADP-bd.
DR InterPro; IPR015815; HIBADH-related.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR Pfam; PF03446; NAD_binding_2; 1.
DR PIRSF; PIRSF000103; HIBADH; 1.
DR SUPFAM; SSF51735; SSF51735; 1.
PE 1: Evidence at protein level;
KW 3D-structure; NADP; Oxidoreductase; Reference proteome; Signal.
FT SIGNAL 1..18
FT /evidence="ECO:0000255"
FT CHAIN 19..295
FT /note="NADPH-dependent reductive aminase"
FT /id="PRO_0000441988"
FT BINDING 6..20
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:Q49A26"
FT MUTAGEN 169
FT /note="D->A,N: Leads to a 30-fold decrease in reductive
FT aminase activity."
FT /evidence="ECO:0000269|Ref.2"
FT MUTAGEN 177
FT /note="Y->A: Leads to a 200-fold decrease in reductive
FT aminase activity."
FT /evidence="ECO:0000269|Ref.2"
FT MUTAGEN 210
FT /note="W->A,S: Displays a dramatic selectivity switch to
FT yield the antipodal (S)-amine products with a variety of
FT amine nucleophiles."
FT /evidence="ECO:0000269|Ref.2"
FT MUTAGEN 240
FT /note="Q->A,S: Displays significant improvements in (R)-
FT selectivity for most substrates."
FT /evidence="ECO:0000269|Ref.2"
FT STRAND 4..8
FT /evidence="ECO:0007829|PDB:5G6R"
FT HELIX 12..23
FT /evidence="ECO:0007829|PDB:5G6R"
FT STRAND 27..31
FT /evidence="ECO:0007829|PDB:5G6R"
FT HELIX 35..38
FT /evidence="ECO:0007829|PDB:5G6R"
FT HELIX 39..43
FT /evidence="ECO:0007829|PDB:5G6R"
FT HELIX 52..57
FT /evidence="ECO:0007829|PDB:5G6R"
FT STRAND 60..64
FT /evidence="ECO:0007829|PDB:5G6R"
FT HELIX 69..77
FT /evidence="ECO:0007829|PDB:5G6R"
FT HELIX 78..82
FT /evidence="ECO:0007829|PDB:5G6R"
FT STRAND 87..90
FT /evidence="ECO:0007829|PDB:5G6R"
FT HELIX 96..107
FT /evidence="ECO:0007829|PDB:5G6R"
FT TURN 108..110
FT /evidence="ECO:0007829|PDB:5G6R"
FT STRAND 112..120
FT /evidence="ECO:0007829|PDB:5G6R"
FT HELIX 122..124
FT /evidence="ECO:0007829|PDB:5G6R"
FT STRAND 131..137
FT /evidence="ECO:0007829|PDB:5G6R"
FT HELIX 139..149
FT /evidence="ECO:0007829|PDB:5G6R"
FT HELIX 150..152
FT /evidence="ECO:0007829|PDB:5G6R"
FT STRAND 153..157
FT /evidence="ECO:0007829|PDB:5G6R"
FT HELIX 164..191
FT /evidence="ECO:0007829|PDB:5G6R"
FT HELIX 200..213
FT /evidence="ECO:0007829|PDB:5G6R"
FT HELIX 215..227
FT /evidence="ECO:0007829|PDB:5G6R"
FT HELIX 237..254
FT /evidence="ECO:0007829|PDB:5G6R"
FT TURN 259..261
FT /evidence="ECO:0007829|PDB:5G6R"
FT HELIX 262..273
FT /evidence="ECO:0007829|PDB:5G6R"
FT HELIX 281..287
FT /evidence="ECO:0007829|PDB:5G6R"
SQ SEQUENCE 295 AA; 31155 MW; 0F3524398534D0AB CRC64;
MSKHIGIFGL GAMGTALAAK YLEHGYKTSV WNRTTAKAIP LVEQGAKLAS TISEGVNAND
LIIICLLNNQ VVEDALRDAL QTLPSKTIVN LTNGTPNQAR KLADFVTSHG ARYIHGGIMA
VPTMIGSPHA VLLYSGESLE LFQSIESHLS LLGMSKYLGT DAGSASLHDL ALLSGMYGLF
SGFLHAVALI KSGQDTSTTA TGLLPLLTPW LSAMTGYLSS IAKQIDDGDY ATQGSNLGMQ
LAGVENIIRA GEEQRVSSQM ILPIKALIEQ AVGEGHGGED LSALIEYFKV GKNVD
