PLP-dependent enzymes as important biocatalysts for the pharmaceutical, chemical and food industries: a structural and mechanistic perspective

Rocha, Juliana F.; Pina, André F.; Sousa, Sérgio F.; Cerqueira, Nuno M. F. S. A.

doi:10.1039/c9cy01210a

Cited by 78 publications

(79 citation statements)

References 102 publications

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“…Despite the tremendous advantage and potential, the widespread application of ω-transaminases for organic synthesis is facing the fundamental challenges of severe by-product inhibition and unfavorable equilibrium 1,[4][5][6][7] . To overcome these challenges, several by-product removal strategies have been developed.…”

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confidence: 99%

Structural studies reveal flexible roof of active site responsible for ω-transaminase CrmG overcoming by-product inhibition

Tang

Zhu

et al. 2020

Commun Biol

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Amine compounds biosynthesis using ω-transaminases has received considerable attention in the pharmaceutical industry. However, the application of ω-transaminases was hampered by the fundamental challenge of severe by-product inhibition. Here, we report that ωtransaminase CrmG from Actinoalloteichus cyanogriseus WH1-2216-6 is insensitive to inhibition from by-product α-ketoglutarate or pyruvate. Combined with structural and QM/MM studies, we establish the detailed catalytic mechanism for CrmG. Our structural and biochemical studies reveal that the roof of the active site in PMP-bound CrmG is flexible, which will facilitate the PMP or by-product to dissociate from PMP-bound CrmG. Our results also show that amino acceptor caerulomycin M (CRM M), but not α-ketoglutarate or pyruvate, can form strong interactions with the roof of the active site in PMP-bound CrmG. Based on our results, we propose that the flexible roof of the active site in PMP-bound CrmG may facilitate CrmG to overcome inhibition from the by-product.

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confidence: 99%

Structural studies reveal flexible roof of active site responsible for ω-transaminase CrmG overcoming by-product inhibition

Tang

Zhu

et al. 2020

Commun Biol

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“…GAD is a pyridoxal 5'-phosphate (PLP) dependent enzyme and as such belongs to the PLP-dependent enzyme superfamily, which contains at seven different folds [67], with GAD from LAB showing the type-I fold of PLP-dependent enzymes [68]. A number of important catalytic reactions including α-and -eliminations, decarboxylation, transamination, racemization and aldol cleavage are catalyzed by various members of this superfamily of enzymes [69]. GAD activity relies on the binding of its co-factor PLP, and belongs to group II of PLP-dependent decarboxylases [70].…”

Section: Glutamate Decarboxylasementioning

confidence: 99%

“…The latter will immediately be released from the enzyme, resulting inactive apoGAD (Figure 2), which can be regenerated to the active GAD-PLP complex when free pyridoxal 5'phosphate is present, thus completing a cycle of inactivation and activation. However, when free PLP is not present, GAD will not be reactive as a function of time and substrate concentration [62,[67][68][69]74,77].…”

Section: Glutamate Decarboxylasementioning

confidence: 99%

A Brief Review on Glutamate Decarboxylase From Lactic Acid Bacteria

Yogeswara¹,

Maneerat²,

Haltrich³

2020

Preprint

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Glutamate decarboxylase (L-glutamate-1-carboxylase, GAD; EC 4.1.1.15) is a pyridoxal 5-phosphate-dependent enzyme, which catalyzes the irreversible α-decarboxylation of L-glutamic acid to γ-aminobutyric acid (GABA) and CO2. The enzyme is widely distributed in eukaryotes as well as prokaryotes, where it – together with its reaction product GABA - fulfils very different physiological functions. The occurrence of gad genes encoding GAD has been shown for many microorganisms, and GABA-producing lactic acid bacteria (LAB) have been a focus of research during recent years. A wide range of traditional foods produced by fermentation based on LAB offer the potential of providing new functional food products enriched with GABA that may offer certain health-benefits. Different GAD enzymes and genes from several strains of LAB have been isolated and characterized recently. GABA-producing LAB, biochemical properties of their GAD enzymes, and possible applications are reviewed here.

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“…The optically pure amines have been used for the synthesis of biologically active compounds ( Koszelewski et al, 2009 ). ω-Transaminases (ω-TAs) are multimeric enzymes and have been investigated for preparation of chiral amines ( Han et al, 2019 ; Rocha et al, 2019 ; Isupov et al, 2019 ; Liao et al, 2019 ). ω-Transaminases are efficient biocatalysts for reductive amination of ketones to corresponding amines ( Kelly et al, 2018 ; Padrosa et al, 2019 ).…”

Section: Introductionmentioning

confidence: 99%

Peptide Bond Formation Between the Hetrosubunits of ω-Transaminase, Alanine Dehydrogenase, and Formate Dehydrogenase Through Subunit Splicing Promoted by Heterodimerization of Leucine Zipper Motifs

Chen

et al. 2020

Front. Bioeng. Biotechnol.

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For the multimeric enzymes R-ω-transaminase (RTA), alanine dehydrogenase (AlaDH), and formate dehydrogenase (FDH), peptide bond formation between the hetrosubunits has been achieved by the intein-mediated in vivo subunit splicing. The subunit ligation is triggered by the heterodimerization of an arginine rich leucine zipper motif with a glutamic acid rich leucine zipper motif. The one-by-one ligation of hetrosubunits constructs the pairing enzymes RTA&AlaDH and AlaDH&FDH. The ligation modes were analyzed based on blue native polyacrylamide gel electrophoresis (BN-PAGE). The spectra of circular dichroism (CD), fluorescence, and two-dimensional FTIR provide information on the secondary structures and stability of the pairing enzymes. The enzymesubstrate interaction was analyzed based on microscale thermophoresis analysis. In contrast to the mixed three enzymes RTA + AlaDH + FDH, the ligated enzymes RTA&AlaDH + AlaDH&FDH exhibited a much larger substrate affinity, higher stability, and significantly enhanced activity.

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PLP-dependent enzymes as important biocatalysts for the pharmaceutical, chemical and food industries: a structural and mechanistic perspective

Abstract: PLP-dependent enzymes described on this review are attractive targets for enzyme engineering towards their application in an industrial biotechnology framework.

Cited by 78 publications

References 102 publications

Structural studies reveal flexible roof of active site responsible for ω-transaminase CrmG overcoming by-product inhibition

Structural studies reveal flexible roof of active site responsible for ω-transaminase CrmG overcoming by-product inhibition

A Brief Review on Glutamate Decarboxylase From Lactic Acid Bacteria

Peptide Bond Formation Between the Hetrosubunits of ω-Transaminase, Alanine Dehydrogenase, and Formate Dehydrogenase Through Subunit Splicing Promoted by Heterodimerization of Leucine Zipper Motifs

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