Crystal structure of a membrane-bound l -amino acid deaminase from Proteus vulgaris

Ju, Yingchen; Shu, Tong; Gao, Yongxiang; Zhao, Wei; Liu, Qi; Gu, Qiong; Xu, Jing; Niu, Liwen; Teng, Maikun; Zhou, Huihao

doi:10.1016/j.jsb.2016.07.008

Cited by 34 publications

(43 citation statements)

References 44 publications

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“…Among them, the reaction catalyzed by ʟ‐AAD does not produce H 2 O 2 , without an extra cofactor and amino receptor, making ʟ‐AAD the most ideal enzyme for α‐keto acids synthesis in the AT1. Two ʟ‐AADs from Proteus vulgaris (Ju et al, ) and Proteus mirabilis (Gourinchas et al, ) were then selected and expressed in E. coli BL21, to obtain the corresponding strains E. coli 02 and 03 (Table ). Between them, ʟ‐AAD from P. vulgaris ( E. coli 02; Pv ‐ʟ‐AAD; 0.042 U/mg cells) was found to be more efficient in the presence of phenol and was thus chosen as the catalyst for the oxidative deamination of ( S )‐α‐ 3 (Figure S8).…”

Section: Resultsmentioning

confidence: 99%

Efficient production of phenylpropionic acids by an amino‐group‐transformation biocatalytic cascade

Wang

Song

et al. 2019

Biotech & Bioengineering

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Phenylpropionic acids are commonly used in the synthesis of pharmaceuticals, cosmetics, and fine chemicals. However, the synthesis of phenylpropionic acids faces the challenges of high cost of substrates and a limited range of products. Here, we present an artificially designed amino-group-transformation biocatalytic process, which uses simple phenols, pyruvate, and ammonia to synthesize diverse phenylpropionic acids. This biocatalytic cascade comprises an amino-group-introduction module and three amino-group-transformation modules, and operates in a modular assembly manner. Escherichia coli catalysts coexpressing enzymes from different modules achieve whole-cell simultaneous one-pot transformations of phenols into the corresponding phenylpropionic acids including (S)-α-amino acids, α-keto acids, (R)-αamino acids, and (R)-β-amino acids. With cofactor recycling, protein engineering, and transformation optimization, four (S)-α-amino acids, four α-keto acids, four (R)-αamino acids, and four (R)-β-amino acids are synthesized with good conversion (68-99%) and high enantioselectivities (>98%). Therefore, the amino-grouptransformation concept provides a universal and efficient tool for synthesizing diverse products.

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Section: Resultsmentioning

confidence: 99%

Efficient production of phenylpropionic acids by an amino‐group‐transformation biocatalytic cascade

Wang

Song

et al. 2019

Biotech & Bioengineering

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“…However, the biocatalyst activity could not reflect the protein expression directly, the expression level of the four high biocatalyst activity mutants (BL21/RBS3, BL21/RBS4, BL21/RBS6, and BL21/RBS8) was analyzed by SDS-PAGE. The L-AAD is linked to the membrane by a peptide and the active center is out of the membrane [27]. So the total cell lysates included the inclusion bodies and active enzyme, but the cell lysate supernatant is just active enzyme because the membrane could not be separated from supernatant by such a low speed centrifugation.…”

Section: Discussionmentioning

confidence: 99%

“…vulgaris is an oxidoreductase that exhibits high substrate specificity. With flavin adenine dinucleotide as a coenzyme, l -AAD catalyzes the deamination of leucine to KIC without generating H 2 O 2 , thereby reducing impacts on host cell growth [11]. Catalysis is proposed to occur via the binding of Proteus l -AADs to the electron transport chain to reduce O 2 to H 2 O [12].…”

Section: Introductionmentioning

confidence: 99%

Tuning the transcription and translation of L-amino acid deaminase in Escherichia coli improves α-ketoisocaproate production from L-leucine

Song

Shin

et al. 2017

PLoS ONE

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α-Ketoisocaproate (KIC) is used widely in the pharmaceutical and nutraceutical industries. In previous studies, we achieved a one-step biosynthesis of KIC from l-leucine, using an Escherichia coli whole-cell biocatalyst expressing an l-amino acid deaminase (l-AAD) from Proteus vulgaris. Herein, we report the fine-tuning of l-AAD gene expression in E. coli BL21 (DE3) at the transcriptional and translational levels to improve the KIC titer. By optimizing the plasmid origin with different copy numbers, modulating messenger RNA structure downstream of the initiation codon, and designing the sequences at the ribosome binding site, we increased biocatalyst activity to 31.77%, 24.89%, and 30.20%, respectively, above that achieved with BL21/pet28a-lad. The highest KIC titers reached 76.47 g·L-1, 80.29 g·L-1, and 81.41 g·L-1, respectively. Additionally, the integration of these three engineering strategies achieved an even higher KIC production of 86.55 g·L-1 and a higher l-leucine conversion rate of 94.25%. The enzyme-engineering strategies proposed herein may be generally applicable to the construction of other biocatalysts.

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“…An in vitro study showed IL-6 and IL-1β production by peritoneal macrophages stimulated with LAAOs, which was dependent on the activation of the Toll-like receptors TLR2 and TLR4. They also reported that LAAOs promoted apop- [28], (b) Proteus vulgaris [29], (c) Rhodococcus opacus [30] and (d) Streptomyces sp. [31].…”

Section: Amino Acid Oxidasesmentioning

confidence: 90%

Therapeutic Uses of Amino Acids

Odia¹,

Esezobor²

2017

Amino Acid - New Insights and Roles in Plant and Animal

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Amino acids, which are the building blocks of peptides and proteins, are indispensable chemicals needed by the body for optimal metabolism and proper body functioning. Classiied as essential, nonessential and conditionally essential, amino acids play vital roles in the body such as in protein synthesis and as precursors in the production of secondary metabolism molecules. Amino acid oxygenases also play vital metabolic roles such as in prevention of diseases; as a result, amino acids and their oxygenases isolated from various organisms are potent candidates in treatment of diseases which include cancers, inlammations, as well as antibacterial agents.

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Crystal structure of a membrane-bound l -amino acid deaminase from Proteus vulgaris

Cited by 34 publications

References 44 publications

Efficient production of phenylpropionic acids by an amino‐group‐transformation biocatalytic cascade

Efficient production of phenylpropionic acids by an amino‐group‐transformation biocatalytic cascade

Tuning the transcription and translation of L-amino acid deaminase in Escherichia coli improves α-ketoisocaproate production from L-leucine

Therapeutic Uses of Amino Acids

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