- Deracemization and stereoinversion to aromatic d-amino acid derivatives with ancestral l-amino acid oxidase
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Enantiomerically pure amino acid derivatives could be foundational compounds for peptide drugs. Deracemization of racemates to l-amino acid derivatives can be achieved through the reaction of evolved d-amino acid oxidase and chemical reductants, whereas deracemization to d-amino acid derivatives has not progressed due to the difficulty associated with the heterologous expression of l-amino acid oxidase (LAAO). In this study, we succeeded in developing an ancestral LAAO (AncLAAO) bearing broad substrate selectivity (13 l-amino acids) and high productivity through an Escherichia coli expression system (50.7 mg/L). AncLAAO can be applied to perform deracemization to d-amino acids in a similar way to deracemization to l-amino acids. In fact, full conversion (>99% ee, d-form) could be achieved for 16 racemates, including nine d,l-Phe derivatives, six d,l-Trp derivatives, and a d,l-phenylglycine. Taken together, we believe that AncLAAO could be a key enzyme to obtain optically pure d-amino acid derivatives in the future.
- Nakano, Shogo,Minamino, Yuki,Hasebe, Fumihito,Ito, Sohei
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p. 10152 - 10158
(2019/10/19)
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- Synthesis of six phenylalanine derivatives and their cell toxicity effect on human colon cancer cell line HT-29
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Some phenylalanine (Phe) derivatives had important roles in pharmacology and may be used as pharmaceutical materials and pharmaceutical intermediates. In order to understand the cell toxicity of phenylalanine derivatives, we synthesized L-4-bromo-phenylalanine (Brp), L-4-iodophenylalanine (Ip), L-4-nitro-phenylalanine (Np), L-4-sulfonic-phenylalanine (Sp), L-4-phosphophenylalanine (Pp) and L-4-amino-phenylalanine (Np). We used mass spectrometry (MS), Infrared Spectroscopy (IR) or hydrogen-1 nuclear magnetic resonance spectrum (1H-NMR), and high performance liquid chromatography (HPLC) to test the correction of these products, MTT assay and Hoechst 33258 staining to detect their cell toxic effect on human colon cancer cell HT-29 (HT-29 cells). The results showed that these products were correct and the cytotoxicity of Pp>Ip>Sp and Np>Brp, Ap almost had no effect on HT-29 cells. In addition, Pp, Ip and Sp induced cell apoptosis, the other three kinds of phenylalanine derivatives induced neither apoptosis nor necrosis.
- Zhao, Qianyi,Xu, Ting,Li, Menghua,Yang, Ying,Hu, Haopeng,Wang, Shupei,Yan, Wenjuan,Chen, Ran,Zhang, Chunyan,Xu, Cunshuan
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p. 466 - 470
(2015/06/22)
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- Distinct metabolites for photoreactive l-phenylalanine derivatives in Klebsiella sp. CK6 isolated from rhizosphere of a wild dipterocarp sapling
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Photoaffinity labeling is a reliable analytical method for biological functional analysis. Three major photophores-aryl azide, benzophenone and trifluoromethyldiazirine- are utilized in analysis. Photophore-bearing L-phenylalanine derivatives, which are used for biological functional analysis, were inoculated into a Klebsiella sp. isolated from the rhizosphere of a wild dipterocarp sapling in Central Kalimantan, Indonesia, under nitrogen-limiting conditions. The proportions of metabolites were quite distinct for each photophore. These results indicated that photophores affected substrate recognition in rhizobacterial metabolic pathways, and differential photoaffinity labeling could be achieved using different photophore-containing L-phenylalanine derivatives.
- Wang, Lei,Hisano, Wataru,Murai, Yuta,Sakurai, Munenori,Muto, Yasuyuki,Ikemoto, Haruka,Okamoto, Masashi,Murotani, Takashi,Isoda, Reika,Kim, Dongyeop,Sakihama, Yasuko,Sitepu, Irnayuli R.,Hashidoko, Yasuyuki,Hatanaka, Yasumaru,Hashimoto, Makoto
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p. 8393 - 8401
(2013/08/23)
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- Identification of new peptide amides as selective cathepsin L inhibitors: The first step towards selective irreversible inhibitors?
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A small library of peptide amides was designed to profile the cathepsin L active site. Within the cathepsin family of cysteine proteases, the first round of selection was on cathepsin L and cathepsin B, and then selected hits were further evaluated for binding to cathepsin K and cathepsin S. Five highly selective sequences with submicromolar affinities towards cathepsin L were identified. An acyloxymethyl ketone warhead was then attached to these sequences. Although these original irreversible inhibitors inactivate cathepsin L, it appears that the nature of the warhead drastically impact the selectivity profile of the resulting covalent inhibitors.
- Torkar, Ana,Lenar?i?, Brigita,Lah, Tamara,Dive, Vincent,Devel, Laurent
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supporting information
p. 2968 - 2973
(2013/06/27)
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- Photoswitching of the Enzymatic Activity of Semisynthetic Ribonuclease S' Bearing Phenylazophenylalanine at a Specific Site
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Photoswitching of the enzymatic activity of ribonuclease S' was successfully carried out by site specific incorporation of phenylazophenylalanine into S-peptide skeleton by semisynthesis.
- Hamachi, Itaru,Hiraoki, Takashi,Yamada, Yasuhiro,Shinkai, Seiji
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p. 704 - 706
(2007/10/03)
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- Photoswitching of the enzymatic activity of semisynthetic ribonuclease S' bearing phenylazophenylalanine at a specific site
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Photos witching of the enzymatic activity of ribonuclease S' was successfully carried out by site specific incorporation of phenylazophenylalanine into S-peptide skeleton by semisynthesis.
- Hamachi, Itaru,Hiraoka, Takashi,Yamada, Yasuhiro,Shinkai, Seiji
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p. 537 - 538
(2007/10/03)
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- Selectivity and Specificity in Substrate Binding to Proteases: Novel Hydrolytic Reactions Catalysed by α-Chymotrypsin Suspended in Organic Solvents with Low Water Content and Mediated by Ammonium Hydrogen Carbonate
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α-Chymotrypsin suspended in organic solvents with low water content catalysed hydrolytic reactions in the presence of ammonium hydrogen carbonate.Molecular modelling studies were carried out and structure-reactivity relationships were established by studying the hydrolysis of amino acid derivatives and analogues.The enzyme was found to be stereoselective with respect to the hydrolysis of L-amino acid derivatives, but no stereoselectivity was observed when α-hydroxy esters were used as substrates.A general procedure for the resolution of aromatic amino acid esters is given.The results are interpreted in terms of molecular modelling based on X-ray crystallographic data and literature data.
- Ricca, Jean-Marc,Crout, David H. G.
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p. 1225 - 1234
(2007/10/02)
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