Characterization of ELP-fused ω-Transaminase and Its Application for the Biosynthesis of β-Amino Acid

Jeon, Hyeongtag; Sarak, Sharad; Lee, Sanghyuk; Bea, Han-Seop; Patil, Mahesh D.; Kim, Geon-Hee; Kim, Byung‐Gee; Won, Jong In; Yun, Hyungdon

doi:10.1007/s12257-018-0268-7

Cited by 4 publications

(2 citation statements)

References 37 publications

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“…β-amino acids are important structural moieties present in numerous natural and synthetic bioactive compounds ( Jeon et al, 2018 ; Kim et al, 2019 ). The efficient synthesis of enantiopure ( S )-4-aminopentoic acid by the biocatalytic system employing HNF encouraged us to check its applicability as a synthetic route for β-amino acids via asymmetric synthesis.…”

Section: Resultsmentioning

confidence: 99%

Dual-function transaminases with hybrid nanoflower for the production of value-added chemicals from biobased levulinic acid

Khobragade,

Giri,

Pagar

et al. 2023

Front. Bioeng. Biotechnol.

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The U.S. Department of Energy has listed levulinic acid (LA) as one of the top 12 compounds derived from biomass. LA has gained much attention owing to its conversion into enantiopure 4-aminopentanoic acid through an amination reaction. Herein, we developed a coupled-enzyme recyclable cascade employing two transaminases (TAs) for the synthesis of (S)-4-aminopentanoic acid. TAs were first utilized to convert LA into (S)-4-aminopentanoic acid using (S)-α-Methylbenzylamine [(S)-α-MBA] as an amino donor. The deaminated (S)-α-MBA i.e., acetophenone was recycled back using a second TAs while using isopropyl amine (IPA) amino donor to generate easily removable acetone. Enzymatic reactions were carried out using different systems, with conversions ranging from 30% to 80%. Furthermore, the hybrid nanoflowers (HNF) of the fusion protein were constructed which afforded complete biocatalytic conversion of LA to the desired (S)-4-aminopentanoic acid. The created HNF demonstrated storage stability for over a month and can be reused for up to 7 sequential cycles. A preparative scale reaction (100 mL) achieved the complete conversion with an isolated yield of 62%. Furthermore, the applicability of this recycling system was tested with different β-keto ester substrates, wherein 18%–48% of corresponding β-amino acids were synthesized. Finally, this recycling system was applied for the biosynthesis of pharmaceutical important drug sitagliptin intermediate ((R)-3-amino-4-(2,4,5-triflurophenyl) butanoic acid) with an excellent conversion 82%.

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Section: Resultsmentioning

confidence: 99%

Dual-function transaminases with hybrid nanoflower for the production of value-added chemicals from biobased levulinic acid

Khobragade,

Giri,

Pagar

et al. 2023

Front. Bioeng. Biotechnol.

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“…As the sidechains of natural amino acids have limited size and functionality, protein space occupied by natural amino acids is also limited. To expand protein space beyond the limit set by nature, several techniques to incorporate noncanonical amino acids into proteins have been developed (Jeon et al, 2018; Johnson, Lu, Van Deventer, & Tirrell, 2010; Liu & Schultz, 2010; Neumann, Wang, Davis, Garcia‐Alai, & Chin, 2010; Wang, Brock, Herberich, & Schultz, 2001; Won, Pagar, Patil, Dawson, & Yun, 2019). There have been recent advances in incorporating noncanonical amino acids into proteins; the design/selection of highly active orthogonal tRNA‐synthetases allowed for the efficient incorporation of a noncanonical amino acid into multiple sites of a target protein (Amiram et al, 2015; Hauf et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

Enhanced production of Dopa‐incorporated mussel adhesive protein using engineered translational machineries

Jeong

Yang

et al. 2020

Biotech & Bioengineering

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Mussel adhesive proteins (MAPs) have great potential as bioglues, particularly in wet conditions. Although in vivo residue‐specific incorporation of 3,4‐dihydroxyphenylalanine (Dopa) in tyrosine‐auxotrophic Escherichia coli cells allows for production of Dopa‐incorporated bioengineered MAPs (dMAPs), the low production yield hinders the practical application of dMAPs. This low production yield of dMAPs is due to low translational activity of a noncanonical amino acid, Dopa, in E. coli cells. Herein, to enhance the production yield of dMAPs, we investigated the coexpression of Dopa‐recognizing tyrosyl‐tRNA synthetases (TyrRSs). To use the Dopa‐specific Methanococcus jannaschii TyrRS (MjTyrRS‐Dopa), we altered the anticodon of tyrosyl‐tRNA amber suppressor into AUA (MjtRNATyrAUA) to recognize a tyrosine codon (AUA). Co‐overexpression of MjTyrRS‐Dopa and MjtRNATyrAUA increased the production yield of Dopa‐incorporated MAP foot protein type 3 (dfp‐3) by 57%. Similarly, overexpression of E. coli TyrRS (EcTyrRS) led to a 72% higher production yield of dfp‐3. Even with coexpression of Dopa‐recognizing TyrRSs, dfp‐3 has a high Dopa incorporation yield (over 90%) compared to ones prepared without TyrRS coexpression.

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The Reductive Amination of Carbonyl Compounds Using Native Amine Dehydrogenase from Laribacter hongkongensis

Lee

Jeon

Giri

et al. 2021

Biotechnol Bioproc E

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Characterization of ELP-fused ω-Transaminase and Its Application for the Biosynthesis of β-Amino Acid

Cited by 4 publications

References 37 publications

Dual-function transaminases with hybrid nanoflower for the production of value-added chemicals from biobased levulinic acid

Dual-function transaminases with hybrid nanoflower for the production of value-added chemicals from biobased levulinic acid

Enhanced production of Dopa‐incorporated mussel adhesive protein using engineered translational machineries

The Reductive Amination of Carbonyl Compounds Using Native Amine Dehydrogenase from Laribacter hongkongensis

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