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Combinatorial approach for improved production of whole-cell 3-aminopropionic acid in recombinant Bacillus megaterium: codon optimization, gene duplication and process optimization

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Abstract

In this study, we aimed to develop a Bacillus megaterium based whole-cell biocatalyst for the bio-production of 3-aminopropionic acid (3-APA). l-aspartate-α-decarboxylases (ADC) (EC: 4.1.1.11) from Escherichia coli, B. megaterium, Corynebacterium glutamicum, and Bacillus subtilis were expressed in B. megaterium. B. subtilis derived ADC (panDBs) exhibited the highest ADC activity of 0.9 ± 0.02 U/mL in recombinant B. megaterium. Combination of codon optimization and gene duplication strategies resulted in 415.56% enhancement of ADC activity compared to panDBs. The culture growth conditions of B. megaterium (BMD-7) for 3-APA production were optimized as follows: inducer concentration, 0.5% (w/v); time of induction, 3 h; induction temperature, 37 °C and post-induction incubation time, 8 h. Improvement of the whole-cell biocatalytic process efficiency, was dealt by optimization of reaction temperature, reaction pH, metal ion additives and l-aspartic acid concentration. Shake flask level experiments yielded an enhanced 3-APA titer (16.18 ± 0.26 g/L) and a yield of 0.89 g/g under optimized conditions viz., 45 °C, pH 6.0 and 20 g/L of l-aspartic acid. This study demonstrates the potential of B. megaterium for 3-APA production and paves the scope for the development of 3-APA producing strains in near future.

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Acknowledgements

This study was financially supported by the Department of Biotechnology, Govt. of India (Grant No. BT/PR15946/NER/95/485/2016). Authors acknowledge the Ministry of Human Resource and Development, New Delhi, India for fellowship.

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Authors and Affiliations

  1. BioPAT Laboratory, Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati, 781039, Assam, India

    Subbi Rami Reddy Tadi, Ganesh Nehru & Senthilkumar Sivaprakasam

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  1. Subbi Rami Reddy Tadi
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  2. Ganesh Nehru
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  3. Senthilkumar Sivaprakasam
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Contributions

SRRT: Conceptualization, Methodology, Investigation, Writing—Original draft of the manuscript. GN: Methodology and Investigation support, Review & Editing, SS: Project administration, Supervision, Writing—Review & Editing.

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Correspondence to Senthilkumar Sivaprakasam.

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Tadi, S.R.R., Nehru, G. & Sivaprakasam, S. Combinatorial approach for improved production of whole-cell 3-aminopropionic acid in recombinant Bacillus megaterium: codon optimization, gene duplication and process optimization. 3 Biotech 11, 333 (2021). https://doi.org/10.1007/s13205-021-02885-7

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