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One-Pot Biosynthesis of 3-Aminopropionic Acid from Fumaric Acid Using Recombinant Bacillus megaterium Containing a Linear Dual-Enzyme Cascade

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Abstract

3-Aminopropionic acid (3-APA) has wide applications in food, cosmetics, pharmaceuticals, chemical, and polymer industries. This present study aimed to develop an eco-friendly whole-cell biocatalytic process for the bio-production of 3-APA from fumaric acid (FA) using Bacillus megaterium. A dual-enzyme cascade route with aspartate-1-decarboxylases (ADC) from Bacillus subtilis and native aspartate ammonia-lyase (AspA) was developed. Divergent catalytic efficiencies between these two enzymes led to an imbalance between both enzyme reactions. In order to coordinate AspA and ADC expression levels, gene mining, optimization, and duplication strategies were employed. Additionally, culture cultivation conditions and biocatalysis process parameters were optimized. A maximum 3-APA titer was obtained (11.68 ± 0.26 g/L) with a yield of 0.78 g/g under the following optimal conditions: 45 °C, pH 6.0, and 15 g/L FA. This study established a biocatalysis process for the production of 3-APA from FA using the whole cells of the recombinant B. megaterium.

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Acknowledgements

The authors acknowledge the Ministry of Human Resource and Development, New Delhi, India, for fellowship. The authors are grateful to the Department of Biosciences and Bioengineering, IIT Guwahati, for providing a platform to conduct research activities.

Funding

This study was financially supported by the Department of Biotechnology, Govt. of India (Grant No—BT/PR15946/NER/95/485/2016).

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Authors and Affiliations

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

    Subbi Rami Reddy Tadi, Ganesh Nehru & Senthilkumar Sivaprakasam

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  1. Subbi Rami Reddy Tadi
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  3. Senthilkumar Sivaprakasam
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Contributions

Subbi Rami Reddy Tadi: conceptualization, methodology, investigation, writing—original draft of the manuscript, review and editing.

Ganesh Nehru: methodology and investigation support, review and editing.

Senthilkumar Sivaprakasam: project administration, supervision, writing—review and editing.

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Correspondence to Senthilkumar Sivaprakasam.

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Tadi, S.R.R., Nehru, G. & Sivaprakasam, S. One-Pot Biosynthesis of 3-Aminopropionic Acid from Fumaric Acid Using Recombinant Bacillus megaterium Containing a Linear Dual-Enzyme Cascade. Appl Biochem Biotechnol 194, 1740–1754 (2022). https://doi.org/10.1007/s12010-021-03783-7

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