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Volume 10
Issue 8
10.3390/foods10081964
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Article

Metabolic Engineering of Pediococcus acidilactici BD16 for Heterologous Expression of Synthetic alaD Gene Cassette and L-Alanine Production in the Recombinant Strain Using Fed-Batch Fermentation

by
Anshula Sharma
1,
Masafumi Noda
2,
Masanori Sugiyama
2,
Baljinder Kaur
1,* and
Ajaz Ahmad
3,*
1
Systems Biology Laboratory, Department of Biotechnology, Punjabi University, Patiala 147002, India
2
Graduate School of Biomedical & Health Sciences, Hiroshima University, Minami-ku, Hiroshima 734-8551, Japan
3
Department of Clinical Pharmacy, College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia
*
Authors to whom correspondence should be addressed.
Academic Editors: Paula Cristina Maia Teixeira and Joana Inês Bastos Barbosa
Foods 2021, 10(8), 1964; https://doi.org/10.3390/foods10081964
Received: 27 June 2021 / Revised: 17 August 2021 / Accepted: 19 August 2021 / Published: 23 August 2021
(This article belongs to the Special Issue Biotechnology Approaches in Food Preservation and Food Safety)
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Abstract

Metabolic engineering substantially aims at the development of more efficient, robust and industrially competitive microbial strains for the potential applications in food, fermentation and pharmaceutical industries. An efficient lab scale bioprocess was developed for high level fermentative production of L-alanine using metabolically engineered Pediococcus acidilactici BD16 (alaD+). Computational biology tools assisted the designing of a synthetic alaD gene cassette, which was further cloned in shuttle vector pLES003 and expressed using an auto-inducible P289 promoter. Further, L-alanine production in the recombinant P. acidilactici BD16 (alaD+) strain was carried out using fed-batch fermentation under oxygen depression conditions, which significantly enhanced L-alanine levels. The recombinant strain expressing the synthetic alaD gene produced 229.12 g/L of L-alanine after 42 h of fed-batch fermentation, which is the second highest microbial L-alanine titer reported so far. After extraction and crystallization, 95% crystal L-alanine (217.54 g/L) was recovered from the culture broth with an enantiomeric purity of 97%. The developed bioprocess using recombinant P. acidilactici BD16 (alaD+) is suggested as the best alternative to chemical-based commercial synthesis of L-alanine for potential industrial applications. View Full-Text
Keywords: amino acid production; bioprocess development; fed-batch fermentation; lactic acid bacteria; L-alanine; metabolic engineering; synthetic gene designing amino acid production; bioprocess development; fed-batch fermentation; lactic acid bacteria; L-alanine; metabolic engineering; synthetic gene designing
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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MDPI and ACS Style

Sharma, A.; Noda, M.; Sugiyama, M.; Kaur, B.; Ahmad, A. Metabolic Engineering of Pediococcus acidilactici BD16 for Heterologous Expression of Synthetic alaD Gene Cassette and L-Alanine Production in the Recombinant Strain Using Fed-Batch Fermentation. Foods 2021, 10, 1964. https://doi.org/10.3390/foods10081964

AMA Style

Sharma A, Noda M, Sugiyama M, Kaur B, Ahmad A. Metabolic Engineering of Pediococcus acidilactici BD16 for Heterologous Expression of Synthetic alaD Gene Cassette and L-Alanine Production in the Recombinant Strain Using Fed-Batch Fermentation. Foods. 2021; 10(8):1964. https://doi.org/10.3390/foods10081964

Chicago/Turabian Style

Sharma, Anshula, Masafumi Noda, Masanori Sugiyama, Baljinder Kaur, and Ajaz Ahmad. 2021. "Metabolic Engineering of Pediococcus acidilactici BD16 for Heterologous Expression of Synthetic alaD Gene Cassette and L-Alanine Production in the Recombinant Strain Using Fed-Batch Fermentation" Foods 10, no. 8: 1964. https://doi.org/10.3390/foods10081964

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