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Engineering Lactococcus lactis as a cell factory for the production of limonene


Shaili, Nurul ‘Aishah and Song, Adelene Ai Lian and Othman, Sarah and Lian, Lionel Aun In and Abdullah, Janna Ong and Abdul Rahim, Raha (2022) Engineering Lactococcus lactis as a cell factory for the production of limonene. Sains Malaysiana, 51 (1). 95 - 105. ISSN 0126-6039


Limonene is a plant monoterpene which contributes significantly to the scent of most essential oils due to its pleasant fragrance. The compound had been reported to have anti-cancer properties against several types of cancer including colorectal cancer. However, the production of this compound in nature is limited because it is produced as a secondary metabolite. To overcome these challenges, Lactococcus lactis was developed as a heterologous host for the production of limonene. A synthesized limonene synthase (LS) from Mentha spicata (mint) was cloned into L. lactis NZ9000. Western blot analysis using mouse IgG His-Tag monoclonal antibody showed successful LS expression by L. lactis at the size of ~55 kDa. GC-MS analysis results showed that limonene production was optimum after 24 h of induction (~8.0 ppm). Metabolic engineering was attempted to enhance the limonene production by overexpression of lactococcal 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) and mevalonate kinase (mvk) genes in the bacterial host. The recombinant L. lactis carrying pNZ:LSMM plasmid successfully enhanced the limonene production to two-fold (~15.1 ppm) after 24 h of induction. The outcomes of this study show the potential of L. lactis to produce plant proteins and bioactive compounds production, which prospectively leads to an oral delivery system for anti-cancer compounds.

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Additional Metadata

Item Type: Article
Divisions: Faculty of Biotechnology and Biomolecular Sciences
Institute of Bioscience
DOI Number: https://doi.org/10.17576/jsm-2022-5101-08
Publisher: Penerbit Universiti Kebangsaan Malaysia
Keywords: Isoprenoid; Lactic acid bacteria; Metabolic engineering; Monoterpene
Depositing User: Ms. Che Wa Zakaria
Date Deposited: 05 Oct 2023 21:04
Last Modified: 05 Oct 2023 21:04
Altmetrics: http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.17576/jsm-2022-5101-08
URI: http://psasir.upm.edu.my/id/eprint/101255
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