Engineering the lactococcal mevalonate pathway for increased sesquiterpene production

Song, Adelene Ai Lian; Abdullah, Janna Ong; Abdullah, Md. Pauzi; Shafee, Norazizah; Othman, Roohaida; Noor, Normah Mohd; Rahim, Raha Abdul

doi:10.1111/1574-6968.12469

Cited by 20 publications

(13 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Mevalonate kinase was identified as a key gene for MK production in MG1363 as overexpression of mvk doubled the amount of MK produced. This result is in accordance with Song et al (2014) who identified mvk to be a metabolic bottleneck of the mevalonate pathway. Overexpression of other mevalonate genes did not result in greatly increased MK levels.…”

Section: Discussionsupporting

confidence: 92%

Engineering Lactococcus lactis for Increased Vitamin K2 Production

Bøe

Holo

2020

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

Cheese produced with Lactococcus lactis is the main source of vitamin K2 in the Western diet. Subclinical vitamin K2 deficiency is common, calling for foods with enhanced vitamin K2 content. In this study we describe analyses of vitamin K2 (menaquinone) production in the lactic acid bacterium L. lactis ssp. cremoris strain MG1363. By cloning and expression from strong promoters we have identified genes and bottlenecks in the biosynthetic pathways leading to the long-chained menaquinones, MK-8 and MK-9. Key genes of the biosynthetic menaquinone pathway were overexpressed, singly or combined, to examine how vitamin K2 production can be enhanced. We observed that the production of the long menaquinone polyprenyl side chain, rather than production of the napthoate ring (1,4-dihydroxy-2-naphtoic acid), limits total menaquinone synthesis. Overexpression of genes causing increased ring formation (menF and menA) led to overproduction of short chained MK-3, while overexpression of other key genes (mvk and llmg_0196) resulted in enhanced full-length MK-9 production. Of two putatively annotated prenyl diphosphate synthases we pinpoint llmg_0196 (preA) to be important for menaquinone production in L. lactis. The genes mvk, preA, menF, and menA were found to be important contributors to menaquinone levels as single overexpression of these genes double and more than triple the total menaquinone content in culture. Combined overexpression of mvk, preA, and menA increased menaquinone levels to a higher level than obtained individually. When the overproducing strains were applied for milk fermentations vitamin K2 content was effectively increased 3-fold compared to the wild type. The results provide a foundation for development of strains to ferment foods with increased functional value i.e., higher vitamin K2 content.

show abstract

Section: Discussionsupporting

confidence: 92%

Engineering Lactococcus lactis for Increased Vitamin K2 Production

Bøe

Holo

2020

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

show abstract

“…Most prokaryotes such as E. coli uses another terpenoid biosynthesis pathway called the 2-C-methyl- d -erythritol-4-phosphate (MEP) pathway, hence almost all metabolic engineering research for heterologous plant terpenoid production have been focussed on the MEP pathway of E. coli or the MVA pathway of yeast. At present, the lactococcal MVA accommodates much room for optimization as initial attempts to metabolically engineer this pathway resulted in more than doubling of sesquiterpenes produced [143]. …”

Section: Lactococcus Lactis As a Cell Factorymentioning

confidence: 99%

A review on Lactococcus lactis: from food to factory

et al. 2017

View full text Add to dashboard Cite

Lactococcus lactis has progressed a long way since its discovery and initial use in dairy product fermentation, to its present biotechnological applications in genetic engineering for the production of various recombinant proteins and metabolites that transcends the heterologous species barrier. Key desirable features of this gram-positive lactic acid non-colonizing gut bacteria include its generally recognized as safe (GRAS) status, probiotic properties, the absence of inclusion bodies and endotoxins, surface display and extracellular secretion technology, and a diverse selection of cloning and inducible expression vectors. This have made L. lactis a desirable and promising host on par with other well established model bacterial or yeast systems such as Escherichia coli, Salmonella cerevisiae and Bacillus subtilis. In this article, we review recent technological advancements, challenges, future prospects and current diversified examples on the use of L. lactis as a microbial cell factory. Additionally, we will also highlight latest medical-based applications involving whole-cell L. lactis as a live delivery vector for the administration of therapeutics against both communicable and non-communicable diseases.

show abstract

“…It also has been proven that cantharidin may derive from sesquiterpenes or be related to the biosynthesis of juvenile hormone. The FPPS is considered to be a key enzyme in the isoprenoid pathway and responsible for synthesizing the common precursor FPP for various essential isoprenoid end products, including sesquiterpenoids . Therefore, the extent of FPPS activity can directly alter the amount of the downstream diverse isoprenoids.…”

Section: Introductionmentioning

confidence: 99%

Cloning and functional analysis of farnesyl pyrophosphate synthase (FPPS) gene from Mylabris cichorii

Zha

Yin

Wang

et al. 2017

Biotech and App Biochem

View full text Add to dashboard Cite

Cantharidin, a defensive terpene compound synthesized by the meloid beetle (Coleoptera, Meloidae), is an important anticancer agent. However, there has been little study done on how this compound synthesized by the beetle. In this paper, a farnesyl pyrophosphate synthase (FPPS) gene, designated McFPPS, was isolated from Mylabris cichorii by reverse transcription PCR based on conserved domains in other organisms. Multiple alignment analysis showed that the deduced amino acids shared >70% homology with FPPSs from other species and contained typically seven conservative regions. Gene expression profile analysis revealed that McFPPS was expressed throughout the tested growth stages of M. cichorii adults, whereas the transcripts accumulated to the highest level at 20 days in male adults while the highest expression level appeared at 15 days in females. Tissue expression pattern analysis showed that McFPPS was expressed constitutively in all tested tissues and a relatively higher expression level in the alimentary canal of males, but no significant tissue difference in the females. For the first time, a RNA interference strategy was employed to induce a greater suppression of McFPPS mRNA, and thus a sharp decrease in the expression levels of downstream genes and the concentration of product. All these results indicated that McFPPS may be directly involved or play an essential role in the biosynthesis of cantharidin.

show abstract

Engineering the lactococcal mevalonate pathway for increased sesquiterpene production

Cited by 20 publications

References 19 publications

Engineering Lactococcus lactis for Increased Vitamin K2 Production

Engineering Lactococcus lactis for Increased Vitamin K2 Production

A review on Lactococcus lactis: from food to factory

Cloning and functional analysis of farnesyl pyrophosphate synthase (FPPS) gene from Mylabris cichorii

Contact Info

Product

Resources

About