Ratio of intracellular precursors concentration and their flux influences hyaluronic acid molecular weight in Streptococcus zooepidemicus and recombinant Lactococcus lactis

Badle, Sneh Sanjay; Jayaraman, Guhan; Ramachandran, K.B.

doi:10.1016/j.biortech.2014.04.027

Cited by 53 publications

(34 citation statements)

References 25 publications

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“…were investigated so far (Liu et al 2011). The ratio of UDP-GlcA and UDP-GlcNAc influences the molecular weight of HA produced by Streptococcus strains (Badle et al 2014), however genetic changes made in this work did not exert such effect.…”

Section: Discussionmentioning

confidence: 59%

Engineering S. equi subsp. zooepidemicus towards concurrent production of hyaluronic acid and chondroitin biopolymers of biomedical interest

et al. 2017

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Glycosaminoglycans, such as hyaluronic acid and chondroitin sulphate, are not only more and more required as main ingredients in cosmeceutical and nutraceutical preparations, but also as active principles in medical devices and pharmaceutical products. However, while biotechnological production of hyaluronic acid is industrially established through fermentation of Streptococcus spp. and recently Bacillus subtilis, biotechnological chondroitin is not yet on the market. A non-hemolytic and hyaluronidase negative S. equi subsp. zooepidemicus mutant strain was engineered in this work by the addition of two E. coli K4 genes, namely kfoA and kfoC, involved in the biosynthesis of chondroitin-like polysaccharide. Chondroitin is the precursor of chondroitin sulphate, a nutraceutical present on the market as anti-arthritic drug, that is lately being studied for its intrinsic bioactivity. In small scale bioreactor batch experiments the production of about 1.46 ± 0.38 g/L hyaluronic acid and 300 ± 28 mg/L of chondroitin with an average molecular weight of 1750 and 25 kDa, respectively, was demonstrated, providing an approach to the concurrent production of both biopolymers in a single fermentation.Electronic supplementary materialThe online version of this article (doi:10.1186/s13568-017-0364-7) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 59%

Engineering S. equi subsp. zooepidemicus towards concurrent production of hyaluronic acid and chondroitin biopolymers of biomedical interest

et al. 2017

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“…A few other studies discuss genetic engineering strategies that confer HA producing capabilities to GRAS organisms that originally did not produce HA [8][9][10][11][12][13][14][15]. Flux analysis studies have predominantly looked at agreeing with conditions observed in the real system, though some of them have furthered to identify bottlenecks in HA production [7,20,50]. However, these studies have not attempted a systematic identification of metabolic engineering strategies using a genome-scale model of the microorganism.…”

Section: Discussionmentioning

confidence: 99%

“…First, we added the reaction catalyzed by HA synthase to match the metabolism of the recombinant strain. Next, we replaced the flux bounds of uptake reactions specified in the original model with those that have been reported for the strain we used [20]. We eliminated gaps and blocked reactions from the model, which resulted in a version of the model specific for the nutrient sources supplied in this study.…”

Section: In Silico Ha Production Using An Adapted L Lactis Gemmentioning

confidence: 99%

“…Accordingly, recombinant L. lactis strains producing HA were previously developed by our group [18,19]. These have been investigated to identify metabolic factors that affect HA yield [20][21][22][23]. However, until now, the entire canvas for metabolic modifications has only included the traditional carbon-backbone based pathway ( Figure 2).…”

Section: Introductionmentioning

confidence: 99%

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Uncovering Novel Pathways for Enhancing Hyaluronan Synthesis in Recombinant Lactococcus lactis: Genome-Scale Metabolic Modeling and Experimental Validation

2019

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Hyaluronan (HA), a glycosaminoglycan with important medical applications, is commercially produced from pathogenic microbial sources. The metabolism of HA-producing recombinant generally regarded as safe (GRAS) systems needs to be more strategically engineered to achieve yields higher than native producers. Here, we use a genome-scale model (GEM) to account for the entire metabolic network of the cell while predicting strategies to improve HA production. We analyze the metabolic network of Lactococcus lactis adapted to produce HA and identify non-conventional strategies to enhance HA flux. We also show experimental verification of one of the predicted strategies. We thus identified an alternate route for enhancement of HA synthesis, originating from the nucleoside inosine, that can function in parallel with the traditionally known route from glucose. Adopting this strategy resulted in a 2.8-fold increase in HA yield. The strategies identified and the experimental results show that the cell is capable of involving a larger subset of metabolic pathways in HA production. Apart from being the first report to use a nucleoside to improve HA production, we demonstrate the role of experimental validation in model refinement and strategy improvisation. Overall, we point out that well-constructed GEMs could be used to derive efficient strategies to improve the biosynthesis of high-value products.

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“…For example, Shih pointed out that poly(γ-L-glutamic acid) with 10 2 kDa to 8 × 10 3 kDa is an excellent viscosity-adding agent, but this property also limits its applicability; in contrast, poly(γ-Lglutamic acid) with 30-50 kDa could be applied as drug delivery for tumor therapy (Bajaj and Singhal 2011;Shih and Van 2001). Hyaluronic acid with a high molecular weight could maintain cell integrity and water content in the extracellular matrix; in contrast, the degraded fragments, oligosaccharide hyaluronic acid, could induce receptor-mediated intracellular signaling (Badle et al 2014;Jia et al 2013;Stern et al 2006). Like the other biopolymers mentioned above, molecular weight is also an important feature for ε-PL.…”

Section: Molecular Weight Of Microbial ɛ-Plmentioning

confidence: 94%

Recent advances in the biotechnological production of microbial poly(ɛ-l-lysine) and understanding of its biosynthetic mechanism

Feng

et al. 2016

Appl Microbiol Biotechnol

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Poly(ɛ-L-lysine) (ɛ-PL) is an unusual biopolymer composed of L-lysine connected between α-carboxyl and ɛ-amino groups. It has been used as a preservative in food and cosmetics industries, drug carrier in medicines, and gene carrier in gene therapy. Modern biotechnology has significantly improved the synthetic efficiency of this novel homopoly(amino acid) on an industrial scale and has expanded its industrial applications. In the latest years, studies have focused on the biotechnological production and understanding the biosynthetic mechanism of microbial ɛ-PL. Herein, this review focuses on the current trends and future perspectives of microbial ɛ-PL. Information on the screening of ɛ-PL-producing strains, fermentative production of ɛ-PL, breeding of high-ɛ-PL-producing strains, genomic data of ɛ-PL-producing strains, biosynthetic mechanism of microbial ɛ-PL, and the control of molecular weight of microbial ɛ-PL is included. This review will contribute to the development of this novel homopoly(amino acid) and serve as a basis of studies on other biopolymers.

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Ratio of intracellular precursors concentration and their flux influences hyaluronic acid molecular weight in Streptococcus zooepidemicus and recombinant Lactococcus lactis

Cited by 53 publications

References 25 publications

Engineering S. equi subsp. zooepidemicus towards concurrent production of hyaluronic acid and chondroitin biopolymers of biomedical interest

Engineering S. equi subsp. zooepidemicus towards concurrent production of hyaluronic acid and chondroitin biopolymers of biomedical interest

Uncovering Novel Pathways for Enhancing Hyaluronan Synthesis in Recombinant Lactococcus lactis: Genome-Scale Metabolic Modeling and Experimental Validation

Recent advances in the biotechnological production of microbial poly(ɛ-l-lysine) and understanding of its biosynthetic mechanism

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