Enhancement of Naringenin Biosynthesis from Tyrosine by Metabolic Engineering of <i>Saccharomyces cerevisiae</i>

Lyu, Xiaomei; Ng, Kuan Rei; Lee, Jie Lin; Mark, Rita; Chen, Wei Ning

doi:10.1021/acs.jafc.7b02507

Cited by 79 publications

(100 citation statements)

References 41 publications

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“…Naringenin is synthesized via the phenylpropanoid pathway, starting from the aromatic amino acids phenylalanine or tyrosine. Yeast strains engineered with the pathway towards naringenin have been described [ 7 , 8 ]. Starting from naringenin, anthocyanidins are biosynthesized by flavanone 3-hydroxylase (F3H; syn.…”

Section: Introductionmentioning

confidence: 99%

Engineering de novo anthocyanin production in Saccharomyces cerevisiae

et al. 2018

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BackgroundAnthocyanins are polyphenolic pigments which provide pink to blue colours in fruits and flowers. There is an increasing demand for anthocyanins, as food colorants and as health-promoting substances. Plant production of anthocyanins is often seasonal and cannot always meet demand due to low productivity and the complexity of the plant extracts. Therefore, a system of on-demand supply is useful. While a number of other (simpler) plant polyphenols have been successfully produced in the yeast Saccharomyces cerevisiae, production of anthocyanins has not yet been reported.ResultsSaccharomyces cerevisiae was engineered to produce pelargonidin 3-O-glucoside starting from glucose. Specific anthocyanin biosynthetic genes from Arabidopsis thaliana and Gerbera hybrida were introduced in a S. cerevisiae strain producing naringenin, the flavonoid precursor of anthocyanins. Upon culturing, pelargonidin and its 3-O-glucoside were detected inside the yeast cells, albeit at low concentrations. A number of related intermediates and side-products were much more abundant and were secreted into the culture medium. To optimize titers of pelargonidin 3-O-glucoside further, biosynthetic genes were stably integrated into the yeast genome, and formation of a major side-product, phloretic acid, was prevented by engineering the yeast chassis. Further engineering, by removing two glucosidases which are known to degrade pelargonidin 3-O-glucoside, did not result in higher yields of glycosylated pelargonidin. In aerated, pH controlled batch reactors, intracellular pelargonidin accumulation reached 0.01 µmol/gCDW, while kaempferol and dihydrokaempferol were effectively exported to reach extracellular concentration of 20 µM [5 mg/L] and 150 µM [44 mg/L], respectively.ConclusionThe results reported in this study demonstrate the proof-of-concept that S. cerevisiae is capable of de novo production of the anthocyanin pelargonidin 3-O-glucoside. Furthermore, while current conversion efficiencies are low, a number of clear bottlenecks have already been identified which, when overcome, have huge potential to enhance anthocyanin production efficiency. These results bode very well for the development of fermentation-based production systems for specific and individual anthocyanin molecules. Such systems have both great scientific value for identifying and characterising anthocyanin decorating enzymes as well as significant commercial potential for the production of, on-demand, pure bioactive compounds to be used in the food, health and even pharma industries.Electronic supplementary materialThe online version of this article (10.1186/s12934-018-0951-6) contains supplementary material, which is available to authorized users.

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Section: Introductionmentioning

confidence: 99%

Engineering de novo anthocyanin production in Saccharomyces cerevisiae

et al. 2018

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“…It was previously shown that the use of 1% sucrose and 1% glycerol as the carbon source instead of 2% glucose could increase the naringenin yield in the Y-28 strains. 6 In order to investigate if the different carbon sources had an effect on the amount of naringenin produced, the six plasmids, which were expressed in the Y-28 strains, were also grown on sucrose and glycerol solution as a carbon source (Table 8). A general increase had been detected in the naringenin yields when sucrose and glycerol were used as the carbon source.…”

Section: Resultsmentioning

confidence: 99%

“…All these limitations add up to an expensive, high energy demanding preparation process, which leaves place for further improvements. 6…”

Section: Introductionmentioning

confidence: 99%

Gene Source Screening as a Tool for Naringenin Production in Engineered Saccharomyces cerevisiae

Mark

Lyu

et al. 2019

ACS Omega

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Flavonoids are plant secondary metabolites with great potential in the food industry. Metabolic engineering of Saccharomyces cerevisiae is a sustainable production technique. However, the current naringenin production yield is low because of inefficient enzymatic activity. Hence, this study uses gene source screening as a tool to identify the best gene source for enzymes such as 4-coumarate: coenzyme ligase (4CL) and chalcone synthase (CHS). For the first time, the 4CL gene from Medicago truncatula and the CHS gene from Vitis vinifera were expressed in S. cerevisiae, and this combination provided the highest yield of naringenin, which was 28-fold higher as compared to the reference strain. The combinations obtained similar performance in the Y-28 strains, where the highest production was 28.68 mg/L. Our results demonstrated that the selection and combination of enzymes from the correct gene source could greatly improve naringenin production. For the future, this could help commercialize flavonoid production, which would result in natural food preservatives and additives.

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“…The resultant strain generated 421.6 mg/L naringenin from tyrosine, which is the highest naringenin production reported so far (Wu et al, 2015). Recently, Lyu et al (2017) tried to improve naringenin titer in S. cerevisiae by dividing the naringenin biosynthetic pathway into three modules: the first module containing TAL, 4CL, CHS, and CHI was applied for naringenin biosynthesis. The second module carrying acetyl-CoA synthase, ATP-citrate lyase, and ACC was used for malonyl-CoA accumulation, whereas the third module harboring the ARO4 K 229L mutant was employed to produce more tyrosine.…”

Section: Flavanonesmentioning

confidence: 99%

“…The second module carrying acetyl-CoA synthase, ATP-citrate lyase, and ACC was used for malonyl-CoA accumulation, whereas the third module harboring the ARO4 K 229L mutant was employed to produce more tyrosine. Engineering and integration of those three modules into the final strain resulted in 90 mg/L of naringenin from glucose (Lyu et al, 2017). In another example, an iterative high-throughput screening method was applied to fine-tune the expression level of naringenin biosynthetic pathway genes.…”

Section: Flavanonesmentioning

confidence: 99%

Metabolic Engineering of Microorganisms for the Production of Flavonoids

Sheng

Sun

Yan

et al. 2020

Front. Bioeng. Biotechnol.

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Flavonoids are a class of secondary metabolites found in plant and fungus. They have been widely used in food, pharmaceutical, and nutraceutical industries owing to their significant biological activities, such as antiaging, antioxidant, anti-inflammatory, and anticancer. However, the traditional approaches for the production of flavonoids including chemical synthesis and plant extraction involved hazardous materials and complicated processes and also suffered from low product titer and yield. Microbial synthesis of flavonoids from renewable biomass such as glucose and xylose has been considered as a sustainable and environmentally friendly method for large-scale production of flavonoids. Recently, construction of microbial cell factories for efficient biosynthesis of flavonoids has gained much attention. In this article, we summarize the recent advances in microbial synthesis of flavonoids including flavanones, flavones, isoflavones, flavonols, flavanols, and anthocyanins. We put emphasis on developing pathway construction and optimization strategies to biosynthesize flavonoids and to improve their titer and yield. Then, we discuss the current challenges and future perspectives on successful strain development for large-scale production of flavonoids in an industrial level.

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Enhancement of Naringenin Biosynthesis from Tyrosine by Metabolic Engineering of Saccharomyces cerevisiae

Cited by 79 publications

References 41 publications

Engineering de novo anthocyanin production in Saccharomyces cerevisiae

Engineering de novo anthocyanin production in Saccharomyces cerevisiae

Gene Source Screening as a Tool for Naringenin Production in Engineered Saccharomyces cerevisiae

Metabolic Engineering of Microorganisms for the Production of Flavonoids

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