Dataset Information

Production and characterization of anthocyanin-rich beer from black wheat by an efficient isolate Saccharomyces cerevisiae CMS12.

ABSTRACT: Beer is the world's third most popular fermented beverage. It is typically made from malted barley. Tropical countries must import barley from temperate countries for brewing, which is an expensive process. Therefore, it is critical to investigate alternative possible substrates for beer production in order to meet the growing demand for high-nutritional-quality beer. The current study involves the creation of a fermented beverage from anthocyanin-rich black wheat with the help of yeast, Saccharomyces cerevisiae CMS12, isolated from fruit waste. Characterization (UV, HPLC, NMR, FTIR, and ICPMS) was then performed, as well as a comparative study with white (amber) wheat beer. Further, process parameters optimization included initial sugar concentration, inoculum size, and pH. Black wheat wort contained 568 mg GAE/L total phenolic content, 4.67 mg/L anthocyanin concentration, 6.8% (v/v) alcohol content, and a pH of 4.04. The sensory analysis revealed that black wheat beer was more acceptable than white wheat beer. The developed fermented beverage has enormous commercialization potential.

SUBMITTER: Singh A

PROVIDER: S-EPMC10090066 | biostudies-literature | 2023 Apr

REPOSITORIES: biostudies-literature

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Production and characterization of anthocyanin-rich beer from black wheat by an efficient isolate Saccharomyces cerevisiae CMS12.

Singh Arshpreet A Singh Saumya S Kansal Sushil K SK Garg Monika M Krishania Meena M

Scientific reports 20230411 1

Beer is the world's third most popular fermented beverage. It is typically made from malted barley. Tropical countries must import barley from temperate countries for brewing, which is an expensive process. Therefore, it is critical to investigate alternative possible substrates for beer production in order to meet the growing demand for high-nutritional-quality beer. The current study involves the creation of a fermented beverage from anthocyanin-rich black wheat with the help of yeast, Sacchar ...[more]

PMID: 37041167

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Project description: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.

Project description:BackgroundGlycyrrhetinic acid (GA) is the most important ingredient in licorice due to its outstanding anti-inflammatory activity and wide application in the medicine and cosmetics industries. Contemporary industrial production of GA by acid hydrolysis of glycyrrhizin which was extracted from Glycyrrhiza plants, is not environment-friendly and devastates farmland since the Glycyrrhiza rhizomes grow up to 10 m underground.ResultsIn this study, GA was produced through metabolically engineering Saccharomyces cerevisiae by introducing the entire heterogeneous biosynthetic pathway of GA. Codon optimized CYP88D6 and CYP72A154, combined with β-AS (β-amyrin synthase encoding gene) and the NADPH-cytochrome P450 reductase gene of Arabidopsis thaliana were introduced into S. cerevisiae. The resulting strain (Y1) produced 2.5 mg/L of β-amyrin and 14 μg/L of GA. The cytochrome b5 from G. uralensis (GuCYB5) was identified and the introduction of this novel GuCYB5 increased the efficiency of GA production by eightfold. The joint utilization of the GuCYB5 gene along with 10 known MVA pathway genes from S. cerevisiae were overexpressed in a stable chromosome integration to achieve higher GA production. Using the combined strategy, GA concentration improved by 40-fold during batch fermentation. The production was further improved to 8.78 mg/L in fed-batch fermentation, which was increased by a factor of nearly 630.ConclusionsThis study first investigated the influence of carbon flux in the upstream module and the introduction of a newly identified GuCYB5 on GA production. The newly identified GuCYB5 was highly effective in improving GA production. An integrated strategy including enzyme discovery, pathway optimization, and fusion protein construction was provided in improving GA production, achieving a 630 fold increase in GA production. The metabolically engineered yeast cell factories provide an alternative approach to glycyrrhetinic acid production, replacing the traditional method of plant extraction.

Dataset Information

Production and characterization of anthocyanin-rich beer from black wheat by an efficient isolate Saccharomyces cerevisiae CMS12.

Publications

Production and characterization of anthocyanin-rich beer from black wheat by an efficient isolate Saccharomyces cerevisiae CMS12.

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