Biochemical characterization of naringenin producing yeast (saccharomyces cerevisiae) strain
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Date
2023
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UMT, Lhr
Abstract
Extensive research has been carried out during the last few decades, providing a detailed account of thousands of discovered phytochemicals and their biological activities that have the potential to be exploited for a wide variety of medicinal purposes. These phytochemicals, which are pharmacologically important for clinical use, primarily consist of polyphenols, followed by terpenoids and alkaloids. Numerous published reports indicate the primary role of phytochemicals proven to possess therapeutic potential against several diseases. However, not all phytochemicals possess significant medicinal properties, and only some of them exhibit viable biological effects. Naringenin, a flavanone found in citrus fruits, is known to improve immunity, repair DNA damage, and scavenge free radicals. Despite the very low bioavailability of naringenin, it is known to exhibit various promising biological properties of medicinal importance, including anti-inflammatory and antioxidant activities. It focuses on
various aspects related to naringenin, particularly its physicochemical properties. Furthermore, various pharmacological activites of naringenin, such as anticancer,
antidiabetic, hepatoprotective, neuroprotective, cardioprotective, nephroprotective and gastroprotective effects, have been discussed along with their mechanisms of action. In our research, yeast was engineered with naringenin pathway genes (TAL, 4CL, CHS, CHI). The production of desired product increased by using different biochemical parameters. I explored various permutations of essential parameters such as pH, temperature, inoculum size, carbon source, nitrogen source, and substrate concentration using Response Surface Methodology (RSM) software. Naringenin compound was extracted from the fermented broth by using ethyl acetate. The extracted compounds was first analyzed by using highthroughput chemicals (sodium Hydroxide, folin, potassium hydroxide, magnesium sulfate, lead oxide and ammonium hydroxide) and subsequently, conducted confirmation test using HPLC. This comprehensive analysis sought to determine the concentration at which our yeast exhibited the highest production of the desired flavonoid, Naringenin. The engineered strain produced naringenin compound which was further optimized with different biochemical parameters. The engineered strain produce maximum compounds when use inoculum size 4%, carbon source 8%, nitrogen 3%, pH 6, temperature 30 oC and subtrate concentration 0.30%. These conditions were confirmed by chemical methods. Among all chemical sodium hydroxide oxide maximally bind
with the naringenin compounds and give bright yellow colour. Other methods which need to be add to increase the desired compounds by adding malonyl-CoA and
overexpression of pathway genes. As one molecule of naringenin requires three molecules of malonyl-CoA, and malonyl-CoA availability represents a bottleneck in
the biosynthesis pathway. The recombinant yeast strain was fed with phenylpropanoid acids and produced naringenin. In conclusion, the production of naringenin using
microbial cell factories, including yeast strains such as Saccharomyces cerevisiae, has shown promise. However, yields remain low, and further optimization is needed to
increase production. There is no specific study on the biochemical characterization of a naringenin -producing S. cerevisiae strain, studies on naringenin biosynthesis in S.
cerevisiae therefore it is the first study to consider biochemical characterization of S. cerevisiae to produce naringenin.