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Improved naringinase double screen plate assay: progress towards the perfect screening
Jitendra D Salunkhe1, Satish V Patil1
1School of Life Sciences, Kavayitri Bahinabai Chaudhari North Maharashtra University, Jalgaon, Maharashtra, India.
Natural Product Research
|September 25, 2023
Summary
A new double screen plate assay improves the identification of microbes producing naringinase, an enzyme with dual α-l-rhamnosidase and β-d-glucosidase activity. This method overcomes limitations of traditional naringin agar screening, reducing false positives for reliable enzyme discovery.
Area of Science:
- Enzymology
- Microbiology
- Biotechnology
Background:
- Naringinase, possessing both α-l-rhamnosidase and β-d-glucosidase activities, is commercially significant.
- Traditional screening using naringin agar yields false positives due to non-specific enzyme activities or alternative carbon source utilization.
- Accurate identification of naringinase-producing microbes is crucial for biotechnological applications.
Purpose of the Study:
- To develop a more reliable screening method for identifying naringinase-producing microorganisms.
- To overcome the limitations of traditional naringin agar screening methods.
- To accurately differentiate true naringinase producers from other microbial contaminants.
Main Methods:
- A novel double screen plate assay was designed utilizing synthetic substrates.
- The assay separately detects β-d-glucosidase and α-l-rhamnosidase activities.
- Enzyme activity is indicated by the formation of a yellow zone resulting from p-nitrophenol production.
Main Results:
- The double screen plate assay effectively distinguishes naringinase producers.
- The intensity of the yellow color zone correlates with the potential for naringinase production.
- This method significantly reduces false positive results compared to traditional assays.
Conclusions:
- The developed double screen plate assay offers a significant improvement for identifying naringinase producers.
- This advancement provides a more reliable and accurate screening tool for enzyme discovery.
- The new method facilitates efficient isolation of microorganisms with valuable naringinase activity.
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