It is well known that Strawberry leaves contain high amounts of diverse phenolic compounds (Kårlund et al. 2014), and also polyphenolic compounds are known to interact with proteins and can inhibit enzymatic activity (Dawra et al. 1988; Suryanarayana et al. 2004).
How can I avoid high levels of polyphenols in strawberry leaf extracts? These high levels of polyphenols interfere with enzymatic activity.
What kind of techniques or protocols do you use to purify raw strawberry leaf extracts and eliminate polyphenols?
Thank you very much!
JD
Dawra, R. K., Makkar, H. P., & Singh, B. (1988). Protein-binding capacity of microquantities of tannins. Analytical Biochemistry, 170(1), 50–53.
Kårlund, A., Salminen, J. P., Koskinen, P., Ahern, J. R., Karonen, M., Tiilikkala, K., & Karjalainen, R. O. (2014). Polyphenols in strawberry (Fragaria× ananassa) leaves induced by plant activators. Journal of agricultural and food chemistry, 62(20), 4592-4600.
Suryanarayana, P., Kumar, P. A., Saraswat, M., Petrash, J. M., & Reddy, G. B. (2004). Inhibition of aldose reductase by tannoid principles of Emblica officinalis: Implications for the prevention of sugar cataract. Molecular Vision, 10, 148–154.
Dear Dr. Franco-Navarro,
We had the same problem with grapevine leaves and berry extracts for transcriptomic and proteomic analysis. We solved it by including 2% (w/v) PVPP (Polyvinylpolypyrrolidone, CAS # 25249-54-1) in extraction buffers (see attached PDF from Lecourieux et al 2010). If this is not efficient enough on your biological samples, you can use more elaborate protocols by purifying your extracts on PVPP packed columns, as described in Ranatunge et al. 2017, International Journal of Analytical Chemistry, Volume 2017, Article ID 7230145.
Hope this'll help you.
Best regards,
E. Gomès.
@ Pascal Dubé
@ J.D. Franco-NavarroYes SPE on N-vinylpyrrolidone-divinylbenzene copolymer would also work nicely, and you can find commercially prepacked cartirdges.
Dear all,
I am pleasantly surprised by the number of responses and the high level of all of them. Thank you very much for the advice brainstorm. We will apply all your suggestions and I will comment again when I'll get results. It is very interesting that of PVPP packed columns.
Hamdy Zahran
: I can not specify the solvent to use for the extracts because I do not use only one, since for each enzyme or product, I need a different kind of solvent. That's why I needed a universal protocol of extraction. Anyway thank you very much.
Depending on the kind of enzyme activity you want to measure, you can add 1% triton X100 in your extraction buffer, and centrifuge, before measuring enzyme activity in the supernatant. Since triton X100 strongly absorb in the UV, you must not dilute too much your plant material in the extrazction buffer, in order that tx100 should be diluted as much as possible in the measurement cuvette. We routinely used Tx100 in conjonction with soluble or insoluble PVP on grape berry , fro enzymatic or proteomic studies, and it really changed our life, (huge increase in activity) when compared to PVP alone. please cite Sauvage et al., aminotransferase in grape. Isolation and characterization of aspartate aminotransferase. Am.J.Enol.Vitic, vol42 n3 1991, pp 209-218
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