Trypsin activity and freeze-thaw stability in the presence of ions and non-ionic surfactants
Samo za registrovane korisnike
2021
Članak u časopisu (Objavljena verzija)
Metapodaci
Prikaz svih podataka o dokumentuApstrakt
Trypsin is a serine protease with important applications such as protein sequencing and tissue dissociation. Preserving protein structure and its activity during freeze-thawing and prolonging its shelf life is one of the most interesting tasks in biochemistry. In the present study, trypsin cryoprotection was achieved by altering buffer composition. Sodium phosphate buffer at pH 8.0 led to pH shift-induced destabilization of trypsin and formation of a molten globule, followed by significant activity loss (about 70%). Potassium phosphate and ammonium bicarbonate buffers at pH 8.0 were used with up to 90% activity recovery rate after 7 freeze-thaw cycles. The addition of non-ionic surfactants Tween 20 and Tween 80 led to up to 99% activity recovery rate. Amide I region changes, corresponding to specific secondary structures in the Fourier transform infrared (FTIR) spectrum, were modest in the case of Tween 20 and Tween 80. On the other hand, the addition of Triton X-100 led to the destabi...lization of α-helicoidal segments of trypsin structure after 7 freeze-thaw cycles but also increased protein substrate availability.
Ključne reči:
Fluorescence / Fourier transform infrared / Freeze-thaw stability / Surfactants / TrypsinIzvor:
Journal of Bioscience and Bioengineering, 2021, 131, 3, 234-240Izdavač:
- Elsevier
Finansiranje / projekti:
- Ministarstvo nauke, tehnološkog razvoja i inovacija Republike Srbije, institucionalno finansiranje - 200168 (Univerzitet u Beogradu, Hemijski fakultet) (RS-MESTD-inst-2020-200168)
DOI: 10.1016/j.jbiosc.2020.10.010
ISSN: 1389-1723
WoS: 000661493400002
Scopus: 2-s2.0-85096181210
URI
https://www.sciencedirect.com/science/article/pii/S1389172320303996http://cherry.chem.bg.ac.rs/handle/123456789/4779
Kolekcije
Institucija/grupa
Hemijski fakultet / Faculty of ChemistryTY - JOUR AU - Vatić, Saša AU - Mirković, Nemanja AU - Milošević, Jelica AU - Jovčić, Branko AU - Polović, Natalija PY - 2021 UR - https://www.sciencedirect.com/science/article/pii/S1389172320303996 UR - http://cherry.chem.bg.ac.rs/handle/123456789/4779 AB - Trypsin is a serine protease with important applications such as protein sequencing and tissue dissociation. Preserving protein structure and its activity during freeze-thawing and prolonging its shelf life is one of the most interesting tasks in biochemistry. In the present study, trypsin cryoprotection was achieved by altering buffer composition. Sodium phosphate buffer at pH 8.0 led to pH shift-induced destabilization of trypsin and formation of a molten globule, followed by significant activity loss (about 70%). Potassium phosphate and ammonium bicarbonate buffers at pH 8.0 were used with up to 90% activity recovery rate after 7 freeze-thaw cycles. The addition of non-ionic surfactants Tween 20 and Tween 80 led to up to 99% activity recovery rate. Amide I region changes, corresponding to specific secondary structures in the Fourier transform infrared (FTIR) spectrum, were modest in the case of Tween 20 and Tween 80. On the other hand, the addition of Triton X-100 led to the destabilization of α-helicoidal segments of trypsin structure after 7 freeze-thaw cycles but also increased protein substrate availability. PB - Elsevier T2 - Journal of Bioscience and Bioengineering T1 - Trypsin activity and freeze-thaw stability in the presence of ions and non-ionic surfactants VL - 131 IS - 3 SP - 234 EP - 240 DO - 10.1016/j.jbiosc.2020.10.010 ER -
@article{ author = "Vatić, Saša and Mirković, Nemanja and Milošević, Jelica and Jovčić, Branko and Polović, Natalija", year = "2021", abstract = "Trypsin is a serine protease with important applications such as protein sequencing and tissue dissociation. Preserving protein structure and its activity during freeze-thawing and prolonging its shelf life is one of the most interesting tasks in biochemistry. In the present study, trypsin cryoprotection was achieved by altering buffer composition. Sodium phosphate buffer at pH 8.0 led to pH shift-induced destabilization of trypsin and formation of a molten globule, followed by significant activity loss (about 70%). Potassium phosphate and ammonium bicarbonate buffers at pH 8.0 were used with up to 90% activity recovery rate after 7 freeze-thaw cycles. The addition of non-ionic surfactants Tween 20 and Tween 80 led to up to 99% activity recovery rate. Amide I region changes, corresponding to specific secondary structures in the Fourier transform infrared (FTIR) spectrum, were modest in the case of Tween 20 and Tween 80. On the other hand, the addition of Triton X-100 led to the destabilization of α-helicoidal segments of trypsin structure after 7 freeze-thaw cycles but also increased protein substrate availability.", publisher = "Elsevier", journal = "Journal of Bioscience and Bioengineering", title = "Trypsin activity and freeze-thaw stability in the presence of ions and non-ionic surfactants", volume = "131", number = "3", pages = "234-240", doi = "10.1016/j.jbiosc.2020.10.010" }
Vatić, S., Mirković, N., Milošević, J., Jovčić, B.,& Polović, N.. (2021). Trypsin activity and freeze-thaw stability in the presence of ions and non-ionic surfactants. in Journal of Bioscience and Bioengineering Elsevier., 131(3), 234-240. https://doi.org/10.1016/j.jbiosc.2020.10.010
Vatić S, Mirković N, Milošević J, Jovčić B, Polović N. Trypsin activity and freeze-thaw stability in the presence of ions and non-ionic surfactants. in Journal of Bioscience and Bioengineering. 2021;131(3):234-240. doi:10.1016/j.jbiosc.2020.10.010 .
Vatić, Saša, Mirković, Nemanja, Milošević, Jelica, Jovčić, Branko, Polović, Natalija, "Trypsin activity and freeze-thaw stability in the presence of ions and non-ionic surfactants" in Journal of Bioscience and Bioengineering, 131, no. 3 (2021):234-240, https://doi.org/10.1016/j.jbiosc.2020.10.010 . .