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Trypsin activity and freeze-thaw stability in the presence of ions and non-ionic surfactants

Authorized Users Only
2021
Authors
Vatić, Saša
Mirković, Nemanja
Milošević, Jelica
Jovčić, Branko
Polović, Natalija
Article (Published version)
Metadata
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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 destabi...lization of α-helicoidal segments of trypsin structure after 7 freeze-thaw cycles but also increased protein substrate availability.

Keywords:
Fluorescence / Fourier transform infrared / Freeze-thaw stability / Surfactants / Trypsin
Source:
Journal of Bioscience and Bioengineering, 2021, 131, 3, 234-240
Publisher:
  • Elsevier
Funding / projects:
  • Ministry of Education, Science and Technological Development, Republic of Serbia, Grant no. 200168 (University of Belgrade, Faculty of Chemistry) (RS-200168)

DOI: 10.1016/j.jbiosc.2020.10.010

ISSN: 1389-1723

WoS: 000661493400002

Scopus: 2-s2.0-85096181210
[ Google Scholar ]
2
URI
https://www.sciencedirect.com/science/article/pii/S1389172320303996
http://cherry.chem.bg.ac.rs/handle/123456789/4779
Collections
  • Publikacije
Institution/Community
Hemijski fakultet
TY  - 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 . .

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