Fourier transform infrared spectroscopy provides an evidence of papain denaturation and aggregation during cold storage
Аутори
Rašković, BrankicaPopović, Milica M.
Ostojić, Sanja B.
Anđelković, Boban D.
Tešević, Vele
Polović, Natalija
Чланак у часопису (Рецензирана верзија)
Метаподаци
Приказ свих података о документуАпстракт
Papain is a cysteine protease with wide substrate specificity and many applications. Despite its widespread applications, cold stability of papain has never been studied. Here, we used differential spectroscopy to monitor thermal denaturation process. Papain was the most stabile from 45 degrees C to 60 degrees C with Delta G degrees(321) of 13.9 +/- 0.3 kJ/mol and T-m value of 84 +/- 1 degrees C. After cold storage, papain lost parts of its native secondary structures elements which gave an increase of 40% of intermolecular beta-sheet content (band maximum detected at frequency of 1621 cm(-1) in Fourier transform infrared (FT-IR) spectrum) indicating the presence of secondary structures necessary for aggregation. The presence of protein aggregates after cold storage was also proven by analytical size exclusion chromatography. After six freeze-thaw cycles around 75% of starting enzyme activity of papain was lost due to cold denaturation and aggregation of unfolded protein. Autoproteolys...is of papain did not cause significant loss of the protein activity. Upon the cold storage, papain underwent structural rearrangements and aggregation that correspond to other cold denatured proteins, rather than autoproteolysis which could have the commercial importance for the growing polypeptide based industry.
Кључне речи:
Papain / FT-IR / Cold stability / Cold denaturation / Cold inactivation / AggregationИзвор:
Spectrochimica Acta. Part A: Molecular and Biomolecular Spectroscopy, 2015, 150, 238-246Издавач:
- Pergamon-Elsevier Science Ltd, Oxford
Финансирање / пројекти:
- Алергени, антитела, ензими и мали физиолошки значајни молекули: дизајн, структура, функција и значај (RS-172049)
Напомена:
- This is the peer-reviewed version of the following article: Rašković, B.; Popović, M.; Ostojić, S.; Ancrossed D Signelković, B.; Tešević, V.; Polović, N. Fourier Transform Infrared Spectroscopy Provides an Evidence of Papain Denaturation and Aggregation during Cold Storage. Spectrochimica Acta - Part A: Molecular and Biomolecular Spectroscopy 2015, 150, 238–246. https://doi.org/10.1016/j.saa.2015.05.061
DOI: 10.1016/j.saa.2015.05.061
ISSN: 1386-1425
PubMed: 26051646
WoS: 000361774900031
Scopus: 2-s2.0-84930935650
Колекције
Институција/група
Hemijski fakultet / Faculty of ChemistryTY - JOUR AU - Rašković, Brankica AU - Popović, Milica M. AU - Ostojić, Sanja B. AU - Anđelković, Boban D. AU - Tešević, Vele AU - Polović, Natalija PY - 2015 UR - https://cherry.chem.bg.ac.rs/handle/123456789/3367 AB - Papain is a cysteine protease with wide substrate specificity and many applications. Despite its widespread applications, cold stability of papain has never been studied. Here, we used differential spectroscopy to monitor thermal denaturation process. Papain was the most stabile from 45 degrees C to 60 degrees C with Delta G degrees(321) of 13.9 +/- 0.3 kJ/mol and T-m value of 84 +/- 1 degrees C. After cold storage, papain lost parts of its native secondary structures elements which gave an increase of 40% of intermolecular beta-sheet content (band maximum detected at frequency of 1621 cm(-1) in Fourier transform infrared (FT-IR) spectrum) indicating the presence of secondary structures necessary for aggregation. The presence of protein aggregates after cold storage was also proven by analytical size exclusion chromatography. After six freeze-thaw cycles around 75% of starting enzyme activity of papain was lost due to cold denaturation and aggregation of unfolded protein. Autoproteolysis of papain did not cause significant loss of the protein activity. Upon the cold storage, papain underwent structural rearrangements and aggregation that correspond to other cold denatured proteins, rather than autoproteolysis which could have the commercial importance for the growing polypeptide based industry. PB - Pergamon-Elsevier Science Ltd, Oxford T2 - Spectrochimica Acta. Part A: Molecular and Biomolecular Spectroscopy T1 - Fourier transform infrared spectroscopy provides an evidence of papain denaturation and aggregation during cold storage VL - 150 SP - 238 EP - 246 DO - 10.1016/j.saa.2015.05.061 ER -
@article{ author = "Rašković, Brankica and Popović, Milica M. and Ostojić, Sanja B. and Anđelković, Boban D. and Tešević, Vele and Polović, Natalija", year = "2015", abstract = "Papain is a cysteine protease with wide substrate specificity and many applications. Despite its widespread applications, cold stability of papain has never been studied. Here, we used differential spectroscopy to monitor thermal denaturation process. Papain was the most stabile from 45 degrees C to 60 degrees C with Delta G degrees(321) of 13.9 +/- 0.3 kJ/mol and T-m value of 84 +/- 1 degrees C. After cold storage, papain lost parts of its native secondary structures elements which gave an increase of 40% of intermolecular beta-sheet content (band maximum detected at frequency of 1621 cm(-1) in Fourier transform infrared (FT-IR) spectrum) indicating the presence of secondary structures necessary for aggregation. The presence of protein aggregates after cold storage was also proven by analytical size exclusion chromatography. After six freeze-thaw cycles around 75% of starting enzyme activity of papain was lost due to cold denaturation and aggregation of unfolded protein. Autoproteolysis of papain did not cause significant loss of the protein activity. Upon the cold storage, papain underwent structural rearrangements and aggregation that correspond to other cold denatured proteins, rather than autoproteolysis which could have the commercial importance for the growing polypeptide based industry.", publisher = "Pergamon-Elsevier Science Ltd, Oxford", journal = "Spectrochimica Acta. Part A: Molecular and Biomolecular Spectroscopy", title = "Fourier transform infrared spectroscopy provides an evidence of papain denaturation and aggregation during cold storage", volume = "150", pages = "238-246", doi = "10.1016/j.saa.2015.05.061" }
Rašković, B., Popović, M. M., Ostojić, S. B., Anđelković, B. D., Tešević, V.,& Polović, N.. (2015). Fourier transform infrared spectroscopy provides an evidence of papain denaturation and aggregation during cold storage. in Spectrochimica Acta. Part A: Molecular and Biomolecular Spectroscopy Pergamon-Elsevier Science Ltd, Oxford., 150, 238-246. https://doi.org/10.1016/j.saa.2015.05.061
Rašković B, Popović MM, Ostojić SB, Anđelković BD, Tešević V, Polović N. Fourier transform infrared spectroscopy provides an evidence of papain denaturation and aggregation during cold storage. in Spectrochimica Acta. Part A: Molecular and Biomolecular Spectroscopy. 2015;150:238-246. doi:10.1016/j.saa.2015.05.061 .
Rašković, Brankica, Popović, Milica M., Ostojić, Sanja B., Anđelković, Boban D., Tešević, Vele, Polović, Natalija, "Fourier transform infrared spectroscopy provides an evidence of papain denaturation and aggregation during cold storage" in Spectrochimica Acta. Part A: Molecular and Biomolecular Spectroscopy, 150 (2015):238-246, https://doi.org/10.1016/j.saa.2015.05.061 . .