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Response surface methodology modelling of an aqueous two-phase system for purification of protease from Penicillium candidum (PCA 1/TT031) under solid state fermentation and its biochemical characterization

Penicillium candidum (PCA 1/TT031) synthesizes different types of extracellular proteases. The objective of this study is to optimize polyethylene glycol (PEG)/citrate based on an aqueous two-phase system (ATPS) and Response Surface Methodology (RSM) to purify protease from Penicillium candidum (PCA...

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Main Authors: Alhelli, Amaal M., Abdul Manap, Mohd Yazid, Mohammed, Abdulkarim Sabo, Mirhosseini, Hamed, Suliman, Eilaf Suliman Khalil, Shad, Zahra, Mohammed, Nameer Khairullah, Meor Hussin, Anis Shobirin
Format: Article
Language:English
Published: Multidisciplinary Digital Publishing Institute 2016
Online Access:http://psasir.upm.edu.my/id/eprint/52872/1/Response%20surface%20methodology%20modelling%20of%20an%20aqueous%20two-phase%20system%20for%20purification%20of%20protease%20from%20Penicillium%20candidum.pdf
http://psasir.upm.edu.my/id/eprint/52872/
https://www.mdpi.com/1422-0067/17/11/1872
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spelling my.upm.eprints.528722022-03-16T00:46:48Z http://psasir.upm.edu.my/id/eprint/52872/ Response surface methodology modelling of an aqueous two-phase system for purification of protease from Penicillium candidum (PCA 1/TT031) under solid state fermentation and its biochemical characterization Alhelli, Amaal M. Abdul Manap, Mohd Yazid Mohammed, Abdulkarim Sabo Mirhosseini, Hamed Suliman, Eilaf Suliman Khalil Shad, Zahra Mohammed, Nameer Khairullah Meor Hussin, Anis Shobirin Penicillium candidum (PCA 1/TT031) synthesizes different types of extracellular proteases. The objective of this study is to optimize polyethylene glycol (PEG)/citrate based on an aqueous two-phase system (ATPS) and Response Surface Methodology (RSM) to purify protease from Penicillium candidum (PCA 1/TT031). The effects of different PEG molecular weights (1500-10,000 g/mol), PEG concentration (9%-20%), concentrations of NaCl (0%-10%) and the citrate buffer (8%-16%) on protease were also studied. The best protease purification could be achieved under the conditions of 9.0% (w/w) PEG 8000, 5.2% NaCl, and 15.9% sodium citrate concentration, which resulted in a one-sided protease partitioning for the bottom phase with a partition coefficient of 0.2, a 6.8-fold protease purification factor, and a yield of 93%. The response surface models displayed a significant (p ≤ 0.05) response which was fit for the variables that were studied as well as a high coefficient of determination (R²). Similarly, the predicted and observed values displayed no significant (p > 0.05) differences. In addition, our enzyme characterization study revealed that Penicillium candidum (PCA 1/TT031) produced a slight neutral protease with a molecular weight between 100 and 140 kDa. The optimal activity of the purified enzyme occurred at a pH of 6.0 and at a temperature of 50 °C. The stability between different pH and temperature ranges along with the effect of chemical metal ions and inhibitors were also studied. Our results reveal that the purified enzyme could be used in the dairy industry such as in accelerated cheese ripening. Multidisciplinary Digital Publishing Institute 2016-11 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/52872/1/Response%20surface%20methodology%20modelling%20of%20an%20aqueous%20two-phase%20system%20for%20purification%20of%20protease%20from%20Penicillium%20candidum.pdf Alhelli, Amaal M. and Abdul Manap, Mohd Yazid and Mohammed, Abdulkarim Sabo and Mirhosseini, Hamed and Suliman, Eilaf Suliman Khalil and Shad, Zahra and Mohammed, Nameer Khairullah and Meor Hussin, Anis Shobirin (2016) Response surface methodology modelling of an aqueous two-phase system for purification of protease from Penicillium candidum (PCA 1/TT031) under solid state fermentation and its biochemical characterization. International Journal of Molecular Science, 17 (11). pp. 1-23. ISSN 1661-6596; ESSN: 1422-0067 https://www.mdpi.com/1422-0067/17/11/1872 10.3390/ijms17111872
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
language English
description Penicillium candidum (PCA 1/TT031) synthesizes different types of extracellular proteases. The objective of this study is to optimize polyethylene glycol (PEG)/citrate based on an aqueous two-phase system (ATPS) and Response Surface Methodology (RSM) to purify protease from Penicillium candidum (PCA 1/TT031). The effects of different PEG molecular weights (1500-10,000 g/mol), PEG concentration (9%-20%), concentrations of NaCl (0%-10%) and the citrate buffer (8%-16%) on protease were also studied. The best protease purification could be achieved under the conditions of 9.0% (w/w) PEG 8000, 5.2% NaCl, and 15.9% sodium citrate concentration, which resulted in a one-sided protease partitioning for the bottom phase with a partition coefficient of 0.2, a 6.8-fold protease purification factor, and a yield of 93%. The response surface models displayed a significant (p ≤ 0.05) response which was fit for the variables that were studied as well as a high coefficient of determination (R²). Similarly, the predicted and observed values displayed no significant (p > 0.05) differences. In addition, our enzyme characterization study revealed that Penicillium candidum (PCA 1/TT031) produced a slight neutral protease with a molecular weight between 100 and 140 kDa. The optimal activity of the purified enzyme occurred at a pH of 6.0 and at a temperature of 50 °C. The stability between different pH and temperature ranges along with the effect of chemical metal ions and inhibitors were also studied. Our results reveal that the purified enzyme could be used in the dairy industry such as in accelerated cheese ripening.
format Article
author Alhelli, Amaal M.
Abdul Manap, Mohd Yazid
Mohammed, Abdulkarim Sabo
Mirhosseini, Hamed
Suliman, Eilaf Suliman Khalil
Shad, Zahra
Mohammed, Nameer Khairullah
Meor Hussin, Anis Shobirin
spellingShingle Alhelli, Amaal M.
Abdul Manap, Mohd Yazid
Mohammed, Abdulkarim Sabo
Mirhosseini, Hamed
Suliman, Eilaf Suliman Khalil
Shad, Zahra
Mohammed, Nameer Khairullah
Meor Hussin, Anis Shobirin
Response surface methodology modelling of an aqueous two-phase system for purification of protease from Penicillium candidum (PCA 1/TT031) under solid state fermentation and its biochemical characterization
author_facet Alhelli, Amaal M.
Abdul Manap, Mohd Yazid
Mohammed, Abdulkarim Sabo
Mirhosseini, Hamed
Suliman, Eilaf Suliman Khalil
Shad, Zahra
Mohammed, Nameer Khairullah
Meor Hussin, Anis Shobirin
author_sort Alhelli, Amaal M.
title Response surface methodology modelling of an aqueous two-phase system for purification of protease from Penicillium candidum (PCA 1/TT031) under solid state fermentation and its biochemical characterization
title_short Response surface methodology modelling of an aqueous two-phase system for purification of protease from Penicillium candidum (PCA 1/TT031) under solid state fermentation and its biochemical characterization
title_full Response surface methodology modelling of an aqueous two-phase system for purification of protease from Penicillium candidum (PCA 1/TT031) under solid state fermentation and its biochemical characterization
title_fullStr Response surface methodology modelling of an aqueous two-phase system for purification of protease from Penicillium candidum (PCA 1/TT031) under solid state fermentation and its biochemical characterization
title_full_unstemmed Response surface methodology modelling of an aqueous two-phase system for purification of protease from Penicillium candidum (PCA 1/TT031) under solid state fermentation and its biochemical characterization
title_sort response surface methodology modelling of an aqueous two-phase system for purification of protease from penicillium candidum (pca 1/tt031) under solid state fermentation and its biochemical characterization
publisher Multidisciplinary Digital Publishing Institute
publishDate 2016
url http://psasir.upm.edu.my/id/eprint/52872/1/Response%20surface%20methodology%20modelling%20of%20an%20aqueous%20two-phase%20system%20for%20purification%20of%20protease%20from%20Penicillium%20candidum.pdf
http://psasir.upm.edu.my/id/eprint/52872/
https://www.mdpi.com/1422-0067/17/11/1872
_version_ 1728052800721518592
score 13.211869

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