Production optimization of a thermostable alkaline and detergent biocompatible protease by Bacillus paramycoides WSA for the green detergent industry

Abstract Industrial applications are evolving to replace chemical catalysts with eco-friendly solutions. This work enhances extracellular protease production from a newly isolated proteolytic Bacillus paramycoides WSA using Response Surface Methodology (RSM). The used quadratic model demonstrated a...

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Main Authors: Wafa A. Alshehri, Sarah A. Alhothifi, Ashjan F. Khalel, Fatimah S. Alqahtani, Bilel Hadrich, Adel Sayari
Format: Article
Language:English
Published: Nature Portfolio 2025-04-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-98094-w
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author Wafa A. Alshehri
Sarah A. Alhothifi
Ashjan F. Khalel
Fatimah S. Alqahtani
Bilel Hadrich
Adel Sayari
author_facet Wafa A. Alshehri
Sarah A. Alhothifi
Ashjan F. Khalel
Fatimah S. Alqahtani
Bilel Hadrich
Adel Sayari
author_sort Wafa A. Alshehri
collection DOAJ
description Abstract Industrial applications are evolving to replace chemical catalysts with eco-friendly solutions. This work enhances extracellular protease production from a newly isolated proteolytic Bacillus paramycoides WSA using Response Surface Methodology (RSM). The used quadratic model demonstrated a good fit with R2 = 80.59%, R2 Adj = 57.94%, a high degree of significance (p = 0.017 < 0.05), and a low RMSE = 0.030 U/mL. Maximum protease activity of 1.346 ± 0.060 U/mL was obtained after 8 h-cultivation at 45 °C and pH 7. The protease activity is maximal at 45 °C and pH 10. It maintains 75% of its activity after 10 min of incubation at 80 °C. Furthermore, the enzyme was found to be stable in an alkaline environment, retaining 89% of its activity after 1 h incubation at pH 12. Several metal ions, surfactants, and inhibitors increased protease performance. Indeed, 110% and 140% protease activity was measured in the presence of 5 mM CoCl2 and CaCl2, respectively. Similarly, 105%, 128%, 135%, 144%, and 160% protease activity were measured in the presence of 0.5% of Tween 20, EDTA, Sodium azide, uric acid, and SDS, respectively. Interestingly, the enzyme activity was enhanced (138%) after 1 h of incubation with 1% Reem detergent. Furthermore, this protease (10 U/mL), in combination with 0.7% detergent, could clean bloodstained fabric after 10 min incubation at 45 °C. Our results suggest that this thermostable, alkaline, and detergent-biocompatible protease could be a promising additive for eco-friendly use in detergent industries.
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spelling doaj-art-8ec3cbf9dcfa4562a4734138fec686df2025-08-20T02:17:50ZengNature PortfolioScientific Reports2045-23222025-04-0115111710.1038/s41598-025-98094-wProduction optimization of a thermostable alkaline and detergent biocompatible protease by Bacillus paramycoides WSA for the green detergent industryWafa A. Alshehri0Sarah A. Alhothifi1Ashjan F. Khalel2Fatimah S. Alqahtani3Bilel Hadrich4Adel Sayari5Department of Biological Sciences, College of Science, University of JeddahDepartment of Biological Sciences, College of Science, University of JeddahDepartment of Biology, University College of Aldarb, Jazan UniversityDepartment of Biology, Faculty of Sciences, University of BishaDepartment of Chemical Engineering, College of Engineering, Imam Mohammad Ibn Saud Islamic University (IMSIU)Department of Biological Sciences, College of Science, University of JeddahAbstract Industrial applications are evolving to replace chemical catalysts with eco-friendly solutions. This work enhances extracellular protease production from a newly isolated proteolytic Bacillus paramycoides WSA using Response Surface Methodology (RSM). The used quadratic model demonstrated a good fit with R2 = 80.59%, R2 Adj = 57.94%, a high degree of significance (p = 0.017 < 0.05), and a low RMSE = 0.030 U/mL. Maximum protease activity of 1.346 ± 0.060 U/mL was obtained after 8 h-cultivation at 45 °C and pH 7. The protease activity is maximal at 45 °C and pH 10. It maintains 75% of its activity after 10 min of incubation at 80 °C. Furthermore, the enzyme was found to be stable in an alkaline environment, retaining 89% of its activity after 1 h incubation at pH 12. Several metal ions, surfactants, and inhibitors increased protease performance. Indeed, 110% and 140% protease activity was measured in the presence of 5 mM CoCl2 and CaCl2, respectively. Similarly, 105%, 128%, 135%, 144%, and 160% protease activity were measured in the presence of 0.5% of Tween 20, EDTA, Sodium azide, uric acid, and SDS, respectively. Interestingly, the enzyme activity was enhanced (138%) after 1 h of incubation with 1% Reem detergent. Furthermore, this protease (10 U/mL), in combination with 0.7% detergent, could clean bloodstained fabric after 10 min incubation at 45 °C. Our results suggest that this thermostable, alkaline, and detergent-biocompatible protease could be a promising additive for eco-friendly use in detergent industries.https://doi.org/10.1038/s41598-025-98094-wAlkaline and thermostable Bacillus paramycoides proteaseResponse surface methodologyProduction optimizationBloodstain enzymatic cleaning
spellingShingle Wafa A. Alshehri
Sarah A. Alhothifi
Ashjan F. Khalel
Fatimah S. Alqahtani
Bilel Hadrich
Adel Sayari
Production optimization of a thermostable alkaline and detergent biocompatible protease by Bacillus paramycoides WSA for the green detergent industry
Scientific Reports
Alkaline and thermostable Bacillus paramycoides protease
Response surface methodology
Production optimization
Bloodstain enzymatic cleaning
title Production optimization of a thermostable alkaline and detergent biocompatible protease by Bacillus paramycoides WSA for the green detergent industry
title_full Production optimization of a thermostable alkaline and detergent biocompatible protease by Bacillus paramycoides WSA for the green detergent industry
title_fullStr Production optimization of a thermostable alkaline and detergent biocompatible protease by Bacillus paramycoides WSA for the green detergent industry
title_full_unstemmed Production optimization of a thermostable alkaline and detergent biocompatible protease by Bacillus paramycoides WSA for the green detergent industry
title_short Production optimization of a thermostable alkaline and detergent biocompatible protease by Bacillus paramycoides WSA for the green detergent industry
title_sort production optimization of a thermostable alkaline and detergent biocompatible protease by bacillus paramycoides wsa for the green detergent industry
topic Alkaline and thermostable Bacillus paramycoides protease
Response surface methodology
Production optimization
Bloodstain enzymatic cleaning
url https://doi.org/10.1038/s41598-025-98094-w
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