Modulation of hide protein resistance to enzymatic hydrolysis by calcium ions: rational design of enzyme-assisted unhairing for high-quality leather production

Abstract Enzymatic unhairing is an environmentally friendly and efficient method for leather processing. However, controlling protease hydrolysis remains challenging, leading to incomplete hair removal and potential grain damage. In modern leather manufacturing, the synergistic application of protea...

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Main Authors: Hao Liu, Xuyang Chen, Juntao Kang, Bi Shi, Yunhang Zeng
Format: Article
Language:English
Published: SpringerOpen 2025-08-01
Series:Collagen and Leather
Subjects:
Online Access:https://doi.org/10.1186/s42825-025-00208-7
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author Hao Liu
Xuyang Chen
Juntao Kang
Bi Shi
Yunhang Zeng
author_facet Hao Liu
Xuyang Chen
Juntao Kang
Bi Shi
Yunhang Zeng
author_sort Hao Liu
collection DOAJ
description Abstract Enzymatic unhairing is an environmentally friendly and efficient method for leather processing. However, controlling protease hydrolysis remains challenging, leading to incomplete hair removal and potential grain damage. In modern leather manufacturing, the synergistic application of proteases, lime, and sulfide is increasingly employed to achieve satisfactory hair-saving unhairing performance. This study investigated the action mechanism of calcium ions in modulating the hydrolysis of hide proteins by proteases and proposed a balanced enzyme-assisted unhairing process. Enzymological and fluorescence spectroscopy analyses revealed that calcium ions could enhance the enzymatic resistance of hide proteins, including noncollagenous proteins and collagen, by binding to them. This enhancement in enzymatic resistance was more pronounced for globular proteins than for collagen fibers. In detail, following the addition of 20 g/L calcium ions, the hydrolytic activity of neutral and alkaline proteases decreased by 66.7% and 57.9% on bovine serum albumin, and by 40.7% and 48.1% on collagen fibers, respectively. Furthermore, the performance of the five unhairing processes was evaluated by varying the sequence of lime and protease application and type of protease used. Results indicated that while calcium ions exerted a protective effect on hide proteins and reduced damage to collagen fibers, they simultaneously hindered the removal of undesired noncollagenous proteins during unhairing. Consequently, the sequential application of lime followed by proteases resulted in the inadequate removal of interfibrillar substances, leading to unsatisfactory leather quality. Additionally, compared with an alkaline protease, a neutral protease was more easily inhibited under alkaline unhairing conditions, posed a lower risk of damage to the hide grain. Therefore, the neutral protease–lime–sodium sulfide unhairing process was chosen as the optimal strategy. This process involves the addition of neutral protease (50 U/g hide) for 60 min, followed by 1.0% lime for 90 min and 2.0% sodium sulfide for 90 min. These findings provide scientific insights for designing a controlled and efficient approach to enzyme-assisted unhairing processes. Graphical Abstract
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series Collagen and Leather
spelling doaj-art-1d90b01c4e904c618cd6235cc3e7538a2025-08-20T03:46:16ZengSpringerOpenCollagen and Leather2097-14192731-69982025-08-017111410.1186/s42825-025-00208-7Modulation of hide protein resistance to enzymatic hydrolysis by calcium ions: rational design of enzyme-assisted unhairing for high-quality leather productionHao Liu0Xuyang Chen1Juntao Kang2Bi Shi3Yunhang Zeng4Key Laboratory of Leather Chemistry and Engineering (Ministry of Education), National Engineering Laboratory for Clean Technology of Leather Manufacture, College of Biomass Science and Engineering, Sichuan UniversityKey Laboratory of Leather Chemistry and Engineering (Ministry of Education), National Engineering Laboratory for Clean Technology of Leather Manufacture, College of Biomass Science and Engineering, Sichuan UniversitySichuan Decision New Material Technology Co., Ltd.Key Laboratory of Leather Chemistry and Engineering (Ministry of Education), National Engineering Laboratory for Clean Technology of Leather Manufacture, College of Biomass Science and Engineering, Sichuan UniversityKey Laboratory of Leather Chemistry and Engineering (Ministry of Education), National Engineering Laboratory for Clean Technology of Leather Manufacture, College of Biomass Science and Engineering, Sichuan UniversityAbstract Enzymatic unhairing is an environmentally friendly and efficient method for leather processing. However, controlling protease hydrolysis remains challenging, leading to incomplete hair removal and potential grain damage. In modern leather manufacturing, the synergistic application of proteases, lime, and sulfide is increasingly employed to achieve satisfactory hair-saving unhairing performance. This study investigated the action mechanism of calcium ions in modulating the hydrolysis of hide proteins by proteases and proposed a balanced enzyme-assisted unhairing process. Enzymological and fluorescence spectroscopy analyses revealed that calcium ions could enhance the enzymatic resistance of hide proteins, including noncollagenous proteins and collagen, by binding to them. This enhancement in enzymatic resistance was more pronounced for globular proteins than for collagen fibers. In detail, following the addition of 20 g/L calcium ions, the hydrolytic activity of neutral and alkaline proteases decreased by 66.7% and 57.9% on bovine serum albumin, and by 40.7% and 48.1% on collagen fibers, respectively. Furthermore, the performance of the five unhairing processes was evaluated by varying the sequence of lime and protease application and type of protease used. Results indicated that while calcium ions exerted a protective effect on hide proteins and reduced damage to collagen fibers, they simultaneously hindered the removal of undesired noncollagenous proteins during unhairing. Consequently, the sequential application of lime followed by proteases resulted in the inadequate removal of interfibrillar substances, leading to unsatisfactory leather quality. Additionally, compared with an alkaline protease, a neutral protease was more easily inhibited under alkaline unhairing conditions, posed a lower risk of damage to the hide grain. Therefore, the neutral protease–lime–sodium sulfide unhairing process was chosen as the optimal strategy. This process involves the addition of neutral protease (50 U/g hide) for 60 min, followed by 1.0% lime for 90 min and 2.0% sodium sulfide for 90 min. These findings provide scientific insights for designing a controlled and efficient approach to enzyme-assisted unhairing processes. Graphical Abstracthttps://doi.org/10.1186/s42825-025-00208-7LeatherUnhairingProteaseCalcium ionsCollagenHide proteins
spellingShingle Hao Liu
Xuyang Chen
Juntao Kang
Bi Shi
Yunhang Zeng
Modulation of hide protein resistance to enzymatic hydrolysis by calcium ions: rational design of enzyme-assisted unhairing for high-quality leather production
Collagen and Leather
Leather
Unhairing
Protease
Calcium ions
Collagen
Hide proteins
title Modulation of hide protein resistance to enzymatic hydrolysis by calcium ions: rational design of enzyme-assisted unhairing for high-quality leather production
title_full Modulation of hide protein resistance to enzymatic hydrolysis by calcium ions: rational design of enzyme-assisted unhairing for high-quality leather production
title_fullStr Modulation of hide protein resistance to enzymatic hydrolysis by calcium ions: rational design of enzyme-assisted unhairing for high-quality leather production
title_full_unstemmed Modulation of hide protein resistance to enzymatic hydrolysis by calcium ions: rational design of enzyme-assisted unhairing for high-quality leather production
title_short Modulation of hide protein resistance to enzymatic hydrolysis by calcium ions: rational design of enzyme-assisted unhairing for high-quality leather production
title_sort modulation of hide protein resistance to enzymatic hydrolysis by calcium ions rational design of enzyme assisted unhairing for high quality leather production
topic Leather
Unhairing
Protease
Calcium ions
Collagen
Hide proteins
url https://doi.org/10.1186/s42825-025-00208-7
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AT juntaokang modulationofhideproteinresistancetoenzymatichydrolysisbycalciumionsrationaldesignofenzymeassistedunhairingforhighqualityleatherproduction
AT bishi modulationofhideproteinresistancetoenzymatichydrolysisbycalciumionsrationaldesignofenzymeassistedunhairingforhighqualityleatherproduction
AT yunhangzeng modulationofhideproteinresistancetoenzymatichydrolysisbycalciumionsrationaldesignofenzymeassistedunhairingforhighqualityleatherproduction