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Heterologous expression of alkaline metalloproteinases in bacillus subtilis sck6 for eco-friendly enzymatic unhairing of goatskins / Shihao Zhang in JOURNAL OF THE AMERICAN LEATHER CHEMISTS ASSOCIATION (JALCA), Vol. CXVII, N° 3 (03/2022)
[article]
Titre : Heterologous expression of alkaline metalloproteinases in bacillus subtilis sck6 for eco-friendly enzymatic unhairing of goatskins Type de document : texte imprimé Auteurs : Shihao Zhang, Auteur ; Ruoshi Zhang, Auteur ; Xiaoguang Li, Auteur ; Zhe Xu, Auteur ; Yongqiang Tian, Auteur Année de publication : 2022 Article en page(s) : p. 113-123 Note générale : Bibliogr. Langues : Américain (ame) Catégories : Caractérisation
Cuirs et peaux -- Propriétés mécaniques
Cuirs et peaux de chèvres
Epilage enzymatique
Matériaux -- Coloration
Métalloprotéinases
Wet-blue (tannage)Peau tannée au chrome (le chrome donne une couleur bleue)Index. décimale : 675 Technologie du cuir et de la fourrure Résumé : In this study, alkaline metalloprotease gene 1067 was cloned from Planococcus halotolerans SCU63T and heterologously expressed in Bacillus subtilis SCK6. Using Luria Bertani (LB) broth medium as the initial medium, the optimal medium was obtained through a series of fermentation and culture optimization (g/L): yeast extract (10), soybean powder (15), urea (20), potassium chloride (6.7), calcium chloride (13.3), NaCl (10). On the basis of the optimal medium, the highest enzymatic activity of 1259.21 U/mL could be obtained by culturing at 30°C for 40 h. with pH 8, inoculation amount of 4% and filling amount of 50 mL. EDTA inhibits protease activity and PMSF promotes it, indicating that it was a metalloprotease rather than a serine protease. The optimum reaction temperature of the protease is 70°C, and the optimum pH is 9. The metal ions Zn 2+, Co2+ and surfactant β-ME, Tween 80 can improve the activity of the protease. The results of unhairing, SEM and staining show that this metalloprotease can completely dehair goatskin. Compared with the conventional chemical method, the goatskin after enzymatic unhairing has softer texture and smoother surface, and there is no obvious damage to the goatskin. Note de contenu : - MATERIALS AND METHODS : Materials - Construction of recombinant plasmid and expression of 1067 in B. subtilis SCK6 - Optimization of fermentations conditions - Purification of protease 1067 - Enzyme assay - Analysis of enzymatic properties - Unhairing function of protease 1067 - Characterization of protease 1067 unhairing
- RESULTS AND DISCUSSION : Construction of recombinant strain and identification of protease activity - Fermentation optimization - Purification and identification of protease 1067 - Effect of temperature on enzyme activity and stability - Effect of pH on enzyme activity and stability - Effect of metal ions on enzyme activity - Effects of chemicals on enzyme activity - Effect of organic solvents on enzyme activity - Analysis of unhairing - SEM and analysis of tissue staining
- Table 1 : Factors and levels for RSM
- Table 2 : ANOVA for quadratic model
- Table 3 : Effect of metal ions on enzyme activity
- Table 4 : Effect of chemicals on enzyme activity
- Table 5 : Effect of solvents on enzyme activity
- table 6 : Characterization of mechanical properties of wet blue skinDOI : https://doi.org/10.34314/jalca.v117i3.4892 En ligne : https://drive.google.com/file/d/1z4VjDZxnKDTJf0c5BtRxJKAwUieQ2jK9/view?usp=drive [...] Format de la ressource électronique : Permalink : https://e-campus.itech.fr/pmb/opac_css/index.php?lvl=notice_display&id=37602
in JOURNAL OF THE AMERICAN LEATHER CHEMISTS ASSOCIATION (JALCA) > Vol. CXVII, N° 3 (03/2022) . - p. 113-123[article]Réservation
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Code-barres Cote Support Localisation Section Disponibilité 23402 - Périodique Bibliothèque principale Documentaires Disponible Heterologous expression of metalloproteinases from planococcus halotolerans SCU63T and eco-friendly enzymatic dehairing of goatskin / Zhang Shihao in JOURNAL OF THE SOCIETY OF LEATHER TECHNOLOGISTS & CHEMISTS (JSLTC), Vol. 105, N° 3 (05-06/2021)
[article]
Titre : Heterologous expression of metalloproteinases from planococcus halotolerans SCU63T and eco-friendly enzymatic dehairing of goatskin Type de document : texte imprimé Auteurs : Zhang Shihao, Auteur ; Ruoshi Zhang, Auteur ; Xiaoguang Li, Auteur ; Zhe Xu, Auteur ; Yongqiang Tian, Auteur Année de publication : 2021 Article en page(s) : p. 132-139 Note générale : Bibliogr. Langues : Anglais (eng) Catégories : Caractérisation
Cuirs et peaux de chèvres
Enzymes microbiennes
Epilage enzymatique
Métalloprotéinases
Microscopie électronique à balayage
Peptidases
pH
Produits chimiques -- Purification
Travail de rivière (cuir)Index. décimale : 675 Technologie du cuir et de la fourrure Résumé : In this study, a metalloproteinase gene from Planococcus halotolerans SCU63 was cloned and expressed in Bacillus sp. SCK6. LB broth medium was used as the initial medium and for a series of fermentation optimisations. The most suitable medium components were as follows (g/l) : yeast extract powder (20), casein (13), tryptone (7), NaCl (10), urea (8). When the inoculation amount was 2%, the filling amount was 100mL, the culture time was 370C, and the pH was 7.0 for 40 hours the protease production reached the highest. The enzyme activity was completely inhibited by EDTA. Surfactants, such as Tween 20, Tween 80, and TritonX-100, could improve the enzyme activity, while SDS could inhibit the enzyme activity. Under protease action, goatskin can be completely depilated at 37oc and 28 hours. Compared with the conventional chemical dehairing, the enzymatic dehairing opened up the goatskin elastic fibre without any hair root in the hair follicle, and the skin was of high quality. Note de contenu : - MATERIALS AND METHODS : Materials - Construction of recombinant plasmid and expression of 1030 in B. subtilis SCK6 - Optimisation of fermentation conditions - Purification of proteases - Determination of enzyme activity - Characterisation of enzymatic properties - Dehairing function of protease 1030 - Analysis of dehairing
- RESULTS : Construction of recombinant plasmid and its expression in SCK6 - Optimisation of fermentation conditions and RSM - Purification and identification of proteases 1030 - Effect of temperature on enzyme activity and stability - Effect of pH on enzyme activity and stability - Effect of NaCl concentration on enzyme activity - Effect of metal ions on enzyme activity - Effect of chemicals on enzyme activity - Effect of organic solvents on enzyme activity - Analysis of enzymatic dehairing - Analysis of SEM and staining
- Table 1 : Factors and levels for RSM
- Table 2 : ANOVA for quadratic model
- Table 3 : Effect of metal ions on enzyme activity
- Table 4 : Effect of chemicals on enzyme activity
- Table 5 : Effect of solvents on enzyme activityEn ligne : https://drive.google.com/file/d/15BWASUeWYefKBb5gBjgfaEyNcAG7bjbz/view?usp=drive [...] Format de la ressource électronique : Permalink : https://e-campus.itech.fr/pmb/opac_css/index.php?lvl=notice_display&id=35833
in JOURNAL OF THE SOCIETY OF LEATHER TECHNOLOGISTS & CHEMISTS (JSLTC) > Vol. 105, N° 3 (05-06/2021) . - p. 132-139[article]Réservation
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Code-barres Cote Support Localisation Section Disponibilité 22745 - Périodique Bibliothèque principale Documentaires Disponible