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Latex peptidases of Calotropis procera for dehairing of leather as an alternative to environmentally toxic sodium sulfide treatment

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Abstract

Dehairing of crude leather is a critical stage performed at the beginning of its processing to obtain industrially useful pieces. Tanneries traditionally apply a chemical process based on sodium sulfide. Since this chemical reactive is environmentally toxic and inefficiently recycled, innovative protocols for reducing or eliminating its use in leather depilation are welcomed. Therefore, latex peptidases from Calotropis procera (CpLP) and Cryptostegia grandiflora (CgLP) were assayed for this purpose. Enzyme activity on substrates representative of skin such as hide powder azure (UHPA), elastin (UE), azocollagen (UAZOCOL), keratin (UK), and epidermis (UEP) was determined, while depilation activity was assayed on cow hide. Only CpLP was active against keratin (13.4 UK) and only CgLP was active against elastin (0.12 UE). CpLP (93.0 UHPA, 403.6 UAZOCOL, 36.3 UEP) showed higher activity against the other substrates than CgLP (47.6 UHPA, 261.5 UAZOCOL, 8.5 UEP). In pilot assays, CpLP (0.05% w/v with sodium sulfite 0.6% w/v as activator) released hairs from cow hide pieces. Macroscopic and microscopic analyses of the hide revealed that the dehairing process was complete and the leather structure was preserved. The proteolytic system of C. procera is a suitable bioresources to be exploited by tanneries.

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References

  1. Durga J, Ranjithkumar A, Ramesh R, Girivasan KTPV, Rose C, Muralidharan C (2015) Replacement of lime with carbohydrases—a successful cleaner process for leather making. J Clean Prod 112:1122–1127

    Article  Google Scholar 

  2. Bhavan S, Rao JR, Nair BU (2008) A potential new commercial method for processing leather to reduce environmental impact. Environ Sci Pollut Res 15:293–295

    Article  Google Scholar 

  3. Dixit S, Yadav A, Dwivedi PD, Das M (2015) Toxic hazards of leather industry and technologies to combat threat: a review. J Clean Prod 87:39–49

    Article  CAS  Google Scholar 

  4. Castilho LR, Medronho RA, Alves TLM (2000) Production and extraction of pectinases obtained by solid state fermentation of agroindustrial residues with Aspergillus niger. Bioresour Technol 71:45–50

    Article  CAS  Google Scholar 

  5. Barbosa O, Ortiz C, Berenguer-Murcia A, Torres R, Rodrigues RC, Fernandez-Lafuente R (2015) Strategies for the one-step immobilization–purification of enzymes as industrial biocatalysts. Biotechnol Adv 33:435–456

    Article  CAS  Google Scholar 

  6. Feijoo-Siota L, Blasco L, Rodríguez-Rama JL, Barros-Velázquez J, Miguel T, Sánchez-Pérez A, Villa TG (2014) Recent patents on microbial proteases for the dairy industry. Recent Adv DNA Gene Seq 8:44–55

    CAS  Google Scholar 

  7. Mateo C, Palomo JM, Fernandez-Lorente G, Guisan JM, Fernandez-Lafuente R (2007) Improvement of enzyme activity, stability and selectivity via immobilization techniques. Enzyme Microb Technol 40:1451–1463

    Article  CAS  Google Scholar 

  8. Jatavathu M, Jatavathu S, Raghavendra Rao MV, Sambasiva Rao KRS (2011) Efficient leather dehairing by bacterial thermostable protease. Int J Biosci Biotechnol 3:11–26

    Google Scholar 

  9. de Souza FR, Gutterres M (2012) Application of enzymes in leather processing: a comparison between chemicals and ecoenzymatic processes. Braz J Chem Eng 29:473–482

    Article  Google Scholar 

  10. Jaouadi NZ, Jaouadi B, Hlima HB, Rekik H, Belhoul M, Hmidi M, Bejar S (2014) Probing the crucial role of Leu31 and Thr33 of the Bacillus pumilus CBS alkaline protease in substrate recognition and enzymatic depilation of animal hide. PLoS One 9:e108367

    Article  Google Scholar 

  11. Briki S, Hamdi O, Landoulsi A (2016) Enzymatic dehairing of goat skins using alkaline protease from Bacillus sp. SB12. Protein Express Purif 121:9–16

    Article  CAS  Google Scholar 

  12. Cantera C, Garro ML, Goya L, Barbeito C, Galarza B (2004) Hair saving unhairing process: part 6. Stratum corneum as a diffusion barrier: chemical–mechanical injury of epidermis. J Soc Leather Technol Chem 88:21–131

    Google Scholar 

  13. Domsalla A, Melzig MF (2008) Occurrence and properties of proteases in plant lattices. Planta Med 74:699–711

    Article  CAS  Google Scholar 

  14. Torres MJ, Trejo SA, Obregón WD, Avilés FX, López LMI, Natalucci CL (2012) Characterization of the proteolytic system present in Vasconcellea quercifolia latex. Planta 236:1471–1484

    Article  CAS  Google Scholar 

  15. Badgujar SB, Mahajan RT (2014) Nivulian-II a new milk clotting cysteine protease of Euphorbia nivulia latex. Int J Biol Macromol 70:391–398

    Article  CAS  Google Scholar 

  16. Raskovic B, Bozovic O, Prodanovic R, Niketic V, Polovic N (2014) Identification, purification and characterization of a novel collagenolytic serine protease from fig (Ficus carica var. Brown Turkey) latex. J Biosci Bioeng 118:622–627

    Article  CAS  Google Scholar 

  17. Rawdkuen S, Jaimakreu M, Benjakul S (2013) Physicochemical properties and tenderness of meat samples using proteolytic extract from Calotropis procera latex. Food Chem 136:909–916

    Article  CAS  Google Scholar 

  18. Mahajan RT, Chaudhari GM (2014) Plant latex as vegetable source for milk clotting enzymes and their use in cheese preparation. Int J Adv Res 2:1173–1181

    CAS  Google Scholar 

  19. Dubey VK, Jagannadham MV (2003) Procerain, a stable cysteine protease from the latex of Calotropis procera. Phytochemistry 62:1057–1071

    Article  Google Scholar 

  20. Singh AN, Shukla AK, Jagannadham MV, Dubey VK (2010) Purification of a novel cysteine protease, procerain B, from Calotropis procera with distinct characteristics compared to procerain. Process Biochem 45:399–406

    Article  CAS  Google Scholar 

  21. Ramos MV, Araújo ES, Jucá TL, Monteiro-Moreira ACO, Vasconcelos IM, Moreira RA, Viana CA, Beltramini LM, Pereira DA, Moreno FB (2013) New insights into the complex mixture of latex cysteine peptidases in Calotropis procera. J Biol Macromol 58:211–219

    Article  CAS  Google Scholar 

  22. Ramos MV, Souza DP, Gomes MTR, Freitas CDT, Carvalho CPS, Júnior PAVR, Salas CE (2014) A phytopathogenic cysteine peptidase from latex of wild rubber vine Cryptostegia grandiflora. Protein J 33:199–209

    Article  CAS  Google Scholar 

  23. Alencar NM, Oliveira JS, Mesquita RO, Lima MW, Vale MR, Etchells JP, Freitas CD, Ramos MV (2006) Pro- and anti-inflammatory activities of the latex from Calotropis procera (Ait.) R.Br. are triggered by compounds fractionated by dialysis. Inflamm Res 55:559–564

    Article  CAS  Google Scholar 

  24. Freitas CDT, Silva MZR, Bruno-Moreno F, Monteiro-Moreira ACO, Moreira RA, Ramos MV (2015) New constitutive latex osmotin-like proteins lacking antifungal activity. Plant Physiol Biochem 96:45–52

    Article  CAS  Google Scholar 

  25. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  CAS  Google Scholar 

  26. López LMI, Sequeiros C, Natalucci CL, Brullo A, Maras B, Barra D, Caffini NO (2000) Purification and characterization of Macrodontain I, a cysteine peptidase from unripe fruits of Pseudananas macrodontes (Morr.) Harms (Bromeliaceae). Protein Expres Purif 18:133–140

    Article  Google Scholar 

  27. Himelbloom BH, Hassan HM (1985) Optimization of the hide powder azure assay for quantitating the protease of Pseudomonas fluorescens. J Microbiol Meth 4:59–66

    Article  CAS  Google Scholar 

  28. Hamood AN, Griswold J, Colmer J (1996) Characterization of elastase-deficient clinical isolates of Pseudomonas aeruginosa. Infect Immun 64:3154–3160

    CAS  Google Scholar 

  29. Scott JA, Untereiner WA (2004) Determination of keratin degradation by fungi using keratin azure. Med Mycol 42:239–246

    Article  CAS  Google Scholar 

  30. Cantera C, Goya L, Galarza B, Garro ML, López LMI (2003) Hair saving unhairing process. Characterization of enzymatic preparations applied in soaking and unhairing processes. J Soc Leather Technol Chem 87:69–77

    CAS  Google Scholar 

  31. de Freitas CDT, de Souza DP, Araújo ES, Cavalheiro MG, Oliveira LS, Ramos MV (2010) Anti-oxidative and proteolytic activities and protein profile of laticifer cells of Cryptostegia grandiflora, Plumeria rubra and Euphorbia tirucalli. Braz J Plant Physiol 22:11–22

    Article  Google Scholar 

  32. Sivasubramanian S, Manohar BM, Puvanakrishnan R (2008) Mechanism of enzymatic dehairing of skins using a bacterial alkaline protease. Chemosphere 70:1025–1034

    Article  CAS  Google Scholar 

  33. Monod M (2008) Secreted proteases from dermatophytes. Mycopathologia 166:285–294

    Article  Google Scholar 

  34. Cordon TC, Jones HW, Clarke ID, Naghski J (1958) Microbial and other enzymes as depilatory agents. Appl Microbiol 6:293–297

    CAS  Google Scholar 

  35. Dayanandan A, Kanagaraj J, Sounderraj L, Govindaraju R, Rajkumar GS (2003) Application of an alkaline protease in leather processing: an ecofriendly approach. J Clean Prod 11:533–536

    Article  Google Scholar 

  36. Jayakumar R, Jayashree S, Annapurna B, Seshadri S (2012) Characterization of thermostable serine alkaline protease from an alkaliphilic strain Bacillus pumilus MCAS8 and its applications. Appl Biochem Biotechnol 168:1849–1866

    Article  CAS  Google Scholar 

Download references

Acknowledgements

L. M. I. López is a member of CONICET Researcher Career; M. E. Errasti is a member of a CICPBA Researcher Career; M. L. Garro is a professional member of CICPBA. The present work was supported by grants from CONICET (PIP 00297), CICPBA (PIT-AP-BA 2016), CAPES-MINCyT agreement (project 017-2014), and the consortium Molecular Biotechnology of Plant Latex. The authors thank Natalia S. Scelsio for their valuable assistance in developing microscopy techniques.

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Authors and Affiliations

  1. Centro de Investigación y Tecnología del Cuero, Comisión de Investigaciones Científicas de la Provincia de Buenos Aires & INTI-Cueros, Gonnet, Buenos Aires, Argentina

    Laura M. I. Lopéz, María E. Errasti, María L. Garro, José E. Martegani & Germán A. Mazzilli

  2. Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Ceará, Fortaleza, Ceará, Brazil

    Carolina A. Viana, Cléverson D. T. Freitas, Rafaela O. da Silva & Márcio V. Ramos

  3. EMBRAPA Agrotropical, Fortaleza, Ceará, Brazil

    Ídila M. S. Araújo

Authors
  1. Laura M. I. Lopéz
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  2. Carolina A. Viana
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  3. María E. Errasti
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  4. María L. Garro
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  5. José E. Martegani
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  6. Germán A. Mazzilli
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  7. Cléverson D. T. Freitas
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  8. Ídila M. S. Araújo
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  9. Rafaela O. da Silva
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  10. Márcio V. Ramos
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Corresponding authors

Correspondence to Laura M. I. Lopéz or Márcio V. Ramos.

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Lopéz, L., Viana, C., Errasti, M. et al. Latex peptidases of Calotropis procera for dehairing of leather as an alternative to environmentally toxic sodium sulfide treatment. Bioprocess Biosyst Eng 40, 1391–1398 (2017). https://doi.org/10.1007/s00449-017-1796-9

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