Abstract
Feather biodegradation is an important premise for efficient resource development and utilization, in which keratinase plays an important role. However, there are few keratinases that combine the high activity, thermal stability, and organic solvent tolerance required for industrialization. This paper reported an efficient feather-degrading Pseudomonas aeruginosa 4–3 isolated from slaughterhouses. After 48 h of fermentation by P. aeruginosa 4–3 in a feather medium at 40 °C, pH 8.0, keratinase was efficiently produced (295.28 ± 5.42 U/mL) with complete feather degradation (95.3 ± 1.5%). Moreover, the keratinase from P. aeruginosa 4–3 showed high optimal temperature (55 °C), good thermal stability, wide pH tolerance, and excellent organic solvent resistance. In addition, P. aeruginosa 4–3–derived aminopeptidases also exhibit excellent thermal stability and organic solvent tolerance. Encouragingly, the reaction of crude keratinase and aminopeptidase with feathers for 8 h resulted in a 78% degradation rate of feathers. These properties make P. aeruginosa 4–3 keratinase and aminopeptidase ideal proteases for potential applications in keratin degradation, as well as provide ideas for the synergistic degradation of keratin by multiple enzymes.
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References
Abdel-Fattah AM, El-Gamal MS, Ismail SA, Emran MA, Hashem AM (2018) Biodegradation of feather waste by keratinase produced from newly isolated Bacillus licheniformis ALW1. J Genet Eng Biotechnol 16:311–318. https://doi.org/10.1016/j.jgeb.2018.05.005
Cahan R, Axelrad I, Safrin M, Ohman DE, Kessler E (2001) A secreted aminopeptidase of Pseudomonas aeruginosa. Identification, primary structure, and relationship to other aminopeptidases. J Biol Chem 276:43645–43652. https://doi.org/10.1074/jbc.M106950200
Chenna R, Sugawara H, Koike T, Lopez R, Gibson TJ, Higgins DG, Thompson JD (2003) Multiple sequence alignment with the Clustal series of programs. Nucleic Acids Res 31:3497–3500. https://doi.org/10.1093/nar/gkg500
da Silva RR (2018) Keratinases as an alternative method designed to solve keratin disposal on the environment: its relevance on agricultural and environmental chemistry. J Agric Food Chem 66:7219–7221. https://doi.org/10.1021/acs.jafc.8b03152
Daroit DJ, Brandelli A (2014) A current assessment on the production of bacterial keratinases. Crit Rev Biotechnol 34:372–384. https://doi.org/10.3109/07388551.2013.794768
Gegeckas A, Simkute A, Gudiukaite R, Citavicius DJ (2018) Characterization and application of keratinolytic paptidases from Bacillus spp. Int J Biol Macromol 113:1206–1213. https://doi.org/10.1016/j.ijbiomac.2018.03.046
Gong JS, Wang Y, Zhang DD, Zhang RX, Su C, Li H, Zhang XM, Xu ZH, Shi JS (2015) Biochemical characterization of an extreme alkaline and surfactant-stable keratinase derived from a newly isolated actinomycete Streptomyces aureofaciens K13. RSC Adv 5:24691–24699. https://doi.org/10.1039/c4ra16423g
Gulmez C, Atakisi O, Dalginli KY, Atakisi E (2018) A novel detergent additive: organic solvent- and thermo-alkaline-stable recombinant subtilisin. Int J Biol Macromol 108:436–443. https://doi.org/10.1016/j.ijbiomac.2017.11.133
Habbeche A, Saoudi B, Jaouadi B, Haberra S, Kerouaz B, Boudelaa M, Badis A, Ladjama A (2014) Purification and biochemical characterization of a detergent-stable keratinase from a newly thermophilic actinomycete Actinomadura keratinilytica strain Cpt29 isolated from poultry compost. J Biosci Bioeng 117:413–421. https://doi.org/10.1016/j.jbiosc.2013.09.006
Hassan MA, Taha TH, Hamad GM, Hashem M, Alamri S, Mostafa YS (2020) Biochemical characterisation and application of keratinase from Bacillus thuringiensis MT1 to enable valorisation of hair wastes through biosynthesis of vitamin B-complex. Int J Biol Macromol 153:561–572. https://doi.org/10.1016/j.ijbiomac.2020.03.032
Jaouadi NZ, Rekik H, Badis A, Trabelsi S, Belhoul M, Yahiaoui AB, Ben Aicha H, Toumi A, Bejar S, Jaouadi B (2013) Biochemical and molecular characterization of a serine keratinase from Brevibacillus brevis US575 with promising keratin-biodegradation and hide-dehairing activities. PLoS ONE 8:e76722. https://doi.org/10.1371/journal.pone.0076722
Kecha M, Benallaoua S, Touzel JP, Bonaly R, Duchiron F (2007) Biochemical and phylogenetic characterization of a novel terrestrial hyperthermophilic archaeon pertaining to the genus Pyrococcus from an Algerian hydrothermal hot spring. Extremophiles 11:65–73. https://doi.org/10.1007/s00792-006-0010-9
Kuo JM, Yang JI, Chen WM, Pan MH, Tsai ML, Lai YJ, Hwang A, Pan BS, Lin CY (2012) Purification and characterization of a thermostable keratinase from Meiothermus sp 140. Int Biodeterior Biodegrad 70:111–116. https://doi.org/10.1016/j.ibiod.2012.02.006
Laane C, Boeren S, Vos K, Veeger C (1987) Rules for optimization of biocatalysis in organic solvents. Biotechnol Bioeng 30:81–87. https://doi.org/10.1002/bit.260300112
Lange L, Huang Y, Busk PK (2016) Microbial decomposition of keratin in nature-a new hypothesis of industrial relevance. Appl Microbiol Biotechnol 100:2083–2096. https://doi.org/10.1007/s00253-015-7262-1
Li S, He B, Bai Z, Ouyang P (2009) A novel organic solvent-stable alkaline protease from organic solvent–tolerant Bacillus licheniformis YP1A. J Mol Catal B-Enzym 56:85–88. https://doi.org/10.1016/j.molcatb.2008.08.001
Lin X, Lee CG, Casale ES, Shih JC (1992) Purification and characterization of a keratinase from a feather-degrading Bacillus licheniformis strain. Appl Environ Microbiol 58:3271–3275. https://doi.org/10.1128/aem.58.10.3271-3275.1992
Liu JZ, Wu SP, Lu Y, Liu Q, Jiao QC, Wang XZ, Zhang HJ (2016) An integrated electrodialysis-biocatalysis-spray-drying process for efficient recycling of keratin acid hydrolysis industrial wastewater. Chem Eng J 302:146–154. https://doi.org/10.1016/j.cej.2016.05.046
Mabrouk MEM (2008) Feather degradation by a new keratinolytic Streptomyces sp MS-2. World J Microbiol Biotechnol 24:2331–2338. https://doi.org/10.1007/s11274-008-9748-9
Nandan A, Nampoothiri KM (2020) Therapeutic and biotechnological applications of substrate specific microbial aminopeptidases. Appl Microbiol Biotechnol 104:5243–5257. https://doi.org/10.1007/s00253-020-10641-9
Peng Z, Mao X, Zhang J, Du G, Chen J (2019) Effective biodegradation of chicken feather waste by co-cultivation of keratinase producing strains. Microb Cell Fact 18:84. https://doi.org/10.1186/s12934-019-1134-9
Peng Z, Xu P, Song Y, Du GC, Zhang J, Chen J (2021) Cysteine-mediated cyclic metabolism drives the microbial degradation of keratin. ACS Sustain Chem Eng 9:9861–9870. https://doi.org/10.1021/acssuschemeng.1c02627
Pillai P, Mandge S, Archana G (2011) Statistical optimization of production and tannery applications of a keratinolytic serine protease from Bacillus subtilis P13. Process Biochem 46:1110–1117. https://doi.org/10.1016/j.procbio.2011.01.030
Preczeski KP, Dalastra C, Czapela FF, Kubeneck S, Scapini T, Camargo AF, Zanivan J, Bonatto C, Stefanski FS, Venturin B, Fongaro G, and Treichel H (2020) Fusarium oxysporum and Aspergillus sp. as keratinase producers using swine hair from agroindustrial residues. Frontiers in Bioengineering and Biotechnology 8.
Rahman RNZRA, Geok LP, Basri M, Salleh AB (2005) An organic solvent-tolerant protease from Pseudomonas aeruginosa strain K. Enzyme Microb Technol 36:749–757. https://doi.org/10.1016/j.enzmictec.2004.12.022
Rai SK, Konwarh R, Mukherjee AK (2009) Purification, characterization and biotechnological application of an alkaline beta-keratinase produced by Bacillus subtilis RM-01 in solid-state fermentation using chicken-feather as substrate. Biochem Eng J 45:218–225. https://doi.org/10.1016/j.bej.2009.04.001
Reddy MR, Reddy KS, Chouhan YR, Bee H, Reddy G (2017) Effective feather degradation and keratinase production by Bacillus pumilus GRK for its application as bio-detergent additive. Bioresour Technol 243:254–263. https://doi.org/10.1016/j.biortech.2017.06.067
Santos SF, Zanette D, Fischer H, Itri R (2003) A systematic study of bovine serum albumin (BSA) and sodium dodecyl sulfate (SDS) interactions by surface tension and small angle X-ray scattering. J Colloid Interface Sci 262:400–408. https://doi.org/10.1016/S0021-9797(03)00109-7
Srivastava B, Khatri M, Singh G, Arya SK (2020) Microbial keratinases: an overview of biochemical characterization and its eco-friendly approach for industrial applications. J Clean Prod 252:119847. https://doi.org/10.1016/j.jclepro.2019.119847
Srivastava B, Singh H, Khatri M, Singh G, Arya SK (2020b) Immobilization of keratinase on chitosan grafted-beta-cyclodextrin for the improvement of the enzyme properties and application of free keratinase in the textile industry. Int J Biol Macromol 165:1099–1110. https://doi.org/10.1016/j.ijbiomac.2020.10.009
Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Mol Biol Evol 30:2725–2729. https://doi.org/10.1093/molbev/mst197
Verma A, Singh H, Anwar S, Chattopadhyay A, Tiwari KK, Kaur S, Dhilon GS (2017) Microbial keratinases: industrial enzymes with waste management potential. Crit Rev Biotechnol 37:476–491. https://doi.org/10.1080/07388551.2016.1185388
Wu YT, Zhou ND, Zhou ZM, Gao XX, Tian YP (2014) A thermo-stable lysine aminopeptidase from Pseudomonas aeruginosa: isolation, purification, characterization, and sequence analysis. J Basic Microbiol 54:1110–1119. https://doi.org/10.1002/jobm.201300752
Yahaya RSR, Normi YM, Phang LY, Ahmad SA, Abdullah JO, Sabri S (2021) Molecular strategies to increase keratinase production in heterologous expression systems for industrial applications. Appl Microbiol Biotechnol 105:3955–3969
Yahaya RSR, Phang LY, Normi YM, Abdullah JO, Ahmad SA, and Sabri S (2022) Feather-degrading Bacillus cereus HD1: genomic analysis and its optimization for keratinase production and feather degradation. Current Microbiology 79.
Yamamura S, Morita Y, Hasan Q, Yokoyama K, Tamiya E (2002) Keratin degradation: a cooperative action of two enzymes from Stenotrophomonas sp. Biochem Biophys Res Commun 294:1138–1143. https://doi.org/10.1016/S0006-291X(02)00580-6
Yusuf I, Ahmad SA, Phang LY, Syed MA, Shamaan NA, Abdul Khalil K, Dahalan FA, Shukor MY (2016) Keratinase production and biodegradation of polluted secondary chicken feather wastes by a newly isolated multi heavy metal tolerant bacterium-Alcaligenes sp. AQ05-001. J Environ Manag 183:182–195. https://doi.org/10.1016/j.jenvman.2016.08.059
Zadymova NM, Yampol’skaya GP, Filatova LY (2006) Interaction of bovine serum albumin with nonionic surfactant Tween 80 in aqueous solutions: complexation and association. Colloid J 68:162–172. https://doi.org/10.1134/S1061933x06020074
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Conceptualization: Xiao-dong Pei and Cheng-Hua Wang. Methodology: Xiao-dong Pei, Fan Li, Shi-Yang Yue, and Cheng-Hua Wang. Formal analysis and investigation: Xiao-Ni Huang, Tian-Tian Gao, Dao-Quan Jiao. Writing—original draft preparation: Xiao-dong Pei, Fan Li and, Cheng-Hua Wang. Writing—review and editing: Shi-Yang Yue, Xiao-Ni Huang, Tian-Tian Gao, and Cheng-Hua Wang.
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Pei, XD., Li, F., Yue, SY. et al. Production and characterization of novel thermo- and organic solvent–stable keratinase and aminopeptidase from Pseudomonas aeruginosa 4–3 for effective poultry feather degradation. Environ Sci Pollut Res 30, 2480–2493 (2023). https://doi.org/10.1007/s11356-022-22367-4
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DOI: https://doi.org/10.1007/s11356-022-22367-4