A Novel Thermostable Keratinase from Deinococcus geothermalis with Potential Application in Feather Degradation
Source abstract
Abstract
Keratinase can specifically attack disulfide bridges in keratin to convert them from complex to simplified forms. Keratinase thermal stability has drawn attention to various biotechnological industries. In this study, a keratinase DgeKer was identified from a slightly thermophilic species, D. geothermalis. The in silico analysis showed that DgeKer is composed of signal peptide, N-terminal propeptide, mature domain, and C-terminal extension. DgeKer and its C-terminal extension-truncated enzyme (DgeKer-C) were cloned and expressed in E. coli. The purified DgeKer and DgeKer-C showed maximum activity at 70 °C and pH 9–The thermal stability assay (60 °C) showed that the half-life value of DgeKer and DgeKer-C were 103.45 min and 169.10 min, respectively. DgeKer and DgeKer-C were stable at the range of pH from 9 to 11 and showed good tolerance to some metal ions, surfactants and organic solvent. Furthermore, DgeKer could degrade feathers at 70 °C for 60 min. However, the medium became turbid with obvious softening of barbules after being treated with DgeKer-C, which might be due to C-terminal extension. In summary, a thermostable keratinase DgeKer with high efficiency degradation of feathers may have great potential in industry.
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Research context
OpenAlex topics: Enzyme Production and Characterization · Phytase and its Applications · Protease and Inhibitor Mechanisms
SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.
Affiliations
School of Life Science and Engineering, Southwest University of Science and Technology, Mianyang 621000, China
Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China
References (47)
Show source-provided references
1. Biodegradation of keratin waste: Theory and practical aspects10.1016/j.wasman.2011.03.024 ↗
2. Using a keratinase to degrade chicken feathers for improved extraction of glucocorticoids10.1016/j.ygcen.2018.10.002 ↗
3. Biochemical characterisation and application of keratinase from Bacillus thuringiensis MT1 to enable valorisation of hair wastes through biosynthesis of vitamin B-complex10.1016/j.ijbiomac.2020.03.032 ↗
4. Coupled action of γ-glutamyl transpeptidase-glutathione and keratinase effectively degrades feather keratin and surrogate prion protein, Sup 35NM10.1016/j.biortech.2012.06.038 ↗
5. In vitro degradation of β-amyloid fibrils by microbial keratinase10.1016/j.trci.2019.03.003 ↗
6. Wool keratin film plasticized by citric acid for food packaging10.1016/j.fpsl.2017.04.004 ↗
7. Progress in Microbial Degradation of Feather Waste10.3389/fmicb.2019.02717 ↗
8. A Review: Biodegradation and Applications of Keratin Degrading Microorganisms and Keratinolytic Enzymes, Focusing on Thermophiles and Thermostable Serine Proteases10.3844/ajassp.2017.1016.1023 ↗
9. The extremophile Anoxybacillus sp. PC2 isolated from Brazilian semiarid region (Caatinga) produces a thermostable keratinase10.1002/jobm.202000186 ↗
10. Subtilisin-like serine protease from hyperthermophilic archaeon Thermococcus kodakaraensis with N- and C-terminal propeptides10.1093/protein/gzp092 ↗
11. The discovery of novel heat-stable keratinases from Meiothermus taiwanensis WR-220 and other extremophiles
12. Extremozymes—biocatalysts with unique properties from extremophilic microorganisms10.1016/j.copbio.2014.04.003 ↗
13. Rapid production of extracellular thermostable alkaline halophilic protease originating from an extreme haloarchaeon, Halobacterium salinarum by recombinant Bacillus subtilis10.1016/j.bcab.2018.06.017 ↗
14. Deinococcus geothermalis sp. nov. and Deinococcus murrayi sp. nov., two extremely radiation-resistant and slightly thermophilic species from hot springs
15. A novel alkaline protease from alkaliphilic Idiomarina sp. C9-1 with potential application for eco-friendly enzymatic dehairing in the leather industry10.1038/s41598-018-34416-5 ↗
16. Efficient keratinase expression via promoter engineering strategies for degradation of feather wastes10.1016/j.enzmictec.2020.109550 ↗
17. Construction of a Rapid Feather-Degrading Bacterium by Overexpression of a Highly Efficient Alkaline Keratinase in Its Parent StrainBacillus amyloliquefaciensK10.1021/acs.jafc.5b04747 ↗
18. Improving the catalytic performance of Proteinase K from Parengyodontium album for use in feather degradation10.1016/j.ijbiomac.2019.11.043 ↗
19. Marine extremophiles: A source of hydrolases for biotechnological applications10.3390/md13041925 ↗
20. Nocardiopsis species as potential sources of diverse and novel extracellular enzymes10.1007/s00253-014-6111-y ↗
21. Efficient Keratinolysis of Poultry Feather Waste by the Halotolerant Keratinase from Salicola Marasensis
22. Extracellular peptidases from Deinococcus radiodurans10.1007/s00792-015-0773-y ↗
23. Pietrow, O., Panek, A., and Synowiecki, J. (2013). Extracellular proteolytic activity of Deinococcus geothermalis. Afr. J. Biotechnol., 12.
24. Zhang, R.-Y., Huang, Y., Qin, W.-J., and Quan, Z.-X. (2020). The complete genome of extracellular protease-producing Deinococcus sp. D7000 isolated from the hadal region of Mariana Trench Challenger Deep. Mar. Genom., 100832.10.1016/j.margen.2020.100832 ↗
25. The flexibility of UV-inducible mutation in Deinococcus ficus as evidenced by the existence of the imuB–dnaE2 gene cassette and generation of superior feather degrading bacteria10.1016/j.micres.2011.02.008 ↗
26. A 38 kDa precursor protein of aqualysin I (a thermophilic subtilisin-type protease) with a C-terminal extended sequence: Its purification and in vitro processing10.1111/j.1365-2958.1996.tb02625.x ↗
27. Thermostable Amylosucrase fromCalidithermus timidusDSM 17022: Insight into Its Characteristics and Tetrameric Conformation10.1021/acs.jafc.9b04023 ↗
28. Characterization of a cloned subtilisin-like serine proteinase from a psychrotrophic Vibrio species10.1046/j.1432-1033.2002.03259.x ↗
29. Gopinath, S.C., Anbu, P., Lakshmipriya, T., Tang, T.-H., Chen, Y., Hashim, U., Ruslinda, A.R., and Arshad, M. (2015). Biotechnological aspects and perspective of microbial keratinase production. BioMed Res. Int., 2015.10.1155/2015/140726 ↗
30. Cloning, heterologous expression and characterization of two keratinases from Stenotrophomonas maltophilia BBE1110.1016/j.procbio.2014.01.009 ↗
31. Shinde, U., and Thomas, G. (2011). Insights from Bacterial Subtilases into the Mechanisms of Intramolecular Chaperone-Mediated Activation of Furin. Proprotein Convertases, Springer.10.1007/978-1-61779-204-5_4 ↗
32. Combining Pro-peptide Engineering and Multisite Saturation Mutagenesis to Improve the Catalytic Potential of Keratinase10.1021/acssynbio.8b00442 ↗
33. Crystal Structure of a Subtilisin Homologue, Tk-SP, from Thermococcus kodakaraensis: Requirement of a C-terminal β-Jelly Roll Domain for Hyperstability10.1016/j.jmb.2010.05.064 ↗
34. Molecular analysis of the gene encoding a cold-adapted halophilic subtilase from deep-sea psychrotolerant bacterium Pseudoalteromonas sp. SM9913: Cloning, expression, characterization and function analysis of the C-terminal PPC domains10.1007/s00792-009-0263-1 ↗
35. A novel type of subtilase from the psychrotolerant bacterium Pseudoalteromonas sp. SM9913: Catalytic and structural properties of deseasin MCP10.1099/mic.0.2007/006056-0 ↗
36. Faculty of 1000 evaluation for New knowledge from old: In silico discovery of novel protein domains in Streptomyces coelicolor10.1186/1471-2180-3-1 ↗
37. Preliminary characterization of extracellular proteolytic enzymes of dermatophytes by chromogenic substrates10.1080/02681218880000261 ↗
38. Biochemical features of microbial keratinases and their production and applications10.1007/s00253-009-2398-5 ↗
39. Keratinase production and keratin degradation by a mutant strain of Bacillus subtilis10.1631/jzus.B061620 ↗
40. Medium optimization for a novel 58kDa dimeric keratinase from Bacillus licheniformis ER-15: Biochemical characterization and application in feather degradation and dehairing of hides10.1016/j.biortech.2010.02.090 ↗
41. Keratinolytic activity of Bacillus megaterium F7-1, a feather-degrading mesophilic bacterium10.1016/j.micres.2007.02.004 ↗
42. A new cold-adapted serine peptidase from Antarctic Lysobacter sp. A03: Insights about enzyme activity at low temperatures10.1016/j.ijbiomac.2017.05.142 ↗
43. Novel serine keratinase from Caldicoprobacter algeriensis exhibiting outstanding hide dehairing abilities10.1016/j.ijbiomac.2016.01.074 ↗
44. Biotransformation of keratin waste to amino acids and active peptides based on cell-free catalysis10.1186/s13068-020-01700-4 ↗
45. Extraction of keratin from waste chicken feathers using sodium sulfide and L-cysteine10.1016/j.procbio.2019.04.010 ↗
46. Revisiting microbial keratinases: Next generation proteases for sustainable biotechnology10.3109/07388551.2012.685051 ↗
47. Functional domains of a zinc metalloprotease from Vibrio vulnificus10.1128/jb.179.23.7606-7609.1997 ↗