Isolation and optimization of keratinase production by Bacillus licheniformis, which produces keratinase enzyme, to break down feathers
Subject Areas : microbiologyParisa Vanaki 1 , Fatemeh Zaboli 2 * , Hami Kaboosi 3
1 - Department of Microbiology, Ayatollah Amoli Branch, Islamic Azad University, Amol, Iran
2 - Department of Microbiology, Ayatollah Amoli Branch, Islamic Azad University, Amol, Iran
3 - Department of Microbiology, Ayatollah Amoli Branch, Islamic Azad University, Amol, Iran
Keywords: Bacillus licheniformis, Optimization, Keratinase, Animal and Poultry Feed.,
Abstract :
Objective: The Keratinase enzyme produced by Bacillus is used to break down raw keratin and produce animal and poultry feed. The purpose of this study is to investigate the optimization of keratinase-producing bacillus to decompose and provide a protein source for animal feed. Materials and methods: This experimental-laboratory study was conducted in 1401 on chicken slaughterhouse samples. Heat treatment, alcohol treatment, and polymerase chain reaction were used to isolate Bacillus licheniformis (B. licheniformis) strains. To optimally check keratinase activity, keratinase was precipitated by ammonium sulfate method. Comparison of the mean changes and standard deviation of keratinase before and after optimization was done with Tukey's one-way analysis of variance and post hoc test and data analysis was done with SPSS-26 (p<0.05). Findings: (85.57 ± 0.64 units/ml) keratinase activity of B. licheniformis strain PVKR15 was higher than other strains. The total keratinase activity after optimization was 800 units/ml and before optimization was 500 units/ml. pH, temperature, heating time, creatine source, and creatine concentration to optimize conditions respectively; 11, 37 degrees Celsius, 72 hours, and azocreatine at a concentration of 10 g/l. There was a significant difference between the keratinase activity of B. licheniformis PVKR15 with pH, temperature, creatine source, creatine concentration, and incubation time (p<0.05). Conclusion: B. licheniformis PVKR15 has a useful and effective role in the growth of animals to break down keratin as a probiotic supplement for livestock and poultry feed
1. Song P, Zhang X, Wang S, Xu W, Wang F, Fu R & et al. Microbial proteases and their applications. Front microbiol. 2023; 14: 1236368.
2. Kotb E, Alabdalall AH, Alsayed MA, Alghamdi AI, Alkhaldi E, AbdulAzeez S, & et al. Isolation, screening, and identification of alkaline protease-producing bacteria and application of the most potent enzyme from Bacillus sp. Mar64. Fermentation. 2023; 9(7): 637.
3. Rahimnahal S, Shams M, Tarrahimofrad H & Mohammadi Y. Analysis to describe the catalytic critical residue of keratinase mojavensis using peptidase inhibitors: A docking-based bioinformatics study. J Basic Res Med Sci. 2020; 7(2): 13-28.
4. Wang Z, Chen Y, Yan M, Li K, Okoye CO, Fang Z, & et al. Research progress on the degradation mechanism and modification of keratinase. Applied Microbiology and Biotechnology. 2023; 107(4): 1003-17.
5. Deivasigamani B & Alagappan K M. Industrial application of keratinase and soluble proteins from feather keratins. J environ biol. 2008; 29(6): 933-936
6. Wang Z, Chen Y, Yan M, Li K, Okoye CO, Fang Z, & et al. Research progress on the degradation mechanism and modification of keratinase. Appl Microbiol Biotechnol. 2023;107(4):1003-17.
7. Abd El Nasser NH, Aly AM, Hanafy SS & Motawe FH. Keratinase production from chicken feather by new Egyptian local Streptomyces Isolates. Egypt J Microbiol. 2024; 58(1): 25-33.
8. Aktayeva S, Baltin K, Kiribayeva A, Akishev Z, Silayev D, Ramankulov Y, & et al. Isolation of Bacillus sp. A5. 3 strain with keratinolytic activity. Biology. 2022; 11(2): 244.
9. Li Q. Progress in microbial degradation of feather waste. Front Microbiol. 2019; 10 (2019): 2717.
10. Mussakhmetov A, Kiribayeva A, Daniyarov A, Bulashev A, Kairov U & Khassenov B. Genome sequence and assembly of the amylolytic Bacillus licheniformis T5 strain isolated from Kazakhstan soil. BMC genomic data. 2024; 25(1): 3.
11. Moridshahi R, Bahreini M, Sharifmoghadam MM, Asoodeh A & Korouzhdehi B. Optimization of keratinase production by a native isolate, Bacillus sp FUM120, using one factor at a time methodology. CMR. 2020; 33(4): 542-554.
12. Vanaki P, Zaboli F, Kaboosi H, Amoli RI & Savadkoohi F. Isolation and identification of keratinolytic probiotic Bucillus licheniformis bacteria from the soil below poultry slaughterhouse waste. Braz J Biol. 2022; 84: e257473.
13. Forbes BA, Sahm DF & Weissfeld AS. Non-branching, catalasepositive, gram-positive bacilli, Bacillus and similar organisms. In: Forbes BA, Sahn DF, Weissfeld AS, editors. Bailey & Scott’s Diagnostic Microbiology. 12th ed. St. Louis, MO: Mosby Elsevier; 2007: p.252, 283-285.
14. Shahbani Zahiri H, Goudarzi Z, Chamani M & Akbari Noghabi K. Isolation and characterization of a PHA-producing bacterial strain: biopolymer production under different growth conditions . NCMBJ. 2011; 1 (3): 23-32.
15. Ashengroph M & Sahami Soltani M. Antimicrobial effects of extracellular copper sulfide nanoparticles synthesized from Bacillus licheniformis. JMW. 2018; 11(3 (36)): 243-257.
16. Ghasemi Y. Shahbazi M. Rasoul-Amini S, Kargar M. Safari A, Kazemi A, & et al. Identification and Characterization of feather – degrading bacteria from keratin-rich wastes. Ann Microbiol. 2012; 62 (2012): 737-744.
17. Mohamed AH, Bakry M, Haroun A, Amara A & Ehab AS. Production and characterization of keratinolytic protease from new wool-degrading Bacillus epecies isolated from egyptian ecosystem. Biomed Res Int. 2013; 2013 (2013): 175012.
18. Mukesh Kumar DJ, Priya P, Nithya Balasundari S, Nandhini Devi GSD, Immaculate Nancy Rebecca A & Kalaichelvan PT. Middle East J Sci Res. 2012; 11(7): 900-907.
19. Mousavi S, Salouti M, Shapoury R & Heidari Z. Optimization of keratinase production for feather degradation by Bacillus subtilis. Jundishapour J Microbiol. 2013: 6(8): e7160.
20. Ahmad Pour F. Yakhchali B & Sadat Musavi, M. Isolation and Identification of a keratinolytic Bacillus cereus and optimization of Keratinase production. JABR. 2016; 3(4): 507-512.
21. Sivakumar T, Shankar T, Vijayabaskar P & Ramasubramanian V. Optimization for keratinase enzyme production using Bacillus thuringiensis TS2. J Plant Sci. 2012; 5(3): 102-109.
22. Surti A & Taral M. Study of optimization, characterization and applications of keratinase produced by a Bacillus strain. JABS. 2024; 18(2): 143-61.
23. Alamnie G, Gessesse A & Andualem B. Production of surfactant-stable keratinolytic protease from B. subtilis ES5 and its application as a detergent additive. Biocatal Agric Biotechnol. 2023; 50 (2023): 102750.
24. Nnolim NE & Nwodo UU. Bacillus sp. CSK2 produced thermostable alkaline keratinase using agro-wastes: keratinolytic enzyme characterization. BMC Biotechnol. 2020; 20(1): 65.