Hydrolysis of Gelatin from Animal Hoof Using Bacterial Gelatinase

Miraf Mekonnen; Selfu Girma; Abay Atnafu

doi:doi:10.11648/j.ijmb.20220703.14

| Peer-Reviewed

Hydrolysis of Gelatin from Animal Hoof Using Bacterial Gelatinase

Miraf Mekonnen, Selfu Girma, Abay Atnafu

Published in International Journal of Microbiology and Biotechnology (Volume 7, Issue 3)

Received: 9 August 2022 Accepted: 30 August 2022 Published: 14 September 2022

Views: Downloads:

Download PDF

Share This Article

Twitter
Linked In
Facebook

Abstract

Enzymatic hydrolysis of gelatin increases its functional, textural, and nutritional characteristics. Gelatinases are enzymes used to hydrolyze gelatin into peptides and amino acids. In recent years gelatin hydrolysates have gained a considerable indication in the applications of food and non-food products. This study aimed to isolate the gelatinase enzyme from bacteria and optimize the reaction condition for gelatin hydrolysis. A total of 100 bacterial isolates were isolated from soil and water samples. All isolates were screened for the production of gelatinase on gelatin-containing media. One isolate was selected for further analysis based on the eminent properties of the enzymatic reaction. The organism can grow under solid-state fermentation and produce gelatinase enzyme with a high activity using wheat bran as a carbon source. Gelatinase was optimally active at a temperature of 50°C and at pH 8. The optimum enzyme production under solid-state fermentation includes an incubation period of 120 h and a moisture content of 66.7%. The isolate produced more enzymes with the addition of peptone as a nitrogen source. The enzyme was used to produce gelatin hydrolysate by hydrolyzing animal hooves. The production process for the gelatin hydrolysis was optimized by varying enzyme concentration and incubation time. The hydrolysis has maximum activity at 4 h incubation period and at a high amount of enzyme concentration. This study indicated that animal waste like hoofs can be converted to useful gelatin hydrolysate using microbial gelatinase and used for various applications.

Published in	International Journal of Microbiology and Biotechnology (Volume 7, Issue 3)
DOI	10.11648/j.ijmb.20220703.14
Page(s)	135-142
Creative Commons	This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.
Copyright	Copyright © The Author(s), 2024. Published by Science Publishing Group

Keywords

Gelatinase, Hooves, Hydrolysis, Enzymes

References

[1]	Mariod AA, Fadul H. Gelatin, source, extraction and industrial applications. Acta Scientiarum Polonorum Technologia Alimentaria. 2013; 12 (2): 135-47.
[2]	Shah H, Yusof F. Gelatin as an ingredient in food and pharmaceutical products: An islamic perspective. Advances in Environmental Biology. 2014 01/01; 8: 774-80.
[3]	Kim S-K, Mendis E. Bioactive compounds from marine processing byproducts - A review. Food Research International. 2006 05/01; 39: 383-93.
[4]	Cavello I, Hours R, Cavalitto S. Enzymatic hydrolysis of gelatin layers of X-Ray films and release of silver particles using keratinolytic serine proteases from Purpureocillium lilacinum LPS # 876. Journal of microbiology and biotechnology. 2013 06/03; 23.
[5]	Qi W, He Z. Enzymatic hydrolysis of protein: Mechanism and kinetic model. Frontiers of Chemistry in China. 2006; 1 (3): 308-14.
[6]	Liu L, Ma M, Cai Z, Yang X, Wang W. Purification and properties of a collagenolytic protease produced by Bacillus cereus MBL13 strain. Food technology and biotechnology. 2010; 48 (2): 151.
[7]	Balan SS, Nethaji R, Sankar S, Jayalakshmi S. Production of gelatinase enzyme from Bacillus spp isolated from the sediment sample of Porto Novo Coastal sites. Asian Pacific Journal of Tropical Biomedicine. 2012; 2 (3): S1811-S6.
[8]	Whaley DN, Dowell VR, Jr., Wanderlinder LM, Lombard GL. Gelatin agar medium for detecting gelatinase production by anaerobic bacteria. J Clin Microbiol. 1982 Aug; 16 (2): 224-9. PubMed PMID: 6288762. Pubmed Central PMCID: PMC272334. Epub 1982/08/01. eng.
[9]	Soemarie YB, Milanda T, Barliana MI. Isolation, Characterization, and Identification Candidate of Probiotic Bacteria Isolated from Wadi Papuyu (Anabas testudineus Bloch.) a Fermented Fish Product from Central Kalimantan, Indonesia. International Journal of Food Science. 2022 2022/05/05; 2022: 4241531.
[10]	Prihanto AA, Firdaus M. Proteolytic and fibrinolytic activities of halophilic lactic acid bacteria from two Indonesian fermented foods. Journal of Microbiology, Biotechnology and Food Sciences. 2013; 2 (5): 2291-3.
[11]	Subramaniyam R, Vimala R. Solid state and submerged fermentation for the production of bioactive substances: a comparative study. Int J Sci Nat. 2012; 3 (3): 480-6.
[12]	Graminha E, Gonçalves A, Pirota R, Balsalobre M, Da Silva R, Gomes E. Enzyme production by solid-state fermentation: Application to animal nutrition. Animal Feed Science and Technology. 2008; 144 (1-2): 1-22.
[13]	Pandey A, Selvakumar P, Soccol CR, Nigam P. Solid state fermentation for the production of industrial enzymes. Current science. 1999: 149-62.
[14]	Gessesse AG, BA. Production of alkalineproteasebyalkaliphilicbacteriaisolatedfromanalkalinesodalake. Biotechnology letters. 1997.
[15]	Rosen H. A Modified Ninhydrin Colorimetric Analysis for Amino Acids. Archives of Biochemistry and Biophysisc. 1957; 67: 10-5. eng.
[16]	Mortezaei M, Dadmehr M, Korouzhdehi B, Hakimi M, Ramshini H. Colorimetric and label free detection of gelatinase positive bacteria and gelatinase activity based on aggregation and dissolution of gold nanoparticles. Journal of Microbiological Methods. 2021; 191: 106349.
[17]	Stack MS, Gray RD. The effect of pH, temperature, and D2O on the activity of porcine synovial collagenase and gelatinase. Archives of biochemistry and biophysics. 1990; 281 (2): 257-63.
[18]	Oliveira ANd, Oliveira LAd, Andrade JS. Production and some properties of crude alkaline proteases of indigenous Central Amazonian rhizobia strains. Brazilian Archives of Biology and Technology. 2010; 53: 1185-95.
[19]	Joseph J, Sasidharan H, Chithira O. Isolation, production and characterisation of novel gelatinase enzyme from Bacillus spp. Int J Sci Res in Biological Sciences Vol. 2018; 5: 6.
[20]	Singh B, Kaur R, Singh K. Characterization of Rhizobium strain isolated from the roots of Trigonella foenumgraecum (fenugreek). African Journal of Biotechnology. 2008; 7 (20).
[21]	Kachlishvili E, Penninckx MJ, Tsiklauri N, Elisashvili V. Effect of nitrogen source on lignocellulolytic enzyme production by white-rot basidiomycetes under solid-state cultivation. World Journal of Microbiology and Biotechnology. 2006; 22 (4): 391-7.
[22]	Kumar SL, Anandhavelu S, Swathy M. Preparation and Characterization of Goat Hoof Keratin/Gelatin/Sodium Alginate Base Biofilm for Tissue Engineering Application. Integrated Ferroelectrics. 2019 2019/10/13; 202 (1): 1-12.
[23]	Alipal J, Pu'ad NASM, Lee TC, Nayan NHM, Sahari NB, Basri H, et al. A review of gelatin: Properties, sources, process, applications, and commercialisation. Materials Today: Proceedings. 2021; 42: 240-50.
[24]	dela Cruz TEE, Torres JMO. Gelatin hydrolysis test protocol. Am Soc Microbiol. 2012.
[25]	Sai-Ut S, Benjakul S, Sumpavapol P. Gelatinolytic enzymes from Bacillus amyloliquefaciens isolated from fish docks: Characteristics and hydrolytic activity. Food science and biotechnology. 2013 08/01; 22: 1015-21.
[26]	Nagano H, To KA. Purification of Collagenase and Specificity of Its Related Enzyme from Bacillus subtilis FS-2. Bioscience, Biotechnology, and Biochemistry. 2000; 64 (1): 181-3.
[27]	Petrova D, Derekova A, Vlahov S. Purification and properties of individual collagenases fromStreptomyces sp. strain 3B. Folia microbiologica. 2006; 51 (2): 93-8.
[28]	Joo H-S, Chang C-S. Production of protease from a new alkalophilic Bacillus sp. I-312 grown on soybean meal: optimization and some properties. Process Biochemistry. 2005; 40 (3-4): 1263-70.
[29]	Pandey A. Solid-state fermentation. Biochemical Engineering Journal. 2003 2003/03/01/; 13 (2): 81-4.
[30]	Gupta R, Beg QK, Khan S, Chauhan B. An overview on fermentation, downstream processing and properties of microbial alkaline proteases. Applied microbiology and biotechnology. 2002 Dec; 60 (4): 381-95. PubMed PMID: 12466877. Epub 2002/12/06. eng.
[31]	Nehete JY, Bhambar RS, Narkhede MR, Gawali SR. Natural proteins: Sources, isolation, characterization and applications. Pharmacognosy reviews. 2013 Jul; 7 (14): 107-16. PubMed PMID: 24347918. Pubmed Central PMCID: PMC3841988. Epub 2013/12/19. eng.
[32]	Khantaphant S, Benjakul S. Comparative study on the proteases from fish pyloric caeca and the use for production of gelatin hydrolysate with antioxidative activity. Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology. 2008; 151 (4): 410-9.

Cite This Article

Plain Text BibTeX RIS

APA Style

Miraf Mekonnen, Selfu Girma, Abay Atnafu. (2022). Hydrolysis of Gelatin from Animal Hoof Using Bacterial Gelatinase. International Journal of Microbiology and Biotechnology, 7(3), 135-142. https://doi.org/10.11648/j.ijmb.20220703.14

Copy | Download

ACS Style

Miraf Mekonnen; Selfu Girma; Abay Atnafu. Hydrolysis of Gelatin from Animal Hoof Using Bacterial Gelatinase. Int. J. Microbiol. Biotechnol. 2022, 7(3), 135-142. doi: 10.11648/j.ijmb.20220703.14

Copy | Download

AMA Style

Miraf Mekonnen, Selfu Girma, Abay Atnafu. Hydrolysis of Gelatin from Animal Hoof Using Bacterial Gelatinase. Int J Microbiol Biotechnol. 2022;7(3):135-142. doi: 10.11648/j.ijmb.20220703.14

Copy | Download

@article{10.11648/j.ijmb.20220703.14,
  author = {Miraf Mekonnen and Selfu Girma and Abay Atnafu},
  title = {Hydrolysis of Gelatin from Animal Hoof Using Bacterial Gelatinase},
  journal = {International Journal of Microbiology and Biotechnology},
  volume = {7},
  number = {3},
  pages = {135-142},
  doi = {10.11648/j.ijmb.20220703.14},
  url = {https://doi.org/10.11648/j.ijmb.20220703.14},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ijmb.20220703.14},
  abstract = {Enzymatic hydrolysis of gelatin increases its functional, textural, and nutritional characteristics. Gelatinases are enzymes used to hydrolyze gelatin into peptides and amino acids. In recent years gelatin hydrolysates have gained a considerable indication in the applications of food and non-food products. This study aimed to isolate the gelatinase enzyme from bacteria and optimize the reaction condition for gelatin hydrolysis. A total of 100 bacterial isolates were isolated from soil and water samples. All isolates were screened for the production of gelatinase on gelatin-containing media. One isolate was selected for further analysis based on the eminent properties of the enzymatic reaction. The organism can grow under solid-state fermentation and produce gelatinase enzyme with a high activity using wheat bran as a carbon source. Gelatinase was optimally active at a temperature of 50°C and at pH 8. The optimum enzyme production under solid-state fermentation includes an incubation period of 120 h and a moisture content of 66.7%. The isolate produced more enzymes with the addition of peptone as a nitrogen source. The enzyme was used to produce gelatin hydrolysate by hydrolyzing animal hooves. The production process for the gelatin hydrolysis was optimized by varying enzyme concentration and incubation time. The hydrolysis has maximum activity at 4 h incubation period and at a high amount of enzyme concentration. This study indicated that animal waste like hoofs can be converted to useful gelatin hydrolysate using microbial gelatinase and used for various applications.},
 year = {2022}
}

Copy | Download

TY - JOUR
T1 - Hydrolysis of Gelatin from Animal Hoof Using Bacterial Gelatinase
AU - Miraf Mekonnen
AU - Selfu Girma
AU - Abay Atnafu
Y1 - 2022/09/14
PY - 2022
N1 - https://doi.org/10.11648/j.ijmb.20220703.14
DO - 10.11648/j.ijmb.20220703.14
T2 - International Journal of Microbiology and Biotechnology
JF - International Journal of Microbiology and Biotechnology
JO - International Journal of Microbiology and Biotechnology
SP - 135
EP - 142
PB - Science Publishing Group
SN - 2578-9686
UR - https://doi.org/10.11648/j.ijmb.20220703.14
AB - Enzymatic hydrolysis of gelatin increases its functional, textural, and nutritional characteristics. Gelatinases are enzymes used to hydrolyze gelatin into peptides and amino acids. In recent years gelatin hydrolysates have gained a considerable indication in the applications of food and non-food products. This study aimed to isolate the gelatinase enzyme from bacteria and optimize the reaction condition for gelatin hydrolysis. A total of 100 bacterial isolates were isolated from soil and water samples. All isolates were screened for the production of gelatinase on gelatin-containing media. One isolate was selected for further analysis based on the eminent properties of the enzymatic reaction. The organism can grow under solid-state fermentation and produce gelatinase enzyme with a high activity using wheat bran as a carbon source. Gelatinase was optimally active at a temperature of 50°C and at pH 8. The optimum enzyme production under solid-state fermentation includes an incubation period of 120 h and a moisture content of 66.7%. The isolate produced more enzymes with the addition of peptone as a nitrogen source. The enzyme was used to produce gelatin hydrolysate by hydrolyzing animal hooves. The production process for the gelatin hydrolysis was optimized by varying enzyme concentration and incubation time. The hydrolysis has maximum activity at 4 h incubation period and at a high amount of enzyme concentration. This study indicated that animal waste like hoofs can be converted to useful gelatin hydrolysate using microbial gelatinase and used for various applications.
VL - 7
IS - 3
ER -

Copy | Download

Author Information

Miraf Mekonnen

Bacterial and Viral Disease Research Directorate, Armauer Hansen Research Institute, Addis Ababa, Ethiopia
Selfu Girma

Mycobacterial Disease Research Directorate, Armauer Hansen Research Institute, Addis Ababa, Ethiopia
Abay Atnafu

Pathology Unit, Armauer Hansen Research Institute, Addis Ababa, Ethiopia

Download PDF

Sections

Plain Text BibTeX RIS

APA Style

Miraf Mekonnen, Selfu Girma, Abay Atnafu. (2022). Hydrolysis of Gelatin from Animal Hoof Using Bacterial Gelatinase. International Journal of Microbiology and Biotechnology, 7(3), 135-142. https://doi.org/10.11648/j.ijmb.20220703.14

Copy | Download

ACS Style

Miraf Mekonnen; Selfu Girma; Abay Atnafu. Hydrolysis of Gelatin from Animal Hoof Using Bacterial Gelatinase. Int. J. Microbiol. Biotechnol. 2022, 7(3), 135-142. doi: 10.11648/j.ijmb.20220703.14

Copy | Download

AMA Style

Miraf Mekonnen, Selfu Girma, Abay Atnafu. Hydrolysis of Gelatin from Animal Hoof Using Bacterial Gelatinase. Int J Microbiol Biotechnol. 2022;7(3):135-142. doi: 10.11648/j.ijmb.20220703.14

Copy | Download

@article{10.11648/j.ijmb.20220703.14,
  author = {Miraf Mekonnen and Selfu Girma and Abay Atnafu},
  title = {Hydrolysis of Gelatin from Animal Hoof Using Bacterial Gelatinase},
  journal = {International Journal of Microbiology and Biotechnology},
  volume = {7},
  number = {3},
  pages = {135-142},
  doi = {10.11648/j.ijmb.20220703.14},
  url = {https://doi.org/10.11648/j.ijmb.20220703.14},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ijmb.20220703.14},
  abstract = {Enzymatic hydrolysis of gelatin increases its functional, textural, and nutritional characteristics. Gelatinases are enzymes used to hydrolyze gelatin into peptides and amino acids. In recent years gelatin hydrolysates have gained a considerable indication in the applications of food and non-food products. This study aimed to isolate the gelatinase enzyme from bacteria and optimize the reaction condition for gelatin hydrolysis. A total of 100 bacterial isolates were isolated from soil and water samples. All isolates were screened for the production of gelatinase on gelatin-containing media. One isolate was selected for further analysis based on the eminent properties of the enzymatic reaction. The organism can grow under solid-state fermentation and produce gelatinase enzyme with a high activity using wheat bran as a carbon source. Gelatinase was optimally active at a temperature of 50°C and at pH 8. The optimum enzyme production under solid-state fermentation includes an incubation period of 120 h and a moisture content of 66.7%. The isolate produced more enzymes with the addition of peptone as a nitrogen source. The enzyme was used to produce gelatin hydrolysate by hydrolyzing animal hooves. The production process for the gelatin hydrolysis was optimized by varying enzyme concentration and incubation time. The hydrolysis has maximum activity at 4 h incubation period and at a high amount of enzyme concentration. This study indicated that animal waste like hoofs can be converted to useful gelatin hydrolysate using microbial gelatinase and used for various applications.},
 year = {2022}
}

Copy | Download