AP19678954 Studying of collagenases for enzymatic hydrolysis of low-value meat industry wastes into protein hydrolysates

Relevance of the project

As the population in Kazakhstan increases, food consumption, including meat, is also rising. According to the Bureau of National Statistics, meat production in the Republic of Kazakhstan is up 4-5% annually and amounted to 2,162,200tonnes in live weight in 2021. At slaughterhouses about 50% of the total weight of the animal carcass represented by bones, skin, blood, inferior by-products remain as low-value secondary raw materials. The utilisation of by-products bears environmental risks and requires additional costs, results in the waste of valuable fodder and edible protein, and causing an overall high cost of meat. It is therefore necessary to strive to recycle them into valuable secondary products.

Most of the by-products of the meat industry consist of tissue, with 25-30% of the total protein mass represented by collagen. However, collagen is one of the most sought-after products on the market of food, cosmetics and biomedicine industries. According to various marketing studies, the global market for collagen in 2021 was estimated to be between $5.03 billion and $7.74 billion, with a projected average annual growth rate of 8.3 to 8.59%, and approximately 20% of the collagen market is in its hydrolyzed form. From an economic point of view, the production of collagen hydrolysates is a promising way of valorising meat waste.

The idea of the project is to find and study bacterial collagenases in order to use them in the enzymatic hydrolysis of slaughterhouse wastes (hides, fetlock joints and tails) into valuable products: food and fodder hydrolysates. Food hydrolyzate of collagen protein is used in sausage production as a source of peptides, amino acids, soluble forms of collagen and as a moisture-retaining filler, and feed hydrolyzate as a source of digestible protein additive. As part of the project, collagenases will be obtained and studied, and their targeted modification will be carried out. The main problem addressed by the project is the recycling of low-value waste products from the meat industry.

The scientific novelty of the project lies in development of directed enzymatic hydrolysis of robust collagen-containing protein substrates using new collagenase enzymes. The practical significance lies in the development of technology for processing low-value and recyclable waste from the meat processing industry into protein hydrolysates of value and used in the food and fodder industries.

Project goal

The aim of the project is to find, modify and study bacterial collagenases for their use in the enzymatic hydrolysis of meat processing wastes and the production of protein hydrolysates.

Expected results

  • Collagenolytic microorganisms will be searched for, isolated, screened on selective media and identified by molecular genetic and proteomic methods.
  • Bacterial collagenases will be isolated, chromatographed and identified.
  • Targeted modification of bacterial collagenases will be carried out.
  • Biochemical properties of native and modified collagenases will be studied.
  • The conditions for hydrolysis of meat processing industry wastes using collagenases will be optimized.

Project leader

Baltin Kairat, candidate of biological sciences, senior researcher. ResearcherID: AAQ-9372-2020, ORCID: 0000-0002-6187-7223, Scopus Author ID: 55437315200

Research team members

Khassenov Bekbolat, full professor, candidate of chemical sciences, head of laboratory. ResearcherID: AAM-8657-2020, ORCID: 0000-0003-4572-948X, Scopus Author ID: 36096620800

Aktayeva Saniya, MSc, senior researcher. ResearcherID: AAR-5133-2020, ORCID: 0000-0001-6346-5866, Scopus Author ID: 57439359000

Akishev Zhiger, MSc, senior researcher. ResearcherID: N-6206-2017, ORCID: 0000-0001-9943-1625, Scopus Author ID: 56674741700

Mussakhmetov Arman, MSc, senior researcher. ResearcherID: AAQ-9945-2020, ORCID: 0000-0002-6182-3487, Scopus Author ID: 57203751227

Sarsen Arailym, MSc, junior researcher. ResearcherID: AED-8089-2022, ORCID: 0000-0002-6071-430X

Shamsiyeva Yuliya, MSc, junior researcher. ResearcherID: AFX-3509-2022, ORCID: 0000-0002-3136-6000

Publications and protection documents

  1. Aktayeva S., Baltin K., Kiribayeva A., Akishev Zh., Silayev D., Ramankulov Ye., Khassenov B. Isolation of Bacillus A5.3 Strain with Keratinolytic Activity // Biology (MDPI). 2022, Vol 11, Issue 2, e244. doi:10.3390/biology11020244 (Q1, IF=5.079, CiteScore=3.3, percentile =79).
  2. Kiribayeva A., Mukanov B., Silayev D., Akishev Zh., Ramankulov Ye., Khassenov B. Cloning, expression, and characterization of a recombinant xylanase from Bacillus T6 // PLoS One. 2022. Vol.17(3). e0202232. doi:10.1371/journal.pone.0202232 (Q2, IF=3.24, CiteScore=5.3, percentile =92).
  3. Aktayeva S., Baltin K., Ramankulov Ye., Khassenov B. Isolation and identification of the keratin degrading bacteria // Journal of Biotechnology. 2019. Vol.305S. P.S30. doi:10.1016/j.jbiotec.2019.05.112 (Q2, IF=3.142, CiteScore=3.09, percentile =74).
  4. KiribayevaK., Mukanov B.A., Baduanova S.D., Silayev D.V., Ramankulov Ye.M., Khassenov B.B. Isolation and study of a recombinant carbohydrase xylanase from Bacillus licheniformis // Eurasian Journal of Applied Biotechnology. 2019. Iss.1. pp.68-72. doi:10.11134/btp.1.2019.8.
  5. Kiribayeva, Ramanculov E., Khassenov B. Constitutive expression of thermostable α-amylase from Bacillus licheniformisin Pichia pastoris // Journal of Biotechnology. 2017. Vol.256S, P.S64. doi:10.1016/j.jbiotec.2017.06.1017 (Q2, IF=3.142, CiteScore=3.09 percentile =74).
  6. Aktayeva, Akishev Zh., Khassenov B. Proteolytic enzymes in cheese making // Eurasian Journal of Applied Biotechnology. Iss.1. 2018. pp. 10-14. doi:10.11134/btp.1.2018.2
  7. Aktayeva S.A., Baltin K.K., Kiribayeva A.K., Ramankulov E.M., Khassenov B.B. Bacterial strain Bacillus licheniformis T7 – producer of proteases and keratinase. Patent of the Republic of Kazakhstan №35357 from 11.2021.

Achieved results

A collection of strains with gelatinase activity was created in the laboratory of genetics and biochemistry of microorganisms. By culturing one of them on depleted nutrient medium, an enzymatic extract was prepared, which showed good collagenase activity in a gelatin zymographic assay. A model experiment was carried out to hydrolyze the scalded putamen joints with the enzymatic extract obtained for 48 hours. It was found that under the selected conditions (1% extract, hydromodule 1:1, 45°C, pH 6.5–7.0), the degree of hydrolysis reached 38–40% after 24 hours (calculated on a dry matter basis). These results are not inferior to the control test, in which a commercially produced enzyme was utilized, and have an advantage over the recognized technology of protein breakdown by protease. The preliminary results obtained show the prospectivity of the chosen direction and require further research.