АР19678941 «Study of biochemical properties and coagulation activity of camel chymosin on mare’s milk and development of its deep processing technology»


The idea of the project is to develop an innovative technology for deep processing of mare’s milk by step-by-step isolation of protein components to obtain new food and pharmaceutical products. The technology of isolation and processing of protein components of mare’s milk will be developed. The main problem that the project is aimed at is related to the resistance of the casein fraction of mare’s milk to known milk-converting enzymes. As a result, there are no effective and inexpensive methods for separating proteins into casein and albumin fractions. The established high coagulation activity of the double-humped camel chymosin in relation to mare’s milk allows for the enzymatic separation of caseins and albumins, followed by the production of new original products: cheeses, highly purified whey proteins, lactoferrin.

Among non-traditional types of milk, mare’s milk seems promising, which is a potential candidate as the basis of various functional products. The increased interest in mare’s milk is due to the unique characteristics of milk and products made from it. The composition of mare’s milk differs significantly from the composition of the milk of the main dairy species: cows, buffaloes, goats and sheep. Productive horse breeding is a traditional branch of agriculture of the Republic of Kazakhstan and can be a promising source of new innovative products. The production of mare’s milk is actively carried out in Kazakhstan, as this product has a high demand in the republic and far beyond its borders.

With proper processing of milk, all its components are used, which brings high economic benefits. Unfortunately, the available cow’s milk processing technologies cannot be applied to mare’s milk due to significant differences in the composition of proteins, fats, minerals and carbohydrates. In terms of proteins, mare’s milk is called albuminous, since the ratio of casein to albuminglobulin fraction in milk is 50.5% to 49.5%, for comparison in cow’s milk this ratio is 78.4% to 21.6%. The lactose content in mare’s milk reaches 7%, which is similar to the lactose content in human breast milk and significantly exceeds the lactose content in cow’s milk (less than 5%). The mineral content of mare’s milk is almost two times lower compared to cow’s milk and is close to the mineral composition of breast milk. In general, according to the totality of characteristics, mare’s milk has a greater similarity with human breast milk than with cow’s milk, which explains the increasing interest in mares’ milk.

The scientific novelty of the project is to study the factors affecting the coagulation, peptidase and proteolytic activity of camel chymosin from C.bactrianus and to study the processes of isolation of whey proteins and lactoferrin without their denaturation while preserving their properties. The practical significance of the project lies in the development of technology for the complex processing of mare’s milk to obtain specific and demanded products

The research results will be in demand in biotechnology, food and processing industries and will have a positive effect on the development of research and human resources.

The aim

The aimof the project is to study the coagulation, peptidase and proteolytic activity of camel chymosin from Camelus bactrianus on mare’s milk and to develop a technology for deep processing of mare’s milk into new innovative products.

Expected results

Recombinant camel chymosin in a preparative amount will be developed and purified. The peptidase activity of camel chymosin on cloned milk k-casein fragments will be determined. The proteolytic activity of camel chymosin on casein, whey proteins and lactoferrin will be determined. The biochemical parameters of the coagulation activity of recombinant camel chymosin on mare’s milk will be studied. The conditions for obtaining a cheese clot from mare’s milk will be optimized. The parameters of isolation and production of lactoalbumins, lactoglobulins and lactoferrin will be optimized. The composition of serum peptides will be determined. The biological properties of serum peptides and lactoferrin will be studied.

Methods of biochemistry, molecular biology, biotechnology will be used

Project Manager

Tursunbekova Annelya E. (Scopus Author ID: 58131481100, Researcher ID: HIG-8491-2022, ORCID: 0000-0002-7536-7451)

Members of the research group

Akishev Zhiger D. (author ID in Scopus: 56674741700, researcher ID: N-6206-2017, ORCHID: 0000-0001-9943-1625)

Kiribayeva Asel K. (Scopus Author ID: 57215499873, ResearcherID: N-6774-2017, ORCHID: 0000-0002-8293-2340)

Aktayeva Sanya A. (Scopus Author ID: 57439359000, ResearcherID: AAR-5133-2020, ORCID: 0000-0001-6346-5866)

Maduakhasova Arina K.  (Researcher ID: JNE-6827-2023, ORCHID: 0009-0005-1460-0802)

Auez Madina K. (Researcher ID: GZG-7318-2022 ORCID: 0000-0001-7159-2793)

Publications and security documents of the project manager and members of the research group related to the project topic

  1. 1. Akishev Zh., Aktayeva S., Abdullayeva A., Baltin K., Kiribayeva A, Mussakhmetov A, Tursunbekova A., Ramankulov Ye., Khassenov B. Obtaining of recombinant camel chymosin and testing its milk-clotting activity on cow’s, goat’s, ewes’, camel’s and mare’s milk // Biology (MDPI). 2022, Vol 11, Issue 11, e1545. https://doi:10.3390/biology11111545   (Q2, Impact Factor 4.2, Cite Score 4.0, Percentile 61).
  2. Akishev ZH.D., Tursunbekova A.E. Hasenov B.B. Molokosvertyvayushchaya aktivnost’ rekombinantnogo verblyuzh’ego himozina // Vestnik KazNU. Seriya biologicheskaya. Tom 90 №1. 2022. S. 39-49. https://doi:10.26577/eb.2022.v90.i1.04.
  3. Akishev Zh., Aktayeva S., Shamsiyeva Yu., Tursunbekova A., Kalemshariv B., Tultabayeva Т., Khassenov B. Milk-clotting activity of recombinant bovine and camel chymosin for cow’s, goat’s and ewes’ milk // Eurasian Journal of Applied Biotechnology, 2023. No. 2. P.61-68. https://doi:10.11134/btp.2.2023.8 (RSCI-0,117).
  4. Akishev Zh., Abdullayeva A., Mussakhmetov A., Bekbayeva A., Tursunbekova A., Kalemshariv B., Tultabayeva Т., Khassenov B. The obtaining of the recombinant camel chymosin by submerge fermentation in the pilot bioreactor // Eurasian Journal of Applied Biotechnology, 2023. No. 1. P.48-55 https://doi:10.11134/btp.1.2023.4 (РИНЦ-0,117).
  5. Akishev Zh., Kiribayeva A., Mussakhmetov A, Baltin K., Ramankulov Ye., Khassenov B. Constitutive expression of Camelus bactrianus prochymosin B in Pichia pastoris // Heliyon. 2021. Volume 7, Issue 5, e07137 https://doi.org/10.1016/j.heliyon.2021.e07137 (Q2, Impact Factor 3.776, CiteScore 2.7, Percentile 75).
  6. Akishev Zh., Abdullaeva A., Khassenov B., Ramankulov Ye. Pilot-scale production of recombinant Camelus bactrianus chymosin B in Pichia (Komagataella) pastoris under constitutive GAP promoter // Journal of Biotechnology. 2018. Vol. 280S. P.S61. https://doi.org/10.1016/j.jbiotec.2018.06.200  (Q2, ImpactFactor 3.142, CiteScore 3.09, Percentile 74)
  7. Akishev ZH.D., Ramankulov E.M., Hasenov B.B. Sposob polucheniya molokosvertyvayushchego fermenta na osnove rekombinantnogo verblyuzh’ego himozina // Patent Respubliki Kazahstan №34896 ot 19.02.2021.
  8. Akishev ZH.D., Abdullaeva A.N., Ramankulov E.M., Hasenov B.B. SHtamm Pichia pastoris GS115/prochymB, produciruyushchij sekretornyj rekombinantnyj prohimozin V iz Bos taurus // Patent Respubliki Kazahstan №33483 ot 25.02.2019.