Multimillion-dollar plastic waste is the cause of global environmental problems. In Kazakhstan, more than 5 million solid household waste is generated annually, of which plastic is 10-15 %. This is a powerful incentive for the creation of domestic, special, environmentally friendly polymer materials, one of which is poly-3-hydroxybutyrate (PHB) and its copolymers.

They are synthesized in the cells of many soil microorganisms as reserve energy substances. The advantages of PHBs are that they are biocompatible with human tissues, biodegradable, have thermoplasticity, optical activity and antioxidant properties. Bioplastics are in high demand and could become a versatile application product. According to the data presented, PHB materials have a high potential as an eco-alternative to plastics based on petrochemical derivatives.

Relevance

Plastic production increases geometrically every year. Over the past 50 years, plastic production has increased 22 times, which has led to the allocation of about 180 billion US dollars in the last decade alone. Currently, 80% of all waste in the world is plastic, which cannot be decomposed for a long time (about 100 years).

In this regard, the development of bioplastics by large–scale manufacturers of biodegradable plastics, such as NatureWorks in the USA, Mitsubishi Chemicals in Japan, and BASF and Novamont in Europe, is being updated. The use of biodegradable plastics is showing a trend of rapid growth worldwide. It increases to 27% every year. In addition, the production of biodegradable plastics themselves is only up to 12%.

Microorganisms produce various types of high molecular weight polyhydroxyalkanoates, and among them, PHB is considered the most common. Chemically, such polyesters are repeating hydroxyacyl monomers, while their general formula is [-O-CH(R) –CH2 -CO-] n. The R = CH3 group is important for the commercialization of these biopolyesters because of its characteristics such as biodegradability and thermal stability. The production of bioplastics proceeds in three directions: by synthesis from monomers, by fermentation by microorganisms and by polymerization of lactic acid derivatives as polylactates. Among the biodegradable plastics, various polyesters, glycolic acid, polymers, polyhydroxybutyrate and its fatty acids, etc. can be distinguished. But, nevertheless, the production of environmentally friendly polymers is not much inferior to the production of petroleum-based plastics.

In addition to thermoplasticity, PHBs have optical activity, antioxidant properties, piezoelectric effect and, most importantly, they are characterized by biodegradability and biocompatibility. Notable amounts of PHB accumulate bacteria from the genera: Alcaligenes, Chromatium, Hyphomicrobium, Methylobacterium, Nocardia, Pseudomonas, Rhizobium, Spirillum, Streptomyces. However, only a few microorganisms are promising for industrial biosynthesis of biopolymer: Bacillus, Alcaligenes, Ralstonia, Azotobacter and Methylobacterium. They are able to accumulate polyesters on available substrates.

Objective

 Project aim is to isolate and identify strains producing polyhydroxybutyrate as a source of biodegradable plastics for household and biomedical purposes.

Expected results

The results of the research will allow us to deepen knowledge on the management of microbial biosynthesis processes and biosynthesis management in various fields of industry, everyday life and medicine. It has a high probability of commercialization of the resulting polymer composite for biomedical needs. There is no active production of PHB in Kazakhstan, therefore, research in this area is new and highly relevant for our country. To date, the topic of studying this project fully meets the requirements and criteria of the green economy of Kazakhstan and is being studied in many countries. The introduction of microbiological producers will significantly expand the potential of biological research and their application in the industry.

Principal investigator

Rysbek A.B. H-index – 1; ResearcherID Q-2325-2017; ORCID 0000-0001-8290-0552; Scopus Author ID 57415916300

Members of the research group

Abeldenov S.K. H-index –4; ResearcherID F-5139-2015; ORCID 0000-0002-6974-9138; Scopus Author ID 56674705400

Turgimbayeva A.M. H-index –2; ResearcherID N-6857-2017; ORCID 0000-0001-7263-1643; Scopus Author ID 57202383621

Zhumabaev A.R. H-index –3; ORCID 0000-0001-9022-0741; Scopus Author ID: 57193544703; SciProfiles: 1937315; Loop profile: 1987375

Kirillov S.O. H-index –1; ResearcherID N-6322-2017; ORCID 0000-0001-6229-5762

Shaizadinova A.M. H-index –2; ORCID 0000-0002-5911-3064; Scopus Author ID: 57224822522

Publications and security documents of the project manager and members of the research group

1. Comparative Characterization and Identification of Poly-3-hydroxybutyrate Producing Bacteria with Subsequent Optimization of Polymer Yield. Rysbek A., Ramankulov Y, Kurmanbayev A, Richert A, Abeldenov S. Polymers. 2022; 14(2): 335. 16 p. https://doi.org/10.3390/polym14020335
2. Microbiological preparation of polyhydroxybutyrate (PHB) when irradiated with ultraviolet light. A. Rysbek, A. Kurmanbayev, U. Jankiewicz, A. Richert. Virtual International Conference “Plant productivity and food safety: Soil science, Microbiology, Agricultural Genetics and Food quality” Nicolaus Copernicus University in Torun, Poland. 15-16.09.2022
3. Obtaining an effective producer of polyhydroxybutyrate (PHB) under ultraviolet irradiation of bacteria of the genus Bacillus and Azotobacter. Rysbek A.B., Kurmanbaev A.A. Collection of materials of the International scientific and practical Conference “Modern Problems of Biotechnology: From Laboratory Research to Production”, 2021, Al-Farabi Kazakh National University, Almaty, Kazakhstan, pp. 218-220.
4. Creation of a collection of microorganisms – destructors of organic substances that are promising for bioremediation of technogenic disturbed lands in Kazakhstan / Baigonusova Zh.A., Rysbek A.B., Kurmanbaev A.A. abstract/ 2nd International Conference «Plants and Microbes: The Future of Biotechnology», Saratov, October 5-9, 2020.
5. Microbiological production of polyhydroxbutyrates from renewable sources. Rysbek A., Kurmanbayev A. Eurasian Journal of Applied Biotechnology No2. (2020) p. 24 – 38.
6. Study Of Morphological And Physiological Properties Of Poly-3-Hydroxybutyrate Producers. Rysbek A.B., Kurmanbaev A.A. Materials of the international scientific conference of students and young scientists “Farabi Alemi”. Almaty, Kazakhstan, April 6-9, 2020 – 341 s
7. The Effects of a Biopreparation on Legume (Pisum Sativum) / Martynenko V.V., Rysbek A.B., Kurmanbayev A.A., Baigonusova Zh.A. Eurasian Journal of Applied Biotechnology No2. (2020) p. 100 – 103Characterization of DNA substrate specificities of apurinic/apyrimidinic endonucleases from Mycobacterium tuberculosis. Abeldenov S, Talhaoui I, Zharkov DO, Ishchenko AA, Ramanculov E, Saparbaev M, Khassenov B. DNA Repair (Amst). 2015 Sep; 33:1-16. doi: 10.1016/j.dnarep.2015.05.007.
8. Thiol-Ene Click Synthesis of Alginate Hydrogels Loaded with Silver Nanoparticles and Cefepime. Akhmetkarimova, Zh.S., Kudaibergen, G.K., Kaukabaeva, G.K., Abeldenov, S.K., & Rysbek, A.B. (2023) Eurasian Journal of Chemistry. https://doi.org/10.31489/2959-0663/2-23-14.
9. Isolation and study of effective biocellulose producers / Nagmetova G., Rysbek A., Kurmanbayev A. Proc. Of Europian Biothecnology Congress / Abstracts / Journal of Biotechnology (2019) p. 33–88. https://doi.org/10.1016/j.jbiotec.2019.05.135.  Q2.
10.  Creation of a collection of active strains of bacteria – destructors of organic matter / Rysbek A. B., Baigonusova Zh. A., Kurmanbaev A.A. 56 “Contribution of young scientists to innovative technologies for agriculture”, dedicated to the 80th anniversary of academician Suleimenov M.K.: Collection of reports, abstracts of the Republican Scientific Conference of Young scientists. – Shortand, 2019.-C. 115-118.
11.  Isolation and investigation of nitrogen-fixing strains of microorganisms promising for the development of a preparation for leguminous crops / Rysbek A. B., Baigonusova Zh. A., Kurmanbaev A.A. 56 “Contribution of young scientists to innovative technologies of agricultural industry”, dedicated to the 80th anniversary of academician Suleimenov M.K.: Collection of reports, abstracts of the Republican Scientific Conference of Young Scientists. – Shortand, 2019.-C. 111-115.

Patents of the research group

1. The bacterium strain of the genus Bacillus aryabhattai Ra 5 is a polyhydroxybutyrate producer used to produce bioplastics. Patent No. 7433 RK 2022/0550.2, 2022.
2. Recombinant strain of Escherichia coli Arctic Express(DE3)RP/papa producing phosphohydrolase AppA Patent No. 30999, 2016;
3. Recombinant Pichia pastoris X-33/AppA strain producing bacterial phytase AppA, Patent No. 32368, 2017;
4. Escherichia coli BL21(DE3)/ptls strain producing thermally stable recombinant DNA polymerase of Thermus thermophilus bacteria, Patent No. 33100, 2018;
5. The strain of hybrid cultured animal cells Mus musculus L. – Mab/1E7-Her-2/ECD is a producer of monoclonal antibodies to the extracellular domain (Her-2/ECD) of the epidermal growth factor receptor Her-2/neu, Patent No. 106734, 2019;
6. The strain of hybrid cultured animal cells Mus Musculus L. is a producer of monoclonal antibodies to the marker of cell proliferation Ki 67, Patent No. 35390, 2021;

Results achieved

2023

5 promising strains of PHB producers were isolated from Lake Kobeytuz, Yerementau district, Akmola region. Their morphological and physiological-biochemical properties were studied. The selected isolates were identified. The design of primers for the amplification of the PHB synthase gene has been prepared.