Alternative protein sources: bacteria Methylococcus capsulatus concentrate, characteristic of composition and biological value
Abstract
A promising growth vector of food protein production in the context of the Russian Federation’s food sovereignty security is the use of microbial synthesis. Taking into consideration the proven promising use of biotechnological processes in the production of alternative protein sources, modern scientific research is focused, among other issues, on improving the technology of obtaining food microbial protein using a variety of substrates and strains-producers, as well as evaluating the consumer properties, food, biological value and safety of such products.
The purpose of the research was to study and comparatively evaluate protein concentrate (PC) from bacteria Methylococcus capsulatus and basic food of animal and plant origin within the development of the technology of optimal in nutritional and biological value PC production.
Material and methods. Analysis of the nutritional and biological value of PC obtained from denucleinized and purified from cell walls biomass of methanoxidizing bacteria Methylococcus capsulatus (strain GSB-15) was carried out on 46 indicators, including estimation of protein content and amino acid composition, fat content and fatty acid composition, ash and moisture. Biological studies based on measuring of net protein ratio / net protein utilization were performed on 28 growing (between 25–50 days of life) male Wistar rats. Rats in the control group (n=14) received a semi-synthetic casein diet with a protein content of ~12% in calories, the test group (n=14) received a diet including an equivalent amount of PC protein. Body weight, feed intake, and fecal and urine nitrogen losses were measured during the experiment. The biological value and digestibility of protein were judged by coefficients of protein efficiency ratio, net protein ratio, true protein digestibility, true protein biological value, true net protein utilization.
Results. The nutritional value study of PC showed high protein content – 69.0%, the share of fat, moisture and ash, accounted for 0.17, 9.5 and 14.4%, respectively. The carbohydrate content was 7.0% (of which mono- and disaccharides were <0.1%). The results of a comparative assessment of Methylococcus capsulatus protein amino acid profile and basic food of animal and plant origin showed a balanced content of the most amino acids, the level of which is comparable with the protein of chicken egg, which is traditionally a standard of quality of complete protein. At the same time, the content of the essential amino acid tryptophan in PC was an order of magnitude lower than in chicken egg protein; the content of this amino acid in PC is comparable with incomplete plant proteins (sunflower, flax, rapeseed). The results of the biological value evaluation of the Methylococcus capsulatus protein in the experiment on rats indicate a relatively low biological value of the microbial synthesis protein, that is caused, most likely, by tryptophan deficiency. Rats of the test group had a significant decrease in body weight gain, feed/protein intake, coefficient of protein efficiency ratio, coefficient of net protein ratio, true protein biological value, true net protein utilization.
Conclusion. The results of a comparative evaluation of PC from methanotrophic bacteria Methylococcus capsulatus denucleinized biomass and basic food of animal and plant origin indicate its relatively high nutritional value. However, the characteristics of this PC sample were not optimal in regard of protein biological value by reason of tryptophan deficiency. A single amino acid deficiency is not a valid argument for not using microbially synthesized protein in human nutrition, considering the capabilities of the modern food industry, including ways to enrich foodstuffs with missing components. In addition, there is every cause to believe that adjusting the hydrolysis technology used in the production of PC will allow to eliminate the essential amino acid loss, thereby increasing the biological value of this product.
Keywords:novel food; alternative sources of protein; microbial synthesis; protein concentrate; Methylococcus capsulatus
Funding. The research was funded by the Russian Science Foundation (project No. 20-76-10014).
Conflict of interest. The authors have no conflict of interest to declare.
Contribution. Concept and design of the study – Tyshko N.V., Sadykova E.O.; collecting and statistical processing the material – Trebukh M.D., Nikitin N.S., Shestakova S.I., Shumakova A.A., Makarenko M.А.; text writing – Sadykova E.O., Tyshko N.V.; editing, approval of the final version of the article, responsibility for the integrity of all parts of the article – all authors.
Acknowledgements. The authors would like to sincerely thank Valentina A. Pashorina, researcher-engineer of the Safety Assessment of Biotechnology and Novel Food Sources Laboratory, for her help in carrying out the research on nutritional and biological values of protein concentrate.
For citation: Sadykova E.O., Trebukh M.D., Nikitin N.S., Shestakova S.I., Shumakova A.A., Makarenko M.А., Tyshko N.V. Alternative protein sources: bacteria Methylococcus capsulatus concentrate, characteristic of composition and biological value. Voprosy pitaniia [Problems of Nutrition]. 2023; 92 (3): 36–44. DOI: https://doi.org/10.33029/0042-8833-2023-92-3-36-44 (in Russian)
References
1. Presidential Decree No. 20 of January 21, 2020 «On Approval of Food Security Doctrine of the Russian Federation». (in Russian)
2. Forecast of Scientific and Technological Development of Russia: 2030. Biotechnology. In: L.M. Gokhberg, M.P. Kirpichnikov (eds). Ministry of Education and Science of the Russian Federation, National Research University Higher School of Economics. Moscow, 2014: 48 p. (in Russian)
3. Orlova N.V., Serova E.V., Nilolaev D.V., Khvorostyanaya A.S., Novikova Yu.A., Yavkina E.V., et al. Development in innovations for agroindustrial sector in Russia. Agriculture 4.0. In: Report to the XXI April International Scientific Conference on the Challenges of Economic and Social Development. Moscow: ID Vysshey shkoly ekonomiki, 2020: 128 p. (in Russian)
4. RAS Academician Natalia Zinov’eva: Genetic technologies are the basis of the country’s food independence: [portal of the Information Agency «Scientific Russia»]. 07.02.2023. URL: https://scientificrussia.ru/articles/akademik-ran-natalia-zinoveva-geneticeskie-tehnologii-lezat-v-osnove-prodovolstvennoj-nezavisimosti-strany (in Russian)
5. Tutelyan V.A., Tyshko N.V., Aksyuk I.N. New food sources: present and future. In: V.A. Tutelyan, D.B. Nikityuk (eds). Nutritiology and Clinical Dietetics: National Guide. Moscow: GEOTAR-Media, 2023: 1008 p. ISBN 978-5-9704-7769-4. (in Russian)
6. Tyshko N.V., Sadykova E.O., Shestakova S.I., Aksyuk I.N. Novel food sources: from GMO to the broadening of Russia’s bioresource base. Voprosy pitaniia [Problems of Nutrition]. 2020; 89 (4): 100–9. DOI: https://doi.org/10.24411/0042-8833-2020-10046 (in Russian)
7. Pokrovsky A.A. (ed.). Biological and medical research on hydrocarbon yeast (1964–1970). Moscow: Nauka, 1972: 468 p. (in Russian)
8. Pokrovskiy A.A. Metabolic aspects of food pharmacology and toxicology. Moscow: Meditsina, 1979: 184 p. (in Russian)
9. Biswas A., Takakuwa F., Yamada S., Matsuda A., Saville R.M., LeBlanc A., et al. Methanotroph (Methylococcus capsulatus, Bath) bacteria meal as an alternative protein source for Japanese yellowtail, Seriola quinqueradiata. Aquaculture. 2020; 529: 735700. DOI: https://doi.org/10.1016/j.aquaculture.2020.735700
10. Chama M.K.H., Liang H., Huang D., Ge X., Ren M., Zhang L., et al. Methanotroph (Methylococcus capsulatus, Bath) as an alternative protein source for genetically improved farmed tilapia (GIFT: Oreochromis niloticus) and its effect on antioxidants and immune response. Aquac Rep. 2021; 21: 100872. DOI: https://doi.org/10.1016/j.aqrep.2021.100872
11. Chen Y., Chi S., Zhang S., Dong X., Yang Q., Liu H., et al. Replacement of fish meal with methanotroph (Methylococcus capsulatus, Bath) bacteria meal in the diets of Pacific white shrimp (Litopenaeus vannamei). Aquaculture. 2021; 541: 736801. DOI: https://doi.org/10.1016/j.aquaculture.2021.736801
12. Guo B., He X., Ge C., Xue M., Wang, J., Longshaw M., et al. A natural gas fermentation bacterial meal (FeedKind®) as a functional alternative ingredient for fishmeal in diet of largemouth bass, Micropterus salmoides. Antioxidants. 2022; 11 (8): 1479. DOI: https://doi.org/10.3390/antiox11081479
13. Jintasataporn O., Chumkam S., Triwutanon S., LeBlanc A., Sawanboonchun J. Effects of a single cell protein (Methylococcus capsulatus, Bath) in pacific white shrimp (Penaeus vannamei) diet on growth performance, survival rate and resistance to Vibrio parahaemolyticus, the causative agent of acute hepatopancreatic necrosis disease. Front Mar Sci. 2021; 8: 7640428. DOI: https://doi.org/10.3389/fmars.2021.764042
14. Lu W., Yu H., Liang Y., Zhai S. Evaluation of methanotroph (Methylococcus capsulatus, Bath) bacteria protein as an alternative to fish meal in the diet of juvenile american eel (Anguilla rostrata). Animals (Basel). 2023; 13 (4): 681. DOI: https://doi.org/10.3390/ani13040681
15. Xu B., Liu Y., Chen K., Wang L., Sagada G., Tegomo A.F., et al. Evaluation of methanotroph (Methylococcus capsulatus, Bath) bacteria meal (FeedKind®) as an alternative protein source for juvenile black sea bream Acanthopagrus schlegelii. Front Mar Sci. 2021; 8: 778301. DOI: https://doi.org/10.3389/fmars.2021.778301
16. Zhang Q., Liang H., Longshaw M., Wang J., Ge X., Zhu J., et al. Effects of replacing fishmeal with methanotroph (Methylococcus capsulatus, Bath) bacteria meal (FeedKind®) on growth and intestinal health status of juvenile largemouth bass (Micropterus salmoides). Fish Shellfish Immunol. 2022; 122: 298–305. DOI: https://doi.org/10.1016/j.fsi.2022.02.008
17. Oshkin I.Y., Khmelenina V.N., But S.Y., Miroshnikov K.K., Belova S.E., Khokhlachev N.S., et al. Analysis of the Complete Genome Sequence of Strain Concept-8, a Novel Representative of the Genus Methylococcus (transl.). Mikrobiologiya [Microbiology]. 2020; 89 (3): 309–17. DOI: https://doi.org/10.1134/S0026261720030131
18. Oshkin I.Y., Danilova O.V., Suleymanov R.Z., Tikhonova E.N., Malakhova T.V., Murashova A.I., et al. Thermotolerant methanotrophic bacteria from sediments of the river Chernaya, Crimea, and assessment of their growth characteristics (transl.). Mikrobiologiya [Microbiology]. 2021; 90 (5): 588–597. DOI: https://doi.org/10.31857/S0026365621050141 (in Russian)
19. Zheleznyakov A.G., Glukhikh S.A., Guzenberg A.S., Romanov S.Yu., Yurgin A.V., Ryabkin A.M. Use of methane in closed-loop life support systems for space missions. Kosmicheskaya tekhnika i tekhnologii [Space-Based Technology and Technologies]. 2021; 4 (35): 78–92. DOI: https://doi.org/10.33950/spacetech-2308-7625-2021-4-78-92 (in Russian)
20. Oshkin I.Y., Suleimanov R.Z., Khmelenina V.N., Mardanov A.V., Pimenov N.V., Dedysh S.N. Complete genome sequence of Methylococcus capsulatus MIR, a methanotroph capable of growth on methanol. Microbiol Resour Announc. 2022; 11 (9): e0054222. DOI: https://doi.org/10.1128/mra.00542-22
21. Tutelyan V.A. Chemical composition and caloric content of Russian food products: Directory. Moscow: DeLi Plyus, 2012: 283 p. ISBN 978-5-905170-20-1. (in Russian)
22. Tyshko N.V., Zhminchenko V.M., Nikitin N.S., Trebukh M.D., Shestakova S.I., Pashorina V.A., Sadykova E.O. The comprehensive studies of Hermetia illucens larvae protein’s biological value. Voprosy pitaniia [Problems of Nutrition]. 2021; 90 (5): 49–58. DOI: https://doi.org/10.33029/0042-8833-2021-90-5-49-58 (in Russian)
23. Tyshko N.V., Zhminchenko V.M., Pashorina V.A., Selyaskin K.E., et al. A comparative assessment of the diet influence on growth and development of rats. Voprosy pitaniia [Problems of Nutrition]. 2011; 80 (5): 30–8. (in Russian)
24. Food chemical composition. Book 2. In: I.M. Skurikhin, M.N. Volgarev M.N. Moscow: Agropromizdat, 1987: 360 p. (in Russian)
25. Stepuro M.V., Khaprova E.N. Comparative assessment of the biological value of proteins of plant raw materials. Izvestiya vuzov. Pischevaya tehnologiya [News of Higher Educational Institutions. Food Technology]. 2010; (4): 34. (in Russian)
26. Kristinsson H.G., Rasco B.A. Fish protein hydrolysates: production, biochemical, and functional properties. Crit Rev Food Sci Nutr. 2000; 40 (1): 43–81. DOI: https://doi.org/10.1080/10408690091189266
27. Amino Acid Content of Foods and Biological Data on Proteins. Rome : FAO, 1981. URL: http://www.fao.org/3/AC854T/AC854T00.htm (date of access October 18, 2022)
28. ILSI. Crop Composition Database Version 4.0. Washington, DC: International Life Science Institute, 2010. URL: www.cropcomposition.org (date of access October 18, 2022)