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Project titleDevelopment of microbial starter cultures for expand the range of bakery products from non-traditional types of flour

Keywordsbakery products, sourdoughs, starter cultures, lactic acid bacteria, yeast, fermented foods

Annotation
Bread and bakery products are physiologically necessary components in the human diet. In Russia, bread is a product of everyday demand. According to the Federal State Statistics Service, bread ranks third in terms of the consumption of basic foodstuffs by the population in the country. Increasing the nutritional and biological value of bakery products is a promising direction in the catering industry. To this end, the recipes include raw materials and additives with high nutritional and biological value. The use of non-traditional types of flour in the production of bakery products (chickpea, buckwheat, green buckwheat, amaranth, flax, etc.) is one of the promising ways to influence the qualitative composition of the diet of the country's population. Such types of flour contain a significant amount of easily digestible macro- and micronutrients. However, the introduction of such raw materials into the recipes of bakery products causes certain difficulties in the technology of their production. This is due to the fact that the new alternative raw material differs significantly from wheat and rye flour in terms of the composition of carbohydrates, acidity, quantity and qualitative composition of fat and protein, which greatly affects the vital processes of microorganisms responsible for the bioconversion of this vegetable raw material in the composition of products, and, as a result, on the quality of test semi-finished products and finished products. One of the main ways to solve this problem is the selection of a unique composition of microbial starter cultures for bakery products from non-traditional types of flour. The development and selection of a unique microbial composition are the basis of the scientific problem of this project. Starter cultures are complex cultures of yeast and bacteria. They significantly influence the characteristic taste, texture and quality of baked goods. The microorganisms present in the sourdough can be isolated from ingredients or selected from existing collections of microorganisms. This opens up a wide range of research opportunities. The scientific novelty of the project lies in the development of a new technology of bakery products using non-traditional types of flour and specialized microbial starter cultures in order to expand the range of bakery products and further scale up the recipes and technologies developed in the laboratory to the industrial level.

Expected results
The implementation of the project involves obtaining a set of scientific results: - native microorganisms of non-traditional raw materials will be isolated and identified, the possibilities of their use in baking will be assessed; - microorganisms will be selected, the composition of microbial starter cultures for products based on non-traditional types of flour; - will be identified metabolites produced by microorganisms that affect the quality of bread; - regularities of growth will be determined and the optimal ratio of microorganisms in starter cultures will be substantiated; - technologies for the production of pure cultures of selected microorganisms and microbial starters based on them will be developed; - recipes and technologies for bakery products based on non-traditional types of flour will be developed; - data will be obtained on the rheological properties of non-traditional types of flour and the effect of microbial starters on them; - applications will be submitted for the deposit of strains in one of the Russian banks of industrially used microorganisms; - a draft regulatory and technical documentation will be developed - technological instructions (TI) and technical specifications (TS) for the developed bakery products; - patent applications will be filed for methods of preparing bakery products based on non-traditional types of flour. The scientific significance for the development of new scientific topics lies in the development of new microbial starter cultures and bakery products based on non-traditional types of flour, which can serve as a start for large-scale studies of starter cultures applicable to non-traditional raw materials in bakery. In addition, the introduction of the developed products and microbial starter cultures at bakery enterprises will expand the range of healthy and safe food products in one of the product categories most in demand by the population of the country. The novelty of the developed solutions will be confirmed by patent applications for methods of preparing bakery products based on non-traditional types of flour. Compliance of the planned results with the world level is ensured by a number of factors, including: - high scientific qualification of the project manager, who has 15 publications in indexed printed publications, including 5 publications in the Scopus and Web of Science databases (h-index 1), 2 patents of the Russian Federation. - compliance of the proposed methods for the development of new technologies, the creation of microbial starter and the study of the properties of the resulting products with the world level of research in these areas Possibility of practical use of the expected results in the social sphere: · The results of the project can be used to provide the population with healthy and safe food products. · The results of the project are applicable not only to meet the needs of people in a healthy diet, but to expand the range of healthy and safe food products in one of the product categories most in demand by the population of the country The possibility of practical use of the expected results in the economy: · The results of the successful completion of the research will be available to both large companies and small food industry enterprises producing bakery products. · Creation of microbial starter cultures and bakery products based on non-traditional types of flour will support the policy of import substitution and resource saving in force on the territory of the Russian Federation. · An increase in the range of bakery products and the consumer's interest in it will lead to an improvement in the quality of nutrition of the country's population and its well-being. · Intensive development of biotechnology fortified and specialized food products may contribute to the isolation of a new market segment aimed at meeting people's need for a healthy diet.

Annotation of the results obtained in 2023
The research revealed information about the composition, properties and microbiome of non-traditional flour types. Literature sources in English and Russian, indexed in Scopus, Web of Science, RSCI for the last 30 years were examined, with priority given to articles published after 2018. A detailed study was conducted on the scientific literature regarding the composition of wheat, lentil, coconut, amaranth, corn, rice flours, as well as flour made from hydrothermally untreated buckwheat. Details were provided on nutritional value, content of various protein fractions, their amino acid composition, the fatty acid composition of lipids, content of carbohydrates of different classes, amylose to amylopectin ratio of respective starches, as well as on the distinctive properties and breadmaking potential of the flours in question. In addition, the available data on the microbiological composition of various non-traditional flours and respective sourdoughs was collected, and the most significant patterns were identified. It was shown that in spontaneous sourdoughs based on the majority of studied flour types lactic acid bacteria are quantitatively dominant, as they displace most of the native microorganisms in flour over the course of fermentation. The group studied the rheological properties of a range of mixtures of wheat flour and non-traditional flours at dosages of 5, 10, 15, 20 and 25%. The effect of non-traditional flours on the water absorption capacity of the mixture, dough stability, dough formation time, degree of softening and baking power, amylolytic activity, as well as elasticity and extensibility of dough semi-finished products were evaluated. The addition of amaranth flour significantly increases the water absorption capacity of the mixture - for instance, addition of 25% of amaranth flour increases this figure by 23%, with a slight increase in the specific volume of bread and significant improvement in water retention, allowing the bread to retain softness for longer. The addition of flour of all other non-traditional types results in the decrease in specific volume, despite the fact that coconut and lentil flours improve water absorption capacity. The addition of over 5% coconut flour, 10% lentil or hydrothermally untreated buckwheat flour, 15% corn flour or 25% brown rice flour is not recommended. The addition of 25% amaranth flour is recommended based on rheological tests conducted. During the course of the project, the isolation of pure cultures from spontaneous sourdoughs based on a wide selection of corn, amaranth, brown rice, coconut, lentil and hydrothermally untreated buckwheat flours was carried out. Pure cultures were isolated using Koch method with inoculation on selective media. The isolated cultures were checked for purity, and if necessary, additional cleaning was carried out by dispersing the cultures into selective environment and re-selecting singular colonies. As a result of initial microbial identification, several yeast and bacterial isolates were selected and submitted for identification to the Center for collective use «Genomic technologies, proteomics and cellular biology» of Federal State Budgetary scientific institution «All-Russian Research Institute for Agricultural Microbiology». Based on the data obtained, among selected and identified microorganisms the yeasts Issatchenkia orientalis YIT1 and Wickerhamomyces anomalus S605T, as well as strains RCAM 04545, RCAM 04544, RCAM 01995 and RCAM 00432 provided from the collections of Federal State Budgetary scientific institution «All-Russian Research Institute for Agricultural Microbiology» and Federal State Autonomous scientific institution «Research institute for Baking Industry» are recommended for the development of sourdoughs. This statement is confirmed by the research on physiological and biochemical properties of the strains by members of the research team, as well as by literature data on the properties of respective yeast species. In addition, analysis of the volatile metabolites of the yeast strains studied by gas chromatography methods showed that the aromatic profiles obtained did not contradict the use of these strains in the composition of the sourdough. Of special interest are the strains Issatchenkia orientalis YIT1 and Wickerhamomyces anomalus S605T characterized by increased accumulation of esters (114±17 and 87±13 mg/l respectively) compared to other researched strains. Based on the results of research on biochemical properties and analysis of qualitative and quantitative composition of organic acids synthesized by studied lactic acid bacteria, the most promising strains were selected for the development of sourdoughs – Lb. plantarum 017BL, Lb. brevis 002BL, Lb. brevis 001BL (BSG), as well as the strain of Leuconostoc mesenteroides (BIT2) selected by the research team. These strains are characterized by intensive acid accumulation, with a high mass fraction of lactic acid. Based on the recommended strains, 16 microbial starters were proposed. During the study the potential for the use of Wickerhamomyces anomalus yeast has been evaluated, the antimicrobial properties of S605T strain in relation to Bacillus subtilis, a ropy bread pathogen, were confirmed, sourdough compositions containing the strain were obtained, with most stable compositions being based on: 1) Lb. plantarum 017BL, Lb. brevis 002BL, Wickerhamomyces anomalus S605T; 2) Lb. plantarum 017BL, Lb. brevis 001BL, Wickerhamomyces anomalus S605T. The sourdoughs based on hydrothermally untreated buckwheat made with the use of those compositions possessed positive organoleptic indicators, while lactic acid bacteria and yeast preserved and developed in the proportions they were inoculated in (quantitatively 1:5:5, Wickerhamomyces anomalus S605T : Lb. plantarum 017BL : Lb.brevis 001BL); the ratio was selected based on literature analysis. At the same time the sourdough sample based on Lb. plantarum 017BL, Lb. brevis 002BL, Wickerhamomyces anomalus S605T has a more distinct and intense aroma,which is why it was recommended for research on other types of flour. As a result of further research into development patterns of microorganisms in sourdough, it was shown that the flour type has a significant effect on the quality of sourdough. For example, on lentil flour the sourdough based on Wickerhamomyces anomalus S605T, Lb. brevis 002BL and Lb. plantarum 017BL has led to the spoilage of the sample. The high water absorption capacity of the coconut flour renders it unusable for soudough fermentation, thereby for the baking products produced with an addition of coconut flour it is recommended to use wheat sourdough. On corn flour, none of the studied sourdoughs have developed a pleasant aromatic profile. At the same time, the sourdough based on brown rice flour had pleasant organoleptic properties; the result confirms the need to create microbial compositions of sourdough considering unique properties of each type of flour. Parts of the current research are described in the thesis work of Sergey Sergeevich Gur’ev «Development of microbial starter culture for bakery products with buckwheat grain flour, untreated hydrothermically» in the speciality: 2.7.1 Biotechnology of Food Products and Medicinal and Biologically Active Substances (Engineering)

Publications

1. Gur'ev S., Ivanova V., Korovyansky V., Novichenko A., Kostin I. Rheological properties of wheat dough with the addition of green buckwheat flour E3S Web of Conferences, Vol. 420, pp. 01010 (year - 2023) https://doi.org/10.1051/e3sconf/202342001010