The importance of much greater thanfunctional foodsmuch less than in the world is increasing, and the procedures for their production are under intense development. The goal of this paper is to optimise the production of a functional food additive based on beetroot juice (Beta vulgaris L.) using brewer's yeast autolysate. In order to improve the nutritive properties of the product and to preserve it, the possibility of beetroot juice fermentation using a Lactobacillus species has been investigated. Comparative investigations of three bacteria cultures (L. plantarum A112, L. acidophilus BGSJ15-3 and L. acidophilus NCDO1748) during fermentation in two media, beetroot juice and a mixture of beetroot juice with an autolysate of brewer's yeast, have been performed. The poorest fermentative activity and growth in both substrates was observed using the L. acidophilus NCDO1748 culture. The two cultures demonstrated better fermentative activity in the mixture of tested substrates, while acidifying activity (production of lactic acid and a decrease in pH) of the L. acidophilus BGSJ15-3 culture was considerably better than that of the L. plantarum A112 culture. L. plantarum A112 culture showed better growth than L. acidophilus BGSJ15-3. From the results obtained, it has been concluded that the L. plantarum A112 and L. acidophilus BGSJ15-3 can be successfully used for fermentation of the mixture of beetroot juice and brewer's yeast autolysate in order to obtain a functional food additive

Baras, Josip

Rakin, M.

Vukašinović-Sekulić, Maja

Food Technology and Biotechnology

The importance of »functional foods« in the world is increasing, and the procedures for their production are under intense development. The goal of this paper is to optimise the production of a functional food additive based on beetroot juice (Beta vulgaris L.) using brewer’s yeast autolysate. In order to improve the nutritive properties of the product and to preserve it, the possibility of beetroot juice fermentation using a Lactobacillus species has been investigated. Comparative investigations of three bacteria cultures (L. plantarum A112, L. acidophilus BGSJ15-3 and L. acidophilus NCDO1748) during fermentation in two media, beetroot juice and a mixture of beetroot juice with an autolysate of brewer´s yeast, have been performed. The poorest fermentative activity and growth in both substrates was observed using the L. acidophilus NCDO1748 culture. The two cultures demonstrated better fermentative activity in the mixture of tested substrates, while acidifying activity (production of lactic acid and a decrease in pH) of the L. acidophilus BGSJ15-3 culture was considerably better than that of the L. plantarum A112 culture. L. plantarum A112 culture showed better growth than L. acidophilus BGSJ15-3. From the results obtained, it has been concluded that the L. plantarum A112 and L. acidophilus BGSJ15-3 can be successfully used for fermentation of the mixture of beetroot juice and brewer’s yeast autolysate in order to obtain a functional food additive

Marica Rakin

Josip Baras

Maja Vukasinovic

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UDC 635.11:663.14:031.23:579.864.1 preliminary communicationISSN 1330-9862(FTB-1249)The Influence of Brewer’s Yeast Autolysate and Lactic AcidBacteria on the Production of a Functional Food AdditiveBased on Beetroot Juice FermentationMarica Rakin*, Josip Baras and Maja VukasinovicFaculty of Technology and Metallurgy, Karnegijeva 4, SCG-11000 Belgrade, Serbia and MontenegroReceived: July 9, 2003Revised version: March 5, 2004Accepted: April 15, 2004SummaryThe importance of »functional foods« in the world is increasing, and the proceduresfor their production are under intense development. The goal of this paper is to optimisethe production of a functional food additive based on beetroot juice (Beta vulgaris L.) usingbrewer’s yeast autolysate. In order to improve the nutritive properties of the product andto preserve it, the possibility of beetroot juice fermentation using a Lactobacillus species hasbeen investigated. Comparative investigations of three bacteria cultures (L. plantarum A112,L. acidophilus BGSJ15-3 and L. acidophilus NCDO1748) during fermentation in two media,beetroot juice and a mixture of beetroot juice with an autolysate of brewer´s yeast, havebeen performed. The poorest fermentative activity and growth in both substrates was ob-served using the L. acidophilus NCDO1748 culture. The two cultures demonstrated betterfermentative activity in the mixture of tested substrates, while acidifying activity (produc-tion of lactic acid and a decrease in pH) of the L. acidophilus BGSJ15-3 culture was consid-erably better than that of the L. plantarum A112 culture. L. plantarum A112 culture showedbetter growth than L. acidophilus BGSJ15-3. From the results obtained, it has been con-cluded that the L. plantarum A112 and L. acidophilus BGSJ15-3 can be successfully used forfermentation of the mixture of beetroot juice and brewer’s yeast autolysate in order to ob-tain a functional food additive.Key words: beetroot, lactic acid fermentation, brewer’s yeast autolysate109M. RAKIN et al.: Beetroot Juice Fermentation, Food Technol. Biotechnol. 42 (2) 109–113 (2004)* Corresponding author; Fax: ++381 11 33 70 387; E-mail: mrakin@Eunet.yu, drmrakin@yahoo.co.ukIntroductionThe aim of the so-called functional foods is to en-sure satisfactory nutritive properties and prevention ofillness, while promoting and extending active humanlife (1–3). In order to obtain these goals, it is necessaryto make substantial modifications in an entire chain offood production, controlling the production accordingto the EU concept of »farm to table« (4–7). Importantcontribution to this goal could be achieved by biologicalprocesses, i.e. biological procedures of food productionand transformation (4,8).As no natural raw material for food production sat-isfies total nutritional requirements, the principle ofcombining two natural raw materials and a bacterial fer-mentation has been applied in this work in order to im-prove nutritional value, the shelf life and the safety ofthe food. In this way, desirable properties have been en-hanced.The goal of this work was to develop the proce-dures for biological processing of beetroot (Beta vulgarisL.) through fermentation using lactic acid bacteria. Beet-root was chosen as a starting substance for the produc-tion of biologically highly valuable food, as there arenumerous publications describing its favourable nutri-tive and protective benefits on humans (9,10).Fermentation using lactic acid bacteria is a wide--spread tradition. However, processing beetroot has notbeen considered in detail. Our previous investigations(11–13) showed that beetroot juice could be successfullyfermented by using lactic acid bacteria, and that success-ful fermentation depended on the selection of bacterialcultures. It has also been established that brewer’s yeastextract favourably promotes fermentative activity of lac-tic acid bacteria, because it is a rich source of nutrientsand biofactors needed by numerous microbial species,and especially lactobacilli (14,15).In this paper we have used a mixture of beetrootjuice and autolysate of brewer’s yeast in equal propor-tion as a starting raw material and subjected this mix-ture to lactic acid fermentation using the Lactobacillusspecies for the sake of improvement of quality and pre-servation. These bacteria were chosen based on our pre-vious investigations, as well as on the fact that they areprobiotics (13,16). The obtained fermented product couldtherefore be catagorised as functional food.Materials and MethodsBacteria culturesFor fermentation of selected substrates the followingcultures have been used: Lactobacillus plantarum A112(Greece), Lactobacillus acidophilus NCDO1748, and a nat-ural isolate, Lactobacillus acidophilus BGSJ15-3, from thecollection of lactic acid bacteria from the Laboratory forMicrobiology of the Faculty of Technology and Metal-lurgy in Belgrade. Tested cultures were propagated inMRS broth. Initial inoculated numbers of viable cells inthe substrates used were 105–107 CFU/mL. Fermenta-tion temperature was 37 °C, while the time of fermenta-tion was 8 h.Fermentation substratesSubstrate S1 was beetroot (Beta vulgaris L.) juice, ob-tained by chopping the fresh vegetable, and separatingthe juice in a juice maker. It was then pasteurised at 70°C for 20 min. Substrate S2 was a mixture of beetrootjuice and brewer’s yeast autolysate prepared in a 1:1mass ratio per dry matter. Brewer’s yeast was first de-bittered with 4 % NaCl, washed (17) and then autolysedat 55 °C for 3 h. Single cultures of L. plantarum A112, L.acidophilus BGSJ15-3 and L. acidophilus NCDO1748 werepropagated in MRS broth. A volume fraction of 2 % ofselected cultures grown in MRS broth was used as ino-culum for the substrates S1 and S2. After the 8-h fermen-tation satisfactory lactic acid production and viable countsin samples were obtained, as described in our previousinvestigations (12,13). A longer time of fermentationwould not be economical for industrial application. Theexperiment was conducted with five combinations ofsubstrates and starter cultures: sample 1 – Substrate S1(beetroot juice) with L. plantarum A112; sample 2 – Sub-strate S1 with L. acidophilus BGSJ15-3; sample 3 – Sub-strate S1 with L. acidophilus NCDO1748; sample 4 – Sub-strate S2 (beetroot juice with autolysate of brewer’s yeast)with L. plantarum A112; sample 5 – Substrate S2 with L.acidophilus BGSJ15-3.AssaysThe progress of fermentation was monitored by de-termination of the number of colony forming units onMRS agar plates using standard decimal dilution me-thod after incubation at 37 °C for 48 h under microaero-philic conditions (18). The following parameters weredetermined over time: change of pH (pH meter), con-centration of lactic acid (potentiometer) (19), sugar con-tent (spectrophotometry, with anthron) (20) and contentof free amino acid nitrogen (spectrophotometric methodwith ninhydrin) (20).The results are expressed as the average values ofthree independent measurements.Results and DiscussionThe data obtained show that the tested bacterial cul-tures, L. plantarum A112, L. acidophilus BGSJ15-3 and L.acidophilus NCDO1748, in two types of substrates, S1 andS2, significantly differ in fermentative activity and growth(Figs. 1-5).L. acidophilus NCDO1748 showed very poor fermen-tative activity in S1. This was shown by the low utilisa-tion of glucose (Fig. 3), small pH drop during the 8-hfermentation (only 0.5 units) and equivalently low pro-duction of lactic acid (Figs. 1 and 2). Assimilation ofsugar and free amino acid nitrogen was also rather low.As the goal of investigation was to attain successfulacidification of beetroot juice (production of lactic acidand decrease of pH value in the tested substrates), thisculture was excluded from further investigations.The other two bacterial cultures exhibited successfulfermentative activity in both tested substrates, and theyshowed differences expressed in all tested parameters.110 M. RAKIN et al.: Beetroot Juice Fermentation, Food Technol. Biotechnol. 42 (2) 109–113 (2004)0 2 4 6 83.03.54.04.55.05.56.06.57.0sample 1sample 2sample 3sample 4sample 5pHFermentation time / hFig. 1. Changes in pH during fermentation in: sample 1 – beet-root juice (S1) with L. plantarum A112; sample 2 – S1 with L.acidophilus BGSJ15-3; sample 3 – S1 with L. acidophilus NCDO1748;sample 4 – beetroot juice and brewer’s yeast autolysate (S2) withL. plantarum A112; sample 5 – S2 with L. acidophilus BGSJ15-3The most successful acidifying activity expressed bythe decrease of pH (from 6 to 4, Fig. 1) and productionof lactic acid (6.6 g/L, Fig. 2) was exhibited by the L.acidophilus BGSJ15-3 culture in substrate S2. This was ac-companied by equivalently high utilisation of sugar fromthe substrate (over 45 %, Fig. 3) and good assimilationof free amino acid nitrogen (24.6 %, Fig. 4). In the samesubstrate, this culture showed considerable growth (1.3log CFU/mL, Fig. 5), while it exhibited poorer fermen-tative activity in substrate S1, where the pH decreasedby 1.6 units (Fig. 1) and the produced lactic acid was 2g/L (Fig. 2). Consequently, utilisation of sugar and freeaminoacid decreased (12.5 %, Fig. 3, and 14.3 %, Fig. 4,respectively).L. plantarum A112 exhibited lower fermentative ac-tivity than L. acidophilus BGSJ15-3 in both tested sub-strates. The decrease in pH of about 1.6 units (Fig. 1)and the production of 2 g/L of lactic acid (Fig. 2) in sub-strate S1 were measured. They were accompanied bylower utilisation of sugar and free amino acids from thesubstrate, which resulted in poor growth of this culture.On the contrary, considering the utilisation of sugar andfree amino acid in substrate S2, this culture exhibited al-most the same activity as the L. acidophilus BGSJ15-3 cul-ture. High utilisation of nutrients resulted in consider-able growth of this culture (2.7 log CFU/mL, Fig. 5).As it can be seen from the data, substrate S2 wasbetter regarding all tested activities of L. plantarum A112and L. acidophilus BGSJ15-3 cultures. The tested culturesshowed similar acidifying activity in substrate S1. Theyare similar in assimilation of sugar and free amino acids,as well as in growth. However, these cultures differedconsiderably in acidifying activity and growth in sub-strate S2. In it, the acidifying activity of L. acidophilusBGSJ15-3, expressed through the production of lacticacid, is 49 % higher (Fig. 2) than that of the L. plantarumA112. On the contrary, L. plantarum A112 in this sub-strate grows better than the L. acidophilus BGSJ15-3 cul-ture by 1.4 log CFU/mL (Fig. 5).By consumption of sugar lactic acid is produced andbacterial biomass, CO2 and energy are created. It is ob-vious that L. acidophilus BGSJ15-3 consumed the sub-111M. RAKIN et al.: Beetroot Juice Fermentation, Food Technol. Biotechnol. 42 (2) 109–113 (2004)0 2 4 6 801234567sample 1sample 2sample 3sample 4sample 5(lacticacid)/(g/L)Fermentation time / hFig. 2. Lactic acid production during fermentation. Samples as inFig. 10 2 4 6 830405060sample 1sample 2sample 3sample 4sample 5(sugars,expresedasglucose)/(g/L)Fermentation time / hFig. 3. Utilisation of sugars (as glucose) during fermentation.Samples as in Fig. 10 2 4 6 8300400500600700sample 1sample 2sample 3sample 4sample 5(freeaminoacidnitrogen)/(mg/L)Fermentation time / hFig. 4. Changes in free amino acid nitrogen during fermentation.Samples as in Fig. 10 2 4 6 856789sample 1sample 2sample 3sample 4sample 5logCFU/(mL)Fermentation time / hFig. 5. Changes in number of colony forming units (CFU) duringfermentation. Samples as in Fig. 1strate for the production of lactic acid (Fig. 2) in the firstplace, while L. plantarum A112 consumed it for thegrowth of cells.The results obtained through investigations in thispaper have confirmed that the extract of brewer’s yeasthas a stimulating effect on acidifying activity and growthof lactic acid bacteria. This is in accordance with ourprevious results and literature data as well (13,21,22).The positive influence of brewer’s yeast autolysate canbe explained by the fact that it contains many nutrientsand biofactors used by most microorganisms, especiallythose like lactic acid bacteria, which are dependant onthe organic sources of nitrogen, and vitamins, particu-larly those from the B group (14,15,23).The product obtained in this investigation can beused as a functional additive in a form of a beverage af-ter pasteurisation, or else it may be dried and used inform of powder or tablets. Our previous investigationsshowed that similar products could be dried using thetechnique of fluidised bed drying, when about 30 % ofbacterial cells of genera Lactobacillus and Bifidobacteriumsurvive (5). Such a product is of a particular importance,as the strains from the above mentioned bacterial spe-cies possess probiotic properties important for humanhealth. Further investigations initiated in this paper haveas their goal an investigation of the fermentation of com-bined beetroot juice and brewer’s yeast autolysate withmixed cultures of L. plantarum A112 and L. acidophilusBGSJ15-3, as well as the investigation of the possibilityof drying the obtained product using the technique offluidised bed drying.ConclusionBased on the results obtained by comparative inves-tigation of fermentative activity of three lactic acid bac-teria cultures, L. plantarum A112, L. acidophilus BGSJ15-3and L. acidophilus NCDO1748, in substrates S1 and S2, thefollowing has been established: good fermentative activ-ity and growth in both substrates were shown by twocultures, L. plantarum A112 and L. acidophilus BGSJ15-3;considerably better results were obtained in substrate S2,which confirms that brewer’s yeast autolysate stimu-lated the activity of those bacterial cultures. Therefore,bacterial cultures L. plantarum A112 and L. acidophilusBGSJ15-3 can be successfully used for fermentation of amixture of beetroot juice and brewer’s yeast autolysatein order to obtain a functional food additive.References1. M. B. Roberfroid, Am. J. Clin. Nutr. 71 (2000) 1660–1664.2. M. B. Roberfroid, Am. J. Clin. Nutr. 71 (2000) 1682–1687.3. E. Sloan, Food Technol. 54 (2000) 33–62.4. S. Dimitrijevic, J. Baras, J. Bojovic, Chemical Industry of Bel-grade, 56 (2002) 113–122.5. J. Baras, S. Dimitrijevic, D. Povrenovic, L. Turubatovic, Bio-technological Processes in Modification of Agricultural Pro-ducts – 6th International Conference, Moscow (2002) 279–298.6. P. Raspor, Croatian Congress of Food Technologists, Biotech-nologists and Nutritionists, Central European Meeting, Opa-tija (2001) FQ/KL 01.7. T. Lovri}, Croatian Congress of Food Technologists, Biotech-nologists and Nutritionists, Central European Meeting, Opa-tija (2001) DTTO, FT/PL 01.8. V. Mari}, Croatian Congress of Food Technologists, Biotechno-logists and Nutritionists, Central European Meeting, Opatija(2001) DTTO, BT/PL 03.9. J. Kaner, S. Harel, R. Granit, J. Agric. Food Chem. 49 (2001)5178–5185.10. G. J. Kapaida, M. A. Azuine, R. Sridhar, Y. Okuda, A. Tsu-ruta, E. Ichiishi, T. Mukainake, M. Takasaki, T. Konoshi-ma, H. Nishino, H. Tokuda, Pharmacol. Res. 47 (2003) 141–148.11. J. Baras, S. Dimitrijevic, M. Rakin, B. Stevovic, Acta Period.Technol. 31 (2000) 609–615.12. J. Baras, M. Rakin, S. Dimitrijevic, B. Stevovic, 2nd Interna-tional Conference of the Chemical Societies of the South-EasternEuropean Countries on Chemical Sciences for Sustainable Deve-lopment, Book of Abstracts, Greece (2000) p. 132.13. S. Dimitrijevic-Brankovic, J. Baras, J. Serb. Chem. Soc. 66(2001) 581–589.14. J. Baras: Optimisation of Manner for Production and Applica-tion of Brewer’s Yeast Autolysate and Hydrolysate, FederalMinistry of Science, Development and Ecology, Belgrade(2000).15. J. Dziezak, Food Technol. 41 (1987) 122–125.16. A. M. P. Gomes, F. X. Malcata, Trends Food Sci. Technol.109 (1999) 139–157.17. B. K. Dwiwedi, D. L. Gibson, J. Inst. Can. Technol. Aliment.3 (1970) 110–112.18. C. H. Collins, P. M. Lynne: Microbiological Methods, Butter-worth, London (1985).19. D. Herbert, P. J. Phipps, R. E. Strange, Methods Enzymol.5B (1971) 265–308.20. J. Trajkovic, M. Miric, J. Baras, S. Siler: Analyses of FoodProducts, Faculty of Technology and Metallurgy, Belgrade(1983).21. A. Aeschlimann, U. von Stockar, Appl. Microbiol. Biotech-nol. 32 (1990) 398–402.22. V. Arastratam, A. Senthuran, K. Balasubramaniam, EnzymeMicrob. Technol. 19 (1996) 482–486.23. F. J. Carr, D. Chill, N. Maida, Crit. Rev. Microbiol. 28 (2002)281–370.112 M. RAKIN et al.: Beetroot Juice Fermentation, Food Technol. Biotechnol. 42 (2) 109–113 (2004)Utjecaj autolizata pivskoga kvasca i laktobacila na vrenje soka cikleza proizvodnju dodatka funkcionalnoj hraniSa`etakBudu}i da funkcionalna hrana u svijetu ima sve ve}e zna~enje i da se intenzivno raz-vijaju postupci za njezinu proizvodnju, svrha je ovoga rada bila doprinos optimiranju pos-tupka proizvodnje dodatka funkcionalnoj hrani na bazi soka cikle (Beta vulgaris L.) koris-te}i autolizat pivskoga kvasca. Da bi se pobolj{ala hranjiva i za{titna svojstva proizvoda teda bi se on mogao konzervirati, ispitana je mogu}nost fermentacije soka cikle bakterijamamlije~no-kiselog vrenja iz roda Lactobacillus. Provedena su komparativna ispitivanja trijubakterijskih kultura L. plantarum A112, L. acidophilus BGSJ15-3 i L. acidophilus NCDO1748,tijekom fermentacije u dvije podloge, u soku cikle te mje{avini soka cikle i autolizata piv-skoga kvasca u jednakim omjerima. Najslabiju fermentativnu aktivnost i rast u oba sup-strata pokazala je kultura L. acidophilus NCDO1748. Ostale su dvije kulture imale boljufermentativnu aktivnost u smjesi ispitivanih supstrata, pri ~emu je acidifikacijska aktivnost(proizvodnja mlije~ne kiseline i snizivanje pH-vrijednosti) kulture L. acidophilus BGSJ15-3bila kudikamo bolja od kulture L. plantarum A112. [to se ti~e rasta navedene su se kulturepona{ale obrnuto. Na osnovi dobivenih rezultata zaklju~eno je da se ispitivane kulture L.plantarum A112 i L. acidophilus BGSJ15-3 mogu uspje{no koristiti za fermentaciju mje{avinesoka cikle i autolizata pivskoga kvasca radi dobivanja dodatka funkcionalnoj hrani.113M. RAKIN et al.: Beetroot Juice Fermentation, Food Technol. Biotechnol. 42 (2) 109–113 (2004)

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The Influence of Brewer’s Yeast Autolysate and Lactic Acid Bacteria on the Production of a Functional Food Additive Based on Beetroot Juice Fermentation

Rakin, Marica

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The influence of brewer's yeast autolysate and lactic acid bacteria on the production of a functional food additive based on beetroot juice fermentation

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The influence of brewer's yeast autolysate and lactic acid bacteria on the production of a functional food additive based on beetroot juice fermentation

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