This study aims to determine the effect of fermentation condition and Saccharomyces strains on the pH, foam property and CO2 concentration of non-alcoholic beer (Ma-al-shaeer). For this, the beer samples were inoculated with four different species of Saccharomyces (Saccharomyces rouxii 70531, S. rouxii 70535, S. ludwigii 3447 and S. cerevisiae 70424) and fermented for 48h in both aerobic and periodic aeration at three different temperatures. Then their pH, CO2 concentration and foam property were analyzed in 12h intervals during 48h fermentation. The results shows that the treatments with 4×107 CFU.ml-1 and periodic aeration at 24˚C showed the greatest decrease in pH, and the treatments with 107 CFU.ml-1 and aerobic-periodic aeration at 4˚C showed the lowest decrease in pH. The highest and lowest amounts of CO2 and foam property were obtained in the treatments with 4×107 CFU.ml-1 inoculation, aerobic condition, and the treatments with 107 CFU.ml-1, periodic aeration, respectively. These results further demonstrated that the highest drop in pH, and the highest ability of producing CO2 and foam were for S. cerevisiae 70424, and the lowest belonged to S. rouxii 70531. The overall outcome of the study points to the fact that physico-chemical properties of Ma-al-shaeer is important from the consumers' point of view. Therefore, S. cerevisiae with 4×107 CFU.ml-1 inoculation and aerobic condition at 4˚C has promising potential for producing Ma-al-shaeer with good physicochemical properties

Malganji, S.H.

Mousavi, S.M.

Razavi, S.H.

Sohrabvandi, S.

English

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   Short communication  APPLIED FOOD BIOTECHNOLOGY, 2015, 2(1): 53-57 Journal's homepage: www.journals.sbmu.ac.ir/afb  pISSN: 2345-5357    The Effect of Saccharomyces Strains and Fermentation Condition on the pH, Foam Property and CO2 Concentration of Non-alcoholic Beer (Ma-al-shaeer) Sohrabvandi, S.1, Malganji, S.H.2, Razavi, S.H. 3٭ , Mousavi, S.M.3 1- Department of Food Technology Research, National Nutrition and Food Technology Research Institute, Faculty of Nutrition Sciences, Food Science and Technology, Shahid Beheshti University of Medical Sciences, Tehran, Iran. 2- Department of Food Science and Technology, Faculty of Food Science and Technology, Islamic Azad University, Khorasgan (Isfahan) Branch, Isfahan, Iran. 3- Department of Food Science, Engineering & Technology, Faculty of Agricultural Engineering and Technology, University of Tehran, Karaj, Iran.   Abstract  Article Info This study aims to determine the effect of fermentation condition and Saccharomyces strains on the pH, foam property and CO2 concentration of non-alcoholic beer (Ma-al-shaeer). For this, the beer samples were inoculated with four different species of Saccharomyces (Saccharomyces rouxii 70531, S. rouxii 70535, S. ludwigii 3447 and S. cerevisiae 70424) and fermented for 48h in both aerobic and periodic aeration at three different temperatures. Then their pH, CO2 concentration and foam property were analyzed in 12h intervals during 48h fermentation. The results shows that the treatments with 4×107 CFU.ml-1 and periodic aeration at 24˚C showed the greatest decrease in pH, and the treatments with 107 CFU.ml-1 and aerobic-periodic aeration at 4˚C showed the lowest decrease in pH. The highest and lowest amounts of CO2 and foam property were obtained in the treatments with 4×107 CFU.ml-1 inoculation, aerobic condition, and the treatments with 107 CFU.ml-1, periodic aeration, respectively. These results further demonstrated that the highest drop in pH, and the highest ability of producing CO2 and foam were for S. cerevisiae 70424, and the lowest belonged to S. rouxii 70531. The overall outcome of the study points to the fact that physico-chemical properties of Ma-al-shaeer is important from the consumers' point of view. Therefore, S. cerevisiae with 4×107 CFU.ml-1 inoculation and aerobic condition at 4˚C has promising potential for producing Ma-al-shaeer with good physicochemical properties.           Article history Received     3 Apr 2014 Revised    21 May 2014 Accepted  28 May 2014   Keywords Fermentation; Physicochemical properties; Saccharomyces; Non-alcoholic beer (Ma-al- shaeer)   Correspondence to Seyed Hadi Razavi, Department of Food Science, Engineering & Technology, Faculty of Agricultural Engineering and Technology, University of Tehran, Karaj, Iran. P.O. Box: 31587-77871. Email: srazavi@ut.ac.ir  1. Introduction Ma-al-shaeer (MAS) is a kind of carbonated beverage (non-alcoholic) based on malt, which contains inorganic salts, hops and CO2 gas (carbonic acid forms), as well as several organic compounds with health benefits [1, 2]. There are some ideal properties of MAS, which would benefit human health such as increasing HDL cholesterol and preventing cardiovascular dis-ease, and inhibitory effect on the growth of cancer cells and osteoporosis [3-6]. It has been determined that organic acids in MAS result in the reduction of uric acid [7, 8]. Furthermore,  high concentration  of  proteins  and  vitamin  B increases the nutritional value of MAS [9]. MAS has some physical properties including CO2 retention and foam stability, and some chemical properties such as acidity, pH and different chemicals. Moreover, this beverage consists of gas (CO2) and liquid phases. Polypeptides, glycoproteins, peptides, poly phenols and dextrin, which are categorized as colloidal, are responsible for production of CO2 and foam head. Glycoproteins contain a polar sector that entraps CO2 bubbles, and a non-polar The effect of Saccharomyces strains and fermentation … 54 Appl Food Biotechnol, Vol. 2, No. 21 (2015)             sector, which creating local high viscosity, which is effective in foam production. It has been indicated that the type of used malt determines the type of glycoproteins, and the latter, stabilizes foam. Dextrin, polypeptide fragments and some gums play an important role  in foam stability [10]. In spite of their ability in extracting protein fragments from malt, these fragments do not have fermentability. Therefore, their ability of foam production is improved by adding glycerol or yeasts, which produce high amount of alcohol sugars. Concentration of 1.5-4% of glycerol has been reported as a sufficient amount for achieving optimum foam body, and desirable flavor [11]. pH of MAS differs from 3.9 to 4.5. Water soluble materials in malt and pH of water will affect wort pH, which is about 5-6. Moreover, organic acids produced by yeasts and carbonic acid gas reduce the pH of the solution. On the other hand, some minerals consumed by yeasts and autolysis of yeast cells have resulted in an increase in pH [10]. Due to the important role of physicochemical properties of MAS in consumer acceptance, the objective of this study was to determine the effect of Saccharomyces strains and fermentation condition on some physic-chemical properties of MAS.  2. Materials and Methods 2.1 Sample preparation Saccharomyces (S.) cerevisiae 70424, S. ludwigii 3447, S. rouxii 70535 and S. rouxii 70531 were supplied by the DSMZ Company (Braunscheig, Germany). Yeasts were inoculated with two different inoculation levels (107 CFU.ml-1 and 4×107 CFU.ml-1) to the wort. Then the wort was fermented for 48h under aerobic and periodic aeration (12h interval) at 4˚C, 12˚C and 24˚C.                                      Figure 1. pH drop during the fermentation (variables F, S, P and N and numbers show 4×107 CFU.ml-1 inoculation, 107 CFU.ml-1 inoculation, periodic aeration, anaerobic condition, and fermentation temperature, respectively). Appl Food Biotechnol, Vol. 2, No. 1 (2015) 55       2.2 Chemical analysis A digital pH meter (Mettler, MA 235, Swit- zerland) was used for pH measurement. CO2 concentration was measured by a pressure gauge (TG, Italy). Head space of the samples was calculated using pressure temperature table related to CO2 at a constant temperature. In order to measure the foam property, the samples were poured into cylinder and their foam property (percentage of foam) was calculated by the following equation: (foam • head height/total height)×100= foam% (ml) 2.3 Statistical analysis The experiments were carried out in triplicate. The results were analyzed by ANOVA Software (SAS 9.1 software Institute Inc., Cary, NC, USA), and expressed as mean±standard deviation.  3. Results and discussion Figure 1 shows the changes in pH during fermentation.   The   highest   drop   in   pH   was  mentioned for FP-24 and FN-24 (4×107 CFU.ml- 1, aerobic-periodic aeration, 24˚C). During the fermentation, the pH of treatments with aerobic and periodic aeration did not show a significant difference. According to FP-4 and FN-4, due to low temperature of fermentation during the yeast metabolism, different types of organic acids are produced. Also nitrogen compounds are consumed by the yeasts in order to grow. Moreover, CO2 is produced during the anaerobic fermentation. Thus, these factors led to a pH drop during the fermentation [10]. In the anaerobic metabolism of yeasts, carboxylic acids are produced by incomplete three carboxylic acids and in the aerobic metabolism, carbohydrates are converts to energy, water and CO2 [11]. It was reported that, S. rouxii 70531 showed the lowest drop in pH, which is due to lower ethanol production of this yeast in comparison to other yeasts during both aerobic and periodic aeration metabolisms. High speed of ethanol production and aerobic yeast metabolism causes a pH drop.    Figure 2. CO2 concentration during the fermentation (variables F, S, P and N and numbers show 4×107 CFU.ml-1 inoculation, 107 CFU.ml-1 inoculation, periodic aeration, anaerobic condition and fermentation temperature, respectively). Sohrabvandi et al.  The effect of Saccharomyces strains and fermentation … 56 Appl Food Biotechnol, Vol. 2, No. 1 (2015)         Figure 3. Foam property during the fermentation (variables F, S, P and N and numbers show 4×107 CFU.ml-1 inoculation, 107 CFU.ml-1 inoculation, periodic aeration, anaerobic condition and fermentation temperature, respectively).    Figure 2 and 3 show the CO2 concentration and foam property of treatments. The highest and lowest concentrations of CO2 and foam property were obtained in the treatments with 4×107 CFU.ml-1 in aerobic condition (FN24), and the treatments with 107 CFU.ml-1 in periodic aeration (SP4), respectively. Fermentation temperature did not show significant difference. S. cerevisiae 70424 and S. rouxii 70531 showed the highest and the lowest ability in producing CO2 and foam, respectively. Based on the results, the amount of ethanol has a reinforcing effect on the treatments, CO2 and foam property. This effect takes place with protein denaturation and appropriate foaming agents in the presence of ethanol [10, 12]. However, in the treatments with high amount of ethanol, CO2 is produced in greater amounts because this gas is the product of alcoholic fermentation [13, 14]. FN-24 and SP-4 showed the highest and lowest amount of CO2 and foam, respectively. The worth noting point is the effect of     ethanol     on     foam     production     and synchronization of ethanol production with CO2, which is produced during the alcoholic fermentation.  4. Conclusions Four species of Saccharomyces were inoculated into MAS, and the samples were fermented for 48h under both aerobic and periodic aerations at three different temperatures. pH, foam property and CO2 concentration were analyzed during the fermentation. The results of this research suggest that the physical properties of MAS such as foam stability and production of CO2 gas play an important role in the consumers' acceptability. Therefore, production of MAS by S. cerevisiae with 4×107 CFU.ml-1 inoculation, fermented at 4˚C under anaerobic condition, is suggested.  References 1. Fillaudeau L, Boissier B,  Moreau A, Blanpain-avet P, Ermolaev S, Jitariouk N, et al.  Investigation  of  rotating  and  vibrating Appd Food Biotechnol, Vol. 2, No. 1 (2015) 57   Sohrabvandi et al.   filtration  for  clarification  of  rough  beer.  J Food Eng. 2007; 80: 206-217. 2. Moll MM: Water. In: Hardwick WA: Handbook of Brewing, Marcel Dekker: New York, 1995: 133-156. 3. Gorinstein S Caspi A, Zemser M, Tarkthenberg S. Comprative content of some phenolics in beer, red and white wines. Nutr Res. 2000; 20: 131-139. 4. Nardini M, Ghiselli A. Determination of free and bound phenolic acids in beer. Food Chem. 2004; 84: 137-143. 5. Ghiselli A, Natella F, Guidi A, Montanari L, Fantozzi P, Scaccini C. Beer increases plasma antioxidant capacity in humans. J Nutr and Biochem. 2000; 11: 76-80. 6. Lapcik O, Hill M, Hampl R, Wahala K, Adlercreutz H. Identification of isoflavonoids in beer. Steroids. 1998; 63: 14-20. 7. Montanari L, Perretti G, Natella F, Guidi A, Fantozzi P. Organic and phenolic acids in beer. Lebensm-wiss. U Technol. 1999; 32: 535-539. 8. Shibano Y, Yomo H, Matsumoto I, Koda H, Suwa Y, Amachi T, et al. Process for manufacturing beer. US Patent. No. 6,013,288. 2000. 9. Pinho O, Ferreira I, Santos L. Method optimization by solid-phase microextraction in combination with gas chromatography with mass spectrometry for analysis of beer volatile fraction. J Chromatogr A. 2006; 1121: 45-153. 10. Hardwick WA. The properties of beer. In: Hardwick WA: Handbook of Brewing, Marcel Dekker: New York, 1995; 551-586. 11. Dziondziak K. Method for the production of low-alcohol or alcohol-free beer. US Patent. No. 4,814,188. 1989. 12. Munroe JH: Fermentation. In: Hardwick WA: Handbook of Brewing, Marcel Dekker: New York, 1995; 323-354. 13. Briggs DE, Boulton CA, Brookes PA, Stevens R. Metabolism of wort by yeast. Brewing science and practice. Elsevier, 2004; 87-92. 14. Lewis MJ, Young TW. Brewing. Chapman and Hall, London, 2002; 45-60. 

The Effect of Saccharomyces Strains and Fermentation Condition on the pH, Foam Property and CO2 Concentration of Non-alcoholic Beer (Ma-al-shaeer)

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