بررسی تاثیر و بهینه‌یابی کمیت فسفات آمونیوم، سولفات پتاسیم و مخمر ساکارومایسز سرویزیه در تولید اسید استیک در فرمانتور ناپیوسته با استفاده از روش سطح پاسخ

نوع مقاله : مقاله پژوهشی

نویسندگان

• مرجان تیموری منش ¹
• هاجر عباسی ²

¹ گروه علوم و صنایع غذایی، دانشکده کشاورزی، واحد اصفهان (خوراسگان)، دانشگاه آزاد اسلامی، اصفهان، ایران

² گروه علوم و صنایع غذایی، دانشکده کشاورزی، واحد اصفهان (خوراسگان)، دانشگاه آزاد اسلامی، اصفهان، ایران.

https://doi.org/10.22067/ifstrj.2021.70315.1043

چکیده

تولید سرکه شامل دو مرحله تخمیر الکلی و اکسیداسیون استیکی می‌باشد. یکی از بارزترین عوامل مؤثر در افزایش راندمان تولید اسید استیک، مواد مغذی موجود در سوبسترا جهت فعالیت بیشتر میکروارگانیسم است. در این پژوهش اثر افزودن فسفات آمونیوم (75/0- صفر گرم)، سولفات پتاسیم (75/0- صفر گرم) و مخمر ساکارومایسز سرویزیه (5/1- صفر گرم) با هدف افزایش راندمان تولید اسید استیک توسط استوباکتر مورد بررسی قرار گرفت. تیمارها با استفاده از روش سطح پاسخ و طرح مرکب مرکزی با چهار نقطه مرکزی ایجاد و پس از سپری شدن مراحل تولید، از نظر اسیدیته اولیه، اسیدیته نهایی، فعالیت، الکل باقی مانده و اندیس اکسیداسیون ارزیابی شدند. شرایط بهینه متغیرهای مستقل مشخص گردید و نتایج حاصل از ارزیابی ویژگی‌های کیفی محصول منتخب (مواد جامد نامحلول، قند احیاء، فنول کل، اسید آسکوربیک، دی‌اکسید گوگرد و فلزات سنگین) با نمونه شاهد مورد مقایسه قرار گرفت. نتایج نشان داد که افزایش غلظت مخمر و فسفات آمونیوم باعث افزایش اسیدیته اولیه و نهایی، فعالیت و اندیس اکسیداسیون و کاهش میزان الکل باقی مانده در محصول می‌شود. اثر افزایش غلظت سولفات پتاسیم خصوصا در سطح بالای مورد بررسی بر اسیدیته اولیه و نهایی، فعالیت و اندیس اکسیداسیون کاهنده ارزیابی گردید. سطوح بهینه متغیرهای مورد بررسی شامل 5/1 گرم مخمر ساکارومایسز سرویزیه، 75/0 گرم فسفات آمونیوم و 38/0 گرم سولفات پتاسیم در نظر گرفته شد. نمونه بهینه تولیدی از نظر اسیدیته اولیه و نهایی، فعالیت، میزان الکل باقی مانده و اندیس اکسیداسیون تفاوت معنی‌داری (05/0p>) با نمونه دارای فعال‌کننده صنعتی (آستازوم) نشان نداد. در حالیکه این نمونه دارای محتوی فنول کل بیشتر (mg/L 5/6) و میزان دی‌اکسید گوگرد (mg/kg 25/6)، سرب (mg/kg 0075/0)، روی (mg/kg 45/0) و مس (mg/kg 225/0) کمتری نسبت به نمونه شاهد بود (05/0p<).

کلیدواژه‌ها

• اسید استیک
• استوباکتر
• ساکارومایسز سرویزیه
• روش سطح پاسخ

عنوان مقاله [English]

Optimization of ammonium phosphate, potassium sulfate and Saccharomyces cerevisiae in the production of acetic acid in a batch fermentor using Response Surface Methodology

نویسندگان [English]

• Marjan Teimorimanesh ¹
• Hajar Abbasi ²

¹ Department of Food Science and Technology, College of Agriculture and Natural Resources, Isfahan (Khorasgan) Branch, Islamic Azad University, Isfahan, Iran.

² Department of Food Science and Technology, College of Agriculture and Natural Resources, Isfahan (Khorasgan) Branch, Islamic Azad University, Isfahan, Iran

چکیده [English]

[1][2]Introduction: Vinegar is an ancient fermented food consumed by human since Babylons period. It is a condiment that produced from various carbohydrate sources by alcoholic and subsequently acetic acid fermentation. Alcoholic fermentation is carried out by Saccharomyces cerevisiae, while the acetic acid fermentation is performed by acetic acid bacteria. Most of the acetic acid bacterial strains are classified in the Acetobacter genus and derived from vinegar factories that are able to oxidize ethanol to acetic acid and some strains over oxidize acetic acid into CO₂ and H₂O (over-oxidation). In acetic acid fermentation, important physical parameters that affect the growth of A.aceti are temperature, aeration and pH. Other most important factors for enhancing production efficiency of acetic acid are the nutrients of substrate for increases microbial activity. Considering the significant role of carbon sources and micronutrients in the fermentation culture to increase the production of acetic acid, in the present study, the effect of adding ammonium phosphate (0- 0.75 g), potassium sulfate (0- 0.75 g) and Saccharomyces cerevisiae (0- 1.5 g), were investigated to increase the production of acetic acid using Acetobacter spp.
 
Materials and methods: Yeast (Saccharomyces cerevisae) was obtained from the Iranian Research Organization for Science and Technology, Department of Biotechnology in Tehran, Iran with code number 5052. All chemicals were from Merck Co. Acetic acid production was performed by Acetobacter spp in a fermenter (Biostat B B.Brun) with a capacity of 10 L which contained 5000 ml of fermentation culture in 9.9 acidity. Mash with a capacity of 3000 ml containing 450 ml of vinegar with 9.9 acidity, 350 ml of ethanol 97%, 2.1 g of dextrose and the rest up to 3000 ml of water was produced and 300 ml of it was injected into the fermenter every 4 hours (in 10 steps). After complete injection, the fermentation operation continued for 9 hours. Agitation speed, aeration rate and temperature in the fermenter were 900 rpm, 50 L/min and 32°C, respectively. Determination of the best conditions for producing acetic acid was performed by Response Surface Methodology (RSM) in the form of central composite design. Independent variables were yeast concentration (0- 1.5 g), concentration of the ammonium phosphate (0- 0.75 g), and the potassium sulfate concentration (0- 0.75 g). RSM models were developed and optimization was done for the highest acidity, activity and oxidation value and the lowest residual alcohol content in the product. Optimal and control samples were examined in terms of qualitative characteristics such as acidity, activity, residual alcohol content, oxidation value, total dissolved solids, reducing sugar, total phenol, ascorbic acid, sulfur dioxide and heavy metals. Comparison of the optimal and control samples was done in a completely randomized design using SAS ver: 9.1 software.
 
Results and discussion: The results showed that increasing the concentration of yeast and ammonium phosphate led to increase the initial and final acidity, activity and oxidation value and decrease the amount of residual alcohol. The effect of increasing potassium sulfate was negative and reduced the initial and final acidity, activity and oxidation value. However, interaction effects of potassium sulfate concentration with other variables at its intermediate levels on reducing the amount of residual alcohol in the product was positive. The optimal levels of the studied variables were determined as 1.5 g of Saccharomyces cerevisiae, 0.75 g of ammonium phosphate and 0.38 g of potassium sulfate. In control sample (without the independent variables), initial and final acidity, activity and oxidation index were lower and the residual alcohol content was higher than the standard level. Therefore, the optimal sample was compared with the control and using industrial activator (Astasome). The optimal sample in terms of initial and final acidity, activity, residual alcohol content and oxidation value had not a significant difference (p> 0.05) with the control sample (containing Astasome). Moreover, the optimal and control samples were not significantly difference in ascorbic acid and reducing sugar (p> 0.05), while the optimal sample in terms of total phenol content, sulfur dioxide and heavy metals like lead, zinc and copper was superior compared to the control sample (p <0.05). Overall, the results of this study indicated the positive effect of yeast, ammonium phosphate, and the potassium sulfate at suitable concentration on qualitative characteristics of acetic acid.
The results revealed that using of appropriate amounts of Saccharomyces cerevisiae, ammonium phosphate and potassium sulfate in substrate as a source of micronutrients improve quality of acetic acid production by Acetobacter spp.
 

کلیدواژه‌ها [English]

• Acetic acid
• Acetobacter
• Saccharomyces cerevisiae
• Response surface methodology

مراجع

1. (2000). Approved Methods of the American Association of Cereal Chemists. 10th ed. Vol 2. American Association of Cereal Chemists, St. Paul, MN; 2000: 02-31.01.
2. Abdolalizadeh, M. S., Vasheghani- Farahani, E., Khodabandeh, M., and Hashemi- Najafabadi, S. (2010). Optimization of Lactic Acid Production Conditions in Batch Fermentation of Whey by Lactobacillus casei. Food Science and Technology (JFST), 7 (2), 96-102.
3. Adachi, O., and Yakushi, T. (2016). Membrane-bound dehydrogenases of acetic acid bacteria. In: Matsushita, K., Toyama, H., Tonouchi, N., and Okamoto-Kainuma, A. editors. Acetic acid bacteria: Ecology and physiology. Tokyo: Springer Japan, pp. 273-97. https://doi.org/10.1007/978-4-431-55933-7_13
4. Aguilar Uscanga, M. G., Escudero Abarca, B. I., Rodriguez, J. G., and Garcia, R. C. (2006). Carbon sources and their effect on growth, acetic acid and ethanol production by brettanomyces bruxellensis in batch culture. Journal of food process Engineering, 30 (1), 13-23. https://doi.org/10.1111/j.1745-4530.2007.00097.x
5. Alben, E., and Erkmen, O. (2004). Production of Citric Acid from a New Substrate, Undersized Semolina, by Aspergillus niger. Food Technol. Biotechnol. 42 (1), 19–22.
6. (2002), Official Methods ofAnalysis of AOAC International, 17th Edn., AOAC International, Maryland.
7. Ataye Salehi, E., Haddad Khodaparast, M. H., Lame, S. H., Habibi Najafi, M. B., and Fatemi, S. H. (2010). Utilization of Date Seed Powder as Supplementary Nutrient in the Alcoholic Fermentation of Cider Vinegar Production. Food Technology and Nutrition, 7 (3), 28-34.
8. Awad, H. M., Diaz, R., Malek, R. A., Othman, N. Z., Aziz, R. A., and EL Enshasy, H. A. (2012). Efficient Production Process for Food Grade Acetic Acid by Acetobacter aceti in Shake Flask and in Bioreactor Cultures. E-Journal of Chemistry, 9 (4), 2275-2286.
9. Balali-Mood, M., Riahi-Zanjani, B., Mahdizadeh, A., Moradi, V., and Fazeli-Bakhtiyari, (2018). Arsenic and Lead Contaminations in Commercial Fruit Juices of Markets in Mashhad, Iran. Iranian Journal of Toxicology, 12, 16-20.
10. Budak, N. H., Aykin, E., Seydim, A.C., Greene, A. K., and Seydim, Z. B. G. (2014). Functional properties of vinegar. Journal of Food Science, 79, 757–764.
11. Cevrimli, B. S., Kariptas, E., and Ciftci, H. (2009). Effects of Fermentation Conditions on Citric Acid Production from Beet Molasses by Aspergillus niger. Asian Journal of Chemistry, 21 (4), 3211-3218.
12. Dahdouh, L., Wisniewski, C., Ricci, J., Vachoud, L., Dornier, M., and Delalonde, M. (2016). Rheological study of orange juices for a better knowledge of their suspended solids interactions at low and high concentration. Journal of Food Engineering, 174, 15-20. https://doi.org/10.1016/j.jfoodeng.2015.11.008
13. Ding , and Tan, T. (2006). L-lactic acid production by Lactobacillus casei fermentation using different fed-batch feeding strategies. Process Biochemistry, 41, 1451-1454. https://doi.org/10.1016/j.procbio.2006.01.014
14. Gorji, M. A., Nahvi Emtiazi, G., and Bidram, H. The study of effective factors in improving the fermentation yield of wheat starch. Pajouhesh and Sazandegi, 74, 79- 89.
15. Hafzan, Y., Saw, J. W. and Fadzilah, I. (2017). Physicochemical properties, total phenolic content, and antioxidant capacity of homemade and commercial date (Phoenix dactylifera ) vinegar. International Food Research Journal, 24 (6), 2557- 2562.
16. Haq, I., Ali, S., Qadeer, M., and Iqbal, J. (2002). Effect of copper ions on mould morphology and citric acid productivity by Aspergillus niger using molasses based media. Process Biochem, 37, 1085-1090. https://doi.org/10.1016/S0032-9592(01)00322-3
17. Institute of Standards and Industrial Research of Iran, Liquid fruit and vegetable products determination of sulphur dioxide content. ISIRI no 4308. 1^st; 2017.
18. Institute of Standards and Industrial Research of Iran, Spice and condiment– Vinegar– Test methods. ISIRI no 1394. 1^st; 2003.
19. Joshi, V. K., Yadav, J., Sharma, R., Joshi, D., and Gupta, R. K. (2016). Effect of Nutrients and Growth Stimulators on Acetic Acid Fermentation using Natural Consortia. International Journal of Food Fermentation Technology, 6 (1), 81-95.
20. Maris, A. J. A., Abbott, D. A., Bellissimi, E., Brink, J., Kuyper, M., Luttik, M. A. H., and Pronk, J. T. (2006). Alcoholic fermentation of carbon sources in biomass hydrolysates by Saccharomyces cerevisiae: Current status. Antonie van Leeuwenhoek, 90, 391–418. https://doi.org/10.1007/s10482-006-9085-7
21. Mazaheri Asadi, M., and Nikkhah Maman, M. (2003). Investigation of citric acid production from date waste using Aspergillus niger. Medical scholar, 11 (47), 57-66.
22. Nadeem, A., Jafri, S. A., Baig, S., Irfan, M., and Syed, Q. (2017). Optimization of Substrate Concentration for Enhanced Citric Acid Production using Aspergillus niger M-101. Pakistan Journal of Scientific and Industrial Research, 52 (5), 275- 277.
23. Nascimento, R. A. d. A., Ferretti, L. P., and Gonçalves, A. R. (2005). Determination of Organic Acids and Ethanol in Commercial Vinegars. Brazilian Journal of Food Technology, 5, 51-6.
24. Ozturk, I., Caliskan, O., Tornuk, F., Ozcan, N., Yalcin, H., Baslar, M., and Sagdic, O. (2015). Antioxidant, antimicrobial, mineral, volatile, physicochemical and microbiological characteristics of traditional home-made Turkish vinegars. LWT– Food Science and Technology, 63, 144–151. https://doi.org/10.1016/j.lwt.2015.03.003
25. Plessi, M. (2003). Vinegar. In Encyclopedia of food Science and nutrition leds B Caballero, Trugo, L.C., and Finge, P. M. Oxford. Academic PP. 5996-6003.
26. Pooja, S., and Soumitra, B. (2013). Optimization of process parameters for vinegar production using banana fermentation. International Journal of Research in Engineering and Technology, 2, 501–514.
27. Rao, P. R., and Reddy, M. K. (2013). Production of Citric Acid by Aspergillus Niger Using Oat Bran as Substrate. International Journal of Chemistry and Chemical Engineering, 3 (3), 181-190.
28. Rashidi, H., and Mazaheri Asadi, M. (2008). Optimization of conditions for citric acid production from date waste using liquid phase method. Microbiological knowledge, 1 (1), 1-11.
29. Robert, W. H. (2006). Microbiology and technology of fermented foods. Blackwell publishing. pp. 397-417.
30. Rosada, K. K. (2018). Enhanced acetic acid production from manalagi apple (Malus sylvestris mill) by mixed cultures of Saccharomyces cerevisiae and Acetobacter aceti in submerged fermentation. Journal of Physics: Conference Series, 1013, 1-7.
31. Sarkar, D., and Das, K. (2017). Optimization of citric acid production from aspergillus niger using pineapple waste as feedstock in submerged fermentation. World Journal of Pharmaceutical Research, 6 (17), 810-818.
32. Sharifi, M., Saeedi Asl, M. R., Haddad Khodaparast, M. H., Jahed, E., Abedinia, A. R., and Sardarian, A. (2015). The investigation of stimulating effects of Kbkab and Mrdasng palm seed powder on Saccharomyces cerevisiae in the apple cider vinegar production. Journal of Innovation in Food Science and Technology, 6 (22), 10-18.
33. Zhang, H., Li, J., Zhang, Q., Zhu, S., Yang, S., and Zhang, Z. (2020). Effect of Substrate Concentration on Photo-Fermentation Bio-Hydrogen Production Process from Starch-Rich Agricultural Leftovers under Oscillation. Sustainability, 12, 1-8.