Use of soybean molasses in the cultivation of Aspergillus niger strain (F-1270) for citric acid synthesis
The high cost of culture media and their import are limiting factors in the production of citric acid. Therefore, the current challenge is to find inexpensive ingredients to create culture media that ensure high yields. More than 80% of citric acid is microbiologically produced. The study aims to investigate the possibility of using soybean molasses to intensify the synthesis of citric acid by Aspergillus niger. In this study, the A. niger strain (F-1270) was investigated for its ability to synthesize citric acid under conditions of modification of the nutrient medium with a carbohydrate-containing component (soybean molasses). During the research, the method of deep cultivation of A. niger in modified nutrient media was used. High-performance liquid chromatography was used to study the effectiveness of the process. Results showed that the studied strain of A. niger can actively grow and produce citric acid in a nutrient medium containing soybean molasses as a carbohydrate source. During the optimization of the biosynthesis process, the recommended values of technological parameters (duration 7 days, temperature 36.0°C, pH 2.0, and content of soybean molasses in the medium 20.0 g/L) were determined at which the yield of citric acid was 49.8 g/L, which is 18% higher than the citric acid yield (42.14 g/L) obtained using a classic culture medium (temperature 26°C, duration 7 days, pH 4.0, sucrose concentration 30.0 g/L). To increase citric acid productivity, it is necessary to improve the performance of A. niger strains by classical mutagenesis and selection.
Babich O, Shipilov G, Budenkova E, Sukhikh S. Use of soybean molasses in the cultivation of Aspergillus niger strain (F-1270) for citric acid synthesis. J Appl Biol Biotech 2026. https://doi.org/10.7324/JABB.2026.272484
1. Ksi??ek E. Citric acid: Properties, microbial production, and applications in industries. Molecules. 2024;29(1):22. https://doi.org/10.3390/molecules29010022
2. Tong Z, Zheng X, Tong Y, Shi YC, Sun J. Systems metabolic engineering for citric acid production by Aspergillus niger in the post-genomic era. Microb Cell Fact. 2019;18:28. https://doi.org/10.1186/s12934-019-1064-6
3. Steiger MG, Rassinger A, Mattanovich D, Sauer M. Engineering of the citrate exporter protein enables high citric acid production in Aspergillus niger. Metab Eng. 2019;52:224-31. https://doi.org/10.1016/j.ymben.2018.12.004
4. Papagianni M. Advances in citric acid fermentation by Aspergillus niger: Biochemical aspects, membrane transport and modeling. Biotechnol Adv. 2007;25(3):244-63. https://doi.org/10.1016/j.biotechadv.2007.01.002
5. Show PL, Oladele KO, Siew QY, Aziz Zakry FA, Lan JC, Ling TC. Overview of citric acid production from Aspergillus niger. Front Life Sci. 2015;8:271-83. https://doi.org/10.1080/21553769.2015.1033653
6. Ali S, Haq I, Qadeer MA, Iqbal J. Production of citric acid by Aspergillus niger using cane molasses in a stirred fermentor. Electron J Biotechnol. 2002;5:125-30. https://doi.org/10.2225/vol5-issue3-fulltext-3
7. Schopf N, Erbino P, Puvogel A. Alternative fuels: Energetic use of liquid by-products from sugar and soy processing. Sugar Tech. 2014;16:333-8. https://doi.org/10.1007/s12355-013-0291-0
8. Gambarato BC, Carvalho AK, De Oliveira F, Da Silva SS, Da Silva ML, Bento HB. Soy molasses: A sustainable resource for industrial biotechnology. Sustainability. 2025;17(12):5667. https://doi.org/10.3390/su17125667
9. Wang ZP, Zhang LL, Liu S, Liu XY, Yu XJ. Whole conversion of soybean molasses into isomaltulose and ethanol by combining enzymatic hydrolysis and successive selective fermentations. Biomolecules. 2019;9(8):353. https://doi.org/10.3390/biom9080353
10. Nile SH, Venkidasamy B, Samynathan R, Nile A, Shao K, Chen T, et al. Soybean processing wastes: Novel insights on their production, extraction of isoflavones, and their therapeutic properties. J Agric Food Chem. 2021;70(23):6849-63. https://doi.org/10.1021/acs.jafc.1c04927
11. Rodrigues BC, De Mello BS, Da Costa Araujo ML, Da Silva GH, Sarti A. Soybean molasses as feedstock for sustainable generation of biomethane using high-rate anaerobic reactor. J Environ Chem Eng. 2021;9:105226. https://doi.org/10.1016/j.jece.2021.105226
12. Fernandes S, Dias B, Belo I, Lopes M. Soybean molasses as a new and low-cost substrate for gluconic acid production by Aspergillus niger. Process Biochem. 2024;146:241-9. https://doi.org/10.1016/j.procbio.2024.07.034
13. Rakita S, Banjac V, Djuragic O, Cheli F, Pinotti L. Soybean molasses in animal nutrition. Animals (Basel). 2021;11(2):514. https://doi.org/10.3390/ani11020514
14. Correa Da Silva AP, Dorigan BS, Da Silva-Neto JM, Rosa-Magri MM, Rossi F, Francisco KR, et al. Soy molasses as culture medium for Bacillus species aiming at plant growth promotion. Fermentation. 2024;10(8):403. https://doi.org/10.3390/fermentation10080403
15. Acosta SB, Marchioro ML, Santos VA, Calegari GC, Lafay CB, Barbosa-Dekker AM, et al. Valorization of soybean molasses as fermentation substrate for the production of microbial exocellular β-glucan. J Polym Environ. 2020;28:2149-60. https://doi.org/10.1007/s10924-020-01758-z
16. Caldeirao L, Tanaka C, Ida E, Spinosa W. Modeling and kinetic study of bio-ethanol production from soy protein concentrate by-product. J Food Sci Technol. 2016;36:369-74. https://doi.org/10.1590/1678-457X.0021
17. Cheng C, Zhou Y, Lin M, Wei P, Yang ST. Polymalic acid fermentation by Aureobasidium pullulans for malic acid production from soybean hull and soy molasses: Fermentation kinetics and economic analysis. Bioresour Technol. 2017;223:166-74. https://doi.org/10.1016/j.biortech.2016.10.042
18. Miranda LCR, Gomes RJ, Mandarino JM, Ida EI, Spinosa WA. Acetic acid fermentation of soybean molasses and characterisation of the produced vinegar. Food Technol Biotechnol. 2020;58:84-90. https://doi.org/10.17113/ftb.58.01.20.6292
19. Rodrigues C, De Souza Vandenberghe LP, Teodoro J, Pandey A, Soccol CR. Improvement on citric acid production in solid-state fermentation by Aspergillus niger LPB BC mutant using citric pulp. Appl Biochem Biotechnol. 2009;158:72-87. https://doi.org/10.1007/s12010-008-8370-5
20. Food and Agriculture Organization of the United Nations (FAO). Global Food Losses and Food Waste -Extent, Causes and Prevention. Available from: https://www.fao.org/3/ai2697e.pdf [Last accessed on 2011 May 04].
21. Khurshid S, Ashraf H, Hussain T, Iqbal M, Qureshi H, Anwar T, et al. Enhanced citric acid production through Aspergillus niger: Insights from fermentation studies using sugarcane molasses. Life. 2024;14(6):756. https://doi.org/10.3390/life14060756
22. Madhusudan C, Rishi CM, Singh M. Citric acid production from cane molasses using submerged fermentation by Aspergillus niger ATCC9142. J Pharm Res. 2010;3(6):1215-22.
23. Amin A, Razu A, Ismail M, Islam TN, Mohasin M. Optimization of Citric Acid Production from Sugarcane Molasses using Aspergillus niger by Submerged Fermentation. Biores Commun. 2024;11(1):1643-50. https://doi.org/10.3329/brc.v11i1.78880
24. Chernysheva EA. Trends and Prospects for Realizing Russia’s Export Potential in the Global Sugar Market. Specialty -08.00.14 -Global Economy. [Dissertation for the Degree of Candidate of Economic Sciences]. Moscow; 2021. p. 198.
25. Kalashnikova O, Pankova E, Sukhikh S, Babich O, Samusev I, Tcibulnikova A, et al. Production of bacterial cellulose using a symbiotic consortium of bacteria and yeast on soybean molasses medium. LWT. 2024;205:116480. https://doi.org/10.1016/j.lwt.2024.116480
26. Order of the Ministry of Agriculture of the Russian Federation. On Approval of Programs for the Promotion and Increase in Export Volumes of Certain Types of Products of the Agro-Industrial Complex, Including Products of Micro-Small and Medium-Sized Enterprises of the Agro-Industrial Complex. Russian: Order of the Ministry of Agriculture of the Russian Federation; 2017.
27. Quan ZX, Jin YS, Yin CR, Lee JJ, Lee ST. Hydrolyzed molasses as an external carbon source in biological nitrogen removal. Bioresour Technol. 2005;96(15):1690-5. https://doi.org/10.1016/j.biortech.2004.12.033
28. Martynets MV, Mezenova NY. Use of soy molasses in the formulation of bakery products of increased nutritional value. Bull Youth Sci. 2018;3(15):9.
29. Sancheti A, Thompson ER, Ju LK. Factors influencing the enzymatic hydrolysis of soy molasses into fermentation feedstock. Enzyme Microb Technol. 2023;170:110302. https://doi.org/10.1016/j.enzmictec.2023.110302
30. GOST 31640-2012. Food. Methods for the Determination of Dry Matter. Vol. 11. Moscow, Standartinform; 2012.
31. GOST R 52304-2005. Beet Molasses. Technical Specifications. Vol. 23. Moscow: IPK Publishing House of Standards; 2005.
32. Mangwanda TW, Mani JS, Johnson JB, Jackson S, McKeown T, Naiker M. Physicochemical and nutritional analysis of molasses for rum fermentation. Biol Life Sci Forum. 2023;26(1):105. https://doi.org/10.3390/Foods2023-15137
33. GOST 13496.15-2016. Feed, Mixed Feed, Feed Raw Materials. Methods for Determining the Mass Fraction of Crude Fat. Vol. 12. Moscow: Standartinform; 2016.
34. Ramesh T, Kalaiselvam M. An experimental study on citric acid production by Aspergillus niger using Gelidiella acerosa as a substrate. Indian J Microbiol. 2011;51:289-93. https://doi.org/10.1007/s12088-011-0066-9
35. Hauka FI, El-Sawah MM, Kassem MM, El-Kady SM. Some Factors Affecting Citric Acid Production from Sugar Cane Molasses by Aspergillus niger. In: The 2nd Arab Mansoura Conference on Food and Dairy Science and Technology; 2005. p. 22-4.
36. Patent 2091485 RU. Aspergillus niger Fungus Strain is a Producer of Citric Acid/P.B. Avchieva V.N. Yashina. The Patent Holder is the State Scientific Research Institute of Biosynthesis of Protein Substances; 1995.
37. Acourene S, Ammouche A. Optimization of ethanol, citric acid, and α-amylase production from date wastes by strains of Saccharomyces cerevisiae, Aspergillus niger, and Candida guilliermondii. J Ind Microbiol Biotechnol. 2012;39(5):759-66. https://doi.org/10.1007/s10295-011-1070-0
38. El-Samragy YA, Khorshid MA, Foda MI, Shehata AE. Effect of fermentation conditions on the production of citric acid from cheese whey by Aspergillus niger. Int J Food Microbiol. 1996;29(2-3):411-6. https://doi.org/10.1016/0168-1605(95)00072-0
39. Pujol-Vila F, Vigués N, Díaz-González M, Munoz-Berbel X, Mas J. Fast and sensitive optical toxicity bioassay based on dual wavelength analysis of bacterial ferricyanide reduction kinetics. Biosens Bioelectron. 2015;67:272-9. https://doi.org/10.1016/j.bios.2014.08.030
40. Utkin DV, Germanchuk VG, Erokhin PS, Spitsyn AN, Shcherbakova SA, Glazkov AN. Application of spectrophotometric analysis methods for detection of microorganisms. Bull Saratov Univ. 2014;14(2):36-7.
41. Rodrigues BC, Mello BS, Araujo ML, Silva GH, Sarti A. Soybean molasses as feedstock for sustainable generation of biomethane using high-rate anaerobic reactor. J Environ Chem Eng. 2021;9:105226. https://doi.org/10.1016/j.jece.2021.105226
42. Mordenti AL, Giaretta E, Campidonico L, Parazza P, Formigoni A. A review regarding the use of molasses in animal nutrition. Animals (Basel). 2021;11:115. https://doi.org/10.3390/ani11010115
43. Mangwanda T, Johnson JB, Mani JS, Jackson S, Chandra S, McKeown T, et al. Processes, challenges and optimisation of rum production from molasses -A contemporary review. Fermentation. 2021;7:21. https://doi.org/10.3390/fermentation7010021
44. Javed S, Asgher M, Sheikh MA, Nawaz H, Jamil A. Enhanced citric acid production by Aspergillus niger EB-3 mutant using an inert solid support in molasses medium. Afr J Biotechnol. 2011;10:11784-91.
45. Yaderets VV, Karpova NV, Glagoleva EV, Shibaeva AS, Dzhavakhiya VV. The optimization of the nutrient medium composition for the submerged cultivation of the Mycolicibacterium neoaurum strain VKM Ac-3067D in a 100 L bioreactor under controlled conditions by mathematical planning. Fermentation. 2025;11:82. https://doi.org/10.3390/fermentation11020082
46. El-Gamal MS, Desouky SE, Abdel-Rahman MA, Khattab AM. Optimization of citric acid production from sugar cane molasses using a fungal isolate, Aspergillus fumigatus Na-1. Egypt J Biomed Sci. 2018;52:1-24.
47. Fan X, Burton R, Zhou Y. Glycerol (byproduct of biodiesel production) as a source for fuels and chemicals-mini review. Open Fuels Ener Sci J. 2010;3:17-22. https://doi.org/10.2174/1876973X01003010017.
48. Rehman A, Ali S, Haq I. Temperature optima for citric acid accumulation by Aspergillus niger. Biotechnology. 2002;1:108-10. https://doi.org/10.3923/biotech.2002.108.110
49. Rodrigues C, De Souza Vandenberghe LP, Teodoro J, Pandey A, Soccol CR. Improvement on citric acid production in solid-state fermentation by Aspergillus niger LPB BC mutant using citric pulp. Appl Biochem Biotechnol. 2009;158:72-87. https://doi.org/10.1007/s12010-008-8370-5
50. Shankar T, Sivakumar T. Optimization of citric acid production using Aspergillus niger isolated from the leaf litter soil of Sathuragiri Hills. Univ J Microbiol Res. 2016;4:79-87. https://doi.org/10.13189/ujmr.2016.040401
51. Salem El-Gamal M, Desouky SES, Abdel-Rahman MA, Mosasad Khattab AR. High-Temperature citric acid production from sugar cane molasses using a newly isolated thermotolerant yeast strain, candida parapsilosis NH-3. International Journal of Advanced Research in Biological Sciences (IJARBS). 2018;30:5(7):187-211.
52. Gallardo-Moreno AM, Garduno E, González-Mart?n ML, Pérez-Giraldo C, Bruque JM, Gómez-Garc?a AC. Analysis of the hydrophobic behaviour of different strains of Candida parapsilosis under two growth temperatures. Colloids Surf B Biointerfaces. 2003;28(2-3):119-26. https://doi.org/10.1016/S0927-7765(02)00145-5
53. De Melo Pereira GV, Maske BL, De Carvalho Neto DP, Karp SG, De Dea Lindner J, Martin JG, et al. What is Candida doing in my food? A review and safety alert on its use as starter cultures in fermented foods. Microorganisms. 2022;10(9):1855. https://doi.org/10.3390/microorganisms10091855
54. Patent 2192460 RU. Aspergillus niger Fungus Strain is a Producer of Citric Acid/N.V. Krasikova T.A. Nikiforov. M. Finko. The Patent Holder is the State Institution All-Russian Scientific Research Institute of Food Flavorings, Acids and Dyes RASN; 2000.
55. Patent 2103346 RU. Aspergillus niger Fungus Strain is a Producer of Citric Acid/A.Y. Vinarov T.E. SidorenkoS.E. Smetanina V.N. Yashina. The Patent Holder is the State Scientific Research Institute of Protein Biosynthesis; 1998.
56. Deme GD, Asfaw BT, Gari MT. Fabrication of citric acid using molasses by Aspergillus niger. J Mod Chem Chem Tech. 2020;11(1):37-58.
57. Khattab AA, Emam DA, Swelim M, Amer MM, Sehim A, Salem AA. Improvement of citric acid production by gamma radiated Aspergillus niger using cane molasses. Arab J Nucl Sci Appl. 2022;55(4):67-78. https://doi.org/10.21608/ajnsa.2022.137400.1580
58. Max B, Salgado JM, Rodríguez N, Cortés S, Converti A, Domínguez JM, et al. Biotechnological production of citric acid. Braz J Microbiol. 2010;41:862-75. https://doi.org/10.1590/S1517-83822010000400005
59. Chergui D, Akretche-Kelfat S, Lamoudi L, Al-Rshaidat M, Boudjelal F, Ait-Amar H. Optimization of citric acid production by Aspergillus niger using two downgraded Algerian date varieties. Saudi J Biol Sci. 2021;28(12):7134-41. https://doi.org/10.1016/j.sjbs.2021.08.013
60. Dhillon GS, Brar SK, Verma M, Tyagi RD. Recent advances in citric acid bio-production and recovery. Food Bioproc Tech. 2011;4(4):505-29. https://doi.org/10.1007/s11947-010-0399-0
61. Pandey A, Soccol CR, Rodriguez-Leon JA, Singh Nee Nigam P. Solid State Fermentation in Biotechnology: Fundamentals and Applications. India: Asiatech Publishers, Inc.; 2021.
Year
Month
Dietary Supplementation of Citric acid (monohydrate) Improves Health Span in Drosophila melanogaster
Komal Panchala, Kesha Patelb , Anand K. TiwariaDagaa (Rastrinoebola argentea) protein hydrolysate as a nitrogen source in microbial culture media
Katherine Pere, Betty Mbatia, Edward Muge, Vitalis W. WekesaIsolation and screening of dye decolorizing bacteria from industrial effluent
Mayur Gahlout, Poonam Chauhan, Hiren Prajapati, Suman Saroj, Poonam NaraleAntibacterial and antifungal activities of leaf and stem of Marsilea minuta L. against selected microbial pathogens
Govindaraj Sabithira, Rajangam UdayakumarRole of effective microorganisms on hematological and biochemical indices of cultured Oreochromis niloticus exposed to lead, copper, and cadmium under temperature variations
Ragab G. Abdel Salam,, Samah M. Bassem, Eman Salah Abdel-Reheim, Mahmoud Abdel-Latif, Giulia Guerriero, Fagr Kh Abdel-Gawad,Biofertilizer science and practice for agriculture and forestry: A review
Sudipta Saha, Debasish Paul, Tika Ram Poudel, Nafis Mahadi Basunia, Tasnimul Hasan, Mahadi Hasan, Bei Li, Rubel Reza, Ahmed Redwan Haque, Md. Abu Hanif, Manobendro Sarker, Nathan James Roberts, Muneer Ahmad Khoso, Haibo Wu, Hai-long ShenBacillus paralicheniformis (OQ202112) - Mediated biodiesel production using groundnut husk: A sustainable approach for bioenergy generation
Roshani Kantibhai Chaudhary, Priti Hemant Patel, Nikita Rajendrasinh Chavda, Vaidehi Kamlesh Patel, Kaushal Kantilal ChaudharyIsolation and screening of potential lignocellulolytic microbes from Phra Nakhon Si Ayutthaya Province
Sunisa Suwannaphan, Paweena Suksaard, Piyangkun Lueangjaroenkit, Suvichark Aroonluk, Phakhwan ThongrakMicrobial resources: Untapped treasure for environmental sustainability
Ajar Nath YadavProduction and Optimization of Protease by Filamentous Fungus Isolated from Paddy Soil in Thiruvarur District Tamilnadu
Sudarkodi Chandrasekaran, Sundar Sundaram Pillai Kumaresan, Murugan ManavalanPurification and characterization of tannase from the local isolate of Aspergillus niger
Saja Taha Yaseen Al-Mraai, Dhia Falih Al-Fekaiki, Alaa Jabbar Abd Al-ManhelAlbizia lebbeck fruit pods as a substrate for the production of lignocellulolytic enzymes by Aspergillus niger
Mamatha Pingili, Shailaja Raj Marla, Ramakrishna RaparlaAn effective and eco-friendly technique for control of post-harvest fungal pathogens of orange (Citrus sinensis) isolated from the distribution chain of Delhi NCR
Gayatri Krishna, Geethu Gopinath, Anupama Sharma AvasthiAmylase production by Aspergillus niger in submerged cultivation using cassava
Muralikandhan Kamaraj, Dhanasekaran SubramaniamComparative transcriptomic analysis of Aspergillus niger cultured in peanut or cashew nut flour based media
Christopher P. Mattison, Brian M. Mack, Jeffrey W. Cary