Citric acid production in bioreactors Mansureh kia lashaky Department of Chemical Engineering University of ferdowsi Overview 1. History of Citric Acid 2. Usage of Citric Acid in various Sectors Globally 3. Methods for production citric acid 4. Citric acid Fermentation techniques in bioreactors 5. Optimization of Citric Acid Fermentation Process by surface fermentation submerged fermentation Statistical Method solid-state fermentation 6. Relations of Citric acid production and morphology of A.niger 7. factors affecting citric acid production 1
History of Citric Acid 1893 Citric acid fermentation was first observed by Wehmer 1916 Currie found that numerous strains of Aspergillus niger produced significant amounts of citric acid 1930 some units were implanted in England, in Soviet Union, and in Germany for the commercial production Usage of Citric Acid in various Sectors Globally used mainly in the pharmaceutical and food industries Industrial applications cosmetics and toiletries, and other Methods for production citric acid Chemical Natural Citric Purification Acid Fermentation (using molass and A.niger) 2
Microbial Production of Citric Acid Microorganisms A large number of microorganisms including fungi and Among the mentioned strains, the fungus bacteria such as Aspergillus. niger has used for commercial Arthrobacter paraffinens , Bacillus licheniformisand Corynebacterium ssp , Aspergillus niger , A . aculeatus , production A.carbonarius , A.awamori , A.foetidus , A. fonse-caeus , A.phoenicis and Penicillium janthinellum and yeasts such as Candida tropicalis , C.oleophila , C.guilliermondii , C . citroformans, Hansenula anamola andYarrowia lipolytica Microbial Production of Citric Acid Substrates Several raw materials such as : hydrocarbons, starchy materials and molasses for commercial citric acid production A variety of agro-industrial residues and by-products 3
Citric acid production techniques fermentation is the most economical and widely used way for synthesis Citric acid production The industrial citric acid production can be carried in three different ways : surface fermentation submerged fermentation solid-state fermentation Citric acid production techniques surface fermentation Surface fermentation is the classic and was the first industrial process for citric production Surface fermentation requires less effort in operation, installation and energy cost The process is carried out in fermentation chambers where a great number of trays is arranged in shelves . 4
Citric acid production techniques submerged fermentation The submerged technique is widely used for citric acid production. advantages : higher productivity and yields lower labour costs lower contamination risk Citric acid production techniques solid-state fermentation Solid- state fermentation, also known by « Koji » process, was first developed in Japan It is the simplest method for citric acid pro-duction 5
Optimization of Citric Acid Fermentation Process by Statistical Method Response Surface Methodology (RSM) was employed for the optimization of the variables. Where x1 = Incubation time x2 = Fermentation temperature x3 = O 2 flow rate Optimization of Citric Acid Fermentation Process by Statistical Method Y = -1463.44326 + 1.99043x 1 + 98.25444x 2 +12.47096x 3 +0.35126x 1 x 2 +0.029100x 1 x 3 – 0.053142x 2 x 3 – 1.16895x 1 2 – 1.63925x 2 2 – 2.87816x 3 2 the significance of each parameter was analyzed using ANOVA table 6
Optimization of Citric Acid Fermentation Process by Statistical Method plot 1 . Response surface curve for citric acid production as function of Incubation time and O 2 flow rate, while Fermentation temperature was fixed at 30 0 C Citric acid production and morphology of A.niger as functions of the mixing intensity in a stirred tank bioreactor Morphology was quantified using an image analysis system consisting of a phase contrast microscope Samples for morphological characterization were taken Samples were immediately fixed with an equal volume of fixative 7
Citric acid production and morphology of A.niger as functions of the mixing intensity in a stirred tank bioreactor Fig 4.shows the characteristic morphology of the fungus at the end (168 h) of the STR run at 500 rpm Citric acid production and morphology of A.niger as functions of the mixing intensity in a stirred tank bioreactor plot 2.of the corresponding convex perimeters of clumps versus stirrer speed in the STR 8
Citric acid production and morphology of A.niger as functions of the mixing intensity in a stirred tank bioreactor Plot 3. of the final citric acid concentrations versus the corresponding convex perimeters of clumps factors affecting citric acid production Nitrogen source pH of fermentation medium Aeration Trace elements 9
Any Question REFERENCES 1. Dhillon, G., Brar, S., Kaur, S., & Verma, M. (2013). Bioproduction and extraction optimization of citric acid from Aspergillus niger by rotating drum type solid-state bioreactor. 2. Soccol, C., Vandenberghe, L., Rodrigues, C., & Pandey, A. (2006). New Perspectives for Citric Acid Production and Application. 3. Beroviè , M., & Popoviæ, M. (2001). Characterisation of Gas Mixed Bioreactors in Submerged Citric Acid Fermentation. 4. Dhillon, G., Kaur Brar, S., Verma, M., & Dayal Tyagi, R. (2010). Recent Advances in Citric Acid Bio-productionand Recovery. 5. KIM , J.-W. (2004 ). OPTIMIZATION OF CITRIC ACID PRODUCTION BY ASPERGILLUS NIGER NRRL 567 IN VARIOUS FERMENTATION SYSTEMS. 10
REFERENCES 6. Kola, A., & Goli, V. (n.d.). Design and Optimization of Citric Acid Fermentation Process by Statistical Method. 7. Papagianni, M., Mattey, M., & Kristiansen, B. (1998). Citric acid production and morphology ofAspergillus nigeras functions of the mixing intensity in a stirred tank and a tubular loop bioreactor. Biochemical Engineering . 8. Ali, H., & Zulkali, M. (2011). Design Aspects of Bioreactors for Solid-state Fermentation. 9. Sikander Ali, S., Haq , I., & Qadeer, A. (2002). Production of citric acid by Aspergillus nigerusing cane molasses in a stirred fermentor. Electronic Journal of Biotechnology . 11
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