                   !  "  # $% 1 Department of Food Engineering, Faculty of Engineering and Architecture, Abant Izzet Baysal University, Golkoy 14280, Bolu, Turkey. 2 Department of Chemical Engineering, Faculty of Engineering, Hacettepe University, Beytepe 06532, Ankara, Turkey. 3 Department of Food Engineering, Faculty of Engineering, Hacettepe University, Beytepe 06532, Ankara, Turkey. *Corresponding author: e/mail: yesim@hacettepe.edu.tr, Phone: +90 312 297 71 12, Fax: +90 312 299 21 23 Citric acid is the most important organic acid produced by fermentation, widely used in food, pharmaceutical and chemical industries. Although   is the traditional producer of citric acid, during the last 30 years the interest of researchers has been attracted by the use of yeasts for citric acid fermentation processes. Among the yeast species,   is known as a potential producer of citric acid. Environmental factors that have been shown to exert an effect on citric acid production are the type and concentration of carbon source of the fermentation medium, nitrogen and phosphate limitations, aeration, trace elements, initial pH and temperature. Besides the regulation of product formation by environmental conditions, strain selection and improvement has become the important factor. The improvement of citric acid producing yeast strains has been carried out by mutagenesis and selection. Because of annual growths in demand of citric acid, using alternative processes and strains for its production are in progress. & citric acid; yeasts;  ; fermentation parameters       Citric acid is a commercially valuable organic acid, widely used in food, pharmaceutical and beverage industries [1]. It is the main additive used in the food industry. Citric acid is widely used to impart a pleasant, tart flavour to foods and beverages. It also contributes to the formulation of many foods as an acidulant, antioxidant, emulsifier or preservative [2, 3]. Among the uses of citric acid, about 70% is used in the food industry, and 10% in the cosmetics and pharmaceuticals [4]. There is a great worldwide demand for citric acid consumption due to its low toxicity when compared with other acidulants [3]. It is reported that the supply of natural citric acid is very limited and the demand can only be satisfied by biotechnological fermentation processes. Citric acid is known as the most important organic acid produced in tonnage by fermentation and is the most exploited biochemical product [3, 5]. The annual production of citric acid was reported as 700 thousand tons in 1993 [6], 1.4 million tons in 2004 [3], and 1.6 million tons in 2008 [7, 8] . A large number of microorganisms have been employed for citric acid production, but a few of them can produce citric acid in industrial scale [3]. It is reported that   is almost exclusively used for industrial scale production of citric acid [5], but during the last 30 years the interest of researchers has been attracted by the use of yeasts as citric acid producers [9]. A number of different strains, mostly belonging to the  () genus have been used for citric acid production, mainly in conventional batch processes, but also in continuous culture, and with immobilized cells [10]. The yeast species which were reported to produce citric acid are;  ()       ,  ,  ,  ,  ,  , and some  species [6, 11]. Among the yeast species,   is known as a potential producer of citric acid [1, 12] and has been developed as a microbial cell factory for citric acid production in recent years [7]. The main advantages of using yeasts are mentioned as follows: Yeasts are characterized by greater resistance to high substrate concentrations than fungi, with comparable conversion rates and have greater tolerance to metal ions that allows the use of less refined substrates. Using yeasts also gives a better process control due to their unicellular nature [13, 14]. It was reported that citric acid production by yeast could be in the future an alternative to   one, especially if the yeast biomass became an additive to animal food and not a by/product [15]. However, the major disadvantage of using yeasts is the simultaneous production of citric and isocitric acids [13, 16]. It is reported that the ratio of citric:isocitric acid can vary between 1:1 to 20:1 according to the yeast strain, carbon source and micronutrient concentration [13]. Selection of a yeast strain with high citric acid production and giving high citric acid:isocitric acid ratios has been reported as the principal step of a citric acid production process. It is reported that more than 90% of the citric acid produced in the world is obtained by fermentation. The industrial citric acid production can be carried out in three different ways: by submerged fermentation, surface fermentation and solid/state fermentation or “Koji” process [3]. It is estimated that about 80% of the world citric acid production is obtained by submerged fermentation in stirred tanks of 40/200 m 3 or larger airlift fermentors of 200/900 m 3 capacity [17]. Submerged fermentation can be carried out in batch, fed batch or continuous systems, although the batch mode is _______________________________________________________________________________________ Current Research, Technology and Education Topics in Applied Microbiology and Microbial Biotechnology A. Méndez-Vilas (Ed.) 1374 ©FORMATEX 2010