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Palm Wine Mixed Culture Fermentation Kinetics

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Palm Wine Mixed Culture Fermentation Kinetics ISSN: 2276-7851 Impact Factor 2012 (UJRI): 0.7799 ICV 2012: 5.88 Palm Wine Mixed Culture Fermentation Kinetics By Opara C.C. Ajoku G. Madumelu N.O. Greener Journal of Physical sciences ISSN: 2276-7851 Vol. 3 (1), pp028-037, January 2013. Research Article Palm Wine Mixed Culture Fermentation Kinetics Opara C.C. Ajoku G. and Madumelu N.O. Department of Chemical Engineering, University of Port Harcourt, Rivers State, Nigeria Email: [email protected], Tel. No: +2348037075314, +2348068868407 Corresponding Author’s Email: [email protected] ABSTRACT In this work, a mathematical model was developed describing the mixed culture fermentation of palm juice based on the growth profiles of the four microorganisms found in the juice. These microorganisms are Yeast, Micrococcus, Lactic acid bacteria and Leuconostoc spp . The mathematical model was simulated using MATLAB which showed the relationship between the substrate concentration and time for the individual microorganisms. Experimental data obtained from literature was used to obtain the growth rates and substrate saturation coefficient using the Monod model. It can be seen that the cell number of Leuconostoc spp . has increased to 1978.3 while the experimental value is 1667.0678 at 24hours. The substrate concentration at 24hrs was found to be 4.968g/100g dry matter while its experimental value is 4.348g/100g dry matter. This shows that there is a reasonable level of accuracy in the model developed . Keywords: Fermentation, kinetics, micro flora, mixed culture, yeast. INTRODUCTION Brief History of Palm Juice Palm-wine is an important alcoholic beverage resulting from the spontaneous fermentation of the sap of the palm, which has been attributed to yeasts and bacteria (Okafor, 1978). Fermented palm-juice is an exudate from palm-tree. Natural palm sap is collected from the oil palm tree_Elaeis guuineenses and the raphia palms, Raphia hookeri and Raphia vinifera , although it could also be obtained from other species of palm trees that are known all over the world. Palm wine is produced when the inflorescence of the palm tree is incised and tapped, which is collected in a calabash that is hung at the base of the incision. Fresh palm-wine is sweet, clear, neutral, colorless juice containing 10-12% sucrose, minimal invest sugar (less than 0.5%), small amounts of proteins, gums and mineral. (Okafor, 1987). Spontaneous fermentation starts immediately the sap is collected and within an hour or two becomes reasonably high in alcohol (up to 4%) (Bassir, 1962). Organisms responsible include S. cerevisiae and schizosaccharomyces pombe, and the bacteria Lactobacillus plantarum and L. mesenteroids (Okafor, 1987). The equations of reaction of fermentation of palm-wine and oxidation of ethanol are represented thus respectively; C6H12 O6 yeast 2C 2H5OH + 2CO 2 Glucose Ethanol Carbon dioxide C2H5OH + O2 Acetobacter CH 3 COOH + H2O Ethanol Oxygen Acetic acid water In the former fermentation process, ethanol is produced as the sugar content diminishes rapidly as it is acted upon by agents of fermentation (microorganisms) while in the latter, ethanol will be on the declining phase as it is converted to acetic acid in conditions of excess oxygen. www.gjournals.org 28 Greener Journal of Physical sciences ISSN: 2276-7851 Vol. 3 (1), pp028-037, January 2013. Mixed Culture Fermentation Mixed culture fermentation is a biochemical process involving two or more microorganisms in a nutrient substance in specially controlled conditions for scientific, medical or commercial purposes (Microsoft Encarta, 2004). The interactions of mixed culture systems are difficult to interpret because there is no good reference point when so many conditions and concentrations may be changing at once. Research Objective The purposes of this research are: - To obtain from literature the microorganisms that proliferates during the mixed-culture fermentation of palm wine. - To determine the process conditions for the fermentation. - To determine the yield coefficients of the respective micro-organisms in the fermentation process with respect to time. - To estimate kinetic parameters of the microorganisms present based on reliable experimental data from literature. - To determine the substrate utilization profile from the start of fermentation (0th hour) to the end of fermentation (72 nd hour) at intervals of two hours. - To write a mathematical model that describes the process, this will be an aid to Biochemical and Chemical Engineers in designing a large-scale fermenter of the process. - To write a computer program that will solve the mathematical model and simulate the process. Microorganisms Responsible for Fermentation Studies on the microbiology of palm wine have shown that palm wine contain both yeast and bacteria (Okafor, 1978). Yeast Okafor (1978) isolated and identified yeasts from palm wine from various parts of Nigeria and concluded that the distribution of yeast in palm wine is not dictated by the type of palm or the locality in which it grows. In his study, all the samples were dominated by saccharomyces and candida irrespective of the type of palm or the area in which it is grown. Bacteria Bacteria encountered in palm wine include Lactobacillus, Leuconostoc, Bacillus, Serratic, Streptococcus, Zymomonas mobilis, Micrococcus, Breviabacterium, Pediacoccus, Corynebacterium, Klebsiella (Bassir, 1962; Faparusi, 1966). SUCCESSION of MICRO FLORA DURING FERMENTATION In this regard, Bassir and Faparusi (1971) reported the presence of Lactobacillus and Leuconostoc in the early stages of fermentation. Okafor (1975) found that while lactic acid bacteria as a group were important, there were no consistent patterns of the distribution of the various lactic . According to him, streptococcus was found throughout the 7days period he carried out his work whereas lactobacillus remained only during the first 3 days of fermentation. Okafor (1975) reported the presence of enterobacteriacae such as serratia and klebsiella early in fermentation. According to him, yeasts and micrococcus seem to occur consistently in many samples of palm wine. Faparusi and Bassir (1971) also reported the same in their investigations. Authorities who have studied the succession of micro flora in palm wine consistently reported the emergence of Acetobacter after about 2 to 3 days of fermentation, at which time; alcohol was present in reasonable quantities (Faparusi and Bassir, 1971; Okafor, 1975). www.gjournals.org 29 Greener Journal of Physical sciences ISSN: 2276-7851 Vol. 3 (1), pp028-037, January 2013. Factors Affecting Rate of Fermentation Despite its complexity, the rate of fermentation is largely dependent upon certain parameters namely yeast, wort composition (i.e. Fermentable filtrates normally nutrients for yeast) and the processing condition such as time, temperature, volume, vessel shape and size, pH, acid, water activity, carbon sources are also complementary variables. These conditions are in general applied to all classes of fermentation, be it batch process or the continuous process. Fermentation Process In fermentation, an accurate mathematical model is a prerequisite for the control, optimization and the simulation of a process. The kinetics for growth is generally modeled as first order. A batch fermentation process such as that for palm wine has a growth rate equation modeled as dX i = µ X (1.1) dt i i Where Xi = concentration (cell number) of the ith organism µi = specific growth rate of the ith organism t = time dS µX = i (1.2) dt Yi Where S = Substrate concentration Y = yield coefficient for individual organism The assumption made in generating this model is simply that the organisms compete for the common substrate without preying on one another. This can be used as an analogy for the mixed culture fermentation of palm juice as there are no serious prey-predator relationships among the organism. The growth rate, µ takes the form µ S µ = i max (1.3) i km + S The equation above is known as the Monod model Where µimax = maximum reaction rate Km = Saturation constant of the micro-organism To solve this set of differential equations, it is important to note that the values of km and µ max must be determined for individual micro-organism. A solution of this will provide an approximate model for this process. METHODOLOGY Mathematical Model Development In developing this model, several assumptions were made so as to simplify this complex process. One of the major assumptions is that the specific growth rate of the cells is exponential. This assumption leads to taking the Monod Model as an approximate model for the growth rate. µ S µ = i , max (1.3) i km + S www.gjournals.org 30 Greener Journal of Physical sciences ISSN: 2276-7851 Vol. 3 (1), pp028-037, January 2013. From the experimental data by Brauman et al. (2002), the following were the cell number and the substrate concentration of the four major micro-organisms common in the mixed culture fermentation of palm juice. These four micro-organisms are Yeast, Micrococcus , Lactic Acid bacteria (LAB) and Leuconostoc spp , (LEU). The result is given below. Table 2.1: Change in Cell number and Substrate Concentrate of the Individual Micro-organism with Time Time (hr) Yeast Micrococcus Lactic Acid Leuconostoc spp Substrate Concentrate Bacteria (LAB) (LEU) (g/100g of dry matter) 0 312316 222246 10000 1584.89 6.5 24 1E+09 1E+07 1E+07 1000.00 6.3 48 1E+12 1.4 E+08 6.31 E+07 1000.00 2.2 60 1E+12 2.9 E+09 2.51 E+08 7328.55 1.6 72 1 E+13 50 E+09 30 E+09 15848.93 1.252 Source: Brauman et al. (2002)
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