Egyptian Sugar Journal, June 2012 Vol.5 : 23- 46 ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ
Physicochemical investigations of Delta beet molasses for the bio-production of ethanol. A. M. Ramadan a*, A. A. Zohrib, , M. M. El-Tabakhc and K. El-Tantawyc a)
Chemistry department, Faculty of Science, Kafr El-Sheikh University, Egypt. b) Botany department, Faculty of Science, Assiut University, Egypt. c) Delta Sugar Company, Egypt.
Abstract Bio-ethanol production through fermentation as a liquid fuel has been accepted technology for many years. Choice of renewable sugar containing substrates as feedstock for yeast fermentation depended on local conditions. Beet molasses as the most available substrate in Egypt was selected for this study. A total of 38 samples of beet molasses of the production lines I and II of the Delta Sugar Company, Egypt were collected during the season of 2010 in order to assessment their ability for bio-ethanol production. Routine physical investigations for the different physical properties of the examined samples demonstrated that there is no appreciable variation of the physical properties of these samples at the beginning of the crush season and its end. As well as the physical properties of these samples were compared with other beet molasses employed for ethanol production. The different chemical investigations which were carried out reflect the following points: i) The present beet molasses have appreciable fermentable sugar content (nearly 97 %). ii) The total nitrogen and amino acids content were in the suitable levels for ethanol production as inferred from analogous previous studies. iii) Although the concentrations of the harmful NO2 and NO3 were apparently high, but these values seem to be in the safe area. As well as the volatile acids concentration were comparable with other fermentable molasses. iv) The elemental phosphorous and its oxide P2O5 contents of the representative samples were suitable for yeast growth. v) The useful trace elements Fe, Zn, Cu, Mn and Mo were present in suitable
Ramadan, et al (2012) Egyptian Sugar 24 Journal,Vol.5 : 23- 46 ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ concentrations, also, the concentrations of the toxic elements such as Cd and Pb were in the safe area. ـــــــــــــــــــــــــــــــــــــــــــــــــــــــ *Author for corresponding: E-mail:
[email protected]
Introduction The natural energy resources such as fossil fuel, petroleum and coal are being utilized at rapid rate and these resources have been estimated to last over a few years. Therefore, alternative energy sources such as ethanol, methane, and hydrogen are being considered. Some biological processes have rendered possible routes for producing ethanol and methane in large volume. A worldwide interest in the utilization of bio-ethanol as an energy source has stimulated studies on the cost and efficiency of industrial processes for ethanol production. Ethanol is one of the most advanced liquid fuels because it is environmental friendly. It is particularly useful in industrial applications because of its relatively high affinity for both water and organic compounds. The composition of other alcohols limits their flexibility as compared to ethanol (Anxo et al., 2008). It is a bio - fuel, which is produced from biomass and wastes. Bio-fuels provide an alternative to fossil fuel dependency and emit fewer pollutants (Carvalho et al., 1993). Although ethanol exhibits various medicinal and industrial uses, the largest single use of ethanol is as a motor fuel and fuel additive. The largest national fuel ethanol industries exist in Brazil and the United States. The Brazilian ethanol industry is based on sugarcane; as of 2004, Brazil produces 14 billion liters annually, enough to replace about 40% of its gasoline demand. Also as a result, they have become 80% independent from foreign oil. Most new cars sold in Brazil are flexible-fuel vehicles that can run on ethanol, gasoline, or any
Physicochemical investigations of Delta beet 25 molasses for the………. ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ blend of the two. The United States fuel ethanol industry is based largely on corn. Thailand, India, China and Japan have now launched their national gasohol policies. The end syrup from the manufacture of sugar was introduced 150 years ago by Achard (1809) in the distilling industry. Balling (1865) gives the beginning of the industrial utilization of beet molasses for alcohol production around 1830. Up to about 1880 the only industrial use of molasses was in the manufacture of alcohol, a process carried out in molasses distilleries. Alcohol is also produced from molasses combined with the manufacture of yeast (yeast aeration spirits). Various processes have been developed for ethanol production but world wide demand of ethanol is generally satisfied by biotechnological fermentation process. A number of organisms including fungi, yeast and bacteria have been screened for ethanol fermentation. Extensive studies have been carried out on the fermentation process of ethanol by these organisms, especially through yeast cells (Bajaj et al., 2001). However, S. cerevisiae remained the organism of choice, which is the same species used for bread making and some wines or beers (Walker et al., 1990; Converti et al., 2003). The alcohol industry, using beet and cane molasses, is concerned with the suitability of these substrates for alcohol fermentation. It is possible to alter greatly the relative yields of alcohol. The previous studies demonstrated that, when molasses comes to be used as a substrate for biochemical transformation, e. g. alcohol fermentation, it was found that it contains substances which can promote, but in some cases also inhibit the growth of microorganisms. However, there are a vital number of factors which govern the alcohol production from molasses, such as the physical and chemical nature of the fermentable molasses, in addition to their suitability for the employed organism. Accordingly, to achieve a successful fermentation process on the beet molasses,
Ramadan, et al (2012) Egyptian Sugar 26 Journal,Vol.5 : 23- 46 ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ it is important to have a pre-picture of the physical and chemical properties of the fermentable molasses samples. Herein we report the physicochemical investigations of samples of the Egyptian beet molasses from the production lines I and II of Delta Sugar Company during the season 2010 to obtain a map of the chemical constituents of these investigated samples to optimize the conditions for the bio-production of ethanol.
Material and methods (a) Molasses samplers: Thirty eight samples of beet roots and other 38 samples of beet molasses were collected from the two production lines I and II of Delta Sugar Company (DSC), Egypt during the production season 2010. Samples represented 19 periods (Table, 1) during the season according to the quality of beet roots. Each sample was a sub-sample of several mixed samples taken every two hours for 24 hours a day through out the selected period. The collected samples were placed in specific sterilized jars and transferred to the laboratory, then kept at 4ºC till analysis.
(b) Analytical methods: The following analytical methods were carrying out according to International Commission for Uniform Methods of Sugar Analysis (ICUMSA) at Delta Sugar Company, Quality Control Department, (Routine and classical Quality Control Analysis): determination of pH, temperature, apparent dry substance (Brix) using a Hydrometer, apparent purity, total sugars as reducing matter, organic matters (calculated mathematically), sulfated & carbonated ashes in addition to the conductivity ash, total dissolved matters, density, viscosity at 85 °Brix, specific gravity & impurities, sodium & potassium using venma, Automation BV Analyzer, sucrose content as well as calcium & magnesium by EDTA titration.
Physicochemical investigations of Delta beet 27 molasses for the………. ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ Particularly in the world of sugar analysis, much work in our own and other laboratories, is directed towards instrumental analysis with atomic absorption and HPLC technique. Much work still remains to be done, particularly with HPLC, in order to achieve the degree of repeatability and reproducibility necessary for a commercial method of analysis. Several chemical investigations were carried out at National Research Center, Cairo as shown below: 1 - Determinations of the following components were performed by using HPLC analysis technique: Sucrose, Glucose, Fructose, Raffinose and the volatile organic acids. 2 - Determination of the minor components in this study (trace elements) was performed by using atomic absorption technique. 3 - Amino acids were determined by using LC3000 amino acid analyzer.
Results and Discussion Chemical investigations of the Egyptian beet roots This study reports the physicochemical investigations of the beet molasses of the production lines I and II of the Delta Sugar Company during the season of 2010. It is well known that the quality of the produced molasses depends greatly on the nature of its original source beet roots. A series of chemical investigations were curried out on beet roots to obtain pre-picture of the chemical nature of the molasses. For this purpose the production season was classified into 19 periods and 38 samples of beet roots were collected and examined. The data in Table (1) reveal that: for potassium and amino nitrogen contents there is no significant variation during the 19 production periods. However, the potassium content seems to be apparently high. On the other hand, a noticeable variation is observed in the case of the values of the quality, the sugar and sodium content. This fact reflects that as the sodium decreases,
Ramadan, et al (2012) Egyptian Sugar 28 Journal,Vol.5 : 23- 46 ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ the sugar and the quality of the beet root increases. This means that there is a reversible relationship between the quality of the beet sugar and the concentration of this alkali metal. This reversible relationship can also be stated for the produced molasses. These inorganic nonsugar materials increase the solubility of sucrose and they are important for the molasses – producing characters. In general, the good quality of beet roots will be reflected in final produced molasses and sugar.
Physical investigations of beet molasses Several studies demonstrated that, the physical properties of the beet molasses are good markers for its chemical characters and quality. For example, the viscosity of beet molasses increases as the alkaline earth metal cations (Ca2+ and Mg2+) content increases (Breitung, 1956). Viscosity further decreases with the removal of sugars at the same density. These physicochemical relationships were established in our cases for Brix, specific gravity and viscosity (Figure, 1). The data in Tables (2) and Figure (1) show that there is a correlation between these physical properties of the investigated samples of molasses and their alkali and alkaline earth metal cations content as well as the content of other chemical constituents.
Brix and color % Brix assay Color % brix assay measures the degree of darkness of color of the molasses. The importance of coloring matters of molasses with respect to their effect on the quality of the produced yeast and their metabolisms is difficult to state exactly. No general statements can be made about the composition and amounts of the molasses coloring matters. This is not surprising in view of the complexity of the pigmenting compounds of molasses, which are chiefly caramel substances, melanoidines and iron phenol compounds which are related
Physicochemical investigations of Delta beet 29 molasses for the………. ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ to the molasses colloids. Molasses from different years and factories seldom have the same color. A light colored molasses may indicate that it contains much SO2 as a result of an intense sulfuring of the juices; this assumption was checked by the chemical analysis. The results of sulfur and sulfite analysis (Table, 3) display that as the sulfur concentration increases, the color % Brix increases. This finding confirms the reliability of the previously stated assumption. On the other hand, it has been reported that certain relationships exist between the colors of crude sugar, white sugar and refinery molasses but no reliable rating of an individual molasses can be derived from the color determination of the molasses. As a rule, very dark molasses must be pretreated; efforts must be made to remove impurities and colloids as well as caramelized and humic-like substances as completely as possible. According to PollaK and Knob (1922), these color materials tend to increase the difficulty of normal fermenting of molasses. The very dark molasses is always a disadvantage from the standpoint of presenting subsidiary problems. Normal procedures cannot produce satisfactory biomass or bio-products from molasses that contain 2.3 to 2.4% caramels.
Chemical investigations It is well known that the yeast growth and the rate of molasses fermentation are governed by a number of factors such as: the nature & content of sugar and other chemical constituents as well as the probable occurrence of deleterious constituents in molasses which include: excessive content of sulfurous acid, nitrates, volatile acids, and toxic trace elements, in addition to the colloidal and suspended matters. Accordingly, a series of chemical analysis were carried out to identify the chemical map of the examined samples of
Ramadan, et al (2012) Egyptian Sugar 30 Journal,Vol.5 : 23- 46 ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ molasses in order to optimize the conditions for achieving a successful fermentation process.
Identification of the nature & sugar content: Nature and sugar content of the molasses are the main key factors for the bio-production of ethanol. Table (4) reports the most important and significant analytical results in this study. The average of total sugar concentrations are 53.8 and 54.01% in the analyzed samples of the production lines I & II, respectively. Most of these sugar levels (nearly 97%) are in the fermentable form. These results reflect that the fermentable sugar content for the bio-production of ethyl alcohol from these molasses samples is appreciable value. The nonfermentable sugar levels are 1.53 and 1.59% in the samples of production lines I& II, respectively (Table, 4). Previously, Honig (1953) reported that the average total sugar levels in beet and cane molasses were 53 and 62%, respectively. Recently, El-Samman (2010) analyzed cane molasses of four sugar factories in Upper Egypt and found that the total sugar% of mixed sample from the four factories was 50.2% and the nonfermentable sugar level in the same sample reached to 4.4%. Due to the vital effect of sugar nature on the fermentation process, a further assessment of sugar content was performed by employing the HPLC technique. For this purpose representative samples of molasses were analyzed and the results obtained are given in Tables (5 & 6). These data are confirm the reported results in Table (4) and reflect the good accordance between the analytical results obtained by the employed analytical traditional method and the more accurate HPLC technique.
Determination of the amino acids: Due to the influence of the amino acids concentration on the fermentation process, a series of Delta beet molasses was analyzed by amino acid analyzer. The results obtained are given in Table (7) and show that glutamic acid was the most prevailing amino acids.
Physicochemical investigations of Delta beet 31 molasses for the………. ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ As shown in Table (7) the average percentages of amino acids of the analyzed molasses samples were 8.94%, and 8.76% for the production lines I and II, respectively. In several studies, the average levels of the amino acids content in molasses were 6 to 7% i.e., around 1/3 of the 20% of the nitrogenous nonsugars of the molasses is present as amino acids. It is will known that amino acids combine with reducing sugars in a number of reactions named Maillard reaction with the formation of melanoidins which are dark colored high molecular mass compounds and increase the viscosity of molasses. This reactions also lead to destruction of reducing sugars and forming additional impurities in molasses. Also, high nitrogen content in molasses, specially amino acids, is considered one of the major causes of rapid decomposition of molasses which stored at ≥ 45ºC.
Determination of nitrate and nitrite: Nitrate and nitrite are the most harmful inorganic nitrogenous components exist in the molasses due to their inhibitory influence on the fermentation process. One of the main constraints for obtaining higher rates of ethanol production is the inhibition of yeast metabolism due to the contamination of molasses with high concentrations of both nitrate and nitrite. Generally in microbial industrial alcohol production, the generation of nitrous gases due to the presence of nitrite becomes pronounced and seriously affects the fermentation efficiency. A considerable loss in ethanol production has been reported for concentrations of nitrite lie in the range 0.004 to 0.001% (Notkina et al., 1975). The analytical result recorded in Table (8) presents the amounts of nitrate and nitrite of the investigated samples of molasses from lines I and II. These analytical results exhibit the highest concentration of nitrite in the molasses at the period from 27/1 to 22/2 especially for line I. This apparent high concentration of nitrite observed for line I at the beginning of the crush
Ramadan, et al (2012) Egyptian Sugar 32 Journal,Vol.5 : 23- 46 ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ season will lead to inhibit the fermentation process. On the other hand, the reported concentrations of NO2 and NO3 at the other periods are in the safe area.
Determination of sulfur and sulfite: If the operating conditions are good about 64 liters of alcohol are obtained from 100 kg of sugar processed molasses (a schematic review of the yields of the principal and by-products resulting from the manufacture of alcohol from molasses). Some of the unfavorable components companied molasses such as SO2 prevent achieving this good yield of ethanol. Honig (1953) stated that the sulfite content in molasses, resulting from the use of SO2 in the sugar manufacturing process, will cause difficulties in the ethanol factory. To obtain a molasses satisfactory with regard to this sulfur dioxide content, not more than 0.003-0.004 % SO2 may be present in the corresponding thin juice or syrup. The significance of the SO2 content of molasses with respect to the manufacture of pressed yeast was later taken into account in the molasses specifications by fixing the maximum allowable content at 0.l5% SO2. The fermentation activity of various kinds of yeast is impeded when the sulfur dioxide content reaches approximately 6.8-9.2 mg/liter. Differences in clarification, dilution and aeration can lead to quite different limiting concentrations of SO2. The experiences gained under specific conditions are not of general validity. Claassen (1940) stated that 0.l25 % SO2 in molasses is not harmful in yeast manufacture. Karczewska (1951) reported that figure may go up to 0.3%, whereas Drews et al. (1955) claimed that 0.02% sulfur dioxide is definitely harmful. The presence of sulfurous acid in molasses does not inevitably lead to difficulties under all conditions. On the other hand, change of color of the yeast and odor of hydrogen sulfide are typical quality disorders likely to make them
Physicochemical investigations of Delta beet 33 molasses for the………. ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ evident when molasses containing sulfurous acid are processed. Such effects, as a rule, can be avoided by minor modifications in the clarification process. The removal of sulfurous acid by the usual hot acid defecation method can be intensified by adding such strong oxidizing agents such as sodium chlorate, converting the sulfites into sulfates. The mixture is thoroughly stirred and heated to 80-95°C. Table (3) presents the analytical results of both sulfur and sulfite of the investigated samples of molasses of lines I and II. These analytical results exhibit the highest concentration of sulfur in the molasses at the selective second two periods (Table, 3). On the other hand, the reported concentrations of sulfur at the other periods are apparently lower. Concerning, the analytical results of sulfite no appreciable variations are observed for the four investigated periods. In general, these recorded amounts of both sulfur and sulfite are almost in the safe concentrations and can be over come by using the treatments mentioned above.
Determination of volatile organic acids: The free volatile acids occurring in molasses are generally present in slight amounts, so that they do not impair the culture of the yeast. Considerable quantities of these volatile fatty acids affect the growth of the yeast unfavorably. Butyric acid content in molasses mash of 0.15% is considered to be the critical level; while greater concentrations of butyric acid led to complete interruption of alcohol fermentation. On the other hand, the lower content of butyric acid is accompanied by an improvement of the fermentation. The normal fermentation rate was attained when the mash contained 0.08% butyric acid. The growth and activity of yeast are very sensitive to the amount of butyric acid exists in molasses. For instance at concentration of 0.005%, the growth and activity of
Ramadan, et al (2012) Egyptian Sugar 34 Journal,Vol.5 : 23- 46 ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ yeast is reduced in the worth, while at higher concentration e. g. at 0.1 %) the fermentation process may completely stop. The great divergence in the fermentation obtained under strictly comparable conditions with ‘normal’ molasses and with one containing 1.23% acetic acid and 0.96% butyric acid is impressive. The employed yeast cannot reach its optimal activity due the presence of these proportions of the volatile acids; i. e. the activity may reduce to about 15 – 20 % of its normal activity in the absence of these acids. There is no satisfactory explanation of the influence of acetic and formic acids on the yeast activity under the conditions for producing pressed yeast. The normal value for the total volatile acid content in molasses is given as 0.1-0.3%. The reported difficulties encountered in the manufacture of yeast and alcohol from molasses led us to detailed investigations, directed especially to the determination of formic, acetic and butyric acid. These investigations were conducted in the laboratories of the National Research Center, Cairo, Egypt and representative samples of Delta beet molasses were analyzed by HPLC technique and the results obtained are given in Tables (9 & 10). The data in Tables (9& 10) denote that formic acid concentrations are slightly higher in the second period (22/3-20/6) then those recorded in the first period (27/1-21/3). Acetic acid levels are fluctuated between 0.57 and 0.64, in all tested samples. High concentrations of butyric acid (0.41% and 0.58%) were observed in all tested samples especially in the second period. However, if the molasses is acidified at boiling, volatile species such as acetic, formic and butyric may completely evolved.
Determination of minor components: The chemical investigations of molasses would be incomplete without accurate determination of the minor components, i.e. those which are present in relatively slight amounts. When
Physicochemical investigations of Delta beet 35 molasses for the………. ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ molasses comes to be used as a substrate for biochemical transformation, e. g. alcohol fermentation, it was found that it contains many elements in very low concentrations, called trace elements which can promote (such as Fe, Cu, Mn, Zn, P and P2O5), but in some cases also inhibit the growth of microorganisms e. g. Pb and Cd. Consequently, a series of the qualitative and quantitative elemental analysis were carried out by using atomic absorption technique to identify the types and amount of some of these trace elements and the results obtained are tabulated in Table (11). The results clarify the type and amount of the detected trace elements exist in representative samples of molasses. These results display that the analyzed samples contain appreciable amounts of the promoted elements such as copper, iron, manganese and zinc, while the toxic heavy metal such as Cd was not detected in most investigated periods. On the other hand, the analytical results (Table, 11) denote that the concentration of Pb seems to be apparently high. Regarding, phosphorus, its measured concentrations, as shown in Table (12), are suitable for promotion of the yeast growth.
Conclusion Several physicochemical investigations of a series of Delta beet molasses samples were carried out. The results obtained revealed that, many chemical inhibitors exist in the investigated samples, e. g. NO2, SO3, volatile organic acids (formic, acetic, and butyric) coloring materials, molasses colloidal and suspended materials. On the other hand, the sugar contents (sucrose, glucose, fructose and raffinose) and the non sugar content, e. g. nitrogenous components were determined. Inorganic contents (Na, K, CaO, MgO, P2O5 and SO3), sulfated ash and carbonated ash were determined. Also, the minor concentrations of the metal ions, such as Zn, Cu, Fe, Pb, Mn, and Cd were assessment. On the other hand, the physical properties, e. g. specific gravity,
Ramadan, et al (2012) Egyptian Sugar 36 Journal,Vol.5 : 23- 46 ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ viscosity, color % Brix, and pH were measured. All results obtained refer to the safe concentration of the harmful chemicals as compared to analogous investigations; in addition the fermentable sugar content is in the promising range for the bio-production of ethanol.
References Achard, F.C. (1809): Die europäische Zuckerfabrikation aus Runkelrüben. J.C. Hinrichs, Leipzig, 14. Cited in Sugar Technology, Beet and Cane Sugar Manufacture", Published with support of the Beet Sugar Development Foundation, Denver, USA, editor Verlag Dr. Albert Bartens KG-Berlin, pp. 209-234. Anxo, M.M., P. Lorenzo, A.V. Jose, M. Jesus and P.G. Maria. (2008): Alcoholic chestnut fermentation in mixed culture. Compatibility criteria between Aspergillus oryzae and Saccharomyces cerevisiae strains. Bioresourc. Technol., 99: 7255-7263. Bajaj, B.K., S. Yousaf and R.I. Thakur. (2001): Selection and characterization of yeasts for desirable fermentation characteristics. Ind. J. Microbiol., 41: 107-110. Balling, C. I. N., (1865). Die Branntweinbrennerei und die Hefeerzeugung, Prag. Cited in Honig. P, (1963): Principles of Sugar Technology, Volume III, editor Elsevier Publishing Company Amsterdam, , pp. 511-693. Bertolini M C, Ernandes J R and Laluce C (1991): New yeast strains for alcoholic fermentation at higher sugar concentration. Biotechnol Lett 13: 197-202. Borzani W, Garab A, Pires M H, Piplovic R, De Ia Higuera, G. A., (1993): Batch ethanol fermentation of molasses: a correlation between the time necessary to complete the fermentation and the initial
Physicochemical investigations of Delta beet 37 molasses for the………. ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ concentrations of sugar and yeast cells. World J Microbial Biotechnol 9: 265-68. Breitung, (1956).
Z. Zuckerind, 6, 185, 254. Cited in Honig. P, (1963):
Principles of Sugar Technology, Volume III, editor Elsevier Publishing Company Amsterdam, , pp. 511-693. Carvalho, J.C.M., E. Aqarone, M.L. Sato, D.A. Brazzach, A. Moraes and H. Borzani. (1993). Fedbatch alcoholic fermentation of sugarcane blackstrap molasses: influence of feeding rate on yeast yield and productivity. Appl. Microbiol. Biotechnol., 38: 596-598. Claassen, (1940).
Die praktische Kristallisation des Zuckers und die
Melassebildung, Magdeburg., Cited in Honig. P, (1963): Principles of Sugar Technology, Volume III, editor Elsevier Publishing Company Amsterdam, , pp. 511-693. Converti, A.S., S. Ami, J.C.M. Sato, M, De-Carvalho and E. Aquarone. (2003): Simplified modeling of fed-batch alcoholic fermentation of sugarcane blackstrap molasses. Biotechnol. Bioengin., 84: 88-95. Drews, B. Just, F.
and Gundermann, K. (1955): Tätigkeitsbericht der
Versuchsanstalt der Hefeindustrie im Institut für Gärungsgewerbe, Berlin p. 26. El-Samman, A.A. (2010): Organic and inorganic constituents analysis of cane molasses and its affect on microbial fermentation industries. Ph. D Thesis, Sugar Technology Research Institute, Assiut University, Assiut, Egypt. Hildebrandt, (1955): Die Rübenzuckerfabrikation, Leipzig. Cited in Honig. P, (1963): Principles of Sugar Technology, Volume III, editor Elsevier Publishing Company Amsterdam, , pp. 511-693.
Ramadan, et al (2012) Egyptian Sugar 38 Journal,Vol.5 : 23- 46 ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ Honig, P. (1953): Principles of sugar technology, volume I, Edition by Elsevier Publishing Company, pp 508-693. Jones, A. M, Thomas K. C. and Inglew W. M., (1994): Ethanolic fermentation of molasses and sugarcane juice using very high gravity technology. J. Agric. Food Chem 42: 1242-1246. Karczewska, (1951): Lebensmittelindustrie, 70., Cited in Honig. P, (1963): Principles of Sugar Technology, Volume III, editor Elsevier Publishing Company Amsterdam, , pp. 511-693. Notkina, L. G., Balyberdina, L. M. and Lavrenchuk, L.D. (1975): The effect of nitrites in baker′s yeast manufacture. Khlebopek. Konditer. Promst. (2) 28-31. Pollak and M. Knob, (1922): Brennerei-Ztg. No. 1497, p. 39. Cited in Honig. P, (1963): Principles of Sugar Technology, Volume III, editor Elsevier Publishing Company Amsterdam, , pp. 511-693. Walker, G.M., A.I. Maynard and C.G.W. Johns. (1990): The importance of magnesium ions in yeast biotechnology. J. Ferment. Technol., 28: 233-240.
Physicochemical investigations of Delta beet 39 molasses for the………. ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ 1.46
Dynamic viscosity
1800
Line II
1600
1.45
1400
Spesific gravity
1.44
1200 1.43 1000 1.42 800
Dynamic viscosity
Specific gravity
1.41 600 1.4
400
79.3
79.3
79.1
79.2
79.4
80
79.8
80.1
80.1
80.2
80.1
80.2
80.1
80.2
80.4
80.6
0
80
1.38
80.4
200
78.8
1.39
Brix
Figure (1): Relationship between Brix, Specific gravity and the Dynamic viscosity of the beet molasses samples of lines I and II.
Table(1): Chemical Analysis of the Egyptian beet roots was used in Delta Sugar Company for the production lines (I & II). Line I No
Period
Sugar
K mmol/
Na mmol/
% 100gm
100gm
Line II Amino N
Quality
Sugar
mmol/
%
%
K mmol/ 100gm
Na mmol/ 100gm
Amino N
Quality
mmol/
%
Ramadan, et al (2012) Egyptian Sugar 40 Journal,Vol.5 : 23- 46 ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ 100gm
100gm
1
27/1- 30/1
15.51
6.4
5.0
3.9
70.6
15.o9
6.2
4.5
3.7
70.1
2
31/1- 4/2
15.67
6.3
4.9
4.1
71.3
15.65
6.5
4.5
4.0
71.1
3
5/2- 8/2
15.90
6.4
4.8
3.9
71.6
16.05
6.5
4.5
4
9/2- 12/2
16.29
6.3
4.5
3.9
73.2
16.61
6.6
4.2
3.6
73.6
5
13/2- 17/2
16.64
6.3
4.4
4.2
73.9
16.96
6.7
4.0
3.8
74.2
6
18/2- 22/2
17.00
6.2
4.0
4.2
75.1
17.24
6.6
3.6
3.9
75.5
7
23/2- 28/2
16.51
6.1
3.9
4.0
75.4
16.98
6.5
3.4
3.5
76.1
8
1/3- 12/3
17.62
6.1
3.4
3.9
77.8
17.53
6.4
3.1
3.5
77.7
9
13/3- 21/3
17.51
6.0
3.1
3.8
78.5
17.50
6.2
2.8
3.5
78.5
10
22/3- 28/3
17.11
5.9
3.0
3.6
78.6
17.56
6.0
2.7
3.3
79.4
11
29/3- 2/4
18.30
6.2
2.8
3.6
79.7
18.08
6.1
2.6
3.5
80.0
12
3/4- 10/4
18.58
5.9
2.8
3.5
80.7
18.51
6.1
2.5
3.5
80.5
13
11/4- 20/4
18.98
5.7
2.6
4.3
81.3
18.83
6.0
2.5
3.5
81.0
14
21/4- 30/4
19.47
5.9
2.6
4.1
81.5
19.28
6.1
2.5
3.5
81.3
15
1/5- 10/5
19.41
6.1
2.5
4.1
81.3
19.39
6.2
2.5
3.8
81.0
16
11/5- 20/5
19.61
6.2
2.5
4.2
81.4
19.43
6.3
2.5
3.7
81.1
17
21/5- 31/5
19.15
6.3
2.6
3.9
80.6
19.11
6.1
2.7
3.6
80.5
18
1/6 -10/6
18.91
6.2
3.3
4.1
79.1
18.88
6.2
3.1
3.7
79.4
19
11/6- 20/6
17.71
6.4
3.6
3.9
76.9
17.87
6.2
3.3
3.9
77.8
20
Average
17.68
6.2
3.5
4.0
77.3
17.86
6.3
3.2
3.6
77.4
3.6
72.3
*Each sample was represented sub-sample from different mixed samples during each period.
Table (2): Physical and chemical properties of the beet molasses samples of line I. Period
Brix
Specific gravity
Dynamic viscosity
pH
Na
K mg/L
CaO mg/L
MgO mg/L
%Sugar
Purity
Physicochemical investigations of Delta beet 41 molasses for the………. ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ mg/L 27/1-30/1
79.4
1.41023
978.6
8.5
0.08
0.20
1.34
4.03
48.0
60.45
31/1-4/2
79.8
1.41288
987.5
9.2
0.07
0.19
1.35
4.07
48.19
60.39
5/2-8/2
79.7
1.41222
984.0
9.3
0.08
0.19
1.41
4.01
48.16
60.43
9/2-12/2
80.0
1.41421
1265.0
9.5
0.08
0.20
1.39
4.08
48.68
60.85
13/2-17/2
80.2
1.41554
1530.1
9.4
0.09
0.16
1.38
3.95
48.68
60.70
18/2-22/2
79.9
1.41355
1400.3
9.1
0.09
0.17
1.36
4.06
47.95
60.01
23/2-28/2
79.6
1.41155
1206.9
8.8
0.08
0.18
1.22
4.12
47.29
59.41
1/3-12/3
79.8
1.41288
914.8
9.2
0.07
0.16
1.11
4.01
47.49
59.51
13/3-21/3
79.7
1.41222
921.1
8.9
0.06
0.16
1.05
4.32
47.43
59.51
22/3-28/3
79.6
1.41155
906.8
9.1
0.05
0.15
0.98
4.46
47.40
59.55
29/3-2/4
79.8
1.41288
958.1
9.0
0.06
0.14
0.97
4.10
47.53
59.56
3/4-10/4
79.9
1.41355
956.9
9.0
0.06
0.16
1.12
4.09
47.61
59.59
11/4-20/4
79.6
1.41155
899.3
9.0
0.08
0.12
1.31
4.07
47.46
59.62
21/4-30/4
79.8
1.41288
901.2
8.9
0.07
0.14
1.21
4.18
47.55
59.59
1/5-10/5
79.8
1.41288
915.5
8.9
0.07
0.18
1.09
3.99
47.64
59.70
11/5-20/5
80.1
1.41488
1309.2
8.8
0.09
0.19
1.58
4.26
47.73
59.59
21/5-31/5
80.2
1.41554
1312.8
8.2
0.08
0.24
1.09
4.31
47.88
59.70
1/6-10/6
80.3
1.41621
1542.1
8.7
0.06
0.23
1.55
4.22
47.99
59.76
11/6-20/6
79.8
1.41550
1530.2
8.1
0.06
0.24
1.49
4.15
46.95
58.83
Average
79.8
1.4133
1127.4
8.9
0.07
0.18
1.26
4.13
47.77
59.83
*Each sample was represented sub-sample from different mixed samples during each period.
Table (3): Analysis of Brix, Color % Brix, sulfur and Sulfite (%) of beet molasses samples of lines and II. Line I No.
Period*
1
Line II
Brix
Color% Brix
S
SO3
Brix
Color% Brix
S
SO3
27/1- 22/2
79.4
28267
0.67
1.68
78.8
40174
0.67
1.68
2
23/2 – 21/3
79.6
31620
0.57
1.43
80.1
44616
0.77
1.93
3
22/3 – 5/6
79.6
33181
3.41
1.53
80.2
42970
3.99
1.98
4
11/6 – 20/6
79.8
32670
2.71
1.78
79.3
43662
2.97
1.43
Average
79.6
31434.5
1.84
1.60
79.6
42855.5
2.10
1.75
*Each sample was represented sub-sample from different mixed samples during each period.
Ramadan, et al (2012) Egyptian Sugar 42 Journal,Vol.5 : 23- 46 ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ Table (4): The percentage (%) of total sugar as reducing sugar, fermentable sugar and nonfermentable sugar of beet molasses samples of lines I and II. Line I No.
Line II
Period* Total
Nonfermentable sugar
sugar
Fermentable
Total
Nonfermentable
Fermentable
sugar
sugar
sugar
sugar
1
27/1-30/1
54.4
0.96
53.44
54.9
1.1
53.8
2
31/1-4/2
54.1
1.2
52.9
54.8
0.95
53.85
3
5/2-8/2
53.9
1.1
52.8
54.9
1.3
53.6
4
9/2-12/2
53.7
0.90
52.8
54.5
1.1
53.4
5
13/2-17/2
54.3
1.6
52.7
55.0
1.4
53.6
6
18/2-22/2
54.2
2.1
52.1
54.3
1.8
52.5
7
23/2-28/2
53.9
1.3
52.6
54.2
1.8
52.4
8
1/3-12/3
53.8
1.0
52.8
54.1
2.3
51.8
9
13/3-21/3
53.2
1.9
51.3
53.8
1.8
52.0
10
22/3-28/3
53.6
1.9
51.7
53.2
1.2
52.0
11
29/3-2/4
53.6
1.3
52.3
54.0
1.6
52.4
12
3/4-10/4
54.0
1.7
52.3
53.8
1.1
52.7
13
11/4-20/4
53.1
1.7
51.4
53.1
1.1
52.0
14
21/4-30/4
53.9
1.2
52.7
53.6
2.0
51.6
15
1/5-10/5
53.6
2.0
51.6
54.1
1.9
52.2
16
11/5-20/5
53.9
2.1
51.8
53.9
1.9
52.0
17
21/5-31/5
53.2
1.7
51.5
53.2
2.1
51.1
18
1/6-10/6
54.1
1.7
52.4
53.2
1.8
51.4
19
11/6-20/6
53.7
1.8
51.9
53.6
1.9
51.7
20
Average
53.8
1.53
52.27
54.01
1.59
52.42
*Each sample was represented sub-sample from different mixed samples during each period.
Physicochemical investigations of Delta beet 43 molasses for the………. ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ Table (5): HPLC analytical results (%) of sucrose, glucose, fructose and raffinose of the beet molasses samples of lines I and II. No.
Period
Sugar
Line I
Line II
1
27/1 - 21/3
Sucrose
48.40
47.60
Glucose
0.30
0.06
Fructose
0.50
0.43
Raffinose
0.82
1.40
Sucrose
47.50
48.70
Glucose
0.07
0.36
Fructose
0.43
0.39
Raffinose
0.18
0.94
2
22/3 – 20/6
Table (6): HPLC analytical results (%) of sucrose, glucose, fructose and raffinose of the beet molasses of mixed samples. Period 27/1 – 20/6
Sugar
% Sugar
Sucrose
48.9
Glucose
0.29
Fructose
0.51
Raffinose
1.70
Ramadan, et al (2012) Egyptian Sugar 44 Journal,Vol.5 : 23- 46 ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ
Table (7): Determination of the percentage (%) of amino acids of the beet molasses of lines I and II. samples Line I No.
Amino acid
Line II
Period*
Period*
27/1-22/2
23/2-21/3
22/3-5/6
27/1-22/2
23/2-21/3
22/3-5/6
1
Glutamic acid
3.467
3.992
3.945
4.088
4.102
2.774
2
Aspartic
0.737
0.641
0.563
0.480
0.549
0.769
3
Threonine
0.104
0.100
0.0474
0.013
0.077
0.0585
4
Serine
0.280
0.146
0.197
0.110
0.103
0.142
5
Glycine
0.174
0.188
0.188
0.232
0.127
0.207
6
Cystin
0.010
0.008
0.00
0.036
0.021
0.00
7
Valine
0.230
0.255
0.327
0.253
0.218
0.400
8
Methionine
0.062
0.022
0.00
0.032
0.040
0.00
9
Isoleucine
0.326
0.401
0.229
0.425
0.358
0.201
10
Leucine
0.366
0.434
0.449
0.480
0.371
0.296
11
Tyrosine
0.395
0.394
0.401
0.540
0.261
0.389
12
Phenylalanine
0.800
0.699
0.810
0.514
0.623
1.438
13
Histidine
0.036
0.147
0.0656
0.276
0.134
0.0689
14
Lysine
0.116
0.115
0.108
0.199
0.073
0.146
15
Arginine
0.071
0.135
0.806
0.161
0.026
0.933
16
Alanine
0.481
0.468
1.070
0.301
0.403
1.084
17
Total
8.55
9.06
9.21
9.01
8.37
8.91
* Each sample was represented sub-sample from different mixed samples during each
period.
Physicochemical investigations of Delta beet 45 molasses for the………. ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ
Table (8):Determination of nitrate and nitrite (ppm) of the beet molasses samples of lines I and II. No.
Period*
Line I
Line II
NO2
NO3
NO2
NO3
1
27/1- 22/2
1169.2
101.4
270.1
132.6
2
23/2 - 21/3
90.0
78.4
87.4
88.1
3
22/3 - 5/6
97.3
89.2
151.0
212.5
4
6/6 - 20/6
18.8
47.0
16.9
50.3
5
Average
343.8
79.0
131.4
120.9
* Each sample was represented sub-sample from different mixed samples during each period.
No.
Period
Organic acid
Line I
ine II
1
27/1-21/3
Formic
0.21
0.25
Acetic
0.64
0.59
Butyric
0.29
0.34
Formic
0.27
0.29
Acetic
0.57
0.61
2
22/3-20/6
Ramadan, et al (2012) Egyptian Sugar 46 Journal,Vol.5 : 23- 46 ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ Butyric
0.41
0.58
Table (9): Assessment the percentage (%) of the volatile organic acids (formic,
Table (10): Determination of volatile organic acid as acetic acid % of the beet molasses samples of lines I and II. No.
Period
Line I
Line 2
1
27/1- 22/2
0.9
1.1
2
23/2 - 21/3
1.3
1.0
3
22/3 - 5/6
0.8
0.8
4
6/6 - 20/6
1.2
0.9
5
Average
1.05
0.95
Table (11): Determination of the heavy metals (ppm) of the beet molasses samples of lines I & II. No.
Period
Pb
Cu
Mn
Zn
Fe
Cd
27/1- 22/2
27.66
5.47
ND
ND
154.65
ND
2
23/2 – 21/3
25.71
7.42
ND
ND
706.17
ND
3
22/3 – 5/6
10.60
1.20
ND
37.70
8.30
0.10
4
6/6 – 20/6
9.70
0.70
13.40
18.80
17.70
1.80
27/1- 22/2
29.49
11.90
ND
ND
237.90
ND
1
1
Lines I
II
Physicochemical investigations of Delta beet 47 molasses for the………. ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ 2
23/2 – 21/3
14.60
3.37
ND
ND
99.47
ND
3
22/3 – 5/6
13.00
1.30
ND
31.30
16.10
0.12
4
6/6 – 20/6
9.50
0.60
10.80
20.50
24.50
1.30
17.54
4.0
12.1
27.08
158.1
0.83
Average
ND = Not detected Table (12): Determination of the percentage (%) of P and P2O5 of representative beet molasses samples of lines I and II. Line I
Line II
No.
Period
P
P2O5
P
P2O5
1
27/1 – 21/3
0.18
0.41
0.22
0.50
2
22/3 – 20/6
0.13
0
0.16
0.37
0.19
0.43
.30 3
Average
0.15
0.35
