PRODUCTION Of NUTRIENT SOURCES FOR RHIZOBIUM

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g/l g/l 1.6 1.4 1.2 1.0 0.8 0.6 0.4 0.2 0.0 0% 0.25% 0.5% 0.75% 10% 0.25% 0.5% 0.75% 1.2 1.0 0.8 0.6 0.4 0.2 Day3 Day4 0.0 1 0%0.25% 0.5%0.75% 0%0.25%0.5%0.75% Day 3 Day 4 Reducing sugars Mannitol in cell lysate Mannitol in cultured medium Figure 52. Effects of various CaCO 3 concentrations (0%, 0.25%, 0.5%, and 0.75%(w/v)) on the production of (a) reducing sugars and (b) mannitol by yeast isolate KAY1 when cultured in the medium containing 2% cassava starch for 4 days at 30°C. (a) (b) ��
�� fermentation, but the incubation products from the medium which containing the calcium carbonate gave the amounts of mannitol lower than the control. Similarly, when cultured in 2% cassava starch medium, at 0% of calcium carbonate gave the higher amounts of mannitol than the medium containing calcium carbonate at the third and the forth days of cultivation. In the same resulted, the control gave the highest amounts of mannitol at the second days of fermentation when cultured in 2% rice starch medium. It was clearly shown that calcium carbonate was not supported the production of mannitol in yeast cell. Calcium carbonate is a classical buffering agent. The addition of calcium carbonate was order to maintain adequate pH values for the enzymatic activity. Additionally calcium had a specific effect on enzyme stability to extreme pH and temperatures (Pintado et al., 1999). Calcium carbonate was used since it was found that under alkaline conditions increased concentrations of glycerol could be obtained (Lie et al., 1991). From the results, 2% Khao-klong starch medium showed the initial pH at 6.0 and pH level was decreased at 5.8 at the first day, but 2% rice starch medium containing calcium carbonate was maintained at pH 6.2-6.4 along the incubation. In 2% cassava starch medium, pH level was 4.4 at the third and the forth day. When added with calcium carbonate, the pH was maintained at pH 7.7-8.2. Similarly, 2% rice starch (Figure 51) gave the same result as 2% cassava starch (Figure 52) and 2% Khao-klong starch medium. The best yield of mannitol gave at the pH 4-5, but when added with calcium carbonate, the pH level was maintained at pH 6.2 or higher than control. It was suggested that in generally, yeast grow well at pH in the range of 3.5 and 5.0 (Kurtzman and Fell, 1998). The optimum pH values for most yeast amylases were found in the 4-6.5 ranges (Verachtert et al., 1990). For the yeast isolate KAY1, its enzymes might performed good activity was at pH 4-6.5. Thus, when calcium carbonate was added, the pH in the medium culture was reached and remained at between 6.0-8.0. In this condition, it could not grow well and amylase activity was reduced. Consequently, the cultivation products such as mannitol was decreased. 3.7.2 Salt-stress condition for sugar alcohol overproduction The fifty millilitres of starch medium containing 2% (w/v) of either cassava starch, rice starch, or Khao-klong starch medium were added with sodium chloride at 0, 3, 5, and 7% (w/v). The
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PRODUCTION OF NUTRIENT SOURCES FOR
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PRODUCTION OF NUTRIENT SOURCES FOR
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CHARDCHAI BUROM : PRODUCTION OF NUT
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CONTENTS Page ABSTRACT (THAI)……
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3.6.1� Determination of the suita
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�� 17� Glycerol concent
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40� Time courses of mannitol prod
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59� Effects of various heat shock
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LIST OF TABLES Table Page ��
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LIST OF ABBREVIATIONS Aw water avai
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CHAPTER I INTRODUCTION For mass cul
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available (Burton, 1965). The molec
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1.1.1� Carbon metabolism in Rhizo
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strains unable to use the carbon so
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Figure 2. The fructose bisphosphate
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1.2�Classification of yeasts Mank
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Table 3. (continued). Class Order F
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Ustilaginales (Oberwinkler, 1987).
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f Cystofilobasidium has thick-walle
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Table 5. (continued). Sugar alcohol
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specific cases, but Brown (1978) pr
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KY 6166 was also studied using larg
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Figure 5. Important pathways of gly
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may be present in different ratios.
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amylase systems of the yeast specie
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Table 7. (continued). EC Recommende
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Table 8. (continued). Species a D.p
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Table 9. (continued). Cell product
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2.1.3 Equipment and other materials
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Spotted either the supernatant of c
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amylase action, the actual reaction
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A) Determination of the suitable an
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B) Salt-stress conditions Fifty mil
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2.2.8 Cultivation of Bradyrhizobium
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The cultured medium of yeast. Quant
Page 71 and 72: intervals for accumulation of gas i
Page 73 and 74: CHAPTER III RESULTS AND DISCUSSION
Page 75 and 76: Glycerol Xylitol Glucose Mannitol S
Page 77 and 78: Glycerol Xylitol Glucose Mannitol S
Page 79 and 80: Table 10. Primary screening of glyc
Page 81 and 82: Table 10. (continued). No. Isolate
Page 83 and 84: Glycerol in cell lysate (g/l) A Gly
Page 85 and 86: average were 4-10 g of glycerol per
Page 87 and 88: were isolated from fresh fruits, Ma
Page 89 and 90: �third day of incubation (Figure
Page 91 and 92: Reducing sugars (g/l) Reducing suga
Page 93 and 94: These results indicated that the co
Page 95 and 96: 3.6.1 Determination of the suitable
Page 97 and 98: g/l 1.8 1.6 1.4 1.2 1.0 0.8 0.6 0.4
Page 99 and 100: Reducing sugars (g/l) 8 6 4 2 0 0 1
Page 101 and 102: Reducing sugars (g/l) 4.0 3.5 3.0 2
Page 103 and 104: g/l 1.4 1.2 1.0 0.8 0.6 0.4 0.2 0.0
Page 105 and 106: g/l g/l g/l 1.5 1.0 0.5 0.0 1.2 1.0
Page 107 and 108: Starch fermentation was conducted i
Page 109 and 110: g/l g/l 7 6 5 4 3 2 1 0 8 6 4 2 0 0
Page 111 and 112: g/l g/l 8 6 4 2 0 8 6 4 2 0 1 2 3 4
Page 113 and 114: g/l g/l 8 6 4 2 0 8 6 4 2 0 0 1 1 2
Page 115 and 116: g/l g/l 1.6 1.4 1.2 1.0 0.8 0.6 0.4
Page 117 and 118: g/l g/l 1.6 1.4 1.2 1.0 0.8 0.6 0.4
Page 119 and 120: The results showed that amounts var
Page 121: g/l g/l 4.0 3.5 3.0 2.5 2.0 1.5 1.0
Page 125 and 126: g/l g/l 2.0 1.6 1.2 0.8 0.4 0.0 3.2
Page 127 and 128: g/l g/l 1.2 1.0 0.8 0.6 0.4 0.2 0.0
Page 129 and 130: �� intracellularly a decreas
Page 131 and 132: g/l g/l 5.0 4.0 3.0 2.0 1.0 0.0 2.5
Page 133 and 134: g/l g/l 2 1.8 1.6 1.4 1.2 1 0.8 0.6
Page 135 and 136: g/L 7 6 5 4 3 2 1 0 1 2 3 4 5 6 7 0
Page 137 and 138: g/l 8 7 6 5 4 3 2 1 0 1 2 3 4 5 6 7
Page 139 and 140: �� effected by heat shock po
Page 141 and 142: Table 15. The comparison of media f
Page 143 and 144: Table 17. Growth of Bradyrhizobium
Page 145 and 146: �� Rhizobium (Stowers, 1985;
Page 147 and 148: �� 3.9 Identification of yea
Page 149 and 150: KAY1 PIY2 PUY4 Y69 LIY2 COY1 FAY2 C
Page 151 and 152: �� assimilations were observ
Page 153 and 154: Table 20. (continued). Yeast strain
Page 155 and 156: Table 20. (continued). 15. Pichia k
Page 157 and 158: �� 3.9.5 Ascospore formation
Page 159 and 160: (a) (b) (c ) (d) (e) �� Figu
Page 161 and 162: (a) (b) (c) (d) (e) (f) �� F
Page 163 and 164: (c) (a) �� Figure 72. Cell m
Page 165 and 166: �� Metshnikowia (Pitt and Mi
Page 167 and 168: CHAPTER IV CONCLUSION This study ex
Page 169 and 170: only a low capital investment. Ther
Page 171 and 172: �� Barnett, J. A., Payne, R.
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�� De Mot, R., and Verachter
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�� Jennings, D. H. (1995). T
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�� Lie, T. A., Muilenbury M.
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�� Onishi, H., and Suzuki, T
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�� Sinclair, N. A., and Stok
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�� Van Rossum, D., Schuurman
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�� Young, J. P. W., and Hauk
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Acetone Methanol Solvent for mobile
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� 300 µg/ml, 200 µg/ml, and 100
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Soluble Starch 20.0 g Distilled wat
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13.�Nitrate broth KNO 3 1.0 g Glu
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If the sample had been diluted befo
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Table 1D. (continued). Source of ye
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E. Additional figures Abs 540 nm (a
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Abs 580 nm 1.4 1.2 1 0.8 0.6 0.4 0.
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Figure 7E. Calibration curves for H
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Figure 9E. Example of HPLC chromato
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Cell/mL 1.0E+09 8.0E+08 6.0E+08 4.0
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PRODUCTION Of NUTRIENT SOURCES FOR RHIZOBIUM

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