PRODUCTION Of NUTRIENT SOURCES FOR RHIZOBIUM

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�� appropriate concentration of inocula which was 24-48 hours olds were inoculated into the starch medium. From the results as shown in Figure 53 and 54, when yeast isolate KAY1 was cultured in 2% Khao-klong starch medium. The maximum yield of mannitol in cell lysate was detected in 5% sodium chloride at 2.837 g/l of mannitol when incubated for 2 days. The reducing sugars disappeared in the 5% and 7% sodium chloride concentrations. Extracellular and intracellular glycerol was found the maximum amounts at the second day of cultivation. In the contrast, when cultured in 2% rice starch medium (Figure 55 (a) and (b)) and 2% cassava starch medium (Figure 56). The yeast cell could not grow well in the 2% starch medium containing sodium chloride, but the reducing sugars and mannitol in cell lysate were observed the highest in the control. The results clearly suggested that the yeast isolate KAY1 could grow well on 2% Khao-klong starch medium containing sodium chloride, but not grow well on 2% rice starch and 2% cassava starch medium containing sodium chloride. From previous study, in Khao-klong starch medium showed the highest yield of mannitol production at the first day of cultivation correlate to the amylase enzyme specific to the substrate. Relatively, at the first day of cultivation the starch was rapidly hydrolyzed, and the reducing sugars was highly absorbed in to the yeast cell as sole carbon source. The high amounts of reducing sugars at the first day of fermentation enough to maintained the energy and converted into the mannitol which protected the cell from salt-stress. Extracellular and intercellular glycerol were detected at the high amounts of 5% and 7% sodium chloride correlate to the previous study from other authors. Van Eck et al., (1993) reported that glycerol was the principle osmolyte in some filamentous fungi, algae, insects, crustaceans and vertebrates, which indicates that there was a selective advantage to organisms using such low molecular mass solutes (Yancey et al., 1982). As on osmolyte, glycerol offers a number of advantages: high glycerol concentrations confer a remarkable degree of protection to enzymes and macromolecular structure (Brown, 1978). From the results in Figure 54, the amounts of glycerol were increased correlated to the increasing concentration of sodium chloride. The ability to retained a greater proportion of a constant total (intra-and extracellular-) amounts of glycerol intracellularly was proposed by Brown (1978) as an essential characteristic of an osmotolerant yeast such as Zygosaccharomyces rouxii, whereas a nonosmotolerant yeast such as S. cerevisiae produced increasing amounts of glycerol and retained
g/l g/l 2.0 1.6 1.2 0.8 0.4 0.0 3.2 2.8 2.4 2.0 1.6 1.2 0.8 0.4 0.0 1 0% 3% 5% 7% 0% 3% 5% 7% Day 1 Day 2 0% 3% 5% 7% 0% 3% 5% 7% 1 Day 1 Day2 Reducing sugars Mannitol in cell lysate Mannitol in cultured medium Figure 53. Effects of various NaCl concentrations (0%, 3%, 5%, and 7% (w/v)) on the production of (a) reducing sugars and (b) mannitol by yeast isolate KAY1 when cultured in the medium containing 2% Khao-klong starch for 2 days at 30°C. (a) (b) ��
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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
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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 and 122: g/l g/l 4.0 3.5 3.0 2.5 2.0 1.5 1.0
Page 123: �� fermentation, but the inc
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.
Page 173 and 174: �� 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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