PRODUCTION Of NUTRIENT SOURCES FOR RHIZOBIUM

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�� In this experiment, heat shock treatment was performed at 45°C for 20 min, a novel method was used for enhanced the sugar alcohol production. It was considered that the method supported the yeast isolate KAY1 to produced a large amounts protein of amylase to converted the starch to high amounts of reducing sugars. Consequently, a large amounts of reducing sugars was uptake into the yeast cell and the high amounts of mannitol were constructed. A residual starch concentration was rapidly decreased, at the same time the maximum yield of sugar alcohol production was appeared. When the cultivation of rhizobia were tested. The adjusted cell lysate medium containing 0.5% mannitol obtained from yeast isolate KAY1 were gave the final number of Bradyrhizobium japonicum USDA 110 and Bradyrhizobium spp. THA 5 cell at 10 8 cells/ml at the sixth day, being almost the same as that found with YMB the standard rhizobia culture medium. In separate experiments, the ability of Bradyrhizobium to grow on fermented sucrose medium was also tested. The good result was obtained with 0.5% glycerol diluted from 5% sucrose prefermented by yeast isolate PIY2 and PUY4 at the second day of fermentation, gave the amounts of bacterial cells at 10 8 - 10 9 cells/ml at the sixth day. In contrast, with the Bradyrhizobium japonicum USDA 110, the number of viable cells gave lower than Bradyrhizobium japonicum spp. THA 5 when cultured in the same medium and compared with the G5 acted as glycerol standard medium. The results presented here clearly demonstrated that mannitol-yeast cell from starch hydrolyzation and glycerol in cell-free sucrose fermentation, can be used for the cultivation of Bradyrhizobium. The identification of yeast isolates were investigated. It was found that the yeast isolate KAY1 most likely belongs to the genus Rhodotorula. Isolates PIY2 and PUY4 were classified in the genera Zygosaccharomyces and Pichia, respectively. However, some fermentation conditions have not been investigated for example pH control, phosphate concentration, effect of aeration, or effect of temperature, and other various nitrogen sources. This study apparently presented rather simple and without operation control. In the future research work, for the isolates KAY1. Increased the large scale production, optimization, and strain improvement should lead to even higher product concentration in the final fermentation broth in the future. In addition, the raw material utilization such as starch fermentation technology which were study in this experiments offered the advantages of being simple and requiring
only a low capital investment. Therefore, by using the utilization of expensive hexoses or mannitol and yeast extract, especially in developing countries, can be reduced. ��
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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
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CHAPTER III RESULTS AND DISCUSSION
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Glycerol Xylitol Glucose Mannitol S
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Glycerol Xylitol Glucose Mannitol S
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Table 10. Primary screening of glyc
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Table 10. (continued). No. Isolate
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Glycerol in cell lysate (g/l) A Gly
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average were 4-10 g of glycerol per
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were isolated from fresh fruits, Ma
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�third day of incubation (Figure
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Reducing sugars (g/l) Reducing suga
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These results indicated that the co
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3.6.1 Determination of the suitable
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g/l 1.8 1.6 1.4 1.2 1.0 0.8 0.6 0.4
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Reducing sugars (g/l) 8 6 4 2 0 0 1
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Reducing sugars (g/l) 4.0 3.5 3.0 2
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g/l 1.4 1.2 1.0 0.8 0.6 0.4 0.2 0.0
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g/l g/l g/l 1.5 1.0 0.5 0.0 1.2 1.0
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Starch fermentation was conducted i
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g/l g/l 7 6 5 4 3 2 1 0 8 6 4 2 0 0
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g/l g/l 8 6 4 2 0 8 6 4 2 0 1 2 3 4
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g/l g/l 8 6 4 2 0 8 6 4 2 0 0 1 1 2
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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 and 124: �� fermentation, but the inc
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: CHAPTER IV CONCLUSION This study ex
Page 171 and 172: �� Barnett, J. A., Payne, R.
Page 173 and 174: �� De Mot, R., and Verachter
Page 175 and 176: �� Jennings, D. H. (1995). T
Page 177 and 178: �� Lie, T. A., Muilenbury M.
Page 179 and 180: �� Onishi, H., and Suzuki, T
Page 181 and 182: �� Sinclair, N. A., and Stok
Page 183 and 184: �� Van Rossum, D., Schuurman
Page 185 and 186: �� Young, J. P. W., and Hauk
Page 187 and 188: Acetone Methanol Solvent for mobile
Page 189 and 190: � 300 µg/ml, 200 µg/ml, and 100
Page 191 and 192: Soluble Starch 20.0 g Distilled wat
Page 193 and 194: 13.�Nitrate broth KNO 3 1.0 g Glu
Page 195 and 196: If the sample had been diluted befo
Page 197 and 198: Table 1D. (continued). Source of ye
Page 212 and 213: E. Additional figures Abs 540 nm (a
Page 214 and 215: Abs 580 nm 1.4 1.2 1 0.8 0.6 0.4 0.
Page 216 and 217: 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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