PRODUCTION Of NUTRIENT SOURCES FOR RHIZOBIUM

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Table 2. Summary of carbon utilization by Rhizobium spp a . Carbon source Fast-growing species Intermediate Slow-growing species Group A Group B Group C Group D Group E Group F Glucose + b + + + + + Fructose + + + + + + Galactose + + + + + + Gluconate N.D. c N.D. + + + + Mannose + + + + + + Mannitol + + + + + + Rhamnose + + + - d N.D. N.D. Dulcitol + ± e - - N.D. N.D. Arabinose + + + + + + Xylose + + + + ± + Raffinose + + + - N.D. N.D. Sucrose + + + + - ± Lactose + + + + ± ± Trehalose + + + - N.D. N.D. Glycerol N.D. N.D. + + + + Pyruvate + N.D. + - ± - Citrate + + - - - - Succinate + N.D. + + ± + Fumarate + N.D. + - N.D. - Malate + N.D. + - - ± aGroup A = Rhizobium meliloti, R. leguminosarum, R. trifolii, R. phaseoli; Group B = fast-growing Rhizobium spp.; Group C = fast-growing Rhizobium japonicum; Group D = intermediate group rhizobia; Group E = Rhizobium japonicum; Group F = cowpea miscellany rhizobia. b + indicates strains able to use the carbon source for growth; N.D. c not determined; d – indicates �
strains unable to use the carbon source for growth; e ± indicates more than one strain able to use the carbon source for growth. Source: Stowers (1985). 1.1.2 Carbon metabolism in free-living rhizobia It is well established that rhizobia possess the Entner-Doudoroff (ED) pathway or the ketogluconate aldolase (Figure 1) and most hexose metabolism proceeds through the ED pathway (Glenn, et al., 1984; Katznelson, 1955; Katznelson, et al., 1957, quoted in Stowers, 1985). A hexose cycle has been shown to operate simultaneously in some slow-growing rhizobia, allowing for the conservation of hexoses for biosynthesis from glyceraldehyde-3-phosphate(G3P). The hexose cycle recycles G3P to glucose-6-phosphate via triose phosphate isomerase, Fructose bisphosphate aldolase, hexose diphosphatase and phosphoglucose isomerase (Moat, 1995). The operation of the Embden-Meyerhof-Parnas (EMP) pathway (Figure 2) in rhizobia appears to be strain-dependent, representing only low levels of activity when observed. Phosphofructokinase activity was not detected in R. trifolii, indicating the lack of the EMP pathway in this species. Phosphofructokinase activity was detected in glucose-grown Rhizobium sp. 32H1, but not with succinate-, gluconate-, mannitol-, or fructose-grown cells (Stowers, 1985). Fructose bisphosphate aldolase, another key enzyme of the EMP pathway, has been demonstrated in rhizobia. This enzyme appears to be involved in the hexose cycle as described above rather than with the EMP pathway. The tricarboxylic acid (TCA) cycle is operational in rhizobia. The operation of the TCA cycle has been confirmed using radiorespirometry (Stowers et al., 1983, quoted in Stowers, 1985). Specifically labeled pyruvate gave the following 14 CO 2 evolution patterns: C-1 > C-2 > C-3. The preferential release of the C-1 (carboxyl carbon) of pyruvate followed by the C-2 (carbonyl carbon) and C-3 (methyl carbon) is indicative of an operational TCA cycle. Central carbon catabolic pathways are outlined for fast- and slow-growing rhizobia in Figures 3 and 4, respectively. Mannitol, the most common carbon source for Rhizobium cultivation, is metabolized via a mannitol dehydrogenase, producing fructose. In R. meliloti, an NAD-linked polyol �
Page 1 and 2: PRODUCTION OF NUTRIENT SOURCES FOR
Page 3 and 4: PRODUCTION OF NUTRIENT SOURCES FOR
Page 5 and 6: CHARDCHAI BUROM : PRODUCTION OF NUT
Page 7 and 8: CONTENTS Page ABSTRACT (THAI)……
Page 9 and 10: 3.6.1� Determination of the suita
Page 11 and 12: �� 17� Glycerol concent
Page 13 and 14: 40� Time courses of mannitol prod
Page 15 and 16: 59� Effects of various heat shock
Page 17 and 18: LIST OF TABLES Table Page ��
Page 19 and 20: LIST OF ABBREVIATIONS Aw water avai
Page 21 and 22: CHAPTER I INTRODUCTION For mass cul
Page 23 and 24: available (Burton, 1965). The molec
Page 25: 1.1.1� Carbon metabolism in Rhizo
Page 29 and 30: Figure 2. The fructose bisphosphate
Page 31 and 32: 1.2�Classification of yeasts Mank
Page 33 and 34: Table 3. (continued). Class Order F
Page 35 and 36: Ustilaginales (Oberwinkler, 1987).
Page 37 and 38: f Cystofilobasidium has thick-walle
Page 39 and 40: Table 5. (continued). Sugar alcohol
Page 41 and 42: specific cases, but Brown (1978) pr
Page 43 and 44: KY 6166 was also studied using larg
Page 45 and 46: Figure 5. Important pathways of gly
Page 47 and 48: may be present in different ratios.
Page 49 and 50: amylase systems of the yeast specie
Page 51 and 52: Table 7. (continued). EC Recommende
Page 53 and 54: Table 8. (continued). Species a D.p
Page 55 and 56: Table 9. (continued). Cell product
Page 57 and 58: 2.1.3 Equipment and other materials
Page 59 and 60: Spotted either the supernatant of c
Page 61 and 62: amylase action, the actual reaction
Page 63 and 64: A) Determination of the suitable an
Page 65 and 66: B) Salt-stress conditions Fifty mil
Page 67 and 68: 2.2.8 Cultivation of Bradyrhizobium
Page 69 and 70: 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
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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
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g/l g/l 1.6 1.4 1.2 1.0 0.8 0.6 0.4
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The results showed that amounts var
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g/l g/l 4.0 3.5 3.0 2.5 2.0 1.5 1.0
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�� fermentation, but the inc
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g/l g/l 2.0 1.6 1.2 0.8 0.4 0.0 3.2
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g/l g/l 1.2 1.0 0.8 0.6 0.4 0.2 0.0
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�� intracellularly a decreas
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g/l g/l 5.0 4.0 3.0 2.0 1.0 0.0 2.5
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g/l g/l 2 1.8 1.6 1.4 1.2 1 0.8 0.6
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g/L 7 6 5 4 3 2 1 0 1 2 3 4 5 6 7 0
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g/l 8 7 6 5 4 3 2 1 0 1 2 3 4 5 6 7
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�� effected by heat shock po
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Table 15. The comparison of media f
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Table 17. Growth of Bradyrhizobium
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�� Rhizobium (Stowers, 1985;
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�� 3.9 Identification of yea
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KAY1 PIY2 PUY4 Y69 LIY2 COY1 FAY2 C
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�� 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
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(a) (b) (c ) (d) (e) �� Figu
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(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
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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
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
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�� Sinclair, N. A., and Stok
Page 183 and 184:
�� 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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Page 201 and 202:
Page 203 and 204:
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Page 207 and 208:
Page 209 and 210:
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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
Page 220:
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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