bioreactor studies of heterologous protein production by ...

More documents

Recommendations

Info

$uWaterloo LaTeX Thesis Template - UWSpace - University of Waterloo$

selected strain higher enzyme productivity was obtained in shake flasks, airairlift. stirred-tank and immobiiized ceii bioreactors; enhanced enzyme production and plasmid stabiiity were observed in the inniobilized aii systems for botb the original and selected strains. Therefore, the conclusions made in this research have a strong bask and the overali accuracy for the study is considered satisfactory. Figure 3.2. Reproducibility Check in Free Suspension Batch Fermentation (Temperature = JO OC, Aeration Rate = 1.0 WM).
-0-Cell(1) +Total CeII (2) 4- Ethanol(1) + Ethanol(2) 4 Enzyme (1) + Enzyme (2) -Fraction of PBC (1) + Fnctlon of PBC (2) Figure 3.3. Reproduabiiîty Check in Free Suspension Continuous Fermentation (Temlperature = 30 OC, Aedon Rate = 1.0 VVM, W o n Rate = 0.1 ho').
Page 1 and 2:
BIOREACTOR STUDIES OF HETEROLOGOUS
Page 3 and 4:
nie University of Waterloo requin%
Page 5 and 6:
ate depended on activities of the p
Page 7 and 8:
Acknowiedgments Fmt of aü, 1 am ve
Page 9 and 10:
2.6. Concluding Rernarks ..........
Page 11 and 12:
5.3.3. Simlilated Results for Prese
Page 13 and 14:
Table 1.1. Table 1.2. Table 1.3. Ta
Page 15 and 16:
Figure 2.1. Figure 3.1. Figure 3.2.
Page 17 and 18:
during Continuous Culture in Airlif
Page 19 and 20:
Figure 5.3. Comparison between Mode
Page 21 and 22:
Cell Systern at Glucose Feed Concen
Page 23 and 24:
a .......... growth ratio (dimensio
Page 25 and 26:
Y, ... ethanol yield for fermentati
Page 27 and 28:
foreign plasmids. One major chalien
Page 29 and 30:
than the denanired, inclusion body
Page 31 and 32:
Table 1.2. Strategies for Enhancing
Page 33 and 34:
Table 1.3. Host Cek, Pfasmids, Gene
Page 35 and 36:
1.4. Immo bilized-Cell Bioreactors
Page 37 and 38:
1. Study the gmwth characteristics,
Page 39 and 40:
plasmid are used; or chromaromal DN
Page 41 and 42:
2.2.1. Genetic Factors 2.2.1.1. Pid
Page 43 and 44: 1- fkquently than a plasmid with Io
Page 45 and 46: Da Silva and Bailey (1991) used the
Page 47 and 48: LEU2 or TRPI is cloned into the aux
Page 49 and 50: eported reduced stabiliity in the s
Page 51 and 52: on metabolic pathways. In the ferme
Page 53 and 54: OC), oniy about 35% of the populati
Page 55 and 56: loss @) is dehned as the probabilit
Page 57 and 58: + s Dilution Rate = p' = CI,- - K*+
Page 59 and 60: concentration couid Iead to coexist
Page 61 and 62: that factors other than immobiiizat
Page 63 and 64: earing and plasmid-tke =Ils. Gel be
Page 65 and 66: productivity irrespective of the mo
Page 67 and 68: inactive and useless. The problern
Page 69 and 70: Aerobic fermentation osing iamnobih
Page 71 and 72: (ranghg h m 1 to 7) on citric acid
Page 73 and 74: celi fraction decreases monotonicai
Page 75 and 76: (1987) model aliows for plasmid ins
Page 77 and 78: affect plasmid stability have ben o
Page 79 and 80: CEtAPTER 3 MATERIALS AND METHODS 3.
Page 81 and 82: stiU deleted hm the plasmid. Such p
Page 83 and 84: 1) Reagent solution preparation was
Page 85 and 86: nonse1ective agar plates, but only
Page 87 and 88: working Liquid volume. The aeration
Page 89 and 90: 3.5. UNnobilized Ceil Culture 35.1.
Page 91 and 92: 3.52 Cd Lmmobiiization Yeast attach
Page 93: 3.6. Experimental Error and Reprodu
Page 97 and 98: * . I I a O 2 4 6 8 10 12 14 Batche
Page 99 and 100: Cell Maso, Glucose and EthanoI(g1L)
Page 101 and 102: 4.2.2. Batch CuIture in Stirreà-Ta
Page 103 and 104: [t ~lucoamylase (ALR) -e Glucoamyla
Page 105 and 106: The finai giucoamyiase concentratio
Page 107 and 108: Bentley and Kompala, 1989; Satyagal
Page 109 and 110: 160 First ExponenUaI Phase 1 -c ALR
Page 111 and 112: Figure 4hl. Pathways of Yeast Intem
Page 113 and 114: fermentation (using nitrogen spargi
Page 115 and 116: 43. Continuous Suspension Culture C
Page 117 and 118: Figure 4.9. Continuous Suspension C
Page 119 and 120: 1 +Enzyme +Fradlan of PBC +CeIl Mas
Page 121 and 122: Figures 4.12 and 4.13 show that by
Page 123 and 124: Fi- 4.13. Cornparison of Glu~iase C
Page 125 and 126: where 6 = p- - p' + pp*, which is a
Page 127 and 128: Figure 4-14. Effeds of Dilution Rat
Page 129 and 130: Impoolsup et ai. (1989a) and Hardji
Page 131 and 132: Figure 4.16. dB/dt versus B Plots a
Page 133 and 134: Figure 4.17. Totai Cell Mas at Diff
Page 135 and 136: another recombinant yeast (impoolsu
Page 137 and 138: other recombinant yeast under nonse
Page 139 and 140: 4.3.4. Effixt of Dilution Rate On R
Page 141 and 142: Figure 4.18- Etlects of Mluiioa Rat
Page 143 and 144: Number of Generations - - - - - - *
Page 145 and 146:
B Table 4.5. Metabolic Pathway Anal
Page 147 and 148:
concentration. But this may be more
Page 149 and 150:
[+ Enzyme (ALR) + Enzyme (STR) +Fra
Page 151 and 152:
necessary for host ce& to grow in t
Page 153 and 154:
Glucoamylase ConcentraUon (unltsll)
Page 155 and 156:
[a Selective Medium A Nonseiective
Page 157 and 158:
5.1 Introduction CHAPTER 5 MODELING
Page 159 and 160:
handle. excess glucose is femented
Page 161 and 162:
The carbon fluxes h m the three met
Page 163 and 164:
53. Modei Simulation in Batch CuItu
Page 165 and 166:
Table 5.1. Summary of Parameters fo
Page 167 and 168:
1 n Glucose o Cell Mass O Ethano1 A
Page 169 and 170:
detennined in the cunent study, the
Page 171 and 172:
Figure 5.4. Cornparison between Mod
Page 173 and 174:
equations are üsted in Appendix G.
Page 175 and 176:
a dilution rate at which the giucoa
Page 177 and 178:
[ o Expetfmental data -Simuiated mt
Page 179 and 180:
The effixts of dilution rate on the
Page 181 and 182:
Ttme (hrs) 1 a Enzyme A Phsrnid-Bea
Page 183 and 184:
It may take several generations for
Page 185 and 186:
understanding of the nature of the
Page 187 and 188:
On the other hand. the net accumula
Page 189 and 190:
Differentiating Equation (6.20) res
Page 191 and 192:
Enyme collcentration m the t>ioreac
Page 193 and 194:
Glucoamylase Concentration (units/L
Page 195 and 196:
50 100 150 200 Time (hrs) 1 O Free
Page 197 and 198:
F i 6.6. Cornparison of Fnx Cd Mars
Page 199 and 200:
. - O 50 100 150 200 250 nme (hrs)
Page 201 and 202:
Nasi et ai.. 1988) and recombinant
Page 203 and 204:
estimated by using Equation (6.24)
Page 205 and 206:
[a lmmobilked System O Free System
Page 207 and 208:
oxidation pathway. W~th increasing
Page 209 and 210:
O 50 100 150 200 250 300 Tirne (hrs
Page 211 and 212:
epeated batch fermentations, the fe
Page 213 and 214:
Glucoamylase Concentration (unitslL
Page 215 and 216:
6) Suitable for repeated-batch and
Page 217 and 218:
ecause glucose fermentation dominat
Page 219 and 220:
cells. Tests of the proposed model
Page 221 and 222:
7.2. Recommendations The foUowing f
Page 223 and 224:
APPENDIX A: PLASMID MAP FOR pGAC9 l
Page 225 and 226:
where Bi (Biot number) = - ks De @
Page 227 and 228:
APPENDK C. EFFECTS OF INITIAL GLUCO
Page 229 and 230:
Figure C.2. Effect of Initial Gluco
Page 231 and 232:
Figure C.4 Effect of Initial Glucos
Page 233 and 234:
APPENDLX D. COMPARISON OF ORIGINAL
Page 235 and 236:
(continue)
Page 237:
(continue)
Page 240 and 241:
(continue)
Page 242 and 243:
E.2. Batch Fermentation with Recomb
Page 244 and 245:
(continue)
Page 246 and 247:
(continue) tfhJ 0.5 10.5 12.75 12.5
Page 248 and 249:
(continue)
Page 250 and 251:
E.3.2. Experimental Data (Parker an
Page 252 and 253:
Run No: CALE (Selected Strain. D =
Page 254 and 255:
Run No: CALR 5 (Original Suain, D =
Page 256 and 257:
Run No: CSTR3 (Selected Suain, D =
Page 258 and 259:
(continue)
Page 260 and 261:
Run No: CICR2 (Selected Suain, D =
Page 262 and 263:
Run No: CiCR4 (Selected Suain, D =
Page 264 and 265:
Eh. Repeated Batch Culture in hmobi
Page 266 and 267:
APPENDIX F. C CODE FOR THE PROPOSED
Page 268 and 269:
* the initial data */ /* gening the
Page 270 and 271:
* getting the founh term in R-K met
Page 272 and 273:
I*Plasrnid-bearing ceU concentratio
Page 274 and 275:
* Func tion C 1 */ îloat C l(float
Page 276 and 277:
APPEMIM G. EXAMPLES OF MAPLE V CODE
Page 278 and 279:
REFERENCES Ataai. MM. and Shuler. M
Page 280 and 281:
Caunt, P.. hpoolsup, A.. and Greenf
Page 282 and 283:
cuitanes of E. coli BZ18@TG 201) wi
Page 284 and 285:
Gabrieisen. O.S. et aL (1990), Effi
Page 286 and 287:
Huang, C-T., Peretti, S.W., and Bry
Page 288 and 289:
Kim, B.G. and Shuler, M.L. (1990a),
Page 290 and 291:
Lee, S.B.. Ryu, D.D.Y., seigel, R,
Page 292 and 293:
Nasri, M., Sayadi. S.. Barbotin, J.
Page 294 and 295:
Pin, S.J. and Kmwski. W.M. (1970),
Page 296 and 297:
Seo, I-H. and Bailey. J.E. (1985).
Page 298 and 299:
Tumer. B.G., Avgerinos G-C.? Melnic
Page 300:
Yu, J. and Tang, X (Editor, 1991),
show all

bioreactor studies of heterologous protein production by ...

Create successful ePaper yourself

Delete template?

Save as template?