Ghasem D. Najafpour. Biochemical Engineering and Biotechnology

Подождите немного. Документ загружается.

414 BIOCHEMICAL ENGINEERING AND BIOTECHNOLOGY

shortened overall process time. Here, the feasibility of the concept of performing cell disrup-

tion and product capture simultaneously was demonstrated for the purification of the G3PDH

from Saccharomyces cerevisiae and L-asparaginase from E. chrysanthemi. From a stage

comparison of conventional purification routes of the respective enzymes with the inte-

grated approach, it is evident that there is a considerable reduction in the number of con-

secutive unit operations in line with operating time-frames and concomitant product losses

(in particular due to the elimination of disruptate accumulation and discrete solid–liquid

separation steps). Because proteins are generally stabilised when adsorbed onto a solid sup-

port, it is advantageous to place an adsorptive step as far upstream as possible. It has been

demonstrated earlier that fluidised bed adsorption is a scaleable operation. In the same way,

bead mills are commercially available from laboratory to industrial scale. Thus, integrated

bead milling and fluidised bed adsorption is a scalable and generic approach to the efficient

recovery of intracellular proteins.

17.11.5 Acknowledgement

We are grateful to Professor Andrew. Lyddiatt, Dr Horst Bierau and the University of

Birmingham, UK, for various aspects of this collaborative work.

REFERENCES

1. Chase, H.A. and Draeger, N.M., J. Chromatogr. 597, 129 (1992).

2. Hjorth, R., Trends Biotechnol. 15, 230 (1997).

FIG. 17.9. Purity comparison (SDS–PAGE) of the conventional purification process and integrated cell disrup-

tion/fluidised bed adsorption.The numbers given in the flow sheet indicate the origin of samples and correspond

to their respective lane numbers. Lanes: M, low molecular weight markers; 1, Erwinia disruptate, 15% biomass

ww/v; 2, eluate CM HyperD LS, fluidised bed; 3, desalted eluate (after dia/ultrafiltration, 30 K MWCO mem-

brane); 4, flow-through, DEAE fixed bed; 5, elution, DEAE fixed bed; 6, eluate CM HyperD LS; 7, CM cellulose

eluate; 8, CM cellulose eluate, final; 9, final commercial product.

Ch017.qxd 10/27/2006 10:52 AM Page 414

ADVANCED DOWNSTREAM PROCESSING IN BIOTECHNOLOGY 415

3. Thommes, J., Halfar, M., Lenz, S. and Kula, M.-R., Biotechnol. Bioengng 45, 205 (1995).

4. Kaufmann, M., Unstable proteins: how to subject them to chromatographic separations for purification proce-

dures. J. Chromatogr. B 699, 347 (1997).

5. Jahanshahi M., Sun, Y., Santos, E., Pacek, A., Teixera, F.T., Nienow, A. and Lyddiatt, A., Biotechnol. Bioengng

J. 80, 201 (2002).

6. Zhang, Z. and Lyddiatt, A., J. Chem. Technol. Biotechnol. 74, 270 (1999).

Ch017.qxd 10/27/2006 10:52 AM Page 415

Appendix

Constants and conversion factors

Power W cal/s kcal/h Btu/s Btu/h lb

⭈ft/s hp

1 W 1 0.239 0.8604 9.478 ⫻ 10

⫺4

3.412 0.7376 1.341 ⫻ 10

⫺3

1 cal/s 4.18 1 3.6 3.966 ⫻ 10

⫺3

14.276 3.086 5.611 ⫻ 10

⫺3

1 kcal/h 1.1622 0.2778 1 1.102 ⫻ 10

⫺3

3.966 0.857 1.559 ⫻ 10

⫺3

1 Btu/s 1055 252 907.78 1 3600 778 1.415

1 Btu/h 0.293 0.07 0.252 2.78 ⫻ 10

⫺4

1 0.216 3.93 ⫻ 10

⫺4

1 lb

⭈ft/s 1.356 0.324 1,167 1.285 ⫻ 10

⫺3

4.63 1 1.818 ⫻ 10

⫺3

1 hp 746 178.2 642 0.707 2540 550 1

Energy J cal Btu lb

⭈ft Kw⭈hhp⭈herg

1 J 1 0.239 9.478⫻ 10

⫺4

0.738 2.778 ⫻ 10

⫺7

3.725 ⫻ 10

⫺7

1 ⫻ 10

1 cal 4.184 1 3.966 ⫻ 10

⫺3

3.086 1.162 ⫻ 10

⫺4

1.559 ⫻ 10

⫺6

4.18 ⫻ 10

1 Btu 1055 252 1 778 2.93 ⫻ 10

⫺4

3.93 ⫻ 10

⫺4

1.055 ⫻ 10

1 lb

⭈ft 1.356 0.324 1.285 ⫻ 10

⫺3

1 3.766⫻ 10

⫺7

5.05 ⫻ 10

⫺7

1.356 ⫻ 10

1 kw⭈h 3.6 ⫻ 10

8.6 ⫻ 10

3412 2.655 ⫻ 10

1 1.341 3.6 ⫻ 10

1 hp⭈h 2.68 ⫻ 10

6.42 ⫻ 10

2.54 ⫻ 10

1.98 ⫻ 10

0.7457 1 2.6845 ⫻

1 erg 1 ⫻ 10

⫺7

2.39 ⫻ 10

⫺8

9.478 ⫻ 10

⫺11

7.376 ⫻ 10

⫺8

2540 3.72 ⫻ 10

⫺4

Physical constants and conversion factors

Ideal gas law constant, R 8.314 J⭈mol

⫺1

⭈K

1.987 cal⭈mol

⫺1

⭈K

82.058 cm

⭈atm⭈mol

⫺1

⭈K

416

Appendix.qxd 10/27/2006 10:50 AM Page 416

Given a quantity Multiply by To get quantity

US gallons 3.785 Litres

Cubic feet 28.316 Litres

Metres 39.37 Inches

Kilograms 2.2046 Pounds

Ponds 453.59 Grams

Poise 0.1 g⭈cm

⫺1

⭈s

⫺1

Centipoise 0.01 poise

Centipoise 0.001 kg⭈m

⫺1

⭈s

⫺1

Centipoise 2.42 lb⭈ft

⫺1

⭈h

⫺1

APPENDIX 417

Appendix.qxd 10/27/2006 10:50 AM Page 417

418

Index

Activated sludge, 30, 37, 44, 180, 312

Adsorption, 171, 185–187

Aeration, 22–23, 44, 72, 84, 142, 148, 163, 181, 265,

269, 312

Agitator power, 26, 29, 42

Airlift, 144, 145, 150, 151, 266, 269, 339

Algae, 5, 208, 332, 333, 339, 340

Antibiotic, 2–4, 9–10, 22, 76, 97, 171, 172, 181–184,

231, 263–270, 290, 332, 395

Arrhenius’ law, 159, 346

Autotrophic, 50

Aspergillus niger, 1, 2, 153, 225, 250, 280–284

Amylase, 6, 10, 170, 405

Acetic acid, 3, 4, 7, 8, 50, 203, 238, 239, 323, 404

Amino acid, 1, 4, 8–9, 12, 76, 188, 333, 335, 339, 340

Acetone, 1, 3, 5

Antifoam, 15, 27, 46, 78, 148, 293

Agitation, 17, 19, 22–24, 26, 28–29, 78, 84, 142–145,

148, 160, 181, 272, 284, 287, 288, 292, 293,

312, 339

Aeration tank, 16, 17, 37, 46–48

Anaerobic, 2–4, 22, 50, 51, 96, 119, 143, 207, 252,

334, 338

Aerobic, 4, 8, 14, 20, 22, 23, 28, 36, 43, 44, 47, 69,

96, 143, 144, 228, 229, 238, 253, 325

Bacillus subtilis, 2, 268–270

Bacillus thuringiensis, 2

Baker’s yeast, 2, 4, 5, 10, 12, 16–18, 20, 144, 149,

193, 225, 293, 329, 403, 410

Batch culture, 23, 51, 57, 81, 83, 84, 90–92, 96, 241,

270, 271

Batch sterilisation, 342–343

Beer, 5, 12, 142, 149, 178, 179, 181, 252, 293

Biochemical oxygen demand (BOD), 37

Biofilm, 199, 200, 208, 224

Biomass, 3, 5, 16, 20, 23, 52, 57, 71, 83, 90, 91,

93, 120, 122, 142, 143, 154, 157, 170, 173,

178, 181, 182, 199, 200, 207, 208, 217,

228, 229, 232, 234, 243, 249, 261, 269,

271, 272, 281, 293, 298–301, 313, 316,

332, 333, 393, 395

Bioreactor, 1, 4, 6, 19, 22–24, 28, 37–40, 45, 51,

69–74, 77–79, 85, 93, 108, 142–151, 153,

154, 159, 171, 208, 223, 272–273, 278, 280,

287, 288, 292–298, 301, 306, 323, 326, 344

Biosensor, 72, 79–80, 85

Biostat, 84, 86, 88, 258, 260, 261, 280, 285, 341

Biosynthesis, 19, 118, 228–230, 237

Bradyrhizobium, 2

Bubble column, 69, 149–150, 293–298

Bubble diameter, 28, 42

Butyric acid, 4, 5

Cake resistance, 174, 175, 197

Candida lipolytica, 338

Candida rugosa, 130

Candida utilis, 2, 230, 338

Carbohydrate, 2, 3, 6, 8, 9, 15–17, 43, 44, 46–48,

207, 228, 230, 231, 237, 244, 252, 261, 269,

285, 334, 336, 342, 395, 405

Catabolism, 2, 3, 19, 230, 244, 341

Catabolite, 144

Cell disruption, 181–182, 392–393, 405, 406, 409–414

Cell dry weight, 15, 17, 19, 51–53, 55–58, 65, 67, 69,

81, 90, 120, 126, 211, 219, 253, 257–258,

261, 269, 284–285

Cell harvesting, 95, 268, 391

Cell load, 207, 221

Cell suspension, 392, 405, 410

Centrifuge, 175–178, 182, 193, 218

Chemolithotroph, 50

Chemostat, 15, 84–89, 94–96, 154, 298–301

Chromatography, 19, 170–172, 187–197, 211, 257,

260, 269, 393–395, 397, 403–405

Citric acid cycle, 50

Citric acid, 1, 4, 181, 185, 250, 280–285

Clostridium acetobutylicum, 2, 3

CMC, 188

COD, 43, 44, 46–48

CODH, 50, 51

Continuous culture, 15, 81, 84–86, 89–91, 93, 96, 230

Control unit, 69, 85

Corn steep liquor, 8, 9, 12, 237, 238, 265

Else_BEB-Naja_Index.qxd 9/19/2006 10:14 AM Page 418

INDEX 419

Corynebacterium glutamicum, 1

Crystallisation, 3, 170, 171, 182, 184, 379

CSTR, 28, 39, 69, 70, 89, 90, 96, 121, 123, 124, 126,

127, 147, 154, 164, 202, 206, 241, 292, 293,

298, 299, 300, 301, 320

Death phase, 58, 83, 91, 270, 271

Dextrin, 10, 188

Dialysis, 170, 354, 357, 406

Dilution rate, 14, 15, 40, 41, 84–86, 90, 91, 93, 96,

124, 154, 155, 157, 219, 226, 227, 287

maximum dilution rate, 86, 121, 125, 165

Dissolved oxygen (DO), 14–17, 20, 22–24, 28, 36,

43, 44, 46–49, 69–72, 74, 75, 79, 85, 144,

223, 272, 281, 290

Double reciprocal plot, 98

Dynamic model, 37, 45, 312

Eadie–Hofstee plot, 111

Electrode, 14, 15, 72, 73, 75–80

Electrodialysis, 351, 353–356, 393

Elemental composition, 228–230

Embden–Myerhof–Parnas pathway (EMP), 3, 207,

244–251

Enzyme inhibitor, 5, 106, 107, 131, 134

Enzyme kinetics, 130–135

Escherichia coli, 2, 8, 200, 225, 229, 230, 397

Ethanol, 1–3, 5, 7, 10, 12, 43, 50, 51, 65, 91, 92, 172,

181, 199, 202, 206–209, 211, 217, 219–221,

225–227, 230, 231, 238–240, 252–255, 257,

260–262, 320–323, 349, 356, 404

Exit gas, 19, 70

Exponential phase, 83, 92, 93, 218

Fed batch, 12, 96, 97, 144, 326, 328

Fick’s law, 366

Filter aid, 173, 174, 182, 236

Filter cake, 7, 173–175, 189, 191, 236, 237, 285, 361,

362, 365, 377

Filtration, 170, 171, 173–175, 180–182, 184, 196,

218, 236, 261, 268–281, 319, 348, 354, 355,

362, 372, 379, 385, 395

Fluidised bed, 392, 395–399, 401, 403–406

adsorption, 395–397

Foam, 77–79, 148, 149, 293, 388

Foaming, 77, 78, 147, 148, 266

Fermentation process, 1, 3, 4, 7, 12, 22, 50, 56, 69, 71,

76, 78, 199, 207, 208, 211, 219, 228, 231,

237, 238, 252, 272, 280, 281, 288, 290, 334

Gas hold-up, 28, 34, 152, 153, 164

Gel filtration, 171

Gel polarization, 365, 367

Glutamic acid, 1, 2, 4, 8, 9

Glycerol, 3, 4, 8, 230, 231, 252

Growth rate, 14, 15, 19, 23, 31, 39, 41, 43, 53, 56,

57, 61–65, 82, 84, 85, 90–93, 97, 107, 120,

132, 154–156, 207, 218, 224, 227, 230,

233, 234, 252–254, 261, 270, 271, 281,

299, 306, 329

Hollow fiber, 130, 359, 363, 369, 371, 372, 374

Immobilised, 130–134, 138, 199, 200, 202, 206–209,

211, 215, 218–221, 223–227, 238, 293, 403,

412

Impeller, 23, 24, 29, 30, 147, 148, 152, 160, 161, 167,

180, 281, 288–293, 323, 392

marine, 29, 162

tip velocity, 160, 288, 290, 317, 319, 331

Insulin, 2, 8, 9

Intracellular enzyme, 171, 180, 200, 208, 218, 397,

409

Kinetic model, 203, 205, 214, 218, 262

Lactic acid, 2, 3, 6, 7, 120, 225, 244, 245, 334

lactic acid bacteria, 2, 3, 7

Lactobacillus bulgaricus, 2, 7

Lag phase, 53, 56, 81–83, 120

Laminar flow, 29, 153, 174, 303, 348, 371

Lineweaver–Burk plot, 98, 108–110, 116, 117, 262,

271

Liquid–liquid extraction, 172, 183

Logistic model, 55, 56

Lyophilisation, 172

Lysine, 1, 8, 202, 339, 340

Malt, 2

Mass transfer coefficient, 20, 21, 24–27, 30–34, 36,

42–46, 49, 59, 60, 61, 64, 66, 72, 151, 160,

164, 223, 277, 288, 289, 295, 297, 303, 306,

308, 310–312, 316, 331, 367

Membrane, 351–354

γ-alumina, 378

anisotropic membrane, 353

ceramic membrane, 353

ceramic, 378

charged membrane, 353

dense membrane, 352

fouling, 376–377

isotropic membranes, 352–353

metal and liquid membranes, 353

microporous membranes, 352

Else_BEB-Naja_Index.qxd 9/19/2006 10:14 AM Page 419

420 INDEX

nonporous membranes, 352

semipermeable membrane, 367

synthetic membranes, 357–360

zirconia, 378

Membrane module, 369–373

Methylophilus methylotrophus, 338

Michaelis–Menten equation, 109, 137

Microfiltration, 357

cross-flow microfiltration, 362–365

Mixed culture, 91, 143, 338, 339

Molasses, 3, 5, 6, 8–10, 12, 226, 237, 238, 252, 265,

280, 281, 283–285

Monod rate equation, 41, 92, 111, 154, 155, 207,

218

Monod kinetic, 121, 218

Morphology, 83, 153, 224, 384, 387

Mutants, 264

NADH, 3, 9, 71, 244

Newtonian fluids, 27, 28, 46, 152, 297

Novobiocin, 395

Optimum pH, 227

Organic acids, 3–5, 50, 51, 76, 172, 199, 203, 288

Oxygen transfer rate, 18, 23, 24, 43–46, 160, 228,

271, 277, 278, 289, 293, 295, 297, 306, 312,

316, 330, 331

Oxygen transport, 22, 33–34, 44, 263

Penicillin, 2, 9, 10, 30, 32, 144, 166, 173, 174, 182,

184, 189, 192, 231–234, 264–269, 278, 306,

315, 319

production, 30, 32, 173, 232

Penicillium chrysogenum, 2, 9, 189, 191, 225,

264–266, 268

Penicillium notatum, 9, 264, 265, 267, 340

Permeate, 351, 354–357, 360, 362, 363, 366,

369–376

Pervaporation, 351, 353, 355–357, 363, 369

Polyamide, 357

Polysaccharide, 4, 152, 179, 208, 224, 347

Polysulphone, 357

Poly-vinyl alcohol (PVA), 382

Power consumption, 29, 143, 288, 290, 306, 315, 317,

329

Power law, 153

Power number, 29, 162, 167, 169, 275, 291, 292, 297,

304, 307, 315, 318, 329

Precipitation, 170, 171, 172, 182, 184, 285, 357, 361,

377

Probe, 14, 15, 23, 24, 70–72, 74, 75, 78–80, 84,

266

Process control, 69, 71

Process integration, 404

Propionic acid, 4, 5, 203

Protein recovery, 404

Protein, 5, 7–9, 12, 14–18, 22, 145, 170, 178,

180–182, 202, 263, 264, 332–334, 336–340,

345–347, 374–377, 390–396, 403–406, 409,

410, 412

Pump, 46, 79, 84, 144, 145, 182, 209, 261, 362,

410

Pumping, 12, 30, 160, 181

Racemic, 130–132, 138

Redox, 69–71, 76–77, 79, 85, 272

Respiration quotient, 19, 21, 22, 69, 71, 228, 229

Respiration, 19, 22, 23, 81

Reverse osmosis, 367–369

Reynolds number, 29, 42, 152, 160–162, 167, 275,

288, 291, 292, 297, 303, 307, 315, 317, 369,

399–402, 407

Rheology, 289

Rhizobium, 2

Rhodospirillum rubrum, 50, 254

Rotary drum filter, 174, 285

Saccharomyces cerevisiae, 2, 12, 17, 200,

206–208, 225, 230, 252, 253, 255, 256,

338, 414

Scale up, 159–161, 170, 177, 197, 207, 287–291, 302,

303, 309, 315, 406, 409

Secondary metabolite, 1, 2, 22, 83, 91, 264, 282

Sedimentation, 171, 178–180, 182, 193, 197

Sensors, 15, 70–72, 79, 80, 144, 258

Sing cell protein (SCP), 5, 14–16, 22, 145, 178,

332

Sol–gel, 378, 379, 381, 384, 385, 387, 388

Solvent extraction, 170–172, 182–185, 268, 270

Spargers, 15, 23, 35, 46, 144, 147–150, 161, 273,

293, 294

Specific death rate, 93, 129, 346

Specific growth rate, 31, 39, 41, 52, 61, 62, 67, 84,

90, 92, 93, 97, 98, 107, 154, 155, 218, 227,

253, 254, 261, 270, 299

Spirulina, 5

Stationary phase, 2, 58, 82, 187, 189, 260, 270

Steady state, 14, 15, 18, 39, 40, 42, 45, 59, 74, 84–86,

88–90, 93, 94, 97, 118, 121, 122, 155, 156,

164, 203, 224, 230, 239, 289, 322, 327, 365,

366, 399, 410

Sterilisation, 6, 10, 69, 74, 76, 144, 149, 261, 265,

272, 342–350, 359, 362, 372, 385

Stirring, 29, 269, 385

Else_BEB-Naja_Index.qxd 9/19/2006 10:14 AM Page 420

INDEX 421

Stoichiometric coefficient, 118, 229, 230, 233, 243,

244, 247

Stoke’s law, 176

Substrate limitation, 97

Superficial velocity, 26, 34, 42, 149, 164,

167, 169, 274, 289, 294, 296, 307,

311, 331

Temperature control, 12, 69, 293, 392, 406

Thermocouples, 73

Tower fermenter, 69, 293

Trace metals, 64, 81, 228, 262, 267, 281, 283, 284,

286, 339

Transmembrane pressure, 361

Tricarboxylic acid (TCA) cycle, 9, 51, 244, 280

Tubular module, 369

Turbidostat, 15, 84, 86, 89

Turbulent flow, 29, 48, 153, 275, 290, 304, 310, 312,

318, 329, 369, 398

Ultrafiltration, 365–367

Vitamin, 2, 5, 7, 10, 64, 85, 184, 188, 228, 261, 339, 347

Void, 34, 187, 188, 203, 209, 218, 364, 377, 401

Volumetric flow rate, 35, 38, 43, 154, 160, 177, 197,

299, 375, 376

Wastewater, 15, 16, 19, 30, 43, 44, 46–48, 143, 144,

149, 178, 233, 234, 293, 313, 325, 379, 385

Whole cell, 6, 71, 79, 199, 200

Yeast, 1–5, 8, 10, 12, 15–18, 20, 118, 144, 149, 175,

178, 179, 193, 207–209, 211, 217–219,

223–225, 230, 255, 262, 270, 272, 280, 281,

283, 293, 294, 329, 332, 333, 335, 338–340,

397, 403–406, 409–413

Yield, 8, 9, 18, 32, 41, 51, 52, 91, 92, 120, 121,

143, 156, 157, 183, 207, 208, 217, 220,

232, 249, 250, 252, 254, 258, 272,

281–284, 287, 290, 299, 394, 405, 406,

409, 410, 412, 413

Zymomonas mobilis, 207, 208, 225, 253

Else_BEB-Naja_Index.qxd 9/19/2006 10:14 AM Page 421

This page intentionally left blank