Practical Methods for Biocatalysis and Biotransformations

Acknowledgements......Page 29
Drug Metabolites and Metabolic Transformations......Page 34
Phase I Metabolic Transformations......Page 37
Phase II Metabolic Transformations......Page 41
Regioselective/Chemoselective Biotransformations......Page 46
Diastereoselective Biotransformations......Page 58
Asymmetric Biocatalysis......Page 62
Kinetic Resolution......Page 63
Dynamic Kinetic Resolution......Page 70
Desymmetrization......Page 73
Asymmetric Ketone Reduction......Page 76
Asymmetrization Using Other Biocatalysts......Page 80
Monophasic Biocatalysis......Page 83
Catalyst Formulation......Page 84
Solvent Engineering......Page 85
Biphasic Biocatalysis......Page 87
Noncovalent Attachment......Page 89
Covalent Attachment and Cross-linking......Page 90
Entrapment......Page 91
Summary, Overview and Future......Page 111
Commercial Enzyme Sources......Page 114
Culture Collections as Sources of Microorganisms......Page 115
Enzyme and Gene Databases, Bioinformatics and the Search for New Enzymes......Page 116
Metagenomics: Sampling DNA Directly from the Environment......Page 118
The Central Dogma of Molecular Biology......Page 122
Enzyme Tools in Molecular Biology......Page 123
The Genetic Code: Nucleotides, Codons and Amino Acids......Page 124
Basic Gene Cloning from DNA Templates......Page 126
PCR......Page 131
Directed Evolution of Enzymes......Page 133
Error-prone PCR......Page 134
DNA Shuffling......Page 135
Combinatorial Methods......Page 136
Rational Enzyme Redesign......Page 137
Overcoming Problems of Codon Bias......Page 138
E. coli as an Expression Host: the Pros and Cons......Page 139
Alternative Host Expression Systems: Bacteria, Yeasts, Filamentous Fungi......Page 140
References and Notes......Page 144
Procedure......Page 145
Analytics......Page 146
Procedure......Page 147
Materials and Equipment......Page 148
Procedure......Page 149
Procedure......Page 152
Procedure......Page 153
Conclusion......Page 154
Procedure......Page 156
Procedure......Page 157
Conclusion......Page 158
Dynamic Kinetic Resolution Synthesis of a Fluorinated Amino Acid Ester Amide by a Continuous Process Lipase-mediated Ethanolysis of an Azalactone......Page 159
Procedure......Page 160
Procedure......Page 161
Conclusion......Page 162
Materials and Equipment......Page 164
Procedure......Page 165
Materials and Equipment......Page 167
Procedure 2: Synthesis of 3-Methoxyphenylpropyl Carbonate......Page 168
Procedure......Page 169
Materials and Equipment......Page 170
Procedure......Page 171
Conclusion......Page 173
Procedure......Page 174
Procedure......Page 175
Conclusion......Page 176
Procedure......Page 179
Procedure......Page 180
Conclusion......Page 181
Procedure......Page 183
Procedure......Page 184
Analytical Results......Page 185
Procedure......Page 186
Conclusion......Page 187
Materials and Equipment......Page 188
Procedure......Page 189
Conclusion......Page 190
One-pot Biocatalytic Synthesis of Methyl S-4-Chloro-3-hydroxybutanoate and Methyl S-4-Cyano-3-hydroxybutanoate......Page 191
Procedure......Page 192
Materials and Equipment......Page 193
Procedure 3: Synthesis of Bz-Arg-Gly-Asp-NH2-OH......Page 194
Conclusion......Page 195
Procedure 1: Synthesis of Acetone O-[Vinyloxycarbonyl]oxime......Page 196
Procedure......Page 197
Conclusion......Page 198
Synthesis of Oxazolidine Esters......Page 199
Enzymatic Synthesis of Novel Urethane Polyesters......Page 200
Materials and Equipment......Page 201
Procedure R. Van Calck and A. Taylor, Unpublished Results......Page 202
Materials and Equipment......Page 203
Materials and Equipment......Page 204
Materials and Equipment......Page 205
Procedure......Page 206
Materials and Equipment......Page 208
Procedure......Page 209
Procedure......Page 210
Conclusion......Page 211
Materials and Equipment......Page 212
Materials and Equipment......Page 213
Materials and Equipment......Page 214
Conclusions......Page 215
Materials and Equipment......Page 216
Procedure 2: Synthesis of Racemic 1-Benzyloxymethyl-1-methyloxirane from 3-Benzyloxy-2-methylpropene......Page 217
Procedure 3: Synthesis of R-3-Benzyloxy-2-methylpropane-1,2-diol from Racemic 1-Benzyloxymethyl-1-methyloxirane......Page 218
Procedure 4: Synthesis of S-1-Benzyloxymethyl-1-methyloxirane 68.1 % ee from R-3-Benzyloxy-2-methylpropane-1,2-diol 68.1 % ee......Page 220
Procedure 5: Synthesis of R-3-Benzyloxy-2-methylpropane-1,2-diol from S-1-Benzyloxymethyl-1-methyloxirane of 68.1 % ee......Page 222
Conclusion......Page 223
Procedure 1: Expression and Purification of a Mutated Halohydrin Dehalogenase HheC-W249F......Page 225
Procedure 2: Synthesis of Methyl S-4-chloro-3-hydroxybutanoate and Methyl S-4-cyano-3-hydroxybutanoate......Page 226
Conclusion......Page 228
Chemoenzymatic Synthesis of 5-Thio-d-xylopyranose......Page 229
Materials and Equipment......Page 230
Conclusion......Page 231
Materials and Equipment......Page 232
Procedure......Page 233
Procedure......Page 234
Procedure......Page 235
Conclusion......Page 236
Procedure......Page 238
Procedure......Page 240
Procedure......Page 241
Conclusion......Page 242
Materials and Equipment......Page 244
Procedure......Page 245
Materials and Equipment......Page 246
Procedure......Page 247
FruA-catalysed One-pot Synthesis of 5-Chloro-d-xylulose from rac-Glycidol......Page 248
Procedure 3: Glucose-isomerase-catalysed Isomerization of 5-Thio-d-xylulofuranose to 5-Thio-d-xylopyranose......Page 249
Conclusion......Page 250
Synthesis of the Acyl Glucuronide of Mycophenolic Acid......Page 252
Procedure A......Page 253
Procedure 2: Glycosynthase-assisted Synthesis of Glycosylated scyllo-Inositols......Page 254
Procedure......Page 255
Conclusion......Page 256
Materials and Equipment......Page 257
Procedure......Page 258
Conclusion......Page 259
Materials and Equipment......Page 263
Conclusion......Page 264
Procedure 1: Laccase-mediated Oxidation of Thiocolchicoside 1......Page 265
Procedure......Page 266
Materials and Equipment......Page 267
Procedure......Page 268
Conclusion......Page 269
Materials and Equipment......Page 271
Procedure......Page 272
Procedure......Page 273
Conclusion......Page 274
Procedure......Page 276
Procedure......Page 277
Conclusion......Page 278
A New R-Hydroxynitrile Lyase from Prunus mume for Asymmetric Synthesis of Cyanohydrins......Page 280
Procedure 2: S-3-Hydroxytetrahydrothiophene-3-carboxylic Acid......Page 281
Materials and Equipment......Page 282
Procedure......Page 283
Procedure......Page 284
Conclusion......Page 285
Materials and Equipment......Page 287
Procedure......Page 288
Conclusion......Page 289
Materials and Equipment......Page 291
Conclusion......Page 292
Materials and Equipment......Page 294
Materials and Equipment......Page 295
Procedure......Page 296
Conclusion......Page 297
Highly Enantioselective and Efficient Synthesis of Methyl R-o-Chloromandelate, Key Intermediate for Clopidogrel Synthesis, with Recombinant Escherichia coli......Page 298
Conclusion......Page 299
Procedure......Page 301
Conclusion......Page 302
Materials and Equipment......Page 303
Materials and Equipment......Page 304
Materials and Equipment......Page 305
Procedure......Page 306
Conclusion......Page 307
Procedure......Page 309
Procedure......Page 310
Conclusion......Page 311
Procedure......Page 313
Conclusion......Page 315
Materials and Equipment......Page 316
Materials and Equipment......Page 317
Conclusion......Page 318
Synthesis of Substituted Derivatives of l-Phenylalanine and of other Non-natural l-Amino Acids Using Engineered Mutants of Phenylalanine Dehydrogenase......Page 320
Procedure......Page 321
Conclusion......Page 322
Procedure......Page 325
Procedure......Page 326
GC Analyses......Page 327
Materials and equipment......Page 328
Conclusion......Page 329
Materials and Equipment......Page 331
Procedure......Page 332
Conclusion......Page 333
Materials and Equipment......Page 335
Procedure......Page 336
Conclusion......Page 337
Materials and Equipment......Page 339
Procedure 2: Enzymatic Synthesis of l-4-Fluorophenylalanine......Page 340
Procedure......Page 341
Conclusion......Page 342
Synthesis of Optically Pure Chiral Lactones by Cyclopentadecanone Monooxygenase-catalyzed Baeyer-Villiger Oxidations......Page 343
Materials and Equipment......Page 344
Conclusion......Page 345
Materials and Equipment......Page 347
2-Deoxy-d-gluconic Acid......Page 348
Conclusion......Page 349
Materials and Equipment......Page 351
Procedure......Page 352
Conclusion......Page 353
Procedure......Page 354
Conclusions......Page 355
Materials and Equipment......Page 357
Procedure......Page 358
Conclusion......Page 359
Procedure......Page 361
Procedure......Page 362
Conclusion......Page 363
Procedure......Page 365
Procedure......Page 366
Conclusion......Page 367
Materials and Equipment......Page 368
Procedure......Page 369
Materials and Equipment......Page 370
Procedure 4: Scale-up Synthesis of S-7-Methyl-2-oxepanone in a 3 L Fermenter......Page 371
Procedure......Page 372
Conclusion......Page 373
Biotransformation of alpha-Bromo and alpha,thinspalpha’-Dibromo Alkanone into alpha-Hydroxyketone and alpha-Diketone by Spirulina platensis......Page 375
Materials and Equipment......Page 376
Procedure 3: Synthesis of 2-Methyl-4-hydroxy-1-tetralone......Page 377
Procedure 1: Cultivation of S. maltophilia CGMCC 1.1788......Page 379
Procedure 2: Synthesis of 5-Hydroxy IMI......Page 380
Conclusion......Page 381
Procedure......Page 383
Materials and Equipment......Page 384
Procedure......Page 385
Materials and Equipment......Page 386
Procedure......Page 387
Conclusion......Page 389
Procedure......Page 391
Conclusion......Page 392
Materials and Equipment......Page 393
Procedure......Page 394
Procedure 2: Synthesis of R-1-Tetralol......Page 395
Procedure......Page 396
Conclusion......Page 398
Materials and Equipment......Page 400
Procedure......Page 401
Conclusion......Page 402
Materials and Equipment......Page 403
Materials and Equipment......Page 404
Materials and Equipment......Page 405
Procedure 4: Biotransformation of Fluorobenzene by Whole Cells of P. mendocina KR1 Expressing T4MO in Tandem with a Cell-free Preparation of Tyrosinase from Mushroom......Page 406
Conclusion......Page 407
Procedure 1: Cultivation of the Seed Culture of Recombinant E. coli JM101 pSPZ10......Page 409
Procedure 2: Cultivation of the Preculture of Recombinant E. coli JM101 pSPZ10......Page 410
Procedure 3: Batch and Fed-batch Cultivation of Recombinant E. coli JM101 pSPZ102......Page 411
Conclusion......Page 412
Procedure 2: Biotransformation of 2-Bromoacetophenone......Page 416
Procedure 3: Biotransformation of 2,6-Dibromo Cyclohexanone......Page 417
Procedure 4: Biotransformation of 2,6-Dibromo Menthone......Page 418
Conclusion......Page 419
Index......Page 421