Index

Absorbants, solvent recovery, 134 Acetate, see also Acetogens

in fermentation, 36-38, 40, 41 industrial production, 178 synthesis of, 157-161

Acetate kinase, 110 Acetic acid: see Acid production Acetoacetate decarboxylase, 202 Acetobacterium, 41 Acetogens

compared to acidogens, 146 Clostridium pfennigii as example of third

group, 150 homoacetogenic clostridia, 150-153 metabolism of

acetyl-CoA autotrophie pathway, 157-162

homoacetogenic clostridia, energetics of, 162-164

purinolytic fermentation, 164-167 properties of, 150-153 purinolytic acetogens, 146, 153-54 saccharolytic acetogens, 146

Acetoin, in fermentation, 41 Acetone-butanol, fermentation process,

129-131 Acetone-butanol-ethanol' (ABE) fermenta­

tion, 105, 122 Acetyl-CoA autotrophie pathway

acetogens, 157-162 scope of, 159

Acidification using CO2, recovery method, 178 Acidogenic phase, and solvent production,

115, 118, 120-124 Acidogens

compared to acetogens, 146 metabolism of

acrylate, 173 amino acids, 175-177 butyrate, 167-171 caproate, 167-171

Acidogens (cont.) metabolism of (cont.)

formate, 173-174 propionate, 171-173 succinate, 174-175

proteolytic clostridia, 156 saccharolytic clostridia, 154-155 species, 147-149

Acid production, industrial, 177-179 factors for consideration, 17'1-178 10w pH and production, 179 major cost, 178 medium used, 179 recovery methods, 178-179

Acrylate, 46, 199 production of, acidogens, 173

Acrylic acid-derived photopolymerizable prepolymers, 204

Acyl-CoA thioesters, enzymes from clostridia, 203

Adenine fermentation, 196 Alanine, in fermentation, 46 Alcohol dehydrogenase, 119

production of, 252-253 Alcohols, in fermentation, 41-42 Aldehyde ketone dehydrogenase, produc­

tion from clostridia, 252-253 Alpha-amylase, production from clostridia,

228-229 Alpha-gludosidase, production from

clostridia, 235 Amine compounds, in fermentation, 52-53 Amino acid fermentation

alanine, 46 amine compounds, 52-53 arginine, 49 aromatic amino acids, 45-46 asparagine, 47 aspartate, 47 branched amino acids, 42, 44-45 cysteine, 46

293

294 INDEX

Amino acid fermentation (cont.) glutamate, 48-49 glutamine, 48-49 glycine, 46 histidine, 48-49 lysine, 50-51 methionine, 46-47 ornithine, 49-50 principal routes of, 44 proline,50 purine, 51-52 pyrimidine, 51-52 serine, 47 threonine, 47

Amino acids, 9, 33 production by acidogens, 175-177 Stickland reaction, 175-176

Aminopeptidase, production from clostridia, 249

Amylolytic enzymes a-amylase, 228-229 a-glucosidase, 235 ß-amylase, 229-230 glucoamylase, 230-232 pullulanase, 232-235

Anaerobic oxidations, 199 Anaerobiosis, and phenotype, 3, 8 Animal diseases

Clostridium peifringens, 275 Clostridium sordellii, 280

Anteiso acids, 45 Antibiotic resistance markers

chloramphenicol (CmR ), 69, 75, 79 erythromycin (EmR), 67, 71-75 kanamycin (KmR), 69, 72, 74 tetracycline (TcR ), 69, 70, 72, 79

Antitoxins botulism, 284 gas gangrene, 283 tetanus, 283-284

Arginine, in fermentation, 49 Arginine deiminase, 204

pathway,49 Aromatic amino acids, in fermentation, 45-

46 Aromatic degradation, fermentative an-

aerobes, 196 Aryl nitro compounds, 206 Asparagine, in fermentation, 47 Asparate, in fermentation, 47 Autolysin, in transformation, 66, 67 Autotrophy

acetogens, 151 growth requirements, 160-161

Bacteremia, 30 ß-amylase, production from clostridia, 229-

230 Betaine, 52, 53 Bile acid bioconversions

from conjugate bile acids, 213-214 dehydroxylation, 215 epimerization of hydroxyl groups, 214-

215 Bioconversions

bile acids from conjugate bile acids, 213-214 dehydroxylation, 215 epimerization of hydroxyl groups,

214-215 bioreductions, 194

background to chiral reductions, 206-207

C=C double bonds, 209-211 C=C double bonds in cyclic com-

pounds, 212 C=O groups, 207-209 e1ectromicrobial reductions, 212-213 factors re1ated to process, 207 productivity number (PN), 207 reductive capacities of clostridia, 205-

206 enzymes from clostridia, 200-204 fermentation

catabolism of hemolyticlheterolytic compounds, 195-197

modified fermentation, 197-200 natural fermentation, 194-195

immobilization and, 193, 204-205 single-step, 194,205 steroids

nuclear dehydrogenation, 216-217 reductions, 215-217 side chain cleavage, 217

Biohydrogenation and clostridia, 210-211 Bioreactors, 35 Bioreductions: see Bioconversions Blepharospasm, botulinum toxin therapy,

285 Botulism: see Clostridium botulinum Branched amino acids, in fermentation, 42,

44-45 Branched pathways, 111 Bulk fermentations: see Bioconversions Butanol

acid production/solvent production switch, 115, 118, 120-124

catabolic pathways used by clostridia, 116-117

Butanol (cont.) chaotrophic effects, 127 fermentation process, 129-131 key factors in production, 121 mutant strains and, 119, 121 pH factors, 123, 125 regulation of production, 115, 118-125

detoxification mechanism, 120 NADH, role of, 118-120, 122, 125

toxicityltolerance, 127-128 Butanol dehydrogenase, 118 Butyraldehyde dehydrogenase, 118, 119 Butyrate

in fermentation, 36-38 fermentation pathway, 168 formation pathway, horn ethanol and

acetate, 170 production of, acidogens, 167-171

Butyrate kinase, 110

C-C bonds, enzymes horn c1ostridia, 202 C-C double bond, reduction, 253 C-C rearrangements, enzymes from

c1ostridia, 200 C=C double bonds

bioreductions, 209-211 in cyclic compounds, 212

c=o groups, bioreductions, 207-209 C-N rearrangements, enzymes from

clostridia, 200-201 Caffeate, 196 Calcium alginate, 204 Calcium-magnesium acetate (CMA), 180 Caproate, production of, acidogens, 167-

171 Carboyex group, 201 Catabolism, hemolytic/heterolytic com-

pounds, 195-197 Cell shape, of clostridia, 27-28 Cellular fatty acids, 10 Cellulases

cellulosomes, 239-243 c1asses of enzymes, in bacteria/fungi, 236 cloning of genes, 238-239 clostridia, types of, 237, 243-245 Clostridium thermocellum, 236-246 crude extracellular enzyme system prop-

erties, 237 endoglucanase activity, 237-238 industrial implications, 245-246 purification of, 237-238

Cellulolytic genes, c1ustering, 92 Cellulose, 235

breakdown, 35

INDEX 295

Cellulosomes, 36, 239-243 cell surface location, 242 cellulose, interaction with, 242 cellulose-binding factor (CBF), 240-241 enzymatic properties, 241-242 noncellulosomal cellulases, 242 physical properties, 240-241

Cell wall composition, of clostridia, 29 Chemotaxonomic properties, 9-11 Chenodeoxycholic acid, 205 Chiral reductions: see Bioreductions Choline, 52 Chopped meat agar, sporulation medium,

28 Chopped meat carbohydrate (CMC), 32 Citrate, in fermentation, 40 Cleavage

C-C bonds, 202 hydrolytic ring, 196-197 side chain, 217

Cloning cellulase genes, 238-239 codon usage, 85-88, 92-93 enzymes, 204 general information, 80 heterologous gene expression, 73, 92-93 localization signals, 93-94 ribosomal binding sites, 88, 89 screening strategies, 83-85 summary, 81-82 transcription signals, 88, 90-92

Cloning protocols, 80, 82-83 screening strategies, 83-85

colony hybridization, 84-85 immunological screening, 84 Southern hybridization, 84, 85

size of gene bank, 82 steps in cloning, 82 vectors used, 82-83

Clostridium, see aLm individual topics and indi-vidual species

acetogens, 146-154 acidogens, 154-156 anaerobic bioconversions, 193 biotechnological interest, 63 criteria for organism classification as, 27 evolutionary aspects, 17-18 fermentation strategy, 35-53 historical view, 2-3 medium normally used, 179 phenotypic aspects, 3-11 phylogenetic aspects, 11-17 properties of, 27-35 solvent production, 105-134

296 INDEX

Clostridium (cont.) sporulation, 193 taxonomic aspects, 18 toxins/phages/bacteriocins, 18-22

Clostridium absonum bile acid epimerization, 214, 215 immobilization, 205

Clostridium aceticum, acetogenic, 146, 150 Clostridium acetobutylicum

(X-amylase production, 228-229 butanol production, 109, 115-123,127-

128 cloning vectors, 78-79 conjugation, 71-72, 73-74 enzymes from, 201, 202, 204 fermentation products, 198-199 formate production, 174 glucoamylase production, 231 immobilization, 205 transformation of, 66-68

Clostridium' acidiurici acetogenic, 146, 153 enzymes from, 203, 204 fermentation products, 196

Clostridium aminovalericum, fermentation products, 194

Clostridium arcticum acidogens, 155 fermentation pattern, 171-173

Clostridium aurantibutyricum, butanol produc­tion, 109, 115

Clostridium barkerii enzymes from, 200 fermentation products, 196

Clostridium beijerinkii, butanol production, 109, 115, 118, 120

Clostridium bifermentans, bile acid epimeriza­tion, 214-215

Clostridium botulinum, 265, 268-272 action/characteristics of toxins, 265, 268-

272 antitoxins, 284 botulinum type C2 , 272 ethanol production, 107 infant botulism, 268, 284 stages of inhibitory action, 270 symptoms of botulism, 268, 272 rherapeutic agents derived from, 284-

286 for blepharospasm, 285 conditions treated, 285-286 preparation of toxin, 286 side effects, 286

Clostridium botulinum (cont.) therapeutic agents derived from (cont.)

for strabismus, 285 toxic types, 268 vaccine against botulism, 282-283 wound botulism, 268

Clostridium butyricum acidogens, 155 (X-amylase production, 228-229 butanol production, 109 electromicrobial reduction, 213 enzymes from, 202, 204 fermentation pattern, 167-170 formate production, 174 immobilization, 205

Clostridium cadaveris butanol production, 109 enzymes from, 202 steroid reduction, 217

Clostridium cellobioparum cellulase, 245 ethanol production, 107

Clostridium cellulovorans acidogens, 154 cellulase, 245

Clostridium chartatbidium, ethanol produc­tion, 107

Clostridium coccoides, succinate production, 174

Clostridium cochlearum acidogens, 156 butanol production, 109

Clostridium cylindrosporum acetogenic, 150, 153 fermentation products, 196

Clostridium difficile, 267, 278-280 action of toxins, 279-280 and antibiotic therapy, 278-279 bile acid epimerization, 215 type A toxin, 279-280 type B toxin, 280

Clostridium felsineum, butanol production, 109

Clostridium formicoaceticum acetogenic, 146, 150, 151, 152 succinate production, 175

Clostridium ghonii, electromicrobial reduc­tion, 213

Clostridium glycolicum, enzymes from, 201, 202

Clostridium histolyticum clostripain production, 248-249 collagenase production, 246-248

Clostridium hydrosulfuricum, enzymes from, 200

Clostridium indolis, ethanol production, 107 Clostridium innocuum, steroid reduction, 216 ClostridiumJ-I, steroid reduction, 216 Clostridium kluyveri

acidogens, 154, 155 diaphorase production, 254 electromicrobial reduction, 213 enoate reductases, 210-211 enzymes from, 203, 204 fermentation pattern, 170-171 formate production, 174 reductive capacity, 205-206

Clostridium leptum, bile acid epimerization, 215

Clostridium lortetii, acidogens, 156 Clostridium magnum, acetogenic, 146, 150,

151 Clostridium malenominatum, acidogens, 156 Clostridium microsporum, enzymes from, 20 I Clostridium multifermentans, pectinase pro-

duction, 250 Clostridium novyi, fermentation pattern,

171-173 Clostridium oceanicum, acidogens, 156 Clostridium paraputrificum

bile acid epimerization, 215 enzymes 1'rom, 203 steroid reduction, 216-217

Clostridium pasteurianum acidogens, 155 butanol production, 109 codon usage, 85-87 electromicrobia1 reduction, 213 enzymes from, 201, 202, 203, 204 immobilization, 205 reductive capacity, 206

Clostridium perfringens, 267, 274-278 action of toxin, 278 animal diseases, 275 bile acids, 215 classification of, 274 conjugation, 69-71 Darmbrand, 275 enzymes 1'rom, 202 exoproteins produced by, 275 food poisoning, type A enterotoxin, 278 gas gangrene, 274 lethallnecrotizing toxins

CL toxin, 276 ß toxin, 276 o toxin, 277

Clostridium peifringens (cant.) lethallnecrotizing toxins (cant.)

E toxin, 276 L toxin, 276-277 ß toxin, 277

INDEX 297

toxins with enzymatic action, 277 open reading frames (ORF), 74-75 pigbel,275 reductive capacity, 206 shuttle vectors, 79-80 symptoms of disease, 274, 275, 278 toxic types, 274, 275 transformation of, 68-69

Clostridium pfennigii acetogenic, 150, 151 fermentation products, 196

Clostridium polysaccharolyticum, acidogens, 154, 155

Clostridium populeti, acidogens, 154 Clostridium propionicum

acidogens, 156 amino acid production, 176 enzymes from, 20 I fermentation pattern, 171-173 fermentation products, 199

Clostridium purinolyticum acetogenic, 150, 153 fermentation products, 196

Clostridium putrefaciens acidogens, 156 succinate production, 174

Clostridium rectum, enzymes from, 202 Clostridium roseum, butanol production, 109 Clostridium rubrum, butanol production, 109 Clostridium saccharolyticum, ethanol produc-

tion, 107 Clostridium septicum, 267, 28 I

enzymes 1'rom, 202 gangrene, 281 treatment, 281

Clostridium sordellii, 280 action 01' toxin, 281 animal diseases, 280 bile acids, 215 electromicrobial red uction, 213 ethanol production, 107 gas gangrene, 280

Clostridium sphenoides enzymes from, 202 ethanol production, 107 fermentation products, 198

Clostridium sporogenes amino acid production, 176, 177

298 INDEX

Clostridium sporogenes (cont.) butanol production, 109 electromicrobial reduction, 213 enzymes from, 200, 201, 202 ethanol production, 107 2-oxo acid synthase production, 254

Clostridium stercorarium, cellulase, 244 Clostridium sticklandii

acidogens, 156 amino acid production, 176, 177 enzymes from, 200, 203

Clostridium subterminale, enzymes from, 200 Clostridium symbiosum, enzymes from, 203 Clostridium tertium, bile acid epimerization,

215 Clostridium tetani, 265, 272-274

action/characteristics of toxin, 272-274 antitoxins, 283-284 corUugation, 72 symptoms of tetanus, 272 vaccine against tetanus, 281-282

Clostridium tetanomorphum amino acid production, 177 butanol production, 109, 115 enzymes from, 202 fermentation products, 196

Clostridium thermoaceticum acetate production, 178 acetogenic, 146, 150, 151, 152 acetyl-CoA pathway, 157-161 acidogens, 154 energetics of, 162-164 enzymes from, 203, 204 fermentation products, 199 reductive capacity, 205

Clostridium thermoamylolyticum, glucoamylase production, 231

Clostridium thermoautotrophicum acetate production, 178 acetogenic, 146, 150 energetics of, 162-164 membrane-associated enzymes, 163

Clostridium thermobutyricum, acidogens, 155 Clostridium thermocellum

cellulase production, 236-246 codon usage, 85-87 enzymes from, 200, 204 ethanol production, 108, 126 fermentation products, 114

Clostridium thermohydrosulfuricum alcohol dehydrogenase, 252 alpha-glucosidase production, 235 enzymes from, 202 ethanol production, 126

Clostridium thermohydrosulfuricum (cont.) fermentation products, 114-115 pullulanase production, 232-234 transformation of, 68

Clostridium thermolacticum, acidogens, 155 Clostridium thermosaccharolyticum

alcohol dehydrogenase, 252 enzymes from, 200 ethanol production, 107-108 fermentation products, 112, 114, 197

Clostridium thermosulfuricum, ethanol produc­tion, 108

Clostridium thermosulfurogenes ß-amylase production, 228-229 ethanol production, 108 pectinase production, 250

Clostridium tyrobutyricum acidogens, 154, 155 butanol production, 109 cyclohexenone reduction, 212 enoate reductases, 210-211 immobilization, 205

Clostridium uracilicum fermentation products, 196 uracil fermentation, 213

Clostripain, production from clostridia, 248-249

Cobamide-coenzyme-dependent pathway, 48

Cobamide-coenzyme-dependent reaction, 50

Codon usage, 85-88, 92-93 Collagenase

inhibitors, 247-248 pH range, 247 production from clostridia, 246-247 type A, B, and C, 247 uses of, 248

Colony hybridization, 84-85 Column filter procedures, 205 Cometabolism, reductive, 199 Congo red assay, 84 Conjugation, types of, 70 C0rUugation methods

cointegrates, formation of, 73-74 conjugal mobilization of nonconjugative

plasmids, 73 conjugal streptococcal plasmids, 71-72 conjugative transposons, 72-73 indigenous conjugative plasmids, 69-71 transfer functions in trans, 74

Continuous culture techniques, solvent pro­duction, 131-132

Creatine, 52, 53

Creatinine, 52, 53 Creatinine desimidase, 203 Cross-flow microfiltration, solvent recovery,

134 Crotonase, 20 I Cyclic compounds, clostridial hydrogena­

tion, 212 Cyclohexenone, 212 Cysteine, in fermentation, 46

D-gluconate dehydratase, 20 I Darmbrand, 274 Decarboxylases, clostridial origin, 202 Decarboxylation, 48 Dehydrogenases, clostridial origin, 203 Dehydroxylation, bile acids, 215 Desmolytic activity, steroids, 217 Dialysis, solvent recovery, 134 Diaminopimelic acid, 9 Dimethylglycine, 52 Dimethylmalate lyase reaction, 52 Dinitrogen fixation, amino acid fermenta-

tion,53 Diol dehydratase, 20 I Diols, in fermentation, 41 Disease and clostridia, see Toxigenic

clostridia and specific d~,erL\e causing species

Distillation after acidification, recovery method, 178

DNA base composition, and phenotype, 8 DNA homologies, phylogenetic hetero­

geneity, 11-12

E. acidaminophilim, 176 Electrodialysis, recovery method, 178-179' Electromicrobial reductions, 212-213

mediators, 212-213 Electron flow, fermentative, diversion of,

199 Electroporation, Clostridium acetobutylicum,

transformation, 67-68 Elimination reactions, enzymes from

clostridia, 20 I Embden-Meyerhof-Parnas pathway, 36,

167 Embden-Meyerhof pathway, 106, 110 Enoate reductase, 205, 210-211, 212

production from clostridia, 253 Entner-Doudoroff pathway, 36, 106 Enzymes

categories of, 227 commercially used enzymes, 228

INDEX 299

Enzymes (cont.) potential of Clostridium in production of,

227 Enzymes of clostridia

(X-amylase, 228-229 (X-glucosidase, 235 aminopeptidase, 249 ß-amylase, 229-230 cellulases

cellulosomes, 239-243 cloning of genes, 84, 238-239 clostridia, types of, 237, 243-245 crude extracellular enzyme system

properties, 237 industrial implications, 245-246 purification of, 237-238

clostripain, 248-249 collagenase, 246-247 glucoamylase, 230-232 intracellular enzymes, 200-204

acyl-CoA thioesters, 203 assay related enzymes, 203-204 C-C bonds, cleavage, 202 C-C rearrangements, 200 C-N rearrangements, 200-201 elimination reactions, 20 I

oxidoreductases, 250-254 aldehyde ketone dehydrogenase, 252-

253 enoate reductase, 253 glycine reductase, 255 NADPH-NADH diaphorase, 254 nitroacryl reductase, 255 novel types, 251 2 oxoacid synthase, 253-254

pectinolytic enzymes, 249-250 pullulanase, 232-235

Epimerization, hydroxyl group of bile acids, 214-215

Ethanol amino acids as substrate, 107 bacterial species producing ethanol, 106 catabolic pathways used by clostridia, 113 clostridia producing ethanol, 106-108 in fermentation, 36-38, 41 fermentation process, 128 regulation of production, 112-115

Clostridium thermocellum, 114 Clostridium thermohydrosulfuricum, 114-

115 Clostridium thermosaccharolyticum, 112,

114 lactate production, 115 NADH, role of, 112, 114-115

300 INDEX

Ethanol (can!.) regulation of production (cont.)

NADPH, role of, 112, 114 oxidoreductases, 112

toxicity Itolerance, 126 Ethanolamine, 52 Ethanolamine deaminase, 201 "Eubacteria," use of term, 13 "Eubacterial," use of term, 13 Eubacterium acidaminophilum, 175 Eubacterium limosum, acetogen, 162 Evolutionary aspects, Clostridium, 17-18

Facial spasm, botulinum toxin therapy, 286 Fatty acid composition, 11 Fatty acid oxidation, 41-42 Fermentation

catabolism of hemolyticlheterolytic com-pounds, 195-197

"distortion" of usual pathway, 198-199 end-products of, 194 initial steps, Clostridium aminovalericum

example, 194-195 modified fermentation, 197-200 natural fermentation, 194-195 reductive cometabolism, 199 substrate level phosphorylation (SLP) re­

actions, 194-195 Fermentation strategy

alcohols, 41-42 amino acids: see Amino acid fermentation dinitrogen fixation, 53 ethanol!acetate/butyrate, as by-products,

36-38 organic acids, 40-41 polysaccharides, degradation, 35-36 solvent production

acetone-butanol, 129-131 continuous fermentation process, 131-

132 ethanol, 128 multistage fermentation process, 135 under stress conditions, 38-40

Ferredoxin, reduced, 46, 47, 52, 110,206 Ferredoxin oxidoreductase (NADH), eth­

anol production, 112, 114-115, 118-120, 122, 125

Ferredoxin promoters, 90-91 Ferulic acid, 33 5S rRNA sequences, phylogenetic hetero­

geneity, 17 Flame ionization detector, II Food poisoning:see Clostridium perfringens

Footprinting,91 Formate

importance of production, 173 production of

acidogens, 173-174 mechanisms in, 173-174

Formate dehydrogenase, 203 Formiminoglycine, 196 Fructose bi phosphate pathway, 110 Fumarate, in fermentation, 40

Gallate, 196 Gangrene, 281; see also Gas gangrene Gas gangrene, 30, 267, 274, 280

antitoxins, 283 Gas-liquid chromatographic (GLC) analy-

ses, product detection, 9 Gas stripping, solvent recovery, 134 GeI-entrapped cell suspensions, 204-205 Gene expression, heterologous, 73, 92-93 Gene pulser, 68 Gene transfer; see also Cloning; Plasmids of

clostridia conjugation

plasmid cointegrates, 73-74 conjugal mobilization of noncon-

jugative plasmids, 73 conjugal streptococcal plasmids, 71-72 conjugative transposons, 72-73 indigenous conjugative plasmids, 69-

71 transfer functions in trans, 74

transformation alkali ne-Tris procedure, 69 e1ectroporation, 67-68 L forms and autoplasts, 68-69 protoplast/autoplast regeneration, 65-

66 protoplast production/regeneration

method, 66-67 Glucoamylase, production from clostridia,

230-232 Glutamate, in fermentation, 48-49 Glutamate dehydrogenase, 203 Glycerol, in fermentation, 41 Glycine, 153, 167, 176,213

in fermentation, 46 Glycine reductase, 205

production from clostridia, 255 Growth factors

nutritional requirements, 32-34 oxygen sensitivity, 30-31, 34 pH,31

Growth factors (cant.)

rate 01' growth, 34 temperature range, 31-32

Guanase, 203 Guanine, fermentation products, 196

Handling/storage, of clostridia, 34-35 Hemolytic compounds, degradation, 195-

196 Heteroduplex analysis, 72 Heterologous gene expression, 73, 92-93 Heterolytic compounds, degradation, 196 Hexose monophosphate pathway, 110 Histidine, in fermentation, 48-49 Homoacetogenic clostridia, 37

energetics of, 162-164 types/ properties of, 150-153

Host-veetor systems, 69, 71-74, 78-80 Human tetanus immunoglobulin (HTI),

284 Hungate teehnique, 34 Hydrocaffeate, 196 Hydrogenation

biohydrogenation ami clostridia, 210-211 cyclic eompounds, 212

H ydrolytic reactions, 49 Hydrolytic ring cleavage, 196-197 H ydroxybenzenoids, 196 Hydroxyglutarate pathway, 48 Hydroxylamine, 206 Hydroxysteroid dehydrogenase (HSDH),

aetivities, bile acid bioconversions, 214-215

Hypoxanthine, 51, 196

L forms, transformation of, 68-69 L-glutamate deearboxylase, 202 L-histidine decarboxylase, 202 L-lysine decarboxylase, 202 L-ornithine decarboxylase, 202

Maize mash, fermentation substrate, 129 Malate, in fermentation, 40 Media for growth, 32

best medium, 179 eost-effeetive medium, 179 ingredients added to, 33 synthetic, 32-33

Membranes, 01' clostridia, 29 Membrane technology, solvent recovery,

134 Mesaconate, 202 Mesophilic enrichment eultures, 40

Metabolie properties, 8-9 Methanol, 199

in fermentation, 41 Methionine, in fermentation, 46-47 Methylcyclohexanone, 212

INDEX 301

Methylene tetrahydrofolate (THF), 47 Methyl glyoxal pathway, 40 Methyl mercury, 29 Methylviologen, 207, 212 Microbial Identification System, 11 Molasses, fermentation substrate, 129 Mole % G + evalues, of clostridia, 28 Molybdate, 33 Motility, of clostridia, 28 mRNA mapping, transcription signals, 88,

90-92 Mutant isolation, 63, 64

Necrotic enteritis, 267 Neurotransmitter, 49 Nicotinic acid, 52

fermentation, 196 fermentation 01', 196-197

Nitrate/nitrite reductase, 206 Nitroacryl reductase, production from

clostridia, 255 Nitro aliphatie compounds, 206 Nuclear dehydrogenation, steroids, 216-

217 Nutritional requirements, growth factors,

32-34

Ocular muscle dysfunction, botulinum toxin therapy, 285-286

Oligonucleotide cataloguing, 28 Organic acids, in fermentation, 40-41 Organic compounds, reductine co-

metabolism, 199 Ornithine, 176

in fermentation, 49-50 Ornitlline mutase reaction, 49 Oxaloacetate, 40 Oxidative phosphorylation, 162 Oxygen sensitivity, growth faetors, 30-31,

34 Oxygen-tolerant speeies, 31

Pathogenic species, 18-22,30; Sfe also

Toxigenic clostridia c1assification of, 18-22

Pathways, solvent production, 110-111 P-cresol, 196 Peetin degradation, 36

302 INDEX

Pectinolytic enzymes categories of, 249 cIostridia producers, 250 production from cIostridia, 249-250

Pelobacter, 41 Peptides, leader peptides, 93-94 Peptidoglycan, 9, 29 Peptone-yeast extract-glucose (PYG), 32 Perstation, solvent recovery, 134 Pervaporation, solvent recovery, 134 pH, growth factors, 31 Phage DNA, in transformation, 66 Phenotypic aspects of cIostridia, 3-11

chemotaxonomic properties, 9-11 circumscription of genus, 3, 8 circumscription of species, 8 metabolic properties, 8-9 species/types, groupings of, 6-7

Phenylalanine, 42, 45 Phenyl lactate dehydratase, 201 Phosphatidylserine decarboxylase, 202 Phylogenetic aspects of cIostridia, 11-17

DNA homologies, use of, 11-12 5S rRNA sequences, 17 rRNA cataloging, 15, 17 rRNA cistron similarities, 12-13 rRNA sequences, 13-15

PigbeI,275 Plasmids of cIostridia, 29

conjugation cointegrates, formation of, 73-74 conjugal streptococcal plasmids, 71-72 indigenous conjugative plasmids, 69-

71 nonconjugative plasmids, 73 transfer functions in trans, 74

indigenous plasmids, 74-75 medical importance, 74 most extensively studied plasmid, 74 open reading frames (ORF), 74-78 replication regions

isolation of replicons, 75-76 pAMß1 replicon, 77-78 pCB 101 replicon, 76-77, 79 pIP404 replicon, 77 shuttle plasmids, use of, 78-80

Polyacrylamide geI eIectrophoresis (PAGE), cIassification method, 9-10

Polyethylene glycol (PEG), 65 Polysaccharides, degradation, 35-36 PreIog's rule, 207 Product recovery, 134, 178, 179 Productivity number (PN), ratio for, 207

Proline, 50, 176 Proline reductase, 205 Promoter sequences

deduced promoter sequences, 90 extended promoter recognition sequence,

91 ferredoxin promoter, 90-91 promoter identification, 91 putative promoter sequences, 91 RNA1 promoter, 90, 91

Properties of cIostridia, 27-35 ceIl shape, 27-28 cell wall composition, 29 gene transfer, 29,66-74, 78-80 growth factors

nutritiona1 requirements, 32-34 oxygen sensitivity, 30-31, 34 pH,31 rate of growth, 34 temperature range, 31-32

handling/storage, 34-35 membranes, 29 mole % G + C values, 28 motility, 28 pathogeneity, 30 plasmids, 29, 74-80 spores, 28

Propionate acry1ate pathway, 172 production of, acidogens, 171-173

Proteinaceous S layer, 29, 69 Proteolytic cIostridia, acidogens,

types/ properties of, 156 Protoplasts, see also Gene transfer

use in genetic analysis, 65-66 Pseudomembranous colitis (PMC): see

Clostridium difficile Pullulanase, production from cIostridia,

232-235 Purine, 51-52, 196 Purine-degrading cIostridia, growth rate, 34 Purinolytic fermentation, acetogens, 164-

167 pathway 0[, 164-166 types/properties of, 146, 153-154

Putrescine, 49 Pyridine nicotinic acid, in fermentation, 42 Pyridoxal phosphate-dependent reaction,

50 Pyrimidine, in fermentation, 51-52 Pyruvate, 37, 46 Pyruvate ferredoxin oxidoreductase, 110 Pyruvate synthesis, 206

Rate of growth, clostridia, 34 Redox reactions, 163 Replicons: see Plasmids, replication regions Res genes, 79 Resorcinol, in fermentation, 41 Reverse osmosis, solvent recovery, 134 Ribosomal binding sites, 88, 89 RNAI promoter, 90, 91 Rod shape, and phenotype, 8 rRNA

cataloguing, 15, 17 cistron similarities, 12-13 sequences, 13-15

Saccharolytic acetogens, types/properties of, 146

Saccharolytie aeidogens, types/properties of, 154-155

Sarcosine, 52, 53 Selenite, 33 Serine, in fermentation, 47 Serine hydroxymethyltransferase, 47 Shuttle vectors, 78-80 Side chain cleavage, steroids, 217 Single-step bioconversions, 194, 205 Solvent production

acetone/butanol/ethanol (ABE) fermenta­tion, 105, 122

acid production/solvent production switch, 115, 118, 120-124

butanol, regulation 01" production, 115, 118-125

butanol-producing species, 108-109 energy-yielding reactions, 110-111 ethanol

catabolic pathways, 113 regulation of production, 112-115

ethanol-producing species, 106-108 fermentation process, 39

acetone-butanol, 129-131 continuous fermentation process, 131-

132 ethanol, 128 multistage fermentation process, 135

fermentation strategies, 110-112 industrial process, 105, 122 instability of, 109 limitations related to, 125, 132, 134 pathways, 110-111 solvent recovery, 132-134 toxicity and tolerance, 125-128

Sporomusa, 29

Sporulation, 28, 193 food poisoning and, 278 problems rclated to, 193 sporulation medium, 28, 35

Steroids bioconversions

INDEX 303

nuclear dehydrogenation, 216-217 reductions, 215-217 side chain cleavage, 217

contraceptive steroids, 216 desmolytic activity, 217

Stickland reaction, 33, 42, 49, 205 basic principle of, 175 as intraspecies redox re action , 176 mixed,199

Strabismus, botulinum toxin therapy, 285 Streptococcal plasmids, conjugation, 71-

72 Substrate level phosphorylation (SLP) reac­

tions, fermentation, 194-195 Succinate

in fermentation, 40 production of, acidogens, 174-175

Sulfite reductase, 206 Superoxide dismutase, 31 Syn elimination, of water, 201 Synthetic media, 32 Syringate, 196

Tartrate, in fermentation, 40 Taurine, 213 Taxonomie aspects, 18 Teichoie acids, 29 Temperature range, growth factors, 31-32 Tetanus: see Clostridium tetani Tetrachloro-I-cyclohexane, 202 Thermophilic amylolytic species, 36 Thermophilie species, 31 Threonine, 47, 173 Threonine aldolase, 47 Toluene, 196 Toxigenic clostridia; see also sperilic sfieei!'s

clostridial antitoxins hotulism, 284 gas gangrene, 283 tetanus, 283-284

Clostridium botulinum, 265, 268-272 Clostridium difficile, 267, 278-280 Clostridium perfringenl, 267, 274-278 Clostridium septicum, 267, 281 Clostridium sordellii, 280 Clostridium tetani, 265, 272-274 histotoxic infections, 267

304 INDEX

Toxigenic clostridia (cont.) prophylactic agents

botulism vaccine, 282-283 tetanus vaccine, 281-282

types of diseases, 266 Toxins/ phages/bacteriocins, 18-22 Toxin therapy: see Clostridium botulinum Transamination reaction, 46 Transcription signals, 88, 90-92

bacterial genes, 91 cellulolytic genes, 92 dyad symmetry elements, 91 promoter sequences, 90-91

Transformation technology alkaline-Tris procedure, 69 electroporation, 67-68 L forms and autoplasts, 68-69 protoplast/autoplast regeneration, 65-66 protoplast production/regeneration meth-

od, 66-67 Transposons, conjugation, 72-73 Trans-vaccenic acid, 210 Trimethylamine, 52, 203 Tryptophan, in fermentation, 45 Tungstate, 33 2-oxoacid synthase, production from

clostridia, 253-254

Two-phase aqueous systems, solvent recov­ery, 134

Tyrosine, in fermentation, 45

Ultrafiltration, solvent recovery, 134 Uracil fermentation, 196,212 Uric acid, 51, 153

fermentation products, 196 Ursodeoxycholic acid, 205

Vaccines botulism vaccine, 282-283 tetanus vaccine, 281-282

Valine, 45 Vanillate, fermentation products, 196 Vectors

cloning vectors, types of, 79 shuttle vectors, 79-80

Viologen dye, 199

Warburg-Dickens pathway, 110 Wound botulism, 268

Xanthine, fermentation products, 196 Xanthine dehydrogenase, 34, 51, 203 Xylan, 36

Zeolites, solvent recovery, 134