Synthetic Methods for Biologically Active Molecules: Exploring the Potential of Bioreductions -- Contents -- Preface -- List of Contributors -- 1 Development of Sustainable Biocatalytic Reduction Processes for Organic Chemists -- 1.1 Introduction -- 1.2 Biocatalytic Reductions of C=O Double Bonds -- 1.2.1 Biocatalytic Reductions of Ketones to Alcohols -- 1.2.2 Biocatalytic Reductions of Aldehydes to Alcohols -- 1.2.3 Biocatalytic Reductions of Carboxylic Acids to Aldehydes -- 1.2.4 Biocatalytic Reductions of Carboxylic Acids to Alcohols -- 1.3 Biocatalytic Reductions of C=C Double Bonds -- 1.4 Biocatalytic Reductions of Imines to Amines -- 1.5 Biocatalytic Reductions of Nitriles to Amines -- 1.6 Biocatalytic Deoxygenation Reactions -- 1.7 Emerging Reductive Biocatalytic Reactions -- 1.8 Reaction Engineering for Biocatalytic Reduction Processes -- 1.9 Summary and Outlook -- References -- 2 Reductases: From Natural Diversity to Established Biocatalysis and to Emerging Enzymatic Activities -- 2.1 Reductases: Natural Occurrence and Context for Biocatalysis -- 2.2 Emerging Cases of Reductases in Biocatalysis -- 2.2.1 Motivation: The Quest for Novel Enzymes and Reactivities -- 2.2.2 Imine Reductases -- 2.2.3 Nitrile Reductases: The Next Member in the Portfolio of Reductases? -- 2.2.4 Other Emerging N-Based Enzymatic Reductions: Nitroalkenes and Oximes -- 2.2.5 From Carboxylic Acids to Alcohols: Biocatalysis -- 2.3 Concluding Remarks -- References -- 3 Synthetic Strategies Based on C=C Bioreductions for the Preparation of Biologically Active Molecules -- 3.1 Introduction -- 3.2 Bioreduction of α,β-Unsaturated Carbonyl Compounds -- 3.2.1 Aldehydes -- 3.2.2 Ketones -- 3.3 Bioreduction of Nitroolefins -- 3.4 Bioreduction of a,b-Unsaturated Carboxylic Acids and Derivatives -- 3.4.1 Monoesters and Lactones -- 3.4.2 Diesters -- 3.4.3 Carboxylic Acids.

3.4.4 Anhydrides and Imides -- 3.5 Bioreduction of a,b-Unsaturated Nitriles -- 3.6 Concluding Remarks -- References -- 4 Synthetic Strategies Based on C=O Bioreductions for the Preparation of Biologically Active Molecules -- 4.1 Introduction -- 4.2 Synthesis of Biologically Active Compounds through C=O Bioreduction -- 4.2.1 Keto Esters -- 4.2.1.1 α-Keto Esters -- 4.2.1.2 β-Keto Esters -- 4.2.1.3 Other Keto Esters -- 4.2.2 Diketones -- 4.2.3 α-Halo Ketones -- 4.2.4 (Hetero)Cyclic Ketones -- 4.2.5 "Bulky-Bulky" Ketones -- 4.2.6 Miscellaneous -- 4.3 Other Strategies to Construct Biologically Active Compounds -- 4.4 Summary and Outlook -- References -- 5 Protein Engineering: Development of Novel Enzymes for the Improved Reduction of C=C Double Bonds -- 5.1 Introduction -- 5.2 The Protein Engineering Process and Employed Mutagenesis Methods -- 5.3 Examples of Rational Design of Old Yellow Enzymes -- 5.4 Evolving Old Yellow Enzymes (OYEs) -- 5.4.1 Evolving OYE1 as a Catalyst in the Stereoselective Reduction of 3-Alkyl-2-cyclohexenone Derivatives and Baylis-Hillman Adducts -- 5.4.2 Evolving the Pentaerythritol Tetranitrate (PETN) Reductase as a Catalyst in the Reduction of α,β-Unsaturated Carbonyl Compounds and E-Nitroolefins -- 5.4.3 Evolving Nicotinamide-Dependent 2-Cyclohexenone Reductase (NCR) from Zymomonas mobilis for the Reduction of α,β-Unsaturated Ketones -- 5.4.4 Evolving the YqjM from Bacillus subtilis for Enhanced Activity, Substrate Scope, and Stereoselectivity in the Reduction of α,β-Unsaturated Ketones -- 5.5 Conclusions and Perspectives -- References -- 6 Protein Engineering: Development of Novel Enzymes for the Improved Reduction of C=O Double Bonds -- 6.1 Introduction -- 6.2 Detailed Characterization of PAR -- 6.2.1 Location of PAR in Styrene Metabolic Pathway -- 6.2.2 Physicochemical Properties of PAR -- 6.2.3 Enzymatic Properties of PAR.

6.2.4 Docking Model Construction of PAR -- 6.3 Detailed Characterization of LSADH -- 6.3.1 Screening of LSADH from Styrene-Assimilating Soil Microorganisms -- 6.3.2 Physicochemical Properties of LSADH -- 6.3.3 Enzymatic Properties of LSADH -- 6.4 Engineering of PAR for Increasing Activity in 2-Propanol/Water Medium -- 6.4.1 Construction of Sar268 Mutant -- 6.4.2 Construction of HAR1 Mutant -- 6.4.3 Characterization of Sar268 and HAR1 -- 6.5 Application of Whole-Cell Biocatalysts Possessing Mutant PARs and LSADH -- 6.5.1 E. coli Whole-Cell Biocatalysts Possessing Mutant PARs and LSADH -- 6.5.2 Application of Immobilized E. coli Whole-Cell Catalysts to Continuous Production of Chiral Alcohol -- 6.5.3 Application of Immobilized E. coli Whole-Cell Catalysts (LASDH) for Regenerating NADH with IPA -- 6.6 Engineering of b-Keto Ester Reductase (KER) for Raising Thermal Stability and Stereoselectivity -- 6.6.1 Enzymatic Properties of KER -- 6.6.2 Engineering of KER and Characterization of Mutant Enzymes -- 6.7 New Approach for Engineering or Obtaining Useful ADHs/ Reductases -- 6.7.1 Engineering the Coenzyme Dependency of Ketol-Acid Reductoisomerase (KARI) -- 6.7.2 Engineering Substrate- and Stereospecificity of Reductases by Structure-Guided Method -- 6.7.3 Engineering Database: Systematic Information of Sequence-Structure-Function -- 6.7.4 Metagenomics -- References -- 7 Synthetic Applications of Aminotransferases for the Preparation of Biologically Active Molecules -- 7.1 Introduction -- 7.1.1 Aminotransferases -- 7.1.2 Transamination Reaction -- 7.1.3 Stereoselectivity of Aminotransferases -- 7.2 Applications -- 7.2.1 Biotransformation Process -- 7.2.2 Biologically Active Molecules -- 7.2.3 Process Economy -- 7.3 Challenges -- 7.3.1 Substrate Specificity -- 7.3.2 Improving Reaction Yield -- 7.3.3 Process Scale-Up -- 7.4 Future Research Needs.

7.5 Conclusions -- References -- 8 Strategies for Cofactor Regeneration in Biocatalyzed Reductions -- 8.1 Introduction: NAD(P)H as the Universal Reductant in Reduction Biocatalysis -- 8.2 The Most Relevant Cofactor Regeneration Approaches - and How to Choose the Most Suitable One -- 8.2.1 Electrochemical Regeneration of NAD(P)H -- 8.2.2 H2 as Reducing Agent -- 8.2.3 Formates as Reducing Agents -- 8.2.4 Phosphites as Stoichiometric Reductants -- 8.2.5 Alcohols as Stoichiometric Reductants -- 8.2.6 Glucose as Stoichiometric Reductant -- 8.3 Coupling the Reduction Reaction to a Regeneration Reaction Producing a Valuable Compound -- 8.4 Avoiding NAD(P)H: Does It Also Mean Avoiding the Challenge? -- 8.5 Conclusions -- References -- 9 Solvent Effects in Bioreductions -- 9.1 Introduction -- 9.2 Solvent Systems for Biocatalytic Reductions -- 9.2.1 Bioreduction in Aqueous Systems -- 9.2.2 Bioreduction in Monophasic Aqueous-Organic Systems -- 9.2.3 Bioreduction in Biphasic Aqueous-Organic Systems -- 9.2.4 Bioreduction in Micro- or Nonaqueous Systems -- 9.2.5 Bioreduction in Nonconventional Media -- 9.2.5.1 Ionic Liquids -- 9.2.5.2 Supercritical Fluids -- 9.2.5.3 Combining ILs and SFs -- 9.2.5.4 Gas-Phase Media -- 9.2.5.5 Reverse Micelles -- 9.3 Solvent Control of Enzyme Selectivity -- 9.4 Concluding Remarks -- References -- 10 Application of In situ Product Removal (ISPR) Technologies for Implementation and Scale-Up of Biocatalytic Reductions -- 10.1 Introduction -- 10.2 Process Requirements for Scale-Up -- 10.3 Bioreduction Process Engineering -- 10.4 In situ Product Removal -- 10.5 Biocatalyst Format -- 10.5.1 Whole-Cell Processes -- 10.5.2 Isolated Enzyme Processes -- 10.6 Selected Examples -- 10.6.1 ISPR with Resins -- 10.6.2 ISPR with Solvent Extraction -- 10.6.3 ISPR with Crystallization -- 10.6.4 Removal of Acetone -- 10.7 Future Outlook.

10.7.1 Protein Engineering -- 10.7.2 Choice of Methods -- 10.7.3 Process Integration -- 10.8 Conclusions -- References -- 11 Bioreductions in Multienzymatic One-Pot and Cascade Processes -- 11.1 Introduction -- 11.2 Coupled Oxidation and Reduction Reactions -- 11.3 Consecutive and Cascade One-Pot Reductions -- 11.4 Cascade Processes, Including Biocatalyzed Reductive Amination Steps -- 11.5 Other Examples of Multienzymatic Cascade Processes, Including Bioreductive Reactions -- References -- 12 Dynamic Kinetic Resolutions Based on Reduction Processes -- 12.1 Introduction -- 12.2 Cyclic Compounds -- 12.3 Acyclic α-Substituted-β-Keto Esters and 2-Substituted-1,3- Diketones -- 12.4 Acyclic Ketones and Aldehydes -- 12.5 Conclusions -- References -- 13 Relevant Practical Applications of Bioreduction Processes in the Synthesis of Active Pharmaceutical Ingredients -- 13.1 Introduction -- 13.2 Ketoreductases -- 13.2.1 Ethyl 4-chloro-3-hydroxybutanoate -- 13.2.2 Atorvastatin -- 13.2.3 Montelukast -- 13.2.4 Ramatroban -- 13.2.5 Ezetimibe -- 13.2.6 Profens -- 13.2.7 Atazanavir -- 13.2.8 Chemokine Receptor Inhibitor -- 13.2.9 Duloxetin -- 13.2.10 6-Hydroxybuspirone -- 13.2.11 LY 300164 -- 13.2.12 Paclitaxel -- 13.3 Ene Reductases -- 13.3.1 Levodione -- 13.3.2 (+)-Dihydrocarvone -- 13.3.3 Butyrolactone - Jasplakinolide and Amphidinolides -- 13.3.4 (R )-Flurbiprofen -- 13.3.5 Ethyl (S )-2-ethoxy-3-(4-methoxyphenyl)propanoate - Tesaglitazar -- 13.3.6 Methyl (Z)-2-bromocrotonate - Antidiabetic Drug Candidates -- 13.3.7 Roche Ester -- 13.3.8 Human Neurokinin-1 Receptor Antagonists -- 13.3.9 Asymmetric Synthesis of Amino Acid Derivatives -- 13.4 Others -- 13.4.1 Amino Acid Dehydrogenase-Catalyzed Processes -- 13.4.1.1 Saxagliptin -- 13.4.1.2 Omapatrilat -- 13.4.1.3 Inogatran -- 13.4.1.4 Corticotropin-releasing Factor-1 (CRF-1) Receptor Antagonist -- 13.4.1.5 AG7088.

13.4.2 Pyrrolo[2,1-c][1,4]benzodiazepines (Antitumor Agents).