Biocatlysis Guide
The Biocatalysis Guide is a simple, double-sided, single-sheet guide to the most used enzyme classes amongst the ACS GCIPR member companies. It is easy to follow for chemists who have not had significant exposure to biocatalysis—showing generic transformations that are available so these can be factored into retrosynthetic analysis. Newly updated in 2026, the Biocatalysis Guide is organized by the number of peer-reviewed reactions at a given scale. While the information on the downloadable PDF is streamlined, we have added substantial literature references below for further guidance. The guide can be downloaded and printed for personal use or display in laboratories
References
Hydrolase
- Development of a Chemoenzymatic Manufacturing Process for Pregabalin. C. A. Martinez et al., Org. Process Res. Dev. 2008, 12, 3, 392–398. 3500 kg scale.
- The Development of a Manufacturing Route for the GPIIb/IIIa Receptor Antagonist SB-214857-A. Part 2: Conversion of the Key Intermediate SB-235349 to SB-214857-A. R. J. Atkins et al., Org. Proc. Res. Dev. 2003, 7, 5, 663–675. 250 kg scale/14kg scale.
- A Practical and Robust Process to Produce SB-214857, Lotrafiban, ((2S)-7-(4,4‘-Bipiperidinylcarbonyl)-2,3,4,5-tetrahydro-4-methyl-3-oxo-1H-1, 4-Benzodiazepine-2-acetic Acid) Utilising an Enzymic Resolution as the Final Step. T. C. Walsgrove et al., Org. Proc. Res. Dev. 2002, 6, 4, 488–491. 80 kg scale.
Ketoreductase
- Development of a Biocatalytic Process as an Alternative to the (−)-DIP-Cl-Mediated Asymmetric Reduction of a Key Intermediate of Montelukast. J. Liang et al., Org. Process Res. Dev. 2010, 14, 1, 193–198. 230 kg scale.
- Development of a First-Generation Stereocontrolled Manufacturing Process of TRPA1 Inhibitor GDC-6599. A. Y. Hong et al., Org. Process Res. Dev. 2024, 28, 6, 2325–2333. 120 kg scale.
- A green-by-design biocatalytic process for atorvastatin intermediate. S. K. Ma et al., Green Chem., 2010, 12, 81-86. >100 kg scale (unpublished)
- Bio- and Chemocatalysis for the Synthesis of Late Stage SAR-Enabling Intermediates for ROMK Inhibitors and MK-7145 for the Treatment of Hypertension and Heart Failure. R. T. Ruck et al., Org. Process Res. Dev. 2021, 25, 3, 405–410. 97 kg scale.
- Chemical and Biochemical Approaches to an Enantiomerically Pure 3,4-Disubstituted Tetrahydrofuran Derivative at a Multikilogram Scale: The Power of KRED. A. Bigot et al., Org. Process Res. Dev. 2024, 28, 12, 4467–4476. 40 kg scale.
- A Chemoenzymatic Route to Chiral Intermediates Used in the Multikilogram Synthesis of a Gamma Secretase Inhibitor. M. Burns et al., Org. Process Res. Dev. 2017, 21, 6, 871–877. 35 kg scale.
- Merging Biocatalysis, Flow, and Surfactant Chemistry: Innovative Synthesis of an FXI (Factor XI) Inhibitor. B. Wu et al., Org. Process Res. Dev. 2020, 24, 11, 2780–2788. 3.5 kg scale.
- Evolution of a Green and Sustainable Manufacturing Process for Belzutifan: Part 5─Chemoenzymatic Diastereoselective Fluorination/DKR. S. D. McCann et al., Org. Process Res. Dev. 2024, 28, 2, 441–450. 1 kg scale.
Transaminase
- A Chemoenzymatic Route to Chiral Intermediates Used in the Multikilogram Synthesis of a Gamma Secretase Inhibitor. M. Burns et al., Org. Process Res. Dev. 2017, 21, 6, 871–877. 25 kg scale.
- Evolution of a Green and Sustainable Manufacturing Process for Belzutifan: Part 5─Chemoenzymatic Diastereoselective Fluorination/DKR. S. D. McCann et al., Org. Process Res. Dev. 2024, 28, 2, 441–450. 12 kg scale.
- Application of Transition-Metal Catalysis, Biocatalysis, and Flow Chemistry as State-of-the-Art Technologies in the Synthesis of LCZ696. X. Gu et al., J. Org. Chem. 2020, 85, 11, 6844–6853. 4.4 kg scale.
- Preclinical Toxicology Supply for a Complex API Enabled by Asymmetric Catalysis and Rapid Chemical Development: IL-17A Inhibitor LY3509754. T. Beauchamp et al., Org. Process Res. Dev. 2025, 29, 3, 889–908. 1.2 kg scale.
- From Chiral Resolution to Diastereoselective Ellman Chemistry to Biocatalysis: Route Evolution for the Efficient Synthesis of the Tetrahydrobenzoazepine Core of BTK Inhibitor BIIB091. C. Li et al., Org. Process Res. Dev. 2023, 27, 8, 1463–1473. 1 kg scale.
- Biocatalytic Asymmetric Synthesis of Chiral Amines from Ketones Applied to Sitagliptin Manufacture. C. K. Saville et al., Science 2010, 329 (5989), 305-309. 1 kg in Sl.
Enereductase
- Rapid Development of a Scalable Reduction of R-Carvone Utilizing Commercially Available Biocatalysts. S. M. McKenna et al., Org. Process Res. Dev. 2025, 29, 6, 1571–1576. 100 kg scale.
- Identification and Implementation of Biocatalytic Transformations in Route Discovery: Synthesis of Chiral 1,3-Substituted Cyclohexanone Building Blocks. T. Hadi et al., Org. Process Res. Dev. 2018, 22, 7, 871–879. 40 kg scale.
- Enantioselective Enzymatic Reduction of Acrylic Acids. C. An et al., Org. Lett. 2020, 22, 21, 8320–8325. 1.6 kg scale.
Nitrilase
- Efficient Chemoenzymatic Synthesis of Optically Active Pregabalin from Racemic Isobutylsuccinonitrile. Q. Zhang et al., Org. Process Res. Dev. 2019, 23, 9, 2042–2049. 300 kg scale.
- Asymmetric Synthesis of Akt Kinase Inhibitor Ipatasertib. C. Han et al., Org. Lett. 2017, 19, 18, 4806–4809. 200 kg scale.
- Enzyme Optimization and Process Development for a Scalable Synthesis of (R)-2-Methoxymandelic Acid. M. E. Scott et al., Org. Process Res. Dev. 2022, 26, 3, 849–858. 40 kg scale.
- Identification and Implementation of Biocatalytic Transformations in Route Discovery: Synthesis of Chiral 1,3-Substituted Cyclohexanone Building Blocks. T. Hadi et al., Org. Process Res. Dev. 2018, 22, 7, 871–879. 20 kg scale.
- Process Development for the Production of (R)-(−)-Mandelic Acid by Recombinant Escherichia coli Cells Harboring Nitrilase from Burkholderia cenocepacia J2315. H. Wang. Et al., Org. Process Res. Dev. 2015, 19, 12, 2012–2016. 3.5 kg scale.
Epoxide Hydrolase
- Development of an Enzymatic Process for the Production of (R)-2-Butyl-2-ethyloxirane. G.-D. Roiban et al., Org. Process Res. Dev. 2017, 21, 9, 1302–1310. 20 g scale.
- Enantioselective Hydrolysis of Racemic and Meso-Epoxides with Recombinant Escherichia coli Expressing Epoxide Hydrolase from Sphingomonas sp. HXN-200: Preparation of Epoxides and Vicinal Diols in High ee and High Concentration. S. Wu et al., ACS Catal. 2013, 3, 4, 752–759. 10 g scale.
- Cascade Biocatalysis for Sustainable Asymmetric Synthesis: From Biobased l-Phenylalanine to High-Value Chiral Chemicals. Y. Zhou et al., Angew. Chem. Int Ed., 2016, 55 (38),11647-11650. 1.7 g scale.
- Enantioselective Bio-Hydrolysis of Various Racemic and meso Aromatic Epoxides Using the Recombinant Epoxide Hydrolase Kau2. W. Zhao et al., Adv. Synth. Catal., 2015, 357 (8), 1895-1908. 1 g scale.
Aldolase
- Preparation of β-hydroxy-α-amino Acid Using Recombinant ᴅ-Threonine Aldolase. S. L. Goldberg et al., Org. Process Res. Dev. 2015, 19, 9, 1308–1316. ~4 kg scale.
- An efficient process for production of ɴ-acetylneuraminic acid using ɴ-acetylneuraminic acid aldolase. M. Mahmoudian et al., Enzyme and Microbial Technology 1997, 20 (5), 393-400. Scale: 817 g limiting reagent.
- Development of an efficient, scalable, aldolase-catalyzed process for enantioselective synthesis of statin intermediates. W. A. Greenberg et al., PNAS, 2004, 101 (16), 5788-5793. 11.1 g scale.
- Design of an in vitro biocatalytic cascade for the manufacture of islatravir. M. A. Huffman et al., Science 2019, 366 (6470), 1255-1259. 8.75 mmole,~ 1.5 g scale (>1 kg unpublished).
- Directed evolution of an industrial biocatalyst: 2-deoxy-D-ribose 5-phosphate aldolase. S. Jennewein et al., Biotechnology Journal 2006, 1 (5), 537-548. Scale: 0.4 g limiting substrate ~ 1.3 g product
- Application of Threonine Aldolases for the Asymmetric Synthesis of α-Quaternary α-Amino Acids. J. Blesl et al., ChemCatChem., 2028, 10 (6), 3453-3458. 19 g scale, but low yield of ~800 mg.
- ᴅ-Serine as a Key Building Block: Enzymatic Process Development and Smart Applications within the Cascade Enzymatic Concept. N. Ocal et al., Org. Process Res. Dev. 2020, 24, 5, 769–775. ~100 mg scale.
Monoamine Oxidase
- Efficient, Chemoenzymatic Process for Manufacture of the Boceprevir Bicyclic [3.1.0]Proline Intermediate Based on Amine Oxidase-Catalyzed Desymmetrization. T. Li et al., J. Am. Chem. Soc. 2012, 134, 14, 6467–6472. 40 g scale (> 1 kg unpublished).
- Enantioselective oxidation of O-methyl-N-hydroxylamines using monoamine oxidase N as catalyst. T.S. C. Eve et al., Chem. Commun., 2007, 1530-1531. 1.2 g scale.
- A Chemo-Enzymatic Route to Enantiomerically Pure Cyclic Tertiary Amines. C. J. Dunsmore et al., J. Am. Chem. Soc. 2006, 128, 7, 2224–2225. 0.4 g scale.
Baeyer-Villiger Monooxygenase
- Development and Scale-Up of an Improved Manufacturing Route to the ATR Inhibitor Ceralasertib. M. A. Graham et al., Org. Process Res. Dev. 2021, 25, 1, 43–56. 64 kg scale.
- Development and Scale-up of a Route to ATR Inhibitor AZD6738. W. R. F. Goundry et al., Org. Process Res. Dev. 2019, 23, 7, 1333–1342. 62 kg scale.
- Baeyer–Villiger Monooxygenase-Mediated Synthesis of Esomeprazole As an Alternative for Kagan Sulfoxidation. Y. K. Bong et al., J. Org. Chem. 2018, 83, 14, 7453–7458. 30 g scale.
- The First 200-L Scale Asymmetric Baeyer−Villiger Oxidation Using a Whole-Cell Biocatalyst. C. V. F. Baldwin et al., Org. Process Res. Dev. 2008, 12, 4, 660–665. 200 L scale.
Amino Acid Deydrogenase
- Preparation of an Amino Acid Intermediate for the Dipeptidyl Peptidase IV Inhibitor, Saxagliptin, using a Modified Phenylalanine Dehydrogenase. R. L. Hanson et al., Adv. Synth. Catal., 2007, 349 (8-9), 1369-1378. 40 kg scale.
- A one-pot system for production of l-2-aminobutyric acid from l-threonine by l-threonine deaminase and a NADH-regeneration system based on l-leucine dehydrogenase and formate dehydrogenase. R. Tao et al., Biotechnol. Lett., 2014, 36, 835–841. 4.6 kg scale.
- Preparation of (S)-1-Cyclopropyl-2-methoxyethanamine by a Chemoenzymatic Route Using Leucine Dehydrogenase. W. L. Parker et al., Org. Process Res. Dev. 2012, 16, 3, 464–469. 50 g scale.
- Asymmetric Synthesis of iso-Boc (S)-2-Amino-8-nonenoic Acid in One Through-Process. J. Park et al., Org. Process Res. Dev. 2016, 20, 1, 76–80. ~15 g scale.
- Biocatalytic synthesis of (2S)-5,5,5-trifluoroleucine and improved resolution into (2S,4S) and (2S,4R) diastereoisomers. H. Biava et al., Tet. Lett., 2013, 54 (28), 3662-3665. 1.5 g scale.
Ammonia Lyase
- Calcium Alginate Bead Immobilization of Cells Containing Tyrosine Ammonia Lyase Activity for Use in the Production of p-Hydroxycinnamic Acid. R. J. Trotman et al., Biotechnol. Progress 2008, 23 (3), 638-644. >4 kg scale.
- Toward a Scalable Synthesis and Process for EMA401, Part III: Using an Engineered Phenylalanine Ammonia Lyase Enzyme to Synthesize a Non-natural Phenylalanine Derivative. L. A. Hardegger et al., Org. Process Res. Dev. 2020, 24, 9, 1763–1771. 2 kg scale.
- Asymmetric Synthesis of (S)-2-Indolinecarboxylic Acid by Combining Biocatalysis and Homogeneous Catalysis. B. de Lange et al., ChemCatChem., 2011, 3 (2), 289-292. 18.1 g scale.
- Cascade Biocatalysis for Sustainable Asymmetric Synthesis: From Biobased ʟ-Phenylalanine to High-Value Chiral Chemicals. Y. Zhou et al., Angew. Chem. Int Ed., 2016, 55 (38),11647-11650. 1.7 g scale.
- Bacterial Anabaena variabilis phenylalanine ammonia lyase: A biocatalyst with broad substrate specificity. S. Lovelock et al., Bioorg. & Med. Chem., 2014, 22 (20), 5555-5557. 1 g scale.
Alcohol Oxidase
- Development of a Biocatalytic Aerobic Oxidation for the Manufacturing Route to Islatravir. M. H. Shaw et al., Org. Process Res. Dev. 2024, 28, 9, 3545–3559. 7.6%w/w of 350 kg soln, ~ 26.6 kg scale.
- Biocatalytic Oxidation in Continuous Flow for the Generation of Carbohydrate Dialdehydes. S. C. Cosgrove et al., ACS Catal. 2019, 9, 12, 11658–11662. 2.1 g scale.
- Design of an in vitro biocatalytic cascade for the manufacture of islatravir. M. A. Huffman et al., Science 2019, 366 (6470), 1255-1259. 8.75 mmole,~ 1.5 g scale.
- Scope and Synthetic Applications of the Aryl-Alcohol Oxidase from Streptomyces hiroshimensis (ShAAO). C. Ascasco-Alegre et al., Org. Lett. 2025, 27, 43, 12086–12091. Low scale but lots of examples.
- Microsome-bound alcohol oxidase catalyzed production of carbonyl compounds from alcohol substrates. A. Kakoti et al., J. Mol. Cat. B: Enzymatic 2012, 78, 98– 104. Low scale but lots of examples.
Iminereductase
- Biocatalytic reductive amination from discovery to commercial manufacturing applied to abrocitinib JAK1 inhibitor. R. Kumar et al., Nat. Catal., 2021, 4, 775–782. 230 kg scale.
- Engineered Imine Reductase for Larotrectinib Intermediate Manufacture. Q. Chen et al., ACS Catal. 2022, 12, 23, 14795–14803. 1.6 kg scale.
- Imine Reductase-Catalyzed Synthesis of a Key Intermediate of Avacopan: Enzymatic Oxidative Kinetic Resolution with Ex Situ Recovery and Dynamic Kinetic Reduction Strategies toward 2,3-Disubstituted Piperidine. Z. J. Pótáriné et al., Org. Process Res. Dev. 2025, 29, 4, 1093–1102. 1 kg scale.
- Chiral synthesis of LSD1 inhibitor GSK2879552 enabled by directed evolution of an imine reductase. N. Schober et al., Nat. Catal., 2019, 2, 909–915. 392 g scale.
- Technical Considerations for Scale-Up of Imine-Reductase-Catalyzed Reductive Amination: A Case Study. A. Bornadel et al., Org. Process Res. Dev. 2019, 23, 6, 1262–1268. 1 g scale.
Nitrile Hydratase
- Nitrile hydratase-mediated conversion of acrylonitrile by Rhodococcus rhodochrous (RS-6). R. Sahu et al., J. Chem. Tech. & Biotech., 2021, 96 (4), 1080-1085. 400 g scale.
- Enzymatic hydrolysis of cyanohydrins with recombinant nitrile hydratase and amidase from hodococcus erythropolis. Ch. Reisinger et al., Biotechnol. Lett., 2004, 26, 1675–1680. 1 g scale.
- High-yield continuous production of nicotinic acid via nitrile hydratase–amidase cascade reactions using cascade CSMRs. L. Cantarella et al., Enz. & Microbial Tech., 2011, 48 (4–5), 345-350. Scale: flow, isolated amount not stated.
Hydroxynitrile Lyase
- ‘Large-Scale Synthesis of (R)-2-Amino-1-(2-furyl)ethanol via a Chemoenzymatic Approach. T.Purkarthofer et al., Org. Process Res. Dev. 2006, 10, 3, 618–621. 1 kg scale.
- Probing o-Diphenylphosphanyl Benzoate (o-DPPB)-Directed C—C Bond Formation: Total Synthesis of Dictyostatin. S. Wünsch et al., Chem. Eur. J., 2015, 21 (6), 2358-2363. 36 g scale.
- A Unified Strategy for the Stereospecific Construction of Propionates and Acetate–Propionates Relying on a Directed Allylic Substitution. T. Reiss et al., Chem. Eur. J., 2009, 15 (26), 6345-6348. 14 g scale.
- Stereoselective Biocatalytic Synthesis of (S)-2-Hydroxy-2-Methylbutyric Acid via Substrate Engineering by Using “Thio-Disguised” Precursors and Oxynitrilase Catalysis. M. H. Fecter et al., Chem. Eur. J., 2007, 13 (12), 3369-3376. 10.8 g scale.
- Chemo-enzymatic synthesis of new ferrocenyl-oxazolidinones and their application as chiral auxiliaries. B. J. Ueberbacher et al., Tet. Asymm., 2008, 19 (7), 838-846. 7 g scale.
- Enzyme and Gold Catalysis: A New Enantioselective Entry into Functionalized 4-Hydroxy-2-pyrrolines. B. Ritzen et al., Synlett 2014, 25(2),270-274. 2.4 g scale.
- Synthesis of Aromatic 1,2-Amino Alcohols Utilizing a Bienzymatic Dynamic Kinetic Asymmetric Transformation. J. Steinreiber et al., Adv. Synth. Catal., 2007, 349 (8-9), 1379-1386. 1.8 g scale.
Amino Acid Oxidase
- Enzymatic Preparation of an (S)-Amino Acid from a Racemic Amino Acid. Y. Chen et al., Org. Process Res. Dev. 2011, 15, 1, 241–248. 15 g substrate (60% potency due to salt and hydrate form) scale.
Halohydrin Dehalogenase
- A green-by-design biocatalytic process for atorvastatin intermediate. S. K. Ma et al., Green Chem., 2010, 12, 81-86. 70 g scale (>100 kg unpublished).
- HHDH-Catalyzed Synthesis of Enantioenriched Fluorinated β-Hydroxy Nitrile─Process Advances through a Reaction Engineering Approach. N. Milčić et al., Ind. Eng. Chem. Res. 2024, 63, 16, 7051–7063. 0.69 g scale.
- A One-Step Biocatalytic Process for (S)-4-Chloro-3-hydroxybutyronitrile using Halohydrin Dehalogenase: A Chiral Building Block for Atorvastatin. N. W. Wan et al., ChemCatChem., 2015, 7 (16), 2446-2450.
Amino Acid Hydroxylase
- Biocatalytic Aerobic Oxidation for Large-Scale Production of trans-3-Hydroxy-l-Proline. K.-J. Xioa et al., Org. Process Res. Dev. 2025, 29, 8, 2076–2085. 465 kg scale.
- Engineering Hydroxylase Activity, Selectivity, and Stability for a Scalable Concise Synthesis of a Key Intermediate to Belzutifan. W. L. Cheung-Lee et al., Angew. Chem. Int. Ed., 2024, 63 (13), e202316133. 1.5 kg scale.
Tryptophan Synthase (TrpB)
- The β-subunit of tryptophan synthase is a latent tyrosine synthase. P. J. Almhjell et al., Nat. Chem. Biol. 2024, 20, 1086–1093. 5 g scale.
- Unlocking Reactivity of TrpB: A General Biocatalytic Platform for Synthesis of Tryptophan Analogues. D. K. Romney et al., J. Am. Chem. Soc. 2017, 139, 10769-10776. 1 g scale.
- Improved Synthesis of 4-Cyanotryptophan and Other Tryptophan Analogues in Aqueous Solvent Using Variants of TrpB from Thermotoga maritima. C. E. Boville et al., J. Org. Chem. 2018, 83, 7447-7452. 0.8 g scale.
Halogenase
- Extended Biocatalytic Halogenation Cascades Involving a Single‐Polypeptide Regeneration System for Diffusible FADH2. N. Montua et al, ChemBioChem 2023, 24 (22), e202300478. 800 mg of product scale.
P450
- Enabling Selective and Sustainable P450 Oxygenation Technology. Production of 4-Hydroxy-α-isophorone on Kilogram Scale. I. Kaluzna et al., Org. Process Res. Dev. 2016, 20, 4, 814–819. 897 g scale.
Unspecific Peroxygenase
- Toward Kilogram-Scale Peroxygenase-Catalyzed Oxyfunctionalization of Cyclohexane. T. Hilberath et al., Org. Process Res. Dev. 2023, 27, 7, 1384–1389. 500 g scale.
- Divergent Oxidation Reactions of E– and Z-Allylic Primary Alcohols by an Unspecific Peroxygenase. J. Li et al., Angew. Chem., Int. Ed., 2025, 64 (12), e202422241. 1.25 g scale.
Carboxylic Acid Reductase
- Computation-driven redesign of an NRPS-like carboxylic acid reductase improves activity and selectivity. K. Shi et al., Sci. Adv. 2024, 10 (48). 2.1 g scale.
- Cell-free in vitro reduction of carboxylates to aldehydes: With crude enzyme preparations to a key pharmaceutical building block. A. Swartz et al., Biotechnol. J., 2021, 16 (4), 2000315. 1.1 g scale.
- Engineering the activity and thermostability of a carboxylic acid reductase in the conversion of vanillic acid to vanillin. Y. Ren et al., Biotechnol. J., 2024, 386, 19-27. 0.5 g scale.
Amide Ligase/Synthetase
- Discovery, characterization and engineering of ligases for amide synthesis. M. Winn et al., Nature 2021, 593, 391–398. Scale: 0.9 g limiting acid.
- Broad Spectrum Enantioselective Amide Bond Synthetase from Streptoalloteichus hindustanus. Q. Tang et al., ACS Catal., 2024, 14, 2, 1021–1029. Scale: 50 mg limiting acid.
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