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11Biocatalytic Synthesis - enzymatic solutions for chemical problems - Enzyme Engineering & Technology - eynzmes designed for industry - Biocatalytic Synthesis - enzymatic solutions for chemical problems - Enzyme Engineering & Technology - eynzmes designed for industry - Biocatalytic Synthesis - enzymatic solutions for chemical problems - Enzyme Engineering & Technology - eynzmes designed for industry - Biocatalytic Synthesis - enzymatic solutions

Biocatalytic Synthesis

enzymatic solutions for chemical problems

Enzyme Engineering & Technology

enzymes designed for industry

science is our success


Contents

Reductive Biotransformations 4-5

Oxidative Biotransformations 6-7

C-C Bond Formation 8-9

C-C Bond Breaking 9

Enzymatic Isomerisation 10

Hydrolysis / Esterification 11-12

Enzymatic Cascades 12-13

Glycozyme Technology 14-15

API Modification 16

In-silico Search for Novel Biocatalysts 17

About acib 18

Contact 19


Reduction of Carboxylic Acids

(CAR = carboxylate reductase, ADH = alcohol dehydrogenase; ChemCatChem 2014, 6, 1089-1095; Biotechnol. J. 2014, 9, 822-843; Chemical Monthly, 2016, 147, 575-578; Adv. Synth. Catal., 2016, in press, DOI: 10.1002/adsc.201600914.)

Reductive Biotransformations

Reduction of Aldehydes and Ketones

(ADH = alcohol dehydrogenase; Angew. Chem., Int. Ed. 2002, 41, 1014; Angew. Chem., Int. Ed. 2008, 47, 714; ChemCatChem 2013, 5, 1744.)

Reduction of C=C-Bonds

(X = electron-withdrawing group; Angew. Chem., Int. Ed. 2007, 46, 3934; Chem. Eur. J. 2012, 18, 10367; Curr. Opin. Chem. Biol. 2007, 11, 203; J. Biotechnol. 2012, 162, 381; Nat. Commun. 2014, 5, 4150.)

Reduction of Imines

(IRED = imine reductase; Adv. Synth. Catal. 2015, 357, 1655.)


Reductive BiotransformationsReductive Amination of Aldehydes and Ketones

(TA = ω-transaminase; Angew. Chem., Int. Ed. 2008, 47, 9337; Angew. Chem., Int. Ed. 2012, 27, 1; Org. Process Res. Dev. 2013, 17, 751; ACS Catal. 2014, 4, 129.)

Structure of an Ene-Reductase


Oxidation of Alcohols / Aldehydes

(ADH = alcohol dehydrogenase; Angew. Chem., Int. Ed. 2002, 41, 1014; Angew. Chem., Int. Ed. 2008, 47, 714; J. Am. Chem. Soc. 2008, 130, 13969; ChemCatChem 2013, 5, 1744.)

C=C-Bond Cleavage

(Angew. Chem., Int. Ed. 2006, 45, 5201; J. Am. Chem. Soc. 2009, 131, 5368; Chem. Commun. 2012, 48, 3303; Adv. Synth. Catal. 2013, 355, 3321.)

Enzymatic De-Amination

(TA = ω-transaminase; AO = α-amino oxidase; Angew. Chem., Int. Ed. 2008, 47, 9337; Angew. Chem., Int. Ed. 2012, 27, 6817; Org. Process Res. Dev. 2013, 17, 751; ACS Catal. 2014, 4, 129.)

Enzymatic De-Alkylation

Oxidative Biotransformations

Enzymatic Hydration

(Angew. Chem. Int. Ed. 2015, 54, 15051.)

(Angew. Chem., Int. Ed. 2013, 52, 2293.)


Oxidative Biotransformations

Enzymatic Hydroxylation

(Nat. Biotechnol. 2002, 20, 1135.)

Baeyer-Villiger Oxidation

(hFMO = human flavin containing monooxygenase; ACS Chem. Biol., 2016, 11, 1039-1048.)

Oxyfunctionalisation of Amino Acids

Oxidative Decarboxylation

(Angew. Chem. Int. Ed. 2015, 54, 8819–8822; Chem. Commun., 2016, 51, 1918; Eur. J. Org. Chem., 2016, 2473–3477.)

(aKG = a-ketoglutarate; AAD = amino acid dioxygenase; Front. Microbiol., 2016, 7, 425.)


Bio-Carboxylation

(Chem. Soc. Rev. 2010, 39, 313; Org. Lett. 2012, 14, 1974; J. Biotechnol. 2013, 168, 264; RSC Adv. 2014, 4, 9673.)

Bio-Friedel-Crafts Acylation

(unpublished results)

Biocatalytic Alkaloid Synthesis

(BBE = berberine bridge enzyme; STR = stricto-sidine synthase; NCS = norcoclaurine synthase; Angew. Chem., Int. Ed. 2011, 50, 1068; Org. Pro-cess Res. Dev. 2013, 17, 751; Angew. Chem., Int. Ed. 2014, 53, 3731.)

Trifluoromethylation

(TFMS = trifluoromethylsulfonic acid; TBHP = tert-butylhydroperoxide; Nat. Commun. 2016, in press, DOI: 10.1038/ncomms13323.)

C-C Bond Formation


Cyanohydrin Synthesis/Henry Reaction

(HNL = hydroxynitrile lyase; Angew. Chem., Int. Ed. 2006, 45, 3454; Adv. Synth. Catal. 2007, 349, 1445; ChemCat-Chem 2015, 7, 325; Angew. Chem., Int. Ed. 2003, 42, 4815; Curr. Biotechnol., 2015, 4, 111-117.)

C-C Bond Formation

Asymmetric Synthesis of Optically Pure α-Substituted Carboxylic Acids

(AMDase = arylmalonate decarboxylase; ChemCatChem, 2016, 8, 916; Appl. Microbiol. Biotechnol., 2016, 20, 8621.)

C-C Bond Breaking


Racemization of Arylpropionates

(APR = arylpropionate racemase; ChemBioChem, 2016, 16, 1943; Cat. Sci. Technol., 2016, 6, 4397.)

Enzymatic Isomerisation

Isomerisations of C=C-Bonds

(ChemBioChem, 2012, 13, 2346–2351.)

Disproportionation: Biocatalytic Cannizzaro Reaction

(ADH = alcohol dehydrogenase; ChemCatChem, 2013, 5, 1744–1748.)


Hydrolysis of Lactams

(ChemCatChem 2014, 6, 2517.)

Bio-Mitsunobu-Inversion

(Angew. Chem., Int. Ed. 2002, 41, 1014; Angew. Chem., Int. Ed. 2005, 44, 6381; Angew. Chem., Int. Ed. 2013, 52, 3277; Eur. J. Org. Chem. 2013, 356; Trends Biotechnol. 2013, 31, 468; Appl. Microbiol. Biotechnol. 2014, 4, 1485.)

C-C Bond Hydrolysis

(Adv. Synth. Catal. 2013, 355, 1677; Adv. Synth. Catal. 2013, 355, 1703; Top. Catal. 2014, 57, 376.)

Hydrolysis / Esterification

Nitrile Hydrolysis

(Tetrahedron 2005, 61, 4249-4260; ChemCatChem 2010, 2, 267-269; Enz. Microb. Technol. 2010, 47, 140-146; OBC 2015, 13, 7803-7812.)


Enantioselective Ester Formation in Aqueous Systems

(AT= acyl transferase; to be published.)

Hydrolysis / Esterification

Enzymatic Phosphorylation

(PPi = phyrophosphate; Eur. J. Org. Chem., 2016, 45–50.)

Chemoenzymatic Preparation of Bio-Based Anti-Oxidants

(PAD = phenolic acid decarboxylase; Angew. Chem., Int. Ed., 2016, in press, DOI: 10.1002/anie.201607777.)

Enzymatic Cascades


Direct Amination of Alcohols

(ADH = alcohol dehydrogenase; TA = ω-transaminase; Angew. Chem., Int. Ed. 2012, 51, 9156; ACS Catal. 2014, 4, 129.)

6-Aminohexanoic Acid from Cyclohexanol

(ADH = alcohol dehydrogenase; BVMO = Baeyer-Villiger monooxygenase, TA = ω-transaminase; Angew. Chem., Int. Ed. 2014, 53, 14153; ACS Catal. 2014, 4, 129.)

Cyclohexylamines from Diketones

(TA = ω-transaminase; Adv. Synth. Catal. 2013, 355, 1703.)

Vinylation of Phenols

(Angew. Chem. Int. Ed. 2015, 45, 10899.)

Enzymatic Cascades


Regioselective Glycosyltransfer

(Biocat. Biotrans. 2010, 28, 10; Pure Appl. Chem. 2013, 85, 1865; PCT Int. Appl. 2015, WO 2015001033.)

Small Molecule Glycosylation

(Angew. Chem., Int. Ed. 2008, 47, 10086; PCT Int. Appl. 2008, WO 2008034158.)

Regioselective Glucosylation – Flavonoid Glycosylation

(Green Chem. 2014, 16, 4417.)

Glycozyme Technology


Nucleotid Sugar Synthesis

(SPase = sucrose phosphorylase; AGP = α-D-glucose 1-phosphate phosphatase; PPase = pyrophosphatase; NTPase = nucleotide transferase; GTP = guanosine 5′-triphosphate; D-Glc = D-glucose; D-Fru = D-fructose; Pi = inorganic phosphate; Adv. Synth. Catal., submitted.)

Glycozyme Technology

Diasterioselecitve Synthesis of Glycosyl Phosphates

(SPase = sucrose phosphorylase; AGP = α -D-glucose 1-phosphate phosphatase; D-Glc = D-glucose; D-Fru = D-fructose; Pi = inorganic phosphate; Angew. Chem.,Int. Ed., 2015, 54,15867.)


Chemo- & Regioselective Oxidation of Soft Nucleophiles

(Chem. Commun. 2012, 48, 6001-6003; ChemCatChem 2014, 6, 1028-1042; Microb. Cell Fact. 2015, 14:82, 1-10; J. Biotechnol. 2016, 235, 3.)

API-Modification with Human Enzymes


In-silico Search for Novel BiocatalystsTraditional screening for novel enzymes requires time-consuming experiments and expensive activity assays in the wet-lab. To reduce costs, the prediction and identification of enzyme func-tionalities is a major challenge of modern bioinformatics. However, the computational annota-tion of proteins proves to be difficult, erroneous and lacks the possibility to identify completely independent novel biocatalysts because they rely on the correlation of (sequence) similarities with the known functions of the template and are bound to find “more of the same”.

Catalophore Search for Novel Enzymes

acib-researchers developed a patented bioinformatics method1 to mine structural databases using three dimensional search templates which cover the arrangement of chemical functio-nal groups or pre-calculated point-clouds representing the “empty space” of active sites. These search templates are termed “catalophores” (i.e. carrier of the catalytic function). The searches are independent of structural or sequence similarities to currently employed enzymes. Therefo-re, these identified enzymes may feature different physico-chemical properties such as stability, selectivity or substrate tolerance.

A successful test-case led to the identification of two “novel” ene-reductases2, by searching with patterns obtained from classical old yellow enzymes. The identified enzymes showed significant conversions on typical old yellow enzyme substrates and even allowed access to enantiomers that could not be obtained using current enzyme portfolio, although the overall sequence and structural similarity are below 10%.

Catalophore used for the identification of the old-yellow enzyme example; a) schematic mechanism of the reaction mechanism; b) 3D active site constella-tion (“catalphore“) ; c) catalophore mo-tif indicating used atom types; d) geo-metrical representation of the search motif used for the database search.

(1. PCT/EP2013/074556, 2014, ACIB GmbH and University of Graz; 2. Nat. Commun. 2014, 5, 4150.)


next step in industrial research

The Austrian Centre of Industrial Biotechnology (acib) is an international research center with 200+ employees based in Austria with with collaborative links to Germany, Slovenia, Italy, Spain and Poland. acib can draw on 25+ ye-ars of experience to apply sophisticated, new methods in industrial production – focused on biocatalysis, enzymes, polymers, (pharmaceutical) protein production & purifica-tion, cell line development, bioinformatics and synthetic biology.

Together with 130+ industrial and scientific partners acib adopts the tools and methods of nature for i) new produc-tion processes & products with improved ecological effici-ency, ii) new production processes with higher economic efficiency, iii) products with higher quality and purity, iv) functional innovative products for everyday use and v) sophisticated computational methods for the health care, pharmaceutical or chemical industry...

acib – science ist our success.

about acib

The competence centre acib is funded in the framework of COMET – Competence Centers for Excellent Technologies – by BMVIT, BMWFW and the provinces of Lower

Austria, Styria, Tyrol and Vienna. The COMET programme is conducted by FFG.


w: www.acib.atw: biocatalysis.uni-graz.at

www.xing.com/companies/acibgmbhwww.linkedin.com/company/acib-gmbhwww.facebook.com/acibgmbh

contact

Dr. Martin Trinker

Research ManagementBusiness Development

t: +43 316 873 9316fax: +43 316 873 9302m: [email protected]

acib GmbHaustrian centre of industrial biotechnologyPetersgasse 148010 GrazAustria

Prof. Kurt Faber

t: +43 316 380 5332m: [email protected]

Prof. Wolfgang Kroutil


Dr. Christoph Winkler


acib GmbHc/o Department of Chemistry, University of Graz8010 GrazAustria


Notes

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