Hoffmann-La Roche Inc.

Roche Molecular Systems, Inc.

Roche Diagnostics Operations, Inc.

Roche Sequencing Solutions, Inc.

Roche Diabetes Care, Inc.

F. Hoffmann-La Roche Ag

Genia Technologies, Inc.

Roche Diagnostics Gmbh

Mara Boenitz-Dulat

Peter Kratzsch

Martin Schatte

Shawn Lee Suko

Aruna Ayer

Eileen Thai

Ilya Lederman

Charles Wayan Schwab

Cleoma Arnold

Colin Alexander McGaw

Barbara Eckert

David Daniel Wunderlich

Bigna Woersdoerfer

Arkadiusz Bibillo

Rainer Schmuck

Tyler O'Brien Shultz

Daniela Beck

Preethi Sarvabhowman

Thomas Streidl

Evan Amato

Herbert Von Der Eltz

Erhard Kopetzki

Carina Horn

Frank Bergmann

Hans Iding

Wolfgang-Reinhold Knappe

Stacy Hunt DuVall

Dennis Wetzl

Alessa Hinzmann

Jessica Laggerbauer

Shawn Suko

Cleoma Renetta Arnold

Tyler O'Brien Schultz

Stacy Hunt Duvall

Reported herein is a sortase comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 11 and that comprises the mutations D101S and K137S.

Sortase

Herein is reported a polypeptide comprising the amino acid sequence of SEQ ID NO: 38 as solederived polypeptide and its use in conjugating polypeptides.

Soluble Sortase A

Herein is reported a method for the enzymatic production of a polypeptide comprising the step of incubating i) a first polypeptide comprising the amino acid sequence LPXTG (SEQ ID NO: 01, wherein X can be any amino acid residue) or LPXTA (SEQ ID NO: 41, wherein X can be any amino acid residue), ii) a second polypeptide that has i) a glycinyl, an alaninyl, or a cysteinyl compound at its N-terminus, or ii) an oligoglycine, or oligoalanine, or a cysteine amino acid residue followed by one to three glycine or alanine amino acid residues at its N-terminus, or iii) a lysine amino acid residue within its 5 N-terminal amino acid residues, and iii) a third polypeptide with sortase A activity, in a deep eutectic solvent and thereby producing a polypeptide.

Transamidation reaction in deep eutectic solvents

Herein is reported a method for the detection of a sortase in a sample, comprising the following steps:

Activity assay for bond forming enzymes

Herein is reported a method for the enzymatic production of a thioester comprising incubating i) a first polypeptide comprising the amino acid sequence LPXTG (SEQ ID NO: 01, wherein X can be any amino acid residue), ii) a second polypeptide that has at its N-terminus a cysteine amino acid residue or is a cysteinyl compound, and iii) a third polypeptide with sortase A activity.

Production of thioesters using sortase

The present invention relates to the use of a protein as a hydroxylase, a host cell comprising the protein, the use of the protein or host cell in the production of hydroxy-pipecolic acid (HPA), a method of production HPA, methods of producing the protein, a mutant protein and a nucleic acid encoding the mutant protein and a vector comprising a nucleic acid encoding the protein.

Proline hydroxylases as well as uses, methods and products involving the same

Described herein is a variant pol6 polymerase having at least one mutation selected from H223, N224, Y225, H227, I295, Y342, T343, I357, S360, L361, I363, S365Q, S366, Y367, P368, D417, E475, Y476, F478, K518, H527, T529, M531, N535, G539, P542, N545, Q546, A547, L549, I550, N552, G553, F558, A596, G603, A610, V615, Y622, C623, D624, I628, Y629, R632, N635, M641, A643, I644, T647, I648, T651, I652, K655, W656, D657, V658, H660, F662, L690 and combinations thereof.

Polymerase variants

The present disclosure provides variant Pol6 polymerase polypeptides, compositions comprising the Pol6 variant polypeptides, and methods for using the variant Pol6 polypeptides for determining the sequencing of nucleic acids, for example, by nanopore sequencing. The variant Pol6 polymerases possess decreased rates of dissociation of template from the polymerase-template complex, which result in increased processivity relative to the parental Pol6 polypeptides from which they are derived.

Pol6 polymerase variants

The present invention relates to a mutant 3-hydroxybutyrate dehydrogenase (3-HBDH) with improved performance relative to the wild-type 3-HBDH, a nucleic acid encoding the mutant 3-HBDH, a cell comprising the mutant 3-HBDH or the nucleic acid, a method of determining the amount or concentration of 3-hydroxybutyrate in a sample, and a device for determining the amount or concentration of 3-hydroxybutyrate in a sample.

Mutant 3-hydroxybutyrate dehydrogenase fromas well as methods and uses involving the same

Described herein is a variant pol6 polymerase having at least one mutation selected from H223, N224, Y225, H227, I295, Y342, T343, I357, S360, L361, I363, S365Q, S366, Y367, P368, D417, E475, Y476, F478, K518, H527, T529, M531, N535, G539, P542, N545, Q546, A547, L549, I550, N552, G553, F558, A596, G603, A610, V615, Y622, C623, D624, I628, Y629, R632, N635, M641, A643, 1644, T647, I648, T651, I652, K655, W656, D657, V658, H660, F662, L690 and combinations thereof.

The present disclosure relates to a mutant 3-hydroxybutyrate dehydrogenase (3-HBDH) with improved performance relative to the wild-type 3-HBDH, a nucleic acid encoding the mutant 3-HBDH, a cell having the mutant 3-HBDH or the nucleic acid, and/or a method of determining the amount or concentration of 3-hydroxybutyrate in a sample. Also disclosed is the use of the mutant 3-HBDH for determining the amount or concentration of 3-hydroxybutyrate in a sample, and a device for determining the amount or concentration of 3-hydroxybutyrate in a sample.

Mutant 3-hydroxybutyrate dehydrogenase from alcaligenes faecalis as well as methods and uses involving the same

Described herein are variant pol6 polymerase having at least having at least two substitutions, wherein at least one of the substitutions is at position S366 of SEQ ID NO: 2. The disclosed polymerases may be coupled to alpha-hemolysin nanopores, and may be used in nanopore-based sequencing apparatuses.

Herein is reported a method for producing an enzymatic conjugation product of two polypeptides comprising incubating of a first polypeptide comprising the amino acid sequence LPXTG (SEQ ID NO: 20, wherein X can be any amino acid residue), a second polypeptide has an oligo-alanine A(m=2 (SEQ ID NO: 26), or 3 (SEQ ID NO: 27), or 4 (SEQ ID NO: 28), or 5 (SEQ ID NO: 29)) amino acid sequence at its N-terminus, a third polypeptide with sortase activity which is derived fromSortase A, and recovering the conjugate from the reaction mixture and thereby producing the enzymatic conjugation product of two polypeptides.

Methods for enzyme mediated polypeptide conjugation

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Tutzing, Germany

Mara Boenitz-Dulat