Thioester-Mediated Biocatalytic Amide Bond Synthesis with In Situ Thiol Recycling

Christian Schnepel; Laura Rodríguez Pérez; Yuqi Yu; Antonio Angelastro; Rachel Heath; Max Lubberink; Francesco Falcioni; Keith Mulholland; Martin A. Hayes; Nicholas Turner; Sabine Flitsch

Thioester-Mediated Biocatalytic Amide Bond Synthesis with In Situ Thiol Recycling

Christian Schnepel, Laura Rodríguez Pérez, Yuqi Yu, Antonio Angelastro, Rachel Heath, Max Lubberink, Francesco Falcioni, Keith Mulholland, Martin A. Hayes, Nicholas Turner, Sabine Flitsch

Research output: Contribution to journal › Article › peer-review

Abstract

Activation of carboxylic acids to thioesters plays an important role in biology. However, biochemical studies and biotechnological applications are hampered by a general lack of access to thioesters, especially those based on HS-Coenzyme A. Here we show that a generic thioester recycling enzyme by exploiting a promiscuous activity of carboxylic acid reductase (CARsr-A). The adenylation domain of CARsr (CARsr-A) catalyses the conversion of a wide range of carboxylic acids to acyl-S-Coenzyme A and other thioesters in good yields. CARsr-A was used in situ as part of a recycling system to regenerate thioesters for acyl-S-Coenzyme A dependent enzymes in one-pot reactions. This concept of thioester recycling was demonstrated with a range of acyltransferases allowing formation of diverse amides and non-native acylation of lysine sidechains in a histone-derived peptide using the epigenetic writer, lysine acetyltransferase HATp300. Overall, these results establish a generic platform for thioester formation towards amide formation and beyond.

Original language	English
Journal	Nature Catalysis
Publication status	Accepted/In press - 21 Oct 2022

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7601_2_merged_1666782482
Accepted author manuscript, 1.63 MB
Embargo ends: 1/01/32

Cite this

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title = "Thioester-Mediated Biocatalytic Amide Bond Synthesis with In Situ Thiol Recycling",

abstract = "Activation of carboxylic acids to thioesters plays an important role in biology. However, biochemical studies and biotechnological applications are hampered by a general lack of access to thioesters, especially those based on HS-Coenzyme A. Here we show that a generic thioester recycling enzyme by exploiting a promiscuous activity of carboxylic acid reductase (CARsr-A). The adenylation domain of CARsr (CARsr-A) catalyses the conversion of a wide range of carboxylic acids to acyl-S-Coenzyme A and other thioesters in good yields. CARsr-A was used in situ as part of a recycling system to regenerate thioesters for acyl-S-Coenzyme A dependent enzymes in one-pot reactions. This concept of thioester recycling was demonstrated with a range of acyltransferases allowing formation of diverse amides and non-native acylation of lysine sidechains in a histone-derived peptide using the epigenetic writer, lysine acetyltransferase HATp300. Overall, these results establish a generic platform for thioester formation towards amide formation and beyond.",

author = "Christian Schnepel and {Rodr{\'i}guez P{\'e}rez}, Laura and Yuqi Yu and Antonio Angelastro and Rachel Heath and Max Lubberink and Francesco Falcioni and Keith Mulholland and Hayes, {Martin A.} and Nicholas Turner and Sabine Flitsch",

year = "2022",

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language = "English",

journal = "Nature Catalysis",

issn = "2520-1158",

publisher = "Springer Nature",

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TY - JOUR

T1 - Thioester-Mediated Biocatalytic Amide Bond Synthesis with In Situ Thiol Recycling

AU - Schnepel, Christian

AU - Rodríguez Pérez, Laura

AU - Yu, Yuqi

AU - Angelastro, Antonio

AU - Heath, Rachel

AU - Lubberink, Max

AU - Falcioni, Francesco

AU - Mulholland, Keith

AU - Hayes, Martin A.

AU - Turner, Nicholas

AU - Flitsch, Sabine

PY - 2022/10/21

Y1 - 2022/10/21

N2 - Activation of carboxylic acids to thioesters plays an important role in biology. However, biochemical studies and biotechnological applications are hampered by a general lack of access to thioesters, especially those based on HS-Coenzyme A. Here we show that a generic thioester recycling enzyme by exploiting a promiscuous activity of carboxylic acid reductase (CARsr-A). The adenylation domain of CARsr (CARsr-A) catalyses the conversion of a wide range of carboxylic acids to acyl-S-Coenzyme A and other thioesters in good yields. CARsr-A was used in situ as part of a recycling system to regenerate thioesters for acyl-S-Coenzyme A dependent enzymes in one-pot reactions. This concept of thioester recycling was demonstrated with a range of acyltransferases allowing formation of diverse amides and non-native acylation of lysine sidechains in a histone-derived peptide using the epigenetic writer, lysine acetyltransferase HATp300. Overall, these results establish a generic platform for thioester formation towards amide formation and beyond.

AB - Activation of carboxylic acids to thioesters plays an important role in biology. However, biochemical studies and biotechnological applications are hampered by a general lack of access to thioesters, especially those based on HS-Coenzyme A. Here we show that a generic thioester recycling enzyme by exploiting a promiscuous activity of carboxylic acid reductase (CARsr-A). The adenylation domain of CARsr (CARsr-A) catalyses the conversion of a wide range of carboxylic acids to acyl-S-Coenzyme A and other thioesters in good yields. CARsr-A was used in situ as part of a recycling system to regenerate thioesters for acyl-S-Coenzyme A dependent enzymes in one-pot reactions. This concept of thioester recycling was demonstrated with a range of acyltransferases allowing formation of diverse amides and non-native acylation of lysine sidechains in a histone-derived peptide using the epigenetic writer, lysine acetyltransferase HATp300. Overall, these results establish a generic platform for thioester formation towards amide formation and beyond.

M3 - Article

SN - 2520-1158

JO - Nature Catalysis

JF - Nature Catalysis

ER -

Thioester-Mediated Biocatalytic Amide Bond Synthesis with In Situ Thiol Recycling

Abstract

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