Metabolic Engineering toward Sustainable Production of Nylon-6

Stefan C. H. J. Turk; Wigard P. Kloosterman; Dennis K. Ninaber; Karin P. A. M. Kolen; Julia Knutova; Erwin Suir; Martin Schurmann; Petronella C. Raemakers-Franken; Monika Muller; Stefaan M. A. de Wildeman; Leonie M. Raamsdonk; Ruud van der Pol; Liang Wu; Margarida F. Temudo; Rob A. M. van der Hoeven; Michiel Akeroyd; Roland E. van der Stoel; Henk J. Noorman; Roel A. L. Bovenberg; Axel C. Trefzer

doi:10.1021/acssynbio.5b00129

Metabolic Engineering toward Sustainable Production of Nylon-6

Stefan C. H. J. Turk^*, Wigard P. Kloosterman, Dennis K. Ninaber, Karin P. A. M. Kolen, Julia Knutova, Erwin Suir, Martin Schurmann, Petronella C. Raemakers-Franken, Monika Muller, Stefaan M. A. de Wildeman, Leonie M. Raamsdonk, Ruud van der Pol, Liang Wu, Margarida F. Temudo, Rob A. M. van der Hoeven, Michiel Akeroyd, Roland E. van der Stoel, Henk J. Noorman, Roel A. L. Bovenberg, Axel C. Trefzer

^*Corresponding author for this work

Groningen Biomolecular Sciences and Biotechnology Institute

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

102 Citations (Scopus)

355 Downloads (Pure)

Abstract

Nylon-6 is a bulk polymer used for many applications. It consists of the non-natural building block 6-aminocaproic acid, the linear form of caprolactam. Via a retro-synthetic approach, two synthetic pathways were identified for the fermentative production of 6-aminocaproic acid. Both pathways require yet unreported novel biocatalytic steps. We demonstrated proof of these bioconversions by in vitro enzyme assays with a set of selected candidate proteins expressed in Escherichia coli. One of the biosynthetic pathways starts with 2-oxoglutarate and contains bioconversions of the ketoacid elongation pathway known from methanogenic archaea. This pathway was selected for implementation in E. coli and yielded 6-aminocaproic acid at levels up to 160 mg/L in lab-scale batch fermentations. The total amount of 6-aminocaproic acid and related intermediates generated by this pathway exceeded 2 g/L in lab-scale fed-batch fermentations, indicating its potential for further optimization toward large-scale sustainable production of nylon-6.

Original language	English
Pages (from-to)	65-73
Number of pages	9
Journal	ACS Synthetic Biology
Volume	5
Issue number	1
DOIs	https://doi.org/10.1021/acssynbio.5b00129
Publication status	Published - Jan-2016

Keywords

nylon-6
caprolactam
metabolic engineering
6-aminocaproic acid
alpha-ketopimelate
adipate
ADIPIC ACID
COENZYME B
CORYNEBACTERIUM-GLUTAMICUM
PROTEIN EXPRESSION
ESCHERICHIA-COLI
BULK CHEMICALS
AMINO-ACIDS
CLONING
BIOSYNTHESIS
IDENTIFICATION

Access to Document

10.1021/acssynbio.5b00129

Metabolic Engineering toward Sustainable Production of Nylon‑6Final publisher's version, 1.93 MBLicence: Taverne

Handle.net

Persistent link

Cite this

Turk, S. C. H. J., Kloosterman, W. P., Ninaber, D. K., Kolen, K. P. A. M., Knutova, J., Suir, E., Schurmann, M., Raemakers-Franken, P. C., Muller, M., de Wildeman, S. M. A., Raamsdonk, L. M., van der Pol, R., Wu, L., Temudo, M. F., van der Hoeven, R. A. M., Akeroyd, M., van der Stoel, R. E., Noorman, H. J., Bovenberg, R. A. L., & Trefzer, A. C. (2016). Metabolic Engineering toward Sustainable Production of Nylon-6. ACS Synthetic Biology, 5(1), 65-73. https://doi.org/10.1021/acssynbio.5b00129

@article{99f0385b58fc41a8872f0fceda626bad,

title = "Metabolic Engineering toward Sustainable Production of Nylon-6",

abstract = "Nylon-6 is a bulk polymer used for many applications. It consists of the non-natural building block 6-aminocaproic acid, the linear form of caprolactam. Via a retro-synthetic approach, two synthetic pathways were identified for the fermentative production of 6-aminocaproic acid. Both pathways require yet unreported novel biocatalytic steps. We demonstrated proof of these bioconversions by in vitro enzyme assays with a set of selected candidate proteins expressed in Escherichia coli. One of the biosynthetic pathways starts with 2-oxoglutarate and contains bioconversions of the ketoacid elongation pathway known from methanogenic archaea. This pathway was selected for implementation in E. coli and yielded 6-aminocaproic acid at levels up to 160 mg/L in lab-scale batch fermentations. The total amount of 6-aminocaproic acid and related intermediates generated by this pathway exceeded 2 g/L in lab-scale fed-batch fermentations, indicating its potential for further optimization toward large-scale sustainable production of nylon-6.",

keywords = "nylon-6, caprolactam, metabolic engineering, 6-aminocaproic acid, alpha-ketopimelate, adipate, ADIPIC ACID, COENZYME B, CORYNEBACTERIUM-GLUTAMICUM, PROTEIN EXPRESSION, ESCHERICHIA-COLI, BULK CHEMICALS, AMINO-ACIDS, CLONING, BIOSYNTHESIS, IDENTIFICATION",

author = "Turk, {Stefan C. H. J.} and Kloosterman, {Wigard P.} and Ninaber, {Dennis K.} and Kolen, {Karin P. A. M.} and Julia Knutova and Erwin Suir and Martin Schurmann and Raemakers-Franken, {Petronella C.} and Monika Muller and {de Wildeman}, {Stefaan M. A.} and Raamsdonk, {Leonie M.} and {van der Pol}, Ruud and Liang Wu and Temudo, {Margarida F.} and {van der Hoeven}, {Rob A. M.} and Michiel Akeroyd and {van der Stoel}, {Roland E.} and Noorman, {Henk J.} and Bovenberg, {Roel A. L.} and Trefzer, {Axel C.}",

year = "2016",

month = jan,

doi = "10.1021/acssynbio.5b00129",

language = "English",

volume = "5",

pages = "65--73",

journal = "ACS Synthetic Biology",

issn = "2161-5063",

publisher = "AMER CHEMICAL SOC",

number = "1",

}

Turk, SCHJ, Kloosterman, WP, Ninaber, DK, Kolen, KPAM, Knutova, J, Suir, E, Schurmann, M, Raemakers-Franken, PC, Muller, M, de Wildeman, SMA, Raamsdonk, LM, van der Pol, R, Wu, L, Temudo, MF, van der Hoeven, RAM, Akeroyd, M, van der Stoel, RE, Noorman, HJ, Bovenberg, RAL & Trefzer, AC 2016, 'Metabolic Engineering toward Sustainable Production of Nylon-6', ACS Synthetic Biology, vol. 5, no. 1, pp. 65-73. https://doi.org/10.1021/acssynbio.5b00129

TY - JOUR

T1 - Metabolic Engineering toward Sustainable Production of Nylon-6

AU - Turk, Stefan C. H. J.

AU - Kloosterman, Wigard P.

AU - Ninaber, Dennis K.

AU - Kolen, Karin P. A. M.

AU - Knutova, Julia

AU - Suir, Erwin

AU - Schurmann, Martin

AU - Raemakers-Franken, Petronella C.

AU - Muller, Monika

AU - de Wildeman, Stefaan M. A.

AU - Raamsdonk, Leonie M.

AU - van der Pol, Ruud

AU - Wu, Liang

AU - Temudo, Margarida F.

AU - van der Hoeven, Rob A. M.

AU - Akeroyd, Michiel

AU - van der Stoel, Roland E.

AU - Noorman, Henk J.

AU - Bovenberg, Roel A. L.

AU - Trefzer, Axel C.

PY - 2016/1

Y1 - 2016/1

N2 - Nylon-6 is a bulk polymer used for many applications. It consists of the non-natural building block 6-aminocaproic acid, the linear form of caprolactam. Via a retro-synthetic approach, two synthetic pathways were identified for the fermentative production of 6-aminocaproic acid. Both pathways require yet unreported novel biocatalytic steps. We demonstrated proof of these bioconversions by in vitro enzyme assays with a set of selected candidate proteins expressed in Escherichia coli. One of the biosynthetic pathways starts with 2-oxoglutarate and contains bioconversions of the ketoacid elongation pathway known from methanogenic archaea. This pathway was selected for implementation in E. coli and yielded 6-aminocaproic acid at levels up to 160 mg/L in lab-scale batch fermentations. The total amount of 6-aminocaproic acid and related intermediates generated by this pathway exceeded 2 g/L in lab-scale fed-batch fermentations, indicating its potential for further optimization toward large-scale sustainable production of nylon-6.

AB - Nylon-6 is a bulk polymer used for many applications. It consists of the non-natural building block 6-aminocaproic acid, the linear form of caprolactam. Via a retro-synthetic approach, two synthetic pathways were identified for the fermentative production of 6-aminocaproic acid. Both pathways require yet unreported novel biocatalytic steps. We demonstrated proof of these bioconversions by in vitro enzyme assays with a set of selected candidate proteins expressed in Escherichia coli. One of the biosynthetic pathways starts with 2-oxoglutarate and contains bioconversions of the ketoacid elongation pathway known from methanogenic archaea. This pathway was selected for implementation in E. coli and yielded 6-aminocaproic acid at levels up to 160 mg/L in lab-scale batch fermentations. The total amount of 6-aminocaproic acid and related intermediates generated by this pathway exceeded 2 g/L in lab-scale fed-batch fermentations, indicating its potential for further optimization toward large-scale sustainable production of nylon-6.

KW - nylon-6

KW - caprolactam

KW - metabolic engineering

KW - 6-aminocaproic acid

KW - alpha-ketopimelate

KW - adipate

KW - ADIPIC ACID

KW - COENZYME B

KW - CORYNEBACTERIUM-GLUTAMICUM

KW - PROTEIN EXPRESSION

KW - ESCHERICHIA-COLI

KW - BULK CHEMICALS

KW - AMINO-ACIDS

KW - CLONING

KW - BIOSYNTHESIS

KW - IDENTIFICATION

U2 - 10.1021/acssynbio.5b00129

DO - 10.1021/acssynbio.5b00129

M3 - Article

C2 - 26511532

SN - 2161-5063

VL - 5

SP - 65

EP - 73

JO - ACS Synthetic Biology

JF - ACS Synthetic Biology

IS - 1

ER -

Metabolic Engineering toward Sustainable Production of Nylon-6

Abstract

Keywords

Access to Document

Handle.net

Fingerprint

Cite this