Metabolic Engineering toward Sustainable Production of Nylon-6.
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.
DSM Biotechnology Center , PO Box 1, 2600 MA Delft, The Netherlands.
Stefan C H J
SC
Turk
DSM Biotechnology Center , PO Box 1, 2600 MA Delft, The Netherlands.|University Medical Center Utrecht , PO Box 85060, 3508 AB Utrecht, The Netherlands.
Wigard P
WP
Kloosterman
DSM Biotechnology Center , PO Box 1, 2600 MA Delft, The Netherlands.|Leiden University Medical Center , Albinusdreef 2, 2333 ZA Leiden, The Netherlands.
Dennis K
DK
Ninaber
DSM Biotechnology Center , PO Box 1, 2600 MA Delft, The Netherlands.
Karin P A M
KP
Kolen
DSM Biotechnology Center , PO Box 1, 2600 MA Delft, The Netherlands.
Julia
J
Knutova
DSM Biotechnology Center , PO Box 1, 2600 MA Delft, The Netherlands.|BioscienZ , Goeseelsstraat 10, 4817 MV Breda, The Netherlands.
Erwin
E
Suir
DSM Innovative Synthesis , PO Box 18, 6160 MD Geleen, The Netherlands.
Martin
M
Schürmann
DSM Innovative Synthesis , PO Box 18, 6160 MD Geleen, The Netherlands.
Petronella C
PC
Raemakers-Franken
DSM Innovative Synthesis , PO Box 18, 6160 MD Geleen, The Netherlands.
Monika
M
Müller
DSM Innovative Synthesis , PO Box 18, 6160 MD Geleen, The Netherlands.
Stefaan M A
SM
de Wildeman
DSM Biotechnology Center , PO Box 1, 2600 MA Delft, The Netherlands.
Leonie M
LM
Raamsdonk
DSM Biotechnology Center , PO Box 1, 2600 MA Delft, The Netherlands.
Ruud
R
van der Pol
DSM Biotechnology Center , PO Box 1, 2600 MA Delft, The Netherlands.
Liang
L
Wu
DSM Biotechnology Center , PO Box 1, 2600 MA Delft, The Netherlands.
Margarida F
MF
Temudo
DSM Biotechnology Center , PO Box 1, 2600 MA Delft, The Netherlands.
Rob A M
RA
van der Hoeven
DSM Biotechnology Center , PO Box 1, 2600 MA Delft, The Netherlands.
Michiel
M
Akeroyd
DSM Fibre Intermediates , Poststraat 1, 6130 AA Sittard, The Netherlands.
Roland E
RE
van der Stoel
DSM Biotechnology Center , PO Box 1, 2600 MA Delft, The Netherlands.
Henk J
HJ
Noorman
DSM Biotechnology Center , PO Box 1, 2600 MA Delft, The Netherlands.|Synthetic Biology and Cell Engineering, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen , 9747 AG Groningen, The Netherlands.
Roel A L
RA
Bovenberg
DSM Biotechnology Center , PO Box 1, 2600 MA Delft, The Netherlands.|Life Technologies, GeneArt , Im Gewerbepark B35, 93059 Regensburg, Germany.
Axel C
AC
Trefzer
D000272:Adipates / Q000378:metabolism; D015119:Aminocaproic Acid / Q000378:metabolism; D061249:Batch Cell Culture Techniques; D002209:Caprolactam / Q000031:analogs & derivatives* / Q000138:chemical synthesis; D002853:Chromatography, Liquid; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005285:Fermentation; D060847:Metabolic Engineering / Q000379:methods*; D055442:Metabolome; D010867:Pimelic Acids / Q000378:metabolism; D011108:Polymers / Q000138:chemical synthesis*; D040901:Proteomics; D053719:Tandem Mass Spectrometry; D014233:Tricarboxylic Acids / Q000378:metabolism
D000272:Adipates; D010867:Pimelic Acids; D011108:Polymers; D014233:Tricarboxylic Acids; C009916:nylon 6; C028143:homocitric acid; D002209:Caprolactam; C029900:adipic acid; D015119:Aminocaproic Acid
6-aminocaproic acid; adipate; caprolactam; metabolic engineering; nylon-6; α-ketopimelate