Imperial College London
Portrait Picture

ProfessorPaulFreemont

Faculty of Medicine, Department of Infectious Disease

Chair in Protein Crystallography
 
 
 
//

Contact

 

+44 (0)20 7594 5327p.freemont

 
 
//

Location

 

259Sir Alexander Fleming BuildingSouth Kensington Campus

//

Summary

 

Publications

Citation

BibTex format

@article{Moore:2021:synbio/ysab021,
author = {Moore, S and Tosi, T and Bell, D and Hleba, Y and Freemont, P and Polizzi, K},
doi = {synbio/ysab021},
journal = {Synthetic Biology},
pages = {1--8},
title = {High-yield ‘one-pot’ biosynthesis of raspberry ketone, a high-value fine chemical},
url = {http://dx.doi.org/10.1093/synbio/ysab021},
volume = {6},
year = {2021}
}
Download

RIS format (EndNote, RefMan)

TY  - JOUR
AB  - Cell-free extract and purified enzyme-based systems provide an attractive solution to study biosynthetic strategies towards a range of chemicals. 4-(4-hydroxyphenyl)-butan-2-one, also known as raspberry ketone, is the major fragrance component of raspberry fruit and is used as a natural additive in the food and sports industry. Current industrial processing of the natural form of raspberry ketone involves chemical extraction with a yield of ~1-4 mg kg-1 of fruit. Due to toxicity, microbial production provides only low yields of up to 5-100 mg L-1. Herein, we report an efficient cell-free strategy to probe a synthetic enzyme pathway that converts either L-tyrosine or the precursor, 4-(4-hydroxyphenyl)-buten-2-one (HBA), into raspberry ketone at up to 100% conversion. As part of this strategy, it is essential to recycle inexpensive cofactors. Specifically, the final enzyme step in the pathway is catalysed by raspberry ketone/zingerone synthase (RZS1), an NADPH-dependent double bond reductase. To relax cofactor specificity towards NADH, the preferred cofactor for cell-free biosynthesis, we identify a variant (G191D) with strong activity with NADH. We implement the RZS1 G191D variant within a ‘one-pot’ cell-free reaction to produce raspberry ketone at high-yield (61 mg L-1), which provides an alternative route to traditional microbial production. In conclusion, our cell-free strategy complements the growing interest in engineering synthetic enzyme cascades towards industrially relevant value-added chemicals.
AU  - Moore,S
AU  - Tosi,T
AU  - Bell,D
AU  - Hleba,Y
AU  - Freemont,P
AU  - Polizzi,K
DO  - synbio/ysab021
EP  - 8
PY  - 2021///
SN  - 2397-7000
SP  - 1
TI  - High-yield ‘one-pot’ biosynthesis of raspberry ketone, a high-value fine chemical
T2  - Synthetic Biology
UR  - http://dx.doi.org/10.1093/synbio/ysab021
UR  - https://academic.oup.com/synbio/advance-article/doi/10.1093/synbio/ysab021/6338191?login=true
UR  - http://hdl.handle.net/10044/1/90949
VL  - 6
ER  -
Download