Engineering Saccharomyces cerevisiae for production of simvastatin.
Natural products
Polyketides
Saccharomyces cerevisiae
Simvastatin
Journal
Metabolic engineering
ISSN: 1096-7184
Titre abrégé: Metab Eng
Pays: Belgium
ID NLM: 9815657
Informations de publication
Date de publication:
01 2019
01 2019
Historique:
received:
07
06
2018
revised:
20
08
2018
accepted:
08
09
2018
pubmed:
15
9
2018
medline:
14
6
2019
entrez:
15
9
2018
Statut:
ppublish
Résumé
Simvastatin is a semisynthetic cholesterol-lowering medication and one of the top-selling statins in the world. Currently, industrial production of simvastatin acid (SVA) is a multistep process starting from the natural product lovastatin. For this reason, there is significant interest in direct production of simvastatin from a microbial host. In this study, six heterologous biosynthetic genes were introduced into Saccharomyces cerevisiae and the acyl-donor dimethylbutyryl-S-methyl mercaptopropionate (DMB-SMMP) was added, resulting in initial production of 0.5 mg/L SVA. Switching the yeast strain from JHY686 to BJ5464-NpgA increased total polyketide production to over 60 mg/L and conversion from dihydromonacolin L acid to monacolin J acid (MJA) was increased from 60% to 90% by tuning the copy number of the P450 lovA. Increasing the media pH to 8.7 led to a further 10-fold increase in SVA production. Optimized chemical lysis of the cell walls in situ after maximum MJA production led to 55 mg/L SVA titer, representing nearly complete conversion from MJA and a 110-fold increase in titer from the initial SVA production strain. The yeast strains developed in this work can be used as an alternative production method for SVA, and the strategies employed can be broadly applied for heterologous production of other fungal polyketides and semisynthetic compounds in yeast.
Identifiants
pubmed: 30213650
pii: S1096-7176(18)30234-9
doi: 10.1016/j.ymben.2018.09.005
pmc: PMC6348118
mid: NIHMS1508392
pii:
doi:
Substances chimiques
Hydroxymethylglutaryl-CoA Reductase Inhibitors
0
Polyketides
0
Cytochrome P-450 Enzyme System
9035-51-2
Lovastatin
9LHU78OQFD
Simvastatin
AGG2FN16EV
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
1-8Subventions
Organisme : NIGMS NIH HHS
ID : T32 GM067555
Pays : United States
Informations de copyright
Copyright © 2018 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.
Références
Appl Microbiol Biotechnol. 2010 Jan;85(4):869-83
pubmed: 19820926
Bioinformatics. 2015 Apr 1;31(7):1120-3
pubmed: 25414360
Chem Biol. 2009 Oct 30;16(10):1064-74
pubmed: 19875080
Nat Chem Biol. 2014 Jun;10(6):431-6
pubmed: 24727900
Science. 1999 May 21;284(5418):1368-72
pubmed: 10334994
Metab Eng. 2018 Jan;45:189-199
pubmed: 29258964
Appl Environ Microbiol. 2005 Mar;71(3):1515-21
pubmed: 15746355
ACS Synth Biol. 2015 May 15;4(5):585-94
pubmed: 25207793
N Biotechnol. 2014 Sep 25;31(5):499-505
pubmed: 25017988
Nat Methods. 2007 Mar;4(3):251-6
pubmed: 17293868
Bioorg Med Chem Lett. 2001 Jun 18;11(12):1527-31
pubmed: 11412974
FEBS Lett. 2015 Jul 8;589(15):1723-1727
pubmed: 26054976
Chembiochem. 2015 Nov;16(17):2420-31
pubmed: 26478227
Yeast. 2005 Apr 30;22(6):431-40
pubmed: 15849781
Nat Methods. 2014 Feb;11(2):122-3
pubmed: 24481216
Nat Protoc. 2007;2(1):31-4
pubmed: 17401334
Metab Eng Commun. 2015 Sep 25;2:117-123
pubmed: 34150514
J Am Chem Soc. 2011 Jun 1;133(21):8078-81
pubmed: 21495633
Appl Environ Microbiol. 2007 Apr;73(7):2054-60
pubmed: 17277201
J Clin Pharm Ther. 2010 Apr;35(2):139-51
pubmed: 20456733
J Biosci Bioeng. 2011 Jan;111(1):10-5
pubmed: 20855233
J Med Chem. 1986 May;29(5):849-52
pubmed: 3634830
Curr Opin Chem Biol. 2012 Apr;16(1-2):117-23
pubmed: 22342766
Mol Genet Genomics. 2002 Jul;267(5):636-46
pubmed: 12172803
J Am Chem Soc. 2009 Jun 24;131(24):8388-9
pubmed: 19530726
Adv Biochem Eng Biotechnol. 2013;131:89-119
pubmed: 23111502
Nat Rev Drug Discov. 2003 Jul;2(7):517-26
pubmed: 12815379
Curr Opin Lipidol. 2013 Jun;24(3):221-6
pubmed: 23571733
Science. 2009 Oct 23;326(5952):589-92
pubmed: 19900898
Curr Opin Plant Biol. 2014 Jun;19:27-34
pubmed: 24709279
Metab Eng. 2014 Sep;25:194-203
pubmed: 25084369
Chem Biol. 2006 Nov;13(11):1161-9
pubmed: 17113998
Biotechnol Bioeng. 2006 Mar 5;93(4):727-36
pubmed: 16245348
J Am Chem Soc. 2015 Nov 4;137(43):13724-7
pubmed: 26469304
Angew Chem Int Ed Engl. 2009;48(26):4688-716
pubmed: 19514004
Proc Natl Acad Sci U S A. 1980 Jul;77(7):3957-61
pubmed: 6933445
Drug Discov Today. 2008 Oct;13(19-20):894-901
pubmed: 18691670
Biotechniques. 2011 May;50(5):325-8
pubmed: 21548894
Cell Syst. 2015 Jul 29;1(1):88-96
pubmed: 27135688
Nat Prod Rep. 2014 Apr;31(4):504-13
pubmed: 24514754
G3 (Bethesda). 2014 Mar 20;4(3):389-98
pubmed: 24374639
Appl Microbiol Biotechnol. 2014 Dec;98(23):9561-78
pubmed: 25359472
Yeast. 2012 Dec;29(12):495-503
pubmed: 23166051
Metab Eng. 2015 Jul;30:96-104
pubmed: 25981946
Anal Biochem. 2000 Oct 15;285(2):205-10
pubmed: 11017703
FASEB J. 2006 Feb;20(2):200-1
pubmed: 16449790
Sci Adv. 2018 Apr 11;4(4):eaar5459
pubmed: 29651464
Yeast. 1998 Jan 30;14(2):115-32
pubmed: 9483801
Angew Chem Int Ed Engl. 2013 Jun 17;52(25):6472-5
pubmed: 23653178
J Am Chem Soc. 2010 Oct 6;132(39):13604-7
pubmed: 20828130