Driving the conversion of phytosterol to 9α-hydroxy-4-androstene-3,17-dione in Mycolicibacterium neoaurum by engineering the supply and regeneration of flavin adenine dinucleotide.
Cofactor regeneration
Flavin adenine dinucleotide
Mycolicibacterium
Steroid synthons
Sterols
Journal
Biotechnology for biofuels and bioproducts
ISSN: 2731-3654
Titre abrégé: Biotechnol Biofuels Bioprod
Pays: England
ID NLM: 9918300888906676
Informations de publication
Date de publication:
08 Jun 2023
08 Jun 2023
Historique:
received:
06
02
2023
accepted:
26
04
2023
medline:
9
6
2023
pubmed:
9
6
2023
entrez:
8
6
2023
Statut:
epublish
Résumé
The conversion of phytosterols to steroid synthons by engineered Mycolicibacteria comprises one of the core steps in the commercial production of steroid hormones. This is a complex oxidative catabolic process, and taking the production of androstenones as example, it requires about 10 equivalent flavin adenine dinucleotide (FAD). As the high demand for FAD, the insufficient supply of FAD may be a common issue limiting the conversion process. We substantiated, using the production of 9α-hydroxy-4-androstene-3,17-dione (9-OHAD) as a model, that increasing intracellular FAD supply could effectively increase the conversion of phytosterols into 9-OHAD. Overexpressing ribB and ribC, two key genes involving in FAD synthesis, could significantly enhance the amount of intracellular FAD by 167.4% and the production of 9-OHAD by 25.6%. Subsequently, styrene monooxygenase NfStyA2B from Nocardia farcinica was employed to promote the cyclic regeneration of FAD by coupling the oxidation of nicotinamide adenine dinucleotide (NADH) to NAD This study highlighted that the cofactor engineering, including the supply and recycling of FAD and NAD
Sections du résumé
BACKGROUND
BACKGROUND
The conversion of phytosterols to steroid synthons by engineered Mycolicibacteria comprises one of the core steps in the commercial production of steroid hormones. This is a complex oxidative catabolic process, and taking the production of androstenones as example, it requires about 10 equivalent flavin adenine dinucleotide (FAD). As the high demand for FAD, the insufficient supply of FAD may be a common issue limiting the conversion process.
RESULTS
RESULTS
We substantiated, using the production of 9α-hydroxy-4-androstene-3,17-dione (9-OHAD) as a model, that increasing intracellular FAD supply could effectively increase the conversion of phytosterols into 9-OHAD. Overexpressing ribB and ribC, two key genes involving in FAD synthesis, could significantly enhance the amount of intracellular FAD by 167.4% and the production of 9-OHAD by 25.6%. Subsequently, styrene monooxygenase NfStyA2B from Nocardia farcinica was employed to promote the cyclic regeneration of FAD by coupling the oxidation of nicotinamide adenine dinucleotide (NADH) to NAD
CONCLUSIONS
CONCLUSIONS
This study highlighted that the cofactor engineering, including the supply and recycling of FAD and NAD
Identifiants
pubmed: 37291661
doi: 10.1186/s13068-023-02331-1
pii: 10.1186/s13068-023-02331-1
pmc: PMC10251532
doi:
Types de publication
Journal Article
Langues
eng
Pagination
98Subventions
Organisme : National Natural Science Foundation of China
ID : 21776075
Organisme : National Natural Science Foundation of China
ID : 32100067
Organisme : Natural Science Foundation of Shanghai
ID : 20ZR1415100
Organisme : National Key Research and Development Program of China
ID : 2021YFC2100300
Informations de copyright
© 2023. The Author(s).
Références
Antonie Van Leeuwenhoek. 2014 Jul;106(1):157-72
pubmed: 24846050
Science. 2013 Mar 8;339(6124):1210-3
pubmed: 23471409
Environ Sci Technol. 2019 Sep 17;53(18):10897-10905
pubmed: 31419125
Microbiol Mol Biol Rev. 2011 Jun;75(2):321-60
pubmed: 21646432
Bioresour Technol. 2019 May;279:209-217
pubmed: 30735930
J Agric Food Chem. 2017 Dec 6;65(48):10520-10525
pubmed: 29131627
Metab Eng. 2009 May;11(3):163-7
pubmed: 19558965
Metab Eng. 2019 Dec;56:97-110
pubmed: 31513889
Microb Cell Fact. 2021 Dec 23;20(1):229
pubmed: 34949197
Microb Cell Fact. 2017 May 22;16(1):89
pubmed: 28532497
J Bacteriol. 2009 Aug;191(15):4996-5009
pubmed: 19482928
J Agric Food Chem. 2017 Jan 25;65(3):626-631
pubmed: 28035826
J Agric Food Chem. 2018 Nov 14;66(45):12141-12150
pubmed: 30362748
Nat Chem Biol. 2022 May;18(5):520-529
pubmed: 35484257
Metab Eng. 2014 Jul;24:181-91
pubmed: 24831710
Microb Cell Fact. 2015 Oct 14;14:163
pubmed: 26463172
Synth Syst Biotechnol. 2021 Dec 06;7(1):453-459
pubmed: 34938904
Biotechnol Bioeng. 2017 Sep;114(9):1928-1936
pubmed: 28498544
Sci Rep. 2016 Feb 22;6:21928
pubmed: 26898409
Microb Cell Fact. 2020 Jan 28;19(1):13
pubmed: 31992309
World J Microbiol Biotechnol. 2022 Aug 17;38(11):191
pubmed: 35974205
Microb Cell Fact. 2022 Oct 20;21(1):218
pubmed: 36266684
Genes (Basel). 2019 Jul 06;10(7):
pubmed: 31284586
Appl Environ Microbiol. 2018 Jul 2;84(14):
pubmed: 29728384
J Biotechnol. 2020 Nov 10;323:341-346
pubmed: 32976867
ChemSusChem. 2022 May 6;15(9):e202102399
pubmed: 35089653
Arch Biochem Biophys. 2010 Aug 1;500(1):74-81
pubmed: 20434429
Molecules. 2019 Oct 25;24(21):
pubmed: 31731395
Microb Cell Fact. 2021 Aug 16;20(1):158
pubmed: 34399754
Microb Cell Fact. 2017 Oct 30;16(1):182
pubmed: 29084539
Front Microbiol. 2018 May 15;9:958
pubmed: 29867863
Enzyme Microb Technol. 2020 Feb;133:109455
pubmed: 31874696