Tuning the Binding Affinity of Heme-Responsive Biosensor for Precise and Dynamic Pathway Regulation.

Bioengineering Biotechnology Metabolic Engineering

Journal

iScience
ISSN: 2589-0042
Titre abrégé: iScience
Pays: United States
ID NLM: 101724038

Informations de publication

Date de publication:
22 May 2020
Historique:
received: 19 10 2019
revised: 21 02 2020
accepted: 13 04 2020
pubmed: 7 5 2020
medline: 7 5 2020
entrez: 7 5 2020
Statut: ppublish

Résumé

Current challenge for dynamic pathway control in metabolic engineering is enabling the components of the artificial regulatory system to be tunable. Here, we designed and built a heme-responsive regulatory system containing a heme biosensor HrtR and CRISPRi to regulate chemicals production while maintaining the intracellular heme homeostasis. A series of engineered biosensors with varied sensitivity and threshold were obtained by semi-rational design with site saturated mutation of HrtR. The modified metabolite-binding affinity of HrtR was confirmed by heme titration and molecular dynamic simulation. Dynamic regulation pattern of the system was validated by the fluctuation of gene expression and intracellular heme concentration. The efficiency of this regulatory system was proved by improving the 5-aminolevulinic acid (ALA) production to 5.35g/L, the highest yield in batch fermentation of Escherichia coli. This system was also successfully used in improving porphobilinogen (PBG) and porphyrins biosynthesis and can be applied in many other biological processes.

Identifiants

DOI: 10.1016/j.isci.2020.101067 PMID: 32371371 PMC: PMC7200772
pubmed: 32371371
pii: S2589-0042(20)30252-2
doi: 10.1016/j.isci.2020.101067
pmc: PMC7200772
pii:
doi:

Types de publication

Journal Article

Langues

eng

Pagination

101067

Informations de copyright

Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of Interests The authors declare no competing interests.

Références

ACS Synth Biol. 2017 Oct 20;6(10):1851-1859
pubmed: 28763198
Biotechnol J. 2013 Jan;8(1):46-58
pubmed: 22890821
Metab Eng. 2015 May;29:217-226
pubmed: 25863265
Dev Neurosci. 1982 Mar-Jun;5(2-3):125-9
pubmed: 6127199
Appl Microbiol Biotechnol. 2016 Jan;100(1):79-90
pubmed: 26521244
Pharmacol Ther. 2006 Aug;111(2):327-45
pubmed: 16513178
Nat Biotechnol. 2012 Mar 25;30(4):354-9
pubmed: 22446695
J Agric Food Chem. 2019 Feb 6;67(5):1478-1483
pubmed: 30644739
Biotechnol Adv. 1995;13(2):247-61
pubmed: 14537822
Metab Eng. 2015 Sep;31:35-43
pubmed: 26142692
Nat Biotechnol. 2013 Feb;31(2):170-4
pubmed: 23334451
Adv Microb Physiol. 2013;62:1-43
pubmed: 23481334
Mol Microbiol. 2007 May;64(4):1090-104
pubmed: 17501930
Sci Adv. 2018 Jan 26;4(1):eaaq1407
pubmed: 29387799
J Biol Chem. 2012 Aug 31;287(36):30755-68
pubmed: 22798069
Nat Biotechnol. 2009 Aug;27(8):760-5
pubmed: 19633654
J Biol Chem. 2017 Oct 13;292(41):16942-16954
pubmed: 28830930
Protein Sci. 2010 Jun;19(6):1137-61
pubmed: 20506125
Structure. 1994 May 15;2(5):415-22
pubmed: 8081757
J Biol Chem. 2010 May 21;285(21):16023-31
pubmed: 20233710
Proc Natl Acad Sci U S A. 2014 Aug 5;111(31):11299-304
pubmed: 25049420
Nat Genet. 2002 May;31(1):64-8
pubmed: 11967538
ACS Synth Biol. 2015 Feb 20;4(2):132-40
pubmed: 24377365
J Bacteriol. 2011 Mar;193(5):1212-21
pubmed: 21217007
Biotechnol J. 2018 Sep;13(9):e1800069
pubmed: 29635744
J Biol Chem. 2012 Feb 10;287(7):4752-8
pubmed: 22084241
Nat Biotechnol. 2014 Dec;32(12):1276-81
pubmed: 25402616
Cell. 2013 Feb 28;152(5):1173-83
pubmed: 23452860
Nat Methods. 2016 Feb;13(2):177-83
pubmed: 26689263
Nat Methods. 2014 May;11(5):508-20
pubmed: 24781324
Nat Commun. 2018 Nov 21;9(1):4917
pubmed: 30464241
J Mol Biol. 2016 Aug 28;428(17):3408-28
pubmed: 27019298
Proc Natl Acad Sci U S A. 1999 Nov 9;96(23):13056-61
pubmed: 10557272
J Biol Chem. 1998 Aug 21;273(34):21669-74
pubmed: 9705301
Metab Eng. 2011 Sep;13(5):492-8
pubmed: 21620993
Free Radic Biol Med. 2000 Jan 15;28(2):289-309
pubmed: 11281297
Science. 1994 Dec 9;266(5191):1713-7
pubmed: 7527589
Sensors (Basel). 2018 Aug 27;18(9):
pubmed: 30150540
Cell. 1981 Oct;26(1 Pt 1):117-22
pubmed: 6799206
Nat Biotechnol. 2003 Jul;21(7):796-802
pubmed: 12778056
Proc Natl Acad Sci U S A. 1997 Oct 14;94(21):11216-20
pubmed: 9326589
Biotechnol Bioeng. 2018 Sep;115(9):2292-2304
pubmed: 29733444

Auteurs

Jian Zhang (J)

State Key Laboratory of Microbial Technology, National Glycoengineering Research Center, Shandong University, Qingdao 266237, P. R. China.

Zhiguo Wang (Z)

Institute of Ageing Research, School of Medicine, Hangzhou Normal University, Hangzhou 311121, P. R. China.

Tianyuan Su (T)

State Key Laboratory of Microbial Technology, National Glycoengineering Research Center, Shandong University, Qingdao 266237, P. R. China.

Huanhuan Sun (H)

State Key Laboratory of Microbial Technology, National Glycoengineering Research Center, Shandong University, Qingdao 266237, P. R. China.

Yuan Zhu (Y)

State Key Laboratory of Microbial Technology, National Glycoengineering Research Center, Shandong University, Qingdao 266237, P. R. China.

Qingsheng Qi (Q)

State Key Laboratory of Microbial Technology, National Glycoengineering Research Center, Shandong University, Qingdao 266237, P. R. China; CAS Key Lab of Biobased Materials, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao 266101, P. R. China. Electronic address: qiqingsheng@sdu.edu.cn.

Qian Wang (Q)

State Key Laboratory of Microbial Technology, National Glycoengineering Research Center, Shandong University, Qingdao 266237, P. R. China. Electronic address: qiqi20011983@gmail.com.

Classifications MeSH