PMID- 34850512
OWN - NLM
STAT- MEDLINE
DCOM- 20220308
LR  - 20220308
IS  - 1439-7633 (Electronic)
IS  - 1439-4227 (Linking)
VI  - 23
IP  - 3
DP  - 2022 Feb 4
TI  - Tailoring Enzyme Stringency Masks the Multispecificity of a Lyngbyatoxin 
      (Indolactam Alkaloid) Nonribosomal Peptide Synthetase.
PG  - e202100574
LID - 10.1002/cbic.202100574 [doi]
AB  - Indolactam alkaloids are activators of protein kinase C (PKC) and are of 
      pharmacological interest for the treatment of pathologies involving PKC 
      dysregulation. The marine cyanobacterial nonribosomal peptide synthetase (NRPS) 
      pathway for lyngbyatoxin biosynthesis, which we previously expressed in E. coli, 
      was studied for its amenability towards the biosynthesis of indolactam variants. 
      Modification of culture conditions for our E. coli heterologous expression host 
      and analysis of pathway products suggested the native lyngbyatoxin pathway NRPS 
      does possess a degree of relaxed specificity. Site-directed mutagenesis of two 
      positions within the adenylation domain (A-domain) substrate-binding pocket was 
      performed, resulting in an alteration of substrate preference between valine, 
      isoleucine, and leucine. We observed relative congruence of in vitro substrate 
      activation by the LtxA NRPS to in vivo product formation. While there was a 
      preference for isoleucine over leucine, the substitution of alternative tailoring 
      domains may unveil the true in vivo effects of the mutations introduced herein.
CI  - (c) 2021 Wiley-VCH GmbH.
FAU - Soeriyadi, Angela H
AU  - Soeriyadi AH
AUID- ORCID: 0000-0003-0064-0051
AD  - School of Biotechnology and Biomolecular Sciences, University of New South Wales, 
      Sydney, 2052, Australia.
FAU - Ongley, Sarah E
AU  - Ongley SE
AUID- ORCID: 0000-0002-1203-7682
AD  - School of Biotechnology and Biomolecular Sciences, University of New South Wales, 
      Sydney, 2052, Australia.
AD  - School of Environmental and Life Sciences, University of Newcastle, Callaghan, 
      2308, Australia.
FAU - Kehr, Jan-Christoph
AU  - Kehr JC
AD  - Department of Microbiology, Institute for Biochemistry and Biology, University of 
      Potsdam, Potsdam-Golm, Germany.
FAU - Pickford, Russel
AU  - Pickford R
AUID- ORCID: 0000-0001-8982-7618
AD  - Bioanalytical Mass Spectrometry Facility, University of New South Wales, Sydney, 
      2052, Australia.
FAU - Dittmann, Elke
AU  - Dittmann E
AUID- ORCID: 0000-0002-7549-7918
AD  - Department of Microbiology, Institute for Biochemistry and Biology, University of 
      Potsdam, Potsdam-Golm, Germany.
FAU - Neilan, Brett A
AU  - Neilan BA
AUID- ORCID: 0000-0001-6113-772X
AD  - School of Biotechnology and Biomolecular Sciences, University of New South Wales, 
      Sydney, 2052, Australia.
AD  - School of Environmental and Life Sciences, University of Newcastle, Callaghan, 
      2308, Australia.
LA  - eng
GR  - Technical University Berlin/
GR  - LP140100642/Australian Research Council/
GR  - Australian Government RTP Scholarship/
GR  - Adrian Lee Travel Scholarship/
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20211210
PL  - Germany
TA  - Chembiochem
JT  - Chembiochem : a European journal of chemical biology
JID - 100937360
RN  - 0 (Lyngbya Toxins)
RN  - EC 6.3.2.- (Peptide Synthases)
RN  - EC 6.3.2.- (non-ribosomal peptide synthase)
SB  - IM
MH  - Lyngbya Toxins/*biosynthesis/chemistry
MH  - Molecular Structure
MH  - Mutagenesis, Site-Directed
MH  - Peptide Synthases/genetics/*metabolism
OTO - NOTNLM
OT  - A domain
OT  - MbtH
OT  - indolactams
OT  - natural products
OT  - teleocidin
EDAT- 2021/12/02 06:00
MHDA- 2022/03/09 06:00
CRDT- 2021/12/01 07:46
PHST- 2021/11/30 00:00 [revised]
PHST- 2021/10/24 00:00 [received]
PHST- 2021/12/02 06:00 [pubmed]
PHST- 2022/03/09 06:00 [medline]
PHST- 2021/12/01 07:46 [entrez]
AID - 10.1002/cbic.202100574 [doi]
PST - ppublish
SO  - Chembiochem. 2022 Feb 4;23(3):e202100574. doi: 10.1002/cbic.202100574. Epub 2021 
      Dec 10.