PMID- 36746911
OWN - NLM
STAT- MEDLINE
DCOM- 20230329
LR  - 20230412
IS  - 1738-8872 (Electronic)
IS  - 1017-7825 (Print)
IS  - 1017-7825 (Linking)
VI  - 33
IP  - 3
DP  - 2023 Mar 28
TI  - Improvement of Bacilysin Production in Bacillus subtilis by CRISPR/Cas9-Mediated 
      Editing of the 5'-Untranslated Region of the bac Operon.
PG  - 410-418
LID - 10.4014/jmb.2209.09035 [doi]
AB  - Bacilysin is a dipeptide antibiotic composed of L-alanine and L-anticapsin 
      produced by certain strains of Bacillus subtilis. Bacilysin is gaining increasing 
      attention in industrial agriculture and pharmaceutical industries due to its 
      potent antagonistic effects on various bacterial, fungal, and algal pathogens. 
      However, its use in industrial applications is hindered by its low production in 
      the native producer. The biosynthesis of bacilysin is mainly based on the 
      bacABCDEF operon. Examination of the sequence surrounding the upstream of the bac 
      operon did not reveal a clear, strong ribosome binding site (RBS). Therefore, in 
      this study, we aimed to investigate the impact of RBS as a potential route to 
      improve bacilysin production. For this, the 5' untranslated region (5'UTR) of the 
      bac operon was edited using the CRISPR/Cas9 approach by introducing a strong 
      ribosome binding sequence carrying the canonical Shine-Dalgarno sequence 
      (TAAGGAGG) with an 8 nt spacing from the AUG start codon. Strong RBS substitution 
      resulted in a 2.87-fold increase in bacilysin production without affecting 
      growth. Strong RBS substitution also improved the mRNA stability of the bac 
      operon. All these data revealed that extensive RBS engineering is a promising key 
      option for enhancing bacilysin production in its native producers.
FAU - Abdulmalek, Hadeel Waleed
AU  - Abdulmalek HW
AD  - Dr. Orhan Ocalgiray Molecular Biology, Biotechnology and Genetics Research Center 
      (ITU-MOBGAM), Istanbul Technical University, Maslak, Istanbul 34469, Turkey.
AD  - Biotechnology Department, Collage of Science, University of Baghdad, Baghdad, 
      Iraq.
FAU - Yazgan-Karatas, Ayten
AU  - Yazgan-Karatas A
AD  - Dr. Orhan Ocalgiray Molecular Biology, Biotechnology and Genetics Research Center 
      (ITU-MOBGAM), Istanbul Technical University, Maslak, Istanbul 34469, Turkey.
AD  - Faculty of Science and Letters, Department of Molecular Biology and Genetics, 
      Istanbul Technical University, Maslak, Istanbul 34469, Turkey.
LA  - eng
PT  - Journal Article
DEP - 20221213
PL  - Korea (South)
TA  - J Microbiol Biotechnol
JT  - Journal of microbiology and biotechnology
JID - 9431852
RN  - 0 (5' Untranslated Regions)
RN  - 90R5Q4BW17 (bacilysin)
RN  - 0 (Dipeptides)
SB  - IM
MH  - *Bacillus subtilis/metabolism
MH  - 5' Untranslated Regions
MH  - *CRISPR-Cas Systems
MH  - Operon
MH  - Dipeptides/genetics
PMC - PMC10084748
OTO - NOTNLM
OT  - Bacillus subtilis
OT  - CRISPR/Cas9
OT  - bacABCDEF
OT  - bacilysin
OT  - ribosome binding site
COIS- Conflict of Interest The authors have no financial conflicts of interest to 
      declare.
EDAT- 2023/02/07 06:00
MHDA- 2023/03/29 06:05
PMCR- 2022/12/13
CRDT- 2023/02/06 23:22
PHST- 2022/09/22 00:00 [received]
PHST- 2022/12/05 00:00 [revised]
PHST- 2022/12/07 00:00 [accepted]
PHST- 2023/03/29 06:05 [medline]
PHST- 2023/02/07 06:00 [pubmed]
PHST- 2023/02/06 23:22 [entrez]
PHST- 2022/12/13 00:00 [pmc-release]
AID - jmb.2209.09035 [pii]
AID - jmb-33-3-410 [pii]
AID - 10.4014/jmb.2209.09035 [doi]
PST - ppublish
SO  - J Microbiol Biotechnol. 2023 Mar 28;33(3):410-418. doi: 10.4014/jmb.2209.09035. 
      Epub 2022 Dec 13.