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Modular metabolic engineering of Bacillus amyloliquefaciens for high-level production of green biosurfactant iturin A
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Modular metabolic engineering of Bacillus amyloliquefaciens for high-level production of green biosurfactant iturin A
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Journal Article
Modular metabolic engineering of Bacillus amyloliquefaciens for high-level production of green biosurfactant iturin A
2024
Overview
As a kind of biosurfactants, iturin A has attracted people’s wide attentions due to their features of biodegradability, environmentally friendly, etc.; however, high production cost limited its extensive application, and the aim of this research wants to improve iturin A production in
Bacillus amyloliquefaciens
. Firstly, dual promoter was applied to strengthen iturin A synthetase expression, and its yield was increased to 1.25 g/L. Subsequently, original 5′-UTRs of downstream genes (
ituA
,
ituB
, and
ituC
) in iturin A synthetase cluster were optimized, which significantly increased mRNA secondary stability, and iturin A yield produced by resultant strain HZ-T3 reached 2.32 g/L. Secondly, synthetic pathway of α-glucosidase inhibitor 1-deoxynojirimycin was blocked to improve substrate corn starch utilization, and iturin A yield was increased by 34.91% to 3.13 g/L. Thirdly, efficient precursor (fatty acids, Ser, and Pro) supplies were proven as the critical role in iturin A synthesis, and 5.52 g/L iturin A was attained by resultant strain, through overexpressing
yngH
,
serC
, and introducing
ocD
. Meanwhile, genes responsible for poly-γ-glutamic acid, extracellular polysaccharide, and surfactin syntheses were deleted, which led to a 30.98% increase of iturin A yield. Finally, lipopeptide transporters were screened, and iturin A yield was increased by 17.98% in SwrC overexpression strain, reached 8.53 g/L, which is the highest yield of iturin A ever reported. This study laid a foundation for industrial production and application development of iturin A, and provided the guidance of metabolic engineering breeding for efficient production of other metabolites synthesized by non-ribosomal peptide synthetase.
Key points
• Optimizing 5′-UTR is an effective tactics to regulate synthetase cluster expression.
• Blocking 1-DNJ synthesis benefited corn starch utilization and iturin A production.
• The iturin A yield attained in this work was the highest yield reported so far.
Publisher
Springer Berlin Heidelberg,Springer,Springer Nature B.V
Subject
/ Applied Genetics and Molecular Biotechnology
/ Bacillus amyloliquefaciens - genetics
/ Bacillus amyloliquefaciens - metabolism
/ Biomedical and Life Sciences
/ Clusters
/ Corn
/ Genes
/ iturin
/ Iturin A
/ Metabolic Engineering - methods
/ Microbial Genetics and Genomics
/ Peptides, Cyclic - biosynthesis
/ Peptides, Cyclic - metabolism
/ Starch
