Modular metabolic engineering of Bacillus amyloliquefaciens for high-level production of green biosurfactant iturin A
© 2024. The Author(s)..
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.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2024 |
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| Erschienen: |
2024 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:108 |
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| Enthalten in: |
Applied microbiology and biotechnology - 108(2024), 1 vom: 27. Apr., Seite 311 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
She, Menglin [VerfasserIn] |
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| Links: |
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| Themen: |
52229-90-0 |
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| Anmerkungen: |
Date Completed 27.04.2024 Date Revised 30.04.2024 published: Electronic Citation Status MEDLINE |
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| doi: |
10.1007/s00253-024-13083-9 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM371655595 |
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| 245 | 1 | 0 | |a Modular metabolic engineering of Bacillus amyloliquefaciens for high-level production of green biosurfactant iturin A |
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| 520 | |a © 2024. The Author(s). | ||
| 520 | |a 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 | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Bacillus amyloliquefaciens | |
| 650 | 4 | |a Iturin A | |
| 650 | 4 | |a Metabolic engineering | |
| 650 | 4 | |a Precursor supply | |
| 650 | 4 | |a Synthetase gene cluster | |
| 650 | 7 | |a iturin A |2 NLM | |
| 650 | 7 | |a 52229-90-0 |2 NLM | |
| 650 | 7 | |a Surface-Active Agents |2 NLM | |
| 650 | 7 | |a Peptides, Cyclic |2 NLM | |
| 650 | 7 | |a Ligases |2 NLM | |
| 650 | 7 | |a EC 6.- |2 NLM | |
| 700 | 1 | |a Zhou, Huijuan |e verfasserin |4 aut | |
| 700 | 1 | |a Dong, Wanrong |e verfasserin |4 aut | |
| 700 | 1 | |a Xu, Yuxiang |e verfasserin |4 aut | |
| 700 | 1 | |a Gao, Lin |e verfasserin |4 aut | |
| 700 | 1 | |a Gao, Jiaming |e verfasserin |4 aut | |
| 700 | 1 | |a Yang, Yong |e verfasserin |4 aut | |
| 700 | 1 | |a Yang, Zhifan |e verfasserin |4 aut | |
| 700 | 1 | |a Cai, Dongbo |e verfasserin |4 aut | |
| 700 | 1 | |a Chen, Shouwen |e verfasserin |4 aut | |
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