Elucidating the regulatory role of the Shine-Dalgarno sequence in structural gene expression of the malolactic enzyme operon using a GFP/mCherry model.
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| Title: | Elucidating the regulatory role of the Shine-Dalgarno sequence in structural gene expression of the malolactic enzyme operon using a GFP/mCherry model. |
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| Authors: | Liu, Zixian1,2 (AUTHOR), Ji, Mingyue1 (AUTHOR), Zhao, Fangkun1 (AUTHOR), Wang, Dandan1 (AUTHOR), Li, Yue1 (AUTHOR), Yi, Yongfeng1 (AUTHOR), Wang, Min3 (AUTHOR) wm1977@126.com, Tan, Jianxin1,2 (AUTHOR) jianxintan@sina.com |
| Source: | World Journal of Microbiology & Biotechnology. Jun2026, Vol. 42 Issue 6, p1-15. 15p. |
| Subjects: | Genetic regulation, Operons, Ribosomes, Fluorescent proteins, Synthetic biology, Bacteria, RNA regulation |
| Abstract: | Malolactic fermentation (MLF) in wine relies on Oenococcus oeni, in which the mleA and mleP genes are translationally coupled. To explore the post-transcriptional regulation involved, we constructed parallel GFP/mCherry reporter systems that mimic both the native overlapping (translationally coupled) structure and an engineered non-overlapping (non-coupled) structure. Through site-directed SD2 mutagenesis coupled with computational prediction of 16 S rRNA hybridization energy (ΔG) and minimum free energy (MFE) of mRNA secondary structure, we quantified the effect of intergenic sequence features on translation efficiency. In the translational coupling system of Escherichia coli, expression ratios (GFP/MCH) varied from 3.58 to 8.58 across variants, while in the non-coupled system, they ranged from 1.54 to 3.09. In non-coupled constructs, downstream gene expression increased under independent SD2 control, consistent with its conserved function. However, in the coupling system, the strongest SD2 (GGAGG) markedly suppressed it, revealing a nonlinear, architecture-specific regulatory logic. The different variants were also expressed in the translational coupling system of Lactococcus lactis, and the expression ratio patterns of the upstream and downstream genes across the variants were consistent with those observed in E. coli. This indicated that SD2 had the same regulatory effect on coupling gene expression ratios in both L. lactis and E. coli. Our findings demonstrate that translational coupling establishes precise protein expression regulation through the integrated effects of upstream translation and local mRNA structure. This work elucidates a key post-transcriptional tuning layer in prokaryotic operons, providing a predictive framework for both the rational engineering of O. oeni to optimize MLF and the design of proportionally controlled multi-gene systems in synthetic biology. [ABSTRACT FROM AUTHOR] |
| Copyright of World Journal of Microbiology & Biotechnology is the property of Springer Nature and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.) | |
| Database: | Engineering Source |
| FullText | Text: Availability: 0 |
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| Header | DbId: egs DbLabel: Engineering Source An: 194750110 AccessLevel: 6 PubType: Academic Journal PubTypeId: academicJournal PreciseRelevancyScore: 0 |
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| Items | – Name: Title Label: Title Group: Ti Data: Elucidating the regulatory role of the Shine-Dalgarno sequence in structural gene expression of the malolactic enzyme operon using a GFP/mCherry model. – Name: Author Label: Authors Group: Au Data: <searchLink fieldCode="AR" term="%22Liu%2C+Zixian%22">Liu, Zixian</searchLink><relatesTo>1,2</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Ji%2C+Mingyue%22">Ji, Mingyue</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Zhao%2C+Fangkun%22">Zhao, Fangkun</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Wang%2C+Dandan%22">Wang, Dandan</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Li%2C+Yue%22">Li, Yue</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Yi%2C+Yongfeng%22">Yi, Yongfeng</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Wang%2C+Min%22">Wang, Min</searchLink><relatesTo>3</relatesTo> (AUTHOR)<i> wm1977@126.com</i><br /><searchLink fieldCode="AR" term="%22Tan%2C+Jianxin%22">Tan, Jianxin</searchLink><relatesTo>1,2</relatesTo> (AUTHOR)<i> jianxintan@sina.com</i> – Name: TitleSource Label: Source Group: Src Data: <searchLink fieldCode="JN" term="%22World+Journal+of+Microbiology+%26+Biotechnology%22">World Journal of Microbiology & Biotechnology</searchLink>. Jun2026, Vol. 42 Issue 6, p1-15. 15p. – Name: Subject Label: Subjects Group: Su Data: <searchLink fieldCode="DE" term="%22Genetic+regulation%22">Genetic regulation</searchLink><br /><searchLink fieldCode="DE" term="%22Operons%22">Operons</searchLink><br /><searchLink fieldCode="DE" term="%22Ribosomes%22">Ribosomes</searchLink><br /><searchLink fieldCode="DE" term="%22Fluorescent+proteins%22">Fluorescent proteins</searchLink><br /><searchLink fieldCode="DE" term="%22Synthetic+biology%22">Synthetic biology</searchLink><br /><searchLink fieldCode="DE" term="%22Bacteria%22">Bacteria</searchLink><br /><searchLink fieldCode="DE" term="%22RNA+regulation%22">RNA regulation</searchLink> – Name: Abstract Label: Abstract Group: Ab Data: Malolactic fermentation (MLF) in wine relies on Oenococcus oeni, in which the mleA and mleP genes are translationally coupled. To explore the post-transcriptional regulation involved, we constructed parallel GFP/mCherry reporter systems that mimic both the native overlapping (translationally coupled) structure and an engineered non-overlapping (non-coupled) structure. Through site-directed SD2 mutagenesis coupled with computational prediction of 16 S rRNA hybridization energy (ΔG) and minimum free energy (MFE) of mRNA secondary structure, we quantified the effect of intergenic sequence features on translation efficiency. In the translational coupling system of Escherichia coli, expression ratios (GFP/MCH) varied from 3.58 to 8.58 across variants, while in the non-coupled system, they ranged from 1.54 to 3.09. In non-coupled constructs, downstream gene expression increased under independent SD2 control, consistent with its conserved function. However, in the coupling system, the strongest SD2 (GGAGG) markedly suppressed it, revealing a nonlinear, architecture-specific regulatory logic. The different variants were also expressed in the translational coupling system of Lactococcus lactis, and the expression ratio patterns of the upstream and downstream genes across the variants were consistent with those observed in E. coli. This indicated that SD2 had the same regulatory effect on coupling gene expression ratios in both L. lactis and E. coli. Our findings demonstrate that translational coupling establishes precise protein expression regulation through the integrated effects of upstream translation and local mRNA structure. This work elucidates a key post-transcriptional tuning layer in prokaryotic operons, providing a predictive framework for both the rational engineering of O. oeni to optimize MLF and the design of proportionally controlled multi-gene systems in synthetic biology. [ABSTRACT FROM AUTHOR] – Name: AbstractSuppliedCopyright Label: Group: Ab Data: <i>Copyright of World Journal of Microbiology & Biotechnology is the property of Springer Nature and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.) |
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| RecordInfo | BibRecord: BibEntity: Identifiers: – Type: doi Value: 10.1007/s11274-026-04999-7 Languages: – Code: eng Text: English PhysicalDescription: Pagination: PageCount: 15 StartPage: 1 Subjects: – SubjectFull: Genetic regulation Type: general – SubjectFull: Operons Type: general – SubjectFull: Ribosomes Type: general – SubjectFull: Fluorescent proteins Type: general – SubjectFull: Synthetic biology Type: general – SubjectFull: Bacteria Type: general – SubjectFull: RNA regulation Type: general Titles: – TitleFull: Elucidating the regulatory role of the Shine-Dalgarno sequence in structural gene expression of the malolactic enzyme operon using a GFP/mCherry model. Type: main BibRelationships: HasContributorRelationships: – PersonEntity: Name: NameFull: Liu, Zixian – PersonEntity: Name: NameFull: Ji, Mingyue – PersonEntity: Name: NameFull: Zhao, Fangkun – PersonEntity: Name: NameFull: Wang, Dandan – PersonEntity: Name: NameFull: Li, Yue – PersonEntity: Name: NameFull: Yi, Yongfeng – PersonEntity: Name: NameFull: Wang, Min – PersonEntity: Name: NameFull: Tan, Jianxin IsPartOfRelationships: – BibEntity: Dates: – D: 01 M: 06 Text: Jun2026 Type: published Y: 2026 Identifiers: – Type: issn-print Value: 09593993 Numbering: – Type: volume Value: 42 – Type: issue Value: 6 Titles: – TitleFull: World Journal of Microbiology & Biotechnology Type: main |
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