دورية أكاديمية
Genetically engineered Lactococcus lactis strain constitutively expresses GABA-producing genes and produces high levels of GABA.
| العنوان: | Genetically engineered Lactococcus lactis strain constitutively expresses GABA-producing genes and produces high levels of GABA. |
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| المؤلفون: | Monteiro MP; Department of Microbiology and Immunology, Emory University School of Medicine, 1510 Clifton Road, RRC 3070, Atlanta, GA 30322, United States., Kohl HM; Department of Biology, Eastern Washington University, 1175 Washington St. Science 226, Cheney, WA 99004, United States., Roullet JB; Department of Pharmacotherapy, Washington State University, 205 East Spokane Falls Boulevard, Spokane, WA 99202, United States., Gibson KM; Department of Pharmacotherapy, Washington State University, 205 East Spokane Falls Boulevard, Spokane, WA 99202, United States., Ochoa-Repáraz J; Department of Biological Sciences, Boise State University, 1910 University Drive, Boise, ID 83725, United States., Castillo AR; Department of Biology, Eastern Washington University, 1175 Washington St. Science 226, Cheney, WA 99004, United States. |
| المصدر: | Letters in applied microbiology [Lett Appl Microbiol] 2024 Jun 03; Vol. 77 (6). |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Oxford University Press Country of Publication: England NLM ID: 8510094 Publication Model: Print Cited Medium: Internet ISSN: 1472-765X (Electronic) Linking ISSN: 02668254 NLM ISO Abbreviation: Lett Appl Microbiol Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: 2023- : Oxford : Oxford University Press Original Publication: Oxford, UK : Published for the Society for Applied Bacteriology by Blackwell Scientific Publications, [c1985- |
| مواضيع طبية MeSH: | Lactococcus lactis*/genetics , Lactococcus lactis*/metabolism , gamma-Aminobutyric Acid*/metabolism , gamma-Aminobutyric Acid*/biosynthesis , Promoter Regions, Genetic* , Glutamate Decarboxylase*/genetics , Glutamate Decarboxylase*/metabolism, Genetic Engineering ; Plasmids/genetics ; Glutamic Acid/metabolism ; Metabolic Engineering ; Bacterial Proteins/genetics ; Bacterial Proteins/metabolism |
| مستخلص: | γ-Aminobutyric acid (GABA) is an inhibitory neurotransmitter of the central nervous system that impacts physical and mental health. Low GABA levels have been documented in several diseases, including multiple sclerosis and depression, and studies suggest that GABA could improve disease outcomes in those conditions. Probiotic bacteria naturally produce GABA and have been engineered to enhance its synthesis. Strains engineered thus far use inducible expression systems that require the addition of exogenous molecules, which complicates their development as therapeutics. This study aimed to overcome this challenge by engineering Lactococcus lactis with a constitutive GABA synthesis gene cassette. GABA synthesizing and transport genes (gadB and gadC) were cloned onto plasmids downstream of constitutive L. lactis promoters [P2, P5, shortened P8 (P8s)] of different strengths and transformed into L. lactis. Fold increase in gadCB expression conferred by these promoters (P2, P5, and P8s) was 322, 422, and 627, respectively, compared to the unmodified strain (P = 0.0325, P8s). GABA synthesis in the highest gadCB expressing strain, L. lactis-P8s-glutamic acid decarboxylase (GAD), was dependent on media supplementation with glutamic acid and significantly higher than the unmodified strain (P < 0.0001, 125 mM, 200 mM glutamic acid). Lactococcus lactis-P8s-GAD is poised for therapeutic testing in animal models of low-GABA-associated disease. (© The Author(s) 2024. Published by Oxford University Press on behalf of Applied Microbiology International.) |
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| معلومات مُعتمدة: | P20GM103408 United States GM NIGMS NIH HHS; R15 NS107743 United States NS NINDS NIH HHS; R15NS107743-01A1 United States NH NIH HHS; C06 RR020533 United States RR NCRR NIH HHS; 0619793 National Science Foundation |
| فهرسة مساهمة: | Keywords: Lactococcus lactis; GABA; therapeutic |
| المشرفين على المادة: | 56-12-2 (gamma-Aminobutyric Acid) EC 4.1.1.15 (Glutamate Decarboxylase) 3KX376GY7L (Glutamic Acid) 0 (Bacterial Proteins) |
| تواريخ الأحداث: | Date Created: 20240530 Date Completed: 20240620 Latest Revision: 20240622 |
| رمز التحديث: | 20240622 |
| مُعرف محوري في PubMed: | PMC11187484 |
| DOI: | 10.1093/lambio/ovae051 |
| PMID: | 38816215 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1472-765X |
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| DOI: | 10.1093/lambio/ovae051 |