دورية أكاديمية
Backbone cyclization of Salmonella typhimurium diaminopropionate ammonia-lyase to enhance the activity and stability.
| العنوان: | Backbone cyclization of Salmonella typhimurium diaminopropionate ammonia-lyase to enhance the activity and stability. |
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| المؤلفون: | He X; College of Biology and Pharmaceutical Engineering, West Anhui University, Lu'an, 237012, PR China., Lin T; School of Life Science, Anhui Agricultural University, Hefei, Anhui, 230036, PR China., Xie Y; School of Life Science, Anhui Agricultural University, Hefei, Anhui, 230036, PR China., Li J; School of Life Science, Anhui Agricultural University, Hefei, Anhui, 230036, PR China., Ge Y; School of Life Science, Anhui Agricultural University, Hefei, Anhui, 230036, PR China., Zhang S; School of Life Science, Anhui Agricultural University, Hefei, Anhui, 230036, PR China., Fan J; School of Life Science, Anhui Agricultural University, Hefei, Anhui, 230036, PR China. Electronic address: fanjun@ahau.edu.cn. |
| المصدر: | Protein expression and purification [Protein Expr Purif] 2024 Jun; Vol. 218, pp. 106447. Date of Electronic Publication: 2024 Feb 16. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Academic Press Country of Publication: United States NLM ID: 9101496 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1096-0279 (Electronic) Linking ISSN: 10465928 NLM ISO Abbreviation: Protein Expr Purif Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Orlando, FL : Academic Press Original Publication: San Diego : Academic Press, c1990- |
| مواضيع طبية MeSH: | Salmonella typhimurium* , Neurotoxins* , Ammonia-Lyases*, Humans ; Cyclization ; Escherichia coli/genetics ; Serine |
| مستخلص: | Diaminopropionate ammonia-lyase transforms D and L isomers of 2,3-diaminopropionate to pyruvate and ammonia. It catalyzes D- and l-serine less effectively. L-2,3-diaminopropionate is a precursor in the biosynthesis of oxalyl diaminopropionate as a neurotoxin in certain legume species. In this work, we cyclized the diaminopropionate ammonia-lyase from Salmonella typhimurium in vitro using the redox-responsive split intein, and identified that backbone cyclization afforded the enzyme with the improved activity, thermal stability and resistance to the exopeptidase proteolysis, different from effects of the incorporated sequence recognized by tobacco vein mottling virus protease at C-terminus. Using analyses of three fluorescent dyes including 8-anilino-1-naphthalenesulfonic acid, N-phenyl-1-naphthylamine, and thioflavin T, the same amounts of the cyclic protein displayed less fluorescence than those of the linear protein upon the heat treatment. The cyclic enzyme displayed the enhanced activity in Escherichia coli cells using the designed novel reporter. In this system, d-serine was added to the culture and transported into the cytoplasm. It was transformed by pre-overexpression of the diaminopropionate ammonia-lyase, and untransformed d-serine was oxidized by the coproduced human d-amino acid oxidase to generate hydrogen peroxide. This oxidant is monitored by the HyPer indicator. The current results presented that the cyclized enzyme could be applied as a better candidate to block the neurotoxin biosynthesis in certain plant species. (Copyright © 2024 Elsevier Inc. All rights reserved.) |
| فهرسة مساهمة: | Keywords: Activity; Backbone cyclization; Diaminopropionate ammonialyase; Fluorescent dyes; In vivo activity assay; Stability |
| المشرفين على المادة: | EC 4.3.1.- (diaminopropionate ammonia-lyase) 0 (Neurotoxins) 452VLY9402 (Serine) EC 4.3.1.- (Ammonia-Lyases) |
| تواريخ الأحداث: | Date Created: 20240218 Date Completed: 20240320 Latest Revision: 20240320 |
| رمز التحديث: | 20240320 |
| DOI: | 10.1016/j.pep.2024.106447 |
| PMID: | 38369031 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1096-0279 |
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| DOI: | 10.1016/j.pep.2024.106447 |