دورية أكاديمية
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Integrated strategies for efficient production of Streptomyces mobaraensis transglutaminase in Komagataella phaffii.

التفاصيل البيبلوغرافية
العنوان: Integrated strategies for efficient production of Streptomyces mobaraensis transglutaminase in Komagataella phaffii.
المؤلفون: Aqeel SM; Science Center for Future Foods, Jiangnan University, 1800 Lihu Avenue, Wuxi, Jiangsu 214122, China; School of Biotechnology and Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Avenue, Wuxi, Jiangsu 214122, China., Abdulqader AA; School of Biotechnology and Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Avenue, Wuxi, Jiangsu 214122, China., Du G; Science Center for Future Foods, Jiangnan University, 1800 Lihu Avenue, Wuxi, Jiangsu 214122, China; School of Biotechnology and Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Avenue, Wuxi, Jiangsu 214122, China; The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Avenue, Wuxi, Jiangsu 214122, China. Electronic address: gcdu@jiangnan.edu.cn., Liu S; Science Center for Future Foods, Jiangnan University, 1800 Lihu Avenue, Wuxi, Jiangsu 214122, China; School of Biotechnology and Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Avenue, Wuxi, Jiangsu 214122, China. Electronic address: liusong@jiangnan.edu.cn.
المصدر: International journal of biological macromolecules [Int J Biol Macromol] 2024 Jul; Vol. 273 (Pt 2), pp. 133113. Date of Electronic Publication: 2024 Jun 15.
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: Elsevier Country of Publication: Netherlands NLM ID: 7909578 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1879-0003 (Electronic) Linking ISSN: 01418130 NLM ISO Abbreviation: Int J Biol Macromol Subsets: MEDLINE
أسماء مطبوعة: Publication: Amsterdam : Elsevier
Original Publication: Guildford, Eng., IPC Science and Technology Press.
مواضيع طبية MeSH: Streptomyces*/genetics , Streptomyces*/enzymology , Transglutaminases*/genetics , Transglutaminases*/metabolism , Transglutaminases*/biosynthesis, Saccharomycetales/genetics ; Saccharomycetales/enzymology ; Saccharomycetales/metabolism ; Fermentation ; Recombinant Proteins/genetics ; Recombinant Proteins/metabolism ; Recombinant Proteins/biosynthesis ; Bacterial Proteins/genetics ; Bacterial Proteins/metabolism ; Bacterial Proteins/biosynthesis ; Kinetics ; Promoter Regions, Genetic
مستخلص: Transglutaminase (TGase) from Streptomyces mobaraensis commonly used to improve protein-based foods due to its unique enzymatic reactions, which imply considerable attention in its production. Recently, TGase exhibit broad market potential in non-food industries. However, achieving efficient synthesis of TGase remains a significant challenge. Herein, we achieved a substantial amount of a fully functional and kinetically stable TGase produced by Komagataella phaffii (Pichia pastoris) using multiple strategies including Geneticin (G418) screening, combinatorial mutations, promoter optimization, and co-expression. The active TGase expression reached a maximum of 10.1 U mL -1 in shake flask upon 96 h of induction, which was 3.8-fold of the wild type. Also, the engineered strain exhibited a 6.4-fold increase in half-life and a 2-fold increase in specific activity, reaching 172.67 min at 60 °C (t 1/2 (60 °C)) and 65.3 U mg -1 , respectively. Moreover, the high-cell density cultivation in 5-L fermenter was also applied to test the productivity at large scale. Following optimization at a fermenter, the secretory yield of TGase reached 47.96 U mL -1 in the culture supernatant. Given the complexity inherent in protein expression and secretion, our research is of great significance and offers a comprehensive guide for improving the production of a wide range of heterologous proteins.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024 Elsevier B.V. All rights reserved.)
فهرسة مساهمة: Keywords: Molecular chaperones; Promoter optimization; Site-directed mutation
المشرفين على المادة: EC 2.3.2.13 (Transglutaminases)
0 (Recombinant Proteins)
0 (Bacterial Proteins)
SCR Organism: Streptomyces mobaraensis; Komagataella phaffii
تواريخ الأحداث: Date Created: 20240617 Date Completed: 20240706 Latest Revision: 20240713
رمز التحديث: 20240713
DOI: 10.1016/j.ijbiomac.2024.133113
PMID: 38885870
قاعدة البيانات: MEDLINE
الوصف
تدمد:1879-0003
DOI:10.1016/j.ijbiomac.2024.133113