دورية أكاديمية
أعرض في EDS

Expression of Bacillus amyloliquefaciens transglutaminase in recombinant E. coli under the control of a bicistronic plasmid system in DO-stat fed-batch bioreactor cultivations.

التفاصيل البيبلوغرافية
العنوان: Expression of Bacillus amyloliquefaciens transglutaminase in recombinant E. coli under the control of a bicistronic plasmid system in DO-stat fed-batch bioreactor cultivations.
المؤلفون: Duarte LS; Biotechnology, Bioprocess, and Biocatalysis Group, Food Science and Technology Institute, Federal University of Rio Grande Do Sul, Av. Bento Gonçalves 9500, PO Box 15090, Porto Alegre, RS, ZC 91501-970, Brazil., Matte CR; Biotechnology, Bioprocess, and Biocatalysis Group, Food Science and Technology Institute, Federal University of Rio Grande Do Sul, Av. Bento Gonçalves 9500, PO Box 15090, Porto Alegre, RS, ZC 91501-970, Brazil., Dall Cortivo PR; Biotechnology, Bioprocess, and Biocatalysis Group, Food Science and Technology Institute, Federal University of Rio Grande Do Sul, Av. Bento Gonçalves 9500, PO Box 15090, Porto Alegre, RS, ZC 91501-970, Brazil., Nunes JES; Centro de Pesquisas Em Biologia Molecular E Funcional (CPBMF), Pontifícia Universidade Católica Do Rio Grande Do Sul (PUCRS), 92A TECNOPUC Building, 4592 Av. Bento Gonçalves, Porto Alegre, ZC 90650-001, Brazil., Barsé LQ; Centro de Pesquisas Em Biologia Molecular E Funcional (CPBMF), Pontifícia Universidade Católica Do Rio Grande Do Sul (PUCRS), 92A TECNOPUC Building, 4592 Av. Bento Gonçalves, Porto Alegre, ZC 90650-001, Brazil., Bizarro CV; Centro de Pesquisas Em Biologia Molecular E Funcional (CPBMF), Pontifícia Universidade Católica Do Rio Grande Do Sul (PUCRS), 92A TECNOPUC Building, 4592 Av. Bento Gonçalves, Porto Alegre, ZC 90650-001, Brazil., Ayub MAZ; Biotechnology, Bioprocess, and Biocatalysis Group, Food Science and Technology Institute, Federal University of Rio Grande Do Sul, Av. Bento Gonçalves 9500, PO Box 15090, Porto Alegre, RS, ZC 91501-970, Brazil. mazayub@ufrgs.br.
المصدر: Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] [Braz J Microbiol] 2021 Sep; Vol. 52 (3), pp. 1225-1233. Date of Electronic Publication: 2021 May 19.
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: Springer International Publishing Country of Publication: Brazil NLM ID: 101095924 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1678-4405 (Electronic) Linking ISSN: 15178382 NLM ISO Abbreviation: Braz J Microbiol Subsets: MEDLINE
أسماء مطبوعة: Publication: 2019- : Switzerland, AG : Springer International Publishing
Original Publication: Rio de Janeiro, RJ, Brasil : Sociedade Brasileira de Microbiologia
مواضيع طبية MeSH: Escherichia coli/*enzymology , Transglutaminases/*biosynthesis, Bacillus amyloliquefaciens/enzymology ; Bacillus amyloliquefaciens/genetics ; Bioreactors ; Culture Media ; Escherichia coli/genetics ; Glucose ; Plasmids/genetics ; Transglutaminases/genetics
مستخلص: We studied the expression of Bacillus amyloliquefaciens transglutaminase cloned in Escherichia coli BL21(DE3)pLysS harboring the plasmid pBAD/3C/bTGase, a bicistronic expression system, in bioreactor cultivation. Batch and fed-batch controlled as DO-stat strategies were employed for the production of the recombinant enzyme. In 30 h-batch cultivations using Terrific broth (TB), 6 g/L of biomass and 3.12 U/mg protein of transglutaminase activity were obtained. DO-stat fed-batch cultivations under the control of oxygen concentration (DO-stat) using TB as medium but fed with glucose allowed the increment in biomass formation (17.5 g/L) and enzyme activity (6.43 U/mg protein ). DO-stat fed-batch using mineral medium (M9) and fed with glucose under the same conditions produced even higher enzymatic activity (9.14 U/mg protein ). The pH effect was investigated, and the best enzymatic activity could be observed at pH 8. In all cultivations, the bicistronic system remained stable, with 100% of plasmid-bearing cells. These results show that E. coli bearing bicistronic plasmid constructs to express recombinant TGase could be cultivated in bioreactors under DO-stat fed-batch using mineral medium and it is a promising strategy in future optimizations to produce this important enzyme.
(© 2021. Sociedade Brasileira de Microbiologia.)
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معلومات مُعتمدة: N. A. CNPq; N. A. CAPES; N. A. FAPERGS
فهرسة مساهمة: Keywords: Bacillus amyloliquefaciens; Bicistronic plasmid system; DO-stat; Fed-batch bioreactor; Food enzymes; Microbial transglutaminase
المشرفين على المادة: 0 (Culture Media)
EC 2.3.2.13 (Transglutaminases)
IY9XDZ35W2 (Glucose)
تواريخ الأحداث: Date Created: 20210519 Date Completed: 20211206 Latest Revision: 20220520
رمز التحديث: 20221213
مُعرف محوري في PubMed: PMC8324676
DOI: 10.1007/s42770-021-00521-3
PMID: 34008152
قاعدة البيانات: MEDLINE
الوصف
تدمد:1678-4405
DOI:10.1007/s42770-021-00521-3