Substitution of Asp29 with Asn29 in the metallochaperone UreE of Streptococcus thermophilus DSM 20617T increases the urease activity and anticipates urea hydrolysis during milk fermentation.

Λεπτομέρειες βιβλιογραφικής εγγραφής
Τίτλος: Substitution of Asp29 with Asn29 in the metallochaperone UreE of Streptococcus thermophilus DSM 20617T increases the urease activity and anticipates urea hydrolysis during milk fermentation.
Συγγραφείς: Arioli S; Department of Food Environmental and Nutritional Sciences (DeFENS), University of Milan, Milan, Italy., Mangieri N; Department of Food Environmental and Nutritional Sciences (DeFENS), University of Milan, Milan, Italy., Zanchetta Y; Department of Food Environmental and Nutritional Sciences (DeFENS), University of Milan, Milan, Italy., Russo P; Department of Food Environmental and Nutritional Sciences (DeFENS), University of Milan, Milan, Italy., Mora D; Department of Food Environmental and Nutritional Sciences (DeFENS), University of Milan, Milan, Italy. Electronic address: diego.mora@unimi.it.
Πηγή: International journal of food microbiology [Int J Food Microbiol] 2024 May 02; Vol. 416, pp. 110684. Date of Electronic Publication: 2024 Mar 16.
Τύπος έκδοσης: Journal Article
Γλώσσα: English
Στοιχεία περιοδικού: Publisher: Elsevier Science Publishers Country of Publication: Netherlands NLM ID: 8412849 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1879-3460 (Electronic) Linking ISSN: 01681605 NLM ISO Abbreviation: Int J Food Microbiol Subsets: MEDLINE
Imprint Name(s): Original Publication: Amsterdam : Elsevier Science Publishers, c1984-
Ιατρικοί όροι (MeSH): Urease*/genetics , Streptococcus thermophilus*/metabolism, Metallochaperones/metabolism ; Carrier Proteins/genetics ; Nickel/metabolism ; Milk/metabolism ; Bacterial Proteins/genetics ; Animals ; Hydrolysis ; Urea ; Fermentation
Περίληψη: Urease operon is highly conserved within the species Streptococcus thermophilus and urease-negative strains are rare in nature. S. thermophilus MIMO1, isolated from commercial yogurt, was previously characterized as urease-positive Ni-dependent strain. Beside a mutation in ureQ, coding for a nickel ABC transporter permease, the strain MIMO1 showed a mutation in ureE gene which code for a metallochaperone involved in Ni delivery to the urease catalytic site. The single base mutation in ureE determined a substitution of Asp29 with Asn29 in the metallochaperone in a conserved protein region not involved in the catalytic activity. With the aim to investigate the role Asp29vs Asn29 substitution in UreE on the urease activity of S. thermophilus, ureE gene of the reference strain DSM 20617T (ureEDSM20617) was replaced by ureE gene of strain MIMO1 (ureEMIMO1) to obtain the recombinant ES3. In-gel detection of urease activity revealed that the substitution of Asp29 with Asn29 in UreE resulted in a higher stability of the enzyme complexes. Moreover, the recombinant ES3 showed higher level of urease activity compared to the wildtype without any detectable increase in the expression level of ureC gene, thus highlighting the role of UreE not only in Ni assembly but also on the level of urease activity. During the growth in milk, the recombinant ES3 showed an anticipated urease activity compared to the wildtype, and analogous milk fermentation performance. The overall data obtained by comparing urease-positive and urease-negative strains/mutants confirmed that urease activity strongly impacts on the milk fermentation process and specifically on the yield of the homolactic fermentation.
(Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)
Competing Interests: Declaration of competing interest None.
Contributed Indexing: Keywords: Milk acidification; Ni; Streptococcus thermophilus; UreE; Urease
Substance Nomenclature: EC 3.5.1.5 (Urease)
0 (Metallochaperones)
0 (Carrier Proteins)
7OV03QG267 (Nickel)
8W8T17847W (Urea)
0 (Bacterial Proteins)
Entry Date(s): Date Created: 20240321 Date Completed: 20240419 Latest Revision: 20240419
Update Code: 20260130
DOI: 10.1016/j.ijfoodmicro.2024.110684
PMID: 38513545
Βάση Δεδομένων: MEDLINE
Περιγραφή
ISSN:1879-3460
DOI:10.1016/j.ijfoodmicro.2024.110684