Random Mutagenesis Applied to Reveal Factors Involved in Oxidative Tolerance and Biofilm Formation in Foodborne Cronobacter malonaticus
Cronobacter species are linked with life-treating diseases in neonates and show strong tolerances to environmental stress. However, the information about factors involved in oxidative tolerance in Cronobacter remains elusive. Here, factors involved in oxidative tolerance in C. malonaticus were ident...
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Frontiers Media S.A.
2019-05-01
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Online Access: | https://www.frontiersin.org/article/10.3389/fmicb.2019.00877/full |
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author | Maofeng Zhang Maofeng Zhang Xiyan Zhang Liaowang Tong Dexin Ou Yaping Wang Jumei Zhang Qingping Wu Yingwang Ye Yingwang Ye |
author_facet | Maofeng Zhang Maofeng Zhang Xiyan Zhang Liaowang Tong Dexin Ou Yaping Wang Jumei Zhang Qingping Wu Yingwang Ye Yingwang Ye |
author_sort | Maofeng Zhang |
collection | DOAJ |
description | Cronobacter species are linked with life-treating diseases in neonates and show strong tolerances to environmental stress. However, the information about factors involved in oxidative tolerance in Cronobacter remains elusive. Here, factors involved in oxidative tolerance in C. malonaticus were identified using a transposon mutagenesis. Eight mutants were successfully screened based on a comparison of the growth of strains from mutant library (n = 215) and wild type (WT) strain under 1.0 mM H2O2. Mutating sites including thioredoxin 2, glutaredoxin 3, pantothenate kinase, serine/threonine protein kinase, pyruvate kinase, phospholipase A, ferrous iron transport protein A, and alanine racemase 2 were successfully identified by arbitrary PCR and sequencing alignment. Furthermore, the comparison about quantity and structure of biofilms formation among eight mutants and WT was determined using crystal violet staining (CVS), scanning electron microscopy (SEM), and confocal laser scanning microscopy (CLSM). Results showed that the biofilms of eight mutants significantly decreased within 48 h compared to that of WT, suggesting that mutating genes play important roles in biofilm formation under oxidative stress. The findings provide valuable information for deeply understanding molecular mechanism about oxidative tolerance of C. malonaticus. |
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spelling | doaj.art-3a25660547f54746a3d6227d38117e682022-12-21T19:12:27ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2019-05-011010.3389/fmicb.2019.00877424709Random Mutagenesis Applied to Reveal Factors Involved in Oxidative Tolerance and Biofilm Formation in Foodborne Cronobacter malonaticusMaofeng Zhang0Maofeng Zhang1Xiyan Zhang2Liaowang Tong3Dexin Ou4Yaping Wang5Jumei Zhang6Qingping Wu7Yingwang Ye8Yingwang Ye9School of Food Science and Engineering, Hefei University of Technology, Hefei, ChinaState Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Institute of Microbiology, Guangzhou, ChinaSchool of Food Science and Engineering, Hefei University of Technology, Hefei, ChinaSchool of Food Science and Engineering, Hefei University of Technology, Hefei, ChinaSchool of Food Science and Engineering, Hefei University of Technology, Hefei, ChinaSchool of Food Science and Engineering, Hefei University of Technology, Hefei, ChinaState Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Institute of Microbiology, Guangzhou, ChinaState Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Institute of Microbiology, Guangzhou, ChinaSchool of Food Science and Engineering, Hefei University of Technology, Hefei, ChinaState Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Institute of Microbiology, Guangzhou, ChinaCronobacter species are linked with life-treating diseases in neonates and show strong tolerances to environmental stress. However, the information about factors involved in oxidative tolerance in Cronobacter remains elusive. Here, factors involved in oxidative tolerance in C. malonaticus were identified using a transposon mutagenesis. Eight mutants were successfully screened based on a comparison of the growth of strains from mutant library (n = 215) and wild type (WT) strain under 1.0 mM H2O2. Mutating sites including thioredoxin 2, glutaredoxin 3, pantothenate kinase, serine/threonine protein kinase, pyruvate kinase, phospholipase A, ferrous iron transport protein A, and alanine racemase 2 were successfully identified by arbitrary PCR and sequencing alignment. Furthermore, the comparison about quantity and structure of biofilms formation among eight mutants and WT was determined using crystal violet staining (CVS), scanning electron microscopy (SEM), and confocal laser scanning microscopy (CLSM). Results showed that the biofilms of eight mutants significantly decreased within 48 h compared to that of WT, suggesting that mutating genes play important roles in biofilm formation under oxidative stress. The findings provide valuable information for deeply understanding molecular mechanism about oxidative tolerance of C. malonaticus.https://www.frontiersin.org/article/10.3389/fmicb.2019.00877/fullCronobacter malonaticusrandom mutagenesisarbitrary PCRoxidative stressbiofilm formation |
spellingShingle | Maofeng Zhang Maofeng Zhang Xiyan Zhang Liaowang Tong Dexin Ou Yaping Wang Jumei Zhang Qingping Wu Yingwang Ye Yingwang Ye Random Mutagenesis Applied to Reveal Factors Involved in Oxidative Tolerance and Biofilm Formation in Foodborne Cronobacter malonaticus Frontiers in Microbiology Cronobacter malonaticus random mutagenesis arbitrary PCR oxidative stress biofilm formation |
title | Random Mutagenesis Applied to Reveal Factors Involved in Oxidative Tolerance and Biofilm Formation in Foodborne Cronobacter malonaticus |
title_full | Random Mutagenesis Applied to Reveal Factors Involved in Oxidative Tolerance and Biofilm Formation in Foodborne Cronobacter malonaticus |
title_fullStr | Random Mutagenesis Applied to Reveal Factors Involved in Oxidative Tolerance and Biofilm Formation in Foodborne Cronobacter malonaticus |
title_full_unstemmed | Random Mutagenesis Applied to Reveal Factors Involved in Oxidative Tolerance and Biofilm Formation in Foodborne Cronobacter malonaticus |
title_short | Random Mutagenesis Applied to Reveal Factors Involved in Oxidative Tolerance and Biofilm Formation in Foodborne Cronobacter malonaticus |
title_sort | random mutagenesis applied to reveal factors involved in oxidative tolerance and biofilm formation in foodborne cronobacter malonaticus |
topic | Cronobacter malonaticus random mutagenesis arbitrary PCR oxidative stress biofilm formation |
url | https://www.frontiersin.org/article/10.3389/fmicb.2019.00877/full |
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