Random Mutagenesis Applied to Reveal Factors Involved in Oxidative Tolerance and Biofilm Formation in Foodborne Cronobacter malonaticus.
Identifieur interne : 000129 ( Main/Exploration ); précédent : 000128; suivant : 000130Random Mutagenesis Applied to Reveal Factors Involved in Oxidative Tolerance and Biofilm Formation in Foodborne Cronobacter malonaticus.
Auteurs : Maofeng Zhang [République populaire de Chine] ; Xiyan Zhang [République populaire de Chine] ; Liaowang Tong [République populaire de Chine] ; Dexin Ou [République populaire de Chine] ; Yaping Wang [République populaire de Chine] ; Jumei Zhang [République populaire de Chine] ; Qingping Wu [République populaire de Chine] ; Yingwang Ye [République populaire de Chine]Source :
- Frontiers in microbiology [ 1664-302X ] ; 2019.
Abstract
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.
DOI: 10.3389/fmicb.2019.00877
PubMed: 31118922
PubMed Central: PMC6504702
Affiliations:
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Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Institute of Microbiology, Guangzhou, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Institute of Microbiology, Guangzhou</wicri:regionArea><placeName><settlement type="city">Jiangmen</settlement><region type="province">Guangdong</region></placeName></affiliation></author><author><name sortKey="Wu, Qingping" sort="Wu, Qingping" uniqKey="Wu Q" first="Qingping" last="Wu">Qingping Wu</name><affiliation wicri:level="3"><nlm:affiliation>State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Institute of Microbiology, Guangzhou, China.</nlm:affiliation><country xml:lang="fr">République 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type="province">Anhui</region></placeName></affiliation><affiliation wicri:level="3"><nlm:affiliation>State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Institute of Microbiology, Guangzhou, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Institute of Microbiology, Guangzhou</wicri:regionArea><placeName><settlement type="city">Jiangmen</settlement><region type="province">Guangdong</region></placeName></affiliation></author><author><name sortKey="Zhang, Xiyan" sort="Zhang, Xiyan" uniqKey="Zhang X" first="Xiyan" last="Zhang">Xiyan Zhang</name><affiliation wicri:level="3"><nlm:affiliation>School of Food Science and Engineering, Hefei University of Technology, Hefei, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>School of Food Science and Engineering, Hefei University of Technology, Hefei</wicri:regionArea><placeName><settlement type="city">Hefei</settlement><region type="province">Anhui</region></placeName></affiliation></author><author><name sortKey="Tong, Liaowang" sort="Tong, Liaowang" uniqKey="Tong L" first="Liaowang" last="Tong">Liaowang Tong</name><affiliation wicri:level="3"><nlm:affiliation>School of Food Science and Engineering, Hefei University of Technology, Hefei, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>School of Food Science and Engineering, Hefei University of Technology, Hefei</wicri:regionArea><placeName><settlement type="city">Hefei</settlement><region type="province">Anhui</region></placeName></affiliation></author><author><name sortKey="Ou, Dexin" sort="Ou, Dexin" uniqKey="Ou D" first="Dexin" last="Ou">Dexin Ou</name><affiliation wicri:level="3"><nlm:affiliation>School of Food Science and Engineering, Hefei University of Technology, Hefei, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>School of Food Science and Engineering, Hefei University of Technology, Hefei</wicri:regionArea><placeName><settlement type="city">Hefei</settlement><region type="province">Anhui</region></placeName></affiliation></author><author><name sortKey="Wang, Yaping" sort="Wang, Yaping" uniqKey="Wang Y" first="Yaping" last="Wang">Yaping Wang</name><affiliation wicri:level="3"><nlm:affiliation>School of Food Science and Engineering, Hefei University of Technology, Hefei, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>School of Food Science and Engineering, Hefei University of Technology, Hefei</wicri:regionArea><placeName><settlement type="city">Hefei</settlement><region type="province">Anhui</region></placeName></affiliation></author><author><name sortKey="Zhang, Jumei" sort="Zhang, Jumei" uniqKey="Zhang J" first="Jumei" last="Zhang">Jumei Zhang</name><affiliation wicri:level="3"><nlm:affiliation>State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Institute of Microbiology, Guangzhou, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Institute of Microbiology, Guangzhou</wicri:regionArea><placeName><settlement type="city">Jiangmen</settlement><region type="province">Guangdong</region></placeName></affiliation></author><author><name sortKey="Wu, Qingping" sort="Wu, Qingping" uniqKey="Wu Q" first="Qingping" last="Wu">Qingping Wu</name><affiliation wicri:level="3"><nlm:affiliation>State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Institute of Microbiology, Guangzhou, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Institute of Microbiology, Guangzhou</wicri:regionArea><placeName><settlement type="city">Jiangmen</settlement><region type="province">Guangdong</region></placeName></affiliation></author><author><name sortKey="Ye, Yingwang" sort="Ye, Yingwang" uniqKey="Ye Y" first="Yingwang" last="Ye">Yingwang Ye</name><affiliation wicri:level="3"><nlm:affiliation>School of Food Science and Engineering, Hefei University of Technology, Hefei, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>School of Food Science and Engineering, Hefei University of Technology, Hefei</wicri:regionArea><placeName><settlement type="city">Hefei</settlement><region type="province">Anhui</region></placeName></affiliation><affiliation wicri:level="3"><nlm:affiliation>State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Institute of Microbiology, Guangzhou, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Institute of Microbiology, Guangzhou</wicri:regionArea><placeName><settlement type="city">Jiangmen</settlement><region type="province">Guangdong</region></placeName></affiliation></author></analytic><series><title level="j">Frontiers in microbiology</title><idno type="ISSN">1664-302X</idno><imprint><date when="2019" type="published">2019</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><i>Cronobacter</i> 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 <i>Cronobacter</i> remains elusive. Here, factors involved in oxidative tolerance in <i>C. malonaticus</i> were identified using a transposon mutagenesis. Eight mutants were successfully screened based on a comparison of the growth of strains from mutant library (<i>n</i> = 215) and wild type (WT) strain under 1.0 mM H<sub>2</sub>O<sub>2</sub>. 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 <i>C. malonaticus</i>.</div></front></TEI><pubmed><MedlineCitation Status="PubMed-not-MEDLINE" Owner="NLM"><PMID Version="1">31118922</PMID><DateRevised><Year>2020</Year><Month>10</Month><Day>01</Day></DateRevised><Article PubModel="Electronic-eCollection"><Journal><ISSN IssnType="Print">1664-302X</ISSN><JournalIssue CitedMedium="Print"><Volume>10</Volume><PubDate><Year>2019</Year></PubDate></JournalIssue><Title>Frontiers in microbiology</Title><ISOAbbreviation>Front Microbiol</ISOAbbreviation></Journal><ArticleTitle>Random Mutagenesis Applied to Reveal Factors Involved in Oxidative Tolerance and Biofilm Formation in Foodborne <i>Cronobacter malonaticus</i>.</ArticleTitle><Pagination><MedlinePgn>877</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.3389/fmicb.2019.00877</ELocationID><Abstract><AbstractText><i>Cronobacter</i> 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 <i>Cronobacter</i> remains elusive. Here, factors involved in oxidative tolerance in <i>C. malonaticus</i> were identified using a transposon mutagenesis. Eight mutants were successfully screened based on a comparison of the growth of strains from mutant library (<i>n</i> = 215) and wild type (WT) strain under 1.0 mM H<sub>2</sub>O<sub>2</sub>. 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 <i>C. malonaticus</i>.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Zhang</LastName><ForeName>Maofeng</ForeName><Initials>M</Initials><AffiliationInfo><Affiliation>School of Food Science and Engineering, Hefei University of Technology, Hefei, China.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Institute of Microbiology, Guangzhou, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Zhang</LastName><ForeName>Xiyan</ForeName><Initials>X</Initials><AffiliationInfo><Affiliation>School of Food Science and Engineering, Hefei University of Technology, Hefei, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Tong</LastName><ForeName>Liaowang</ForeName><Initials>L</Initials><AffiliationInfo><Affiliation>School of Food Science and Engineering, Hefei University of Technology, Hefei, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Ou</LastName><ForeName>Dexin</ForeName><Initials>D</Initials><AffiliationInfo><Affiliation>School of Food Science and Engineering, Hefei University of Technology, Hefei, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Wang</LastName><ForeName>Yaping</ForeName><Initials>Y</Initials><AffiliationInfo><Affiliation>School of Food Science and Engineering, Hefei University of Technology, Hefei, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Zhang</LastName><ForeName>Jumei</ForeName><Initials>J</Initials><AffiliationInfo><Affiliation>State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Institute of Microbiology, Guangzhou, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Wu</LastName><ForeName>Qingping</ForeName><Initials>Q</Initials><AffiliationInfo><Affiliation>State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Institute of Microbiology, Guangzhou, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Ye</LastName><ForeName>Yingwang</ForeName><Initials>Y</Initials><AffiliationInfo><Affiliation>School of Food Science and Engineering, Hefei University of Technology, Hefei, China.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Institute of Microbiology, Guangzhou, China.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2019</Year><Month>05</Month><Day>01</Day></ArticleDate></Article><MedlineJournalInfo><Country>Switzerland</Country><MedlineTA>Front Microbiol</MedlineTA><NlmUniqueID>101548977</NlmUniqueID><ISSNLinking>1664-302X</ISSNLinking></MedlineJournalInfo><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">Cronobacter malonaticus</Keyword><Keyword MajorTopicYN="N">arbitrary PCR</Keyword><Keyword MajorTopicYN="N">biofilm formation</Keyword><Keyword MajorTopicYN="N">oxidative stress</Keyword><Keyword MajorTopicYN="N">random mutagenesis</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2019</Year><Month>01</Month><Day>19</Day></PubMedPubDate><PubMedPubDate 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name="Anhui"><name sortKey="Zhang, Maofeng" sort="Zhang, Maofeng" uniqKey="Zhang M" first="Maofeng" last="Zhang">Maofeng Zhang</name></region><name sortKey="Ou, Dexin" sort="Ou, Dexin" uniqKey="Ou D" first="Dexin" last="Ou">Dexin Ou</name><name sortKey="Tong, Liaowang" sort="Tong, Liaowang" uniqKey="Tong L" first="Liaowang" last="Tong">Liaowang Tong</name><name sortKey="Wang, Yaping" sort="Wang, Yaping" uniqKey="Wang Y" first="Yaping" last="Wang">Yaping Wang</name><name sortKey="Wu, Qingping" sort="Wu, Qingping" uniqKey="Wu Q" first="Qingping" last="Wu">Qingping Wu</name><name sortKey="Ye, Yingwang" sort="Ye, Yingwang" uniqKey="Ye Y" first="Yingwang" last="Ye">Yingwang Ye</name><name sortKey="Ye, Yingwang" sort="Ye, Yingwang" uniqKey="Ye Y" first="Yingwang" last="Ye">Yingwang Ye</name><name sortKey="Zhang, Jumei" sort="Zhang, Jumei" uniqKey="Zhang J" first="Jumei" last="Zhang">Jumei Zhang</name><name sortKey="Zhang, Maofeng" sort="Zhang, Maofeng" uniqKey="Zhang M" first="Maofeng" last="Zhang">Maofeng Zhang</name><name sortKey="Zhang, Xiyan" sort="Zhang, Xiyan" uniqKey="Zhang X" first="Xiyan" last="Zhang">Xiyan Zhang</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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