Optimizing Protein-Glutaminase Expression in Bacillus subtilis
Protein-glutaminase (PG) is a promising protein deaminase. It only hydrolyzes the side chain amido groups of protein-bound to generate ammonia and protein-L-glutamic acid and does not catalyze any other undesirable changes in protein structures. Deamidation of proteins via PG can influence the solub...
Gespeichert in:
| Veröffentlicht in: | Current microbiology 2021-05, Vol.78 (5), p.1752-1762 |
|---|---|
| Hauptverfasser: | , , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 1762 |
|---|---|
| container_issue | 5 |
| container_start_page | 1752 |
| container_title | Current microbiology |
| container_volume | 78 |
| creator | Ouyang, Xiaoying Liu, Yingjie Qu, Ruidan Tian, Min Yang, Ting Zhu, Rui Gao, Hongliang Jin, Mingfei Huang, Jing |
| description | Protein-glutaminase (PG) is a promising protein deaminase. It only hydrolyzes the side chain amido groups of protein-bound to generate ammonia and protein-L-glutamic acid and does not catalyze any other undesirable changes in protein structures. Deamidation of proteins via PG can influence the solubility, emulsification, foaming, and gelation properties of proteins, which are important properties for some food proteins. Therefore, there is great potential for the application of PG in the food industry. PG is derived from
Chryseobacterium proteolyticum
(
C. proteolyticum
); however, wild strains are difficult to industrialize because of their low levels of enzyme production. In this article, we studied different strategies for PG expression in
B. subtilis.
Results showed that PG produced from
C. proteolyticum
could be successfully secreted in
B. subtilis WB800N
, and actively secreted in
B. subtilis 168
(
BS168
) or
DB403
containing a pro-peptide (pro-PG). The secreted PG from
B. subtilis WB800N
was inactive unless digested by exogenous proteases, such as trypsin, alkaline protease, and neutral protease. However, active PG was secreted by the self-processing of
BS168 and DB403.
The specific activity of purified PG reached 20.9 U/mg. PG showed maximum activity at pH 5.5, 55 °C and more than 80% of PG activity was retained within a range of pH 3.5–6.5. When Cbz-Gln-Gly was used as the substrate, PG activity was 31.1
±
0.9 μM min
−1
mg
−1
. Mg
2+
, Ca
2+
, and Zn
2+
stabilized and even activated PG activity. These strategies concerning PG expression in
B. subtilis
and the enzymatic properties of PG provide efficient alternatives for PG research and contribute to the industrial-scale production of PG. |
| doi_str_mv | 10.1007/s00284-021-02404-0 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_journals_2519567031</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2503445288</sourcerecordid><originalsourceid>FETCH-LOGICAL-c375t-e0f79a9acc2fdade39ced090b91bd234742b52a42d4fc1104c1d0c5b9f92b6483</originalsourceid><addsrcrecordid>eNp9kE1LAzEQhoMotlb_gAdZ8OJldSbJdjcXQUv9gEI96Dlks9mSsh812QX115u6VcGDhzCBPPNO5iHkFOESAdIrD0AzHgPFcDiE2x4ZI2c0BiFwn4yBcRZn0wRH5Mj7NQBSAXhIRoylHBCTMblebjpb2w_brKIn13bGNvF91Xeqto3yJpq_bZzx3rZNZJvoVmlbVb2PfJ93trL-mByUqvLmZFcn5OVu_jx7iBfL-8fZzSLWLE262ECZCiWU1rQsVGGY0KYAAbnAvKCMp5zmCVWcFrzUiMA1FqCTXJSC5lOesQm5GHI3rn3tje9kbb02VaUa0_Ze0iTsyhOabdHzP-i67V0TfhcoFMk0BYaBogOlXeu9M6XcOFsr9y4R5NauHOzKYFd-2ZUQms520X1em-Kn5VtnANgA-PDUrIz7nf1P7Cf6i4Rs</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2519567031</pqid></control><display><type>article</type><title>Optimizing Protein-Glutaminase Expression in Bacillus subtilis</title><source>MEDLINE</source><source>Springer journals</source><creator>Ouyang, Xiaoying ; Liu, Yingjie ; Qu, Ruidan ; Tian, Min ; Yang, Ting ; Zhu, Rui ; Gao, Hongliang ; Jin, Mingfei ; Huang, Jing</creator><creatorcontrib>Ouyang, Xiaoying ; Liu, Yingjie ; Qu, Ruidan ; Tian, Min ; Yang, Ting ; Zhu, Rui ; Gao, Hongliang ; Jin, Mingfei ; Huang, Jing</creatorcontrib><description>Protein-glutaminase (PG) is a promising protein deaminase. It only hydrolyzes the side chain amido groups of protein-bound to generate ammonia and protein-L-glutamic acid and does not catalyze any other undesirable changes in protein structures. Deamidation of proteins via PG can influence the solubility, emulsification, foaming, and gelation properties of proteins, which are important properties for some food proteins. Therefore, there is great potential for the application of PG in the food industry. PG is derived from
Chryseobacterium proteolyticum
(
C. proteolyticum
); however, wild strains are difficult to industrialize because of their low levels of enzyme production. In this article, we studied different strategies for PG expression in
B. subtilis.
Results showed that PG produced from
C. proteolyticum
could be successfully secreted in
B. subtilis WB800N
, and actively secreted in
B. subtilis 168
(
BS168
) or
DB403
containing a pro-peptide (pro-PG). The secreted PG from
B. subtilis WB800N
was inactive unless digested by exogenous proteases, such as trypsin, alkaline protease, and neutral protease. However, active PG was secreted by the self-processing of
BS168 and DB403.
The specific activity of purified PG reached 20.9 U/mg. PG showed maximum activity at pH 5.5, 55 °C and more than 80% of PG activity was retained within a range of pH 3.5–6.5. When Cbz-Gln-Gly was used as the substrate, PG activity was 31.1
±
0.9 μM min
−1
mg
−1
. Mg
2+
, Ca
2+
, and Zn
2+
stabilized and even activated PG activity. These strategies concerning PG expression in
B. subtilis
and the enzymatic properties of PG provide efficient alternatives for PG research and contribute to the industrial-scale production of PG.</description><identifier>ISSN: 0343-8651</identifier><identifier>EISSN: 1432-0991</identifier><identifier>DOI: 10.1007/s00284-021-02404-0</identifier><identifier>PMID: 33740115</identifier><language>eng</language><publisher>New York: Springer US</publisher><subject>Alkaline protease ; Ammonia ; Bacillus subtilis - genetics ; Bacterial Proteins - genetics ; Biomedical and Life Sciences ; Biotechnology ; Calcium ; Calcium ions ; Chryseobacterium ; Emulsification ; Foaming ; Food industry ; Food processing industry ; Glutamic acid ; Glutaminase ; Life Sciences ; Magnesium ; Microbiology ; pH effects ; Protease ; Proteins ; Solubility ; Substrates ; Trypsin ; Zinc</subject><ispartof>Current microbiology, 2021-05, Vol.78 (5), p.1752-1762</ispartof><rights>The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2021</rights><rights>The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2021.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c375t-e0f79a9acc2fdade39ced090b91bd234742b52a42d4fc1104c1d0c5b9f92b6483</citedby><cites>FETCH-LOGICAL-c375t-e0f79a9acc2fdade39ced090b91bd234742b52a42d4fc1104c1d0c5b9f92b6483</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00284-021-02404-0$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00284-021-02404-0$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27924,27925,41488,42557,51319</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/33740115$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ouyang, Xiaoying</creatorcontrib><creatorcontrib>Liu, Yingjie</creatorcontrib><creatorcontrib>Qu, Ruidan</creatorcontrib><creatorcontrib>Tian, Min</creatorcontrib><creatorcontrib>Yang, Ting</creatorcontrib><creatorcontrib>Zhu, Rui</creatorcontrib><creatorcontrib>Gao, Hongliang</creatorcontrib><creatorcontrib>Jin, Mingfei</creatorcontrib><creatorcontrib>Huang, Jing</creatorcontrib><title>Optimizing Protein-Glutaminase Expression in Bacillus subtilis</title><title>Current microbiology</title><addtitle>Curr Microbiol</addtitle><addtitle>Curr Microbiol</addtitle><description>Protein-glutaminase (PG) is a promising protein deaminase. It only hydrolyzes the side chain amido groups of protein-bound to generate ammonia and protein-L-glutamic acid and does not catalyze any other undesirable changes in protein structures. Deamidation of proteins via PG can influence the solubility, emulsification, foaming, and gelation properties of proteins, which are important properties for some food proteins. Therefore, there is great potential for the application of PG in the food industry. PG is derived from
Chryseobacterium proteolyticum
(
C. proteolyticum
); however, wild strains are difficult to industrialize because of their low levels of enzyme production. In this article, we studied different strategies for PG expression in
B. subtilis.
Results showed that PG produced from
C. proteolyticum
could be successfully secreted in
B. subtilis WB800N
, and actively secreted in
B. subtilis 168
(
BS168
) or
DB403
containing a pro-peptide (pro-PG). The secreted PG from
B. subtilis WB800N
was inactive unless digested by exogenous proteases, such as trypsin, alkaline protease, and neutral protease. However, active PG was secreted by the self-processing of
BS168 and DB403.
The specific activity of purified PG reached 20.9 U/mg. PG showed maximum activity at pH 5.5, 55 °C and more than 80% of PG activity was retained within a range of pH 3.5–6.5. When Cbz-Gln-Gly was used as the substrate, PG activity was 31.1
±
0.9 μM min
−1
mg
−1
. Mg
2+
, Ca
2+
, and Zn
2+
stabilized and even activated PG activity. These strategies concerning PG expression in
B. subtilis
and the enzymatic properties of PG provide efficient alternatives for PG research and contribute to the industrial-scale production of PG.</description><subject>Alkaline protease</subject><subject>Ammonia</subject><subject>Bacillus subtilis - genetics</subject><subject>Bacterial Proteins - genetics</subject><subject>Biomedical and Life Sciences</subject><subject>Biotechnology</subject><subject>Calcium</subject><subject>Calcium ions</subject><subject>Chryseobacterium</subject><subject>Emulsification</subject><subject>Foaming</subject><subject>Food industry</subject><subject>Food processing industry</subject><subject>Glutamic acid</subject><subject>Glutaminase</subject><subject>Life Sciences</subject><subject>Magnesium</subject><subject>Microbiology</subject><subject>pH effects</subject><subject>Protease</subject><subject>Proteins</subject><subject>Solubility</subject><subject>Substrates</subject><subject>Trypsin</subject><subject>Zinc</subject><issn>0343-8651</issn><issn>1432-0991</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>8G5</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>GUQSH</sourceid><sourceid>M2O</sourceid><recordid>eNp9kE1LAzEQhoMotlb_gAdZ8OJldSbJdjcXQUv9gEI96Dlks9mSsh812QX115u6VcGDhzCBPPNO5iHkFOESAdIrD0AzHgPFcDiE2x4ZI2c0BiFwn4yBcRZn0wRH5Mj7NQBSAXhIRoylHBCTMblebjpb2w_brKIn13bGNvF91Xeqto3yJpq_bZzx3rZNZJvoVmlbVb2PfJ93trL-mByUqvLmZFcn5OVu_jx7iBfL-8fZzSLWLE262ECZCiWU1rQsVGGY0KYAAbnAvKCMp5zmCVWcFrzUiMA1FqCTXJSC5lOesQm5GHI3rn3tje9kbb02VaUa0_Ze0iTsyhOabdHzP-i67V0TfhcoFMk0BYaBogOlXeu9M6XcOFsr9y4R5NauHOzKYFd-2ZUQms520X1em-Kn5VtnANgA-PDUrIz7nf1P7Cf6i4Rs</recordid><startdate>20210501</startdate><enddate>20210501</enddate><creator>Ouyang, Xiaoying</creator><creator>Liu, Yingjie</creator><creator>Qu, Ruidan</creator><creator>Tian, Min</creator><creator>Yang, Ting</creator><creator>Zhu, Rui</creator><creator>Gao, Hongliang</creator><creator>Jin, Mingfei</creator><creator>Huang, Jing</creator><general>Springer US</general><general>Springer Nature B.V</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QL</scope><scope>7T7</scope><scope>7TK</scope><scope>7TM</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>8G5</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>GUQSH</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2O</scope><scope>M7N</scope><scope>M7P</scope><scope>MBDVC</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>20210501</creationdate><title>Optimizing Protein-Glutaminase Expression in Bacillus subtilis</title><author>Ouyang, Xiaoying ; Liu, Yingjie ; Qu, Ruidan ; Tian, Min ; Yang, Ting ; Zhu, Rui ; Gao, Hongliang ; Jin, Mingfei ; Huang, Jing</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c375t-e0f79a9acc2fdade39ced090b91bd234742b52a42d4fc1104c1d0c5b9f92b6483</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>Alkaline protease</topic><topic>Ammonia</topic><topic>Bacillus subtilis - genetics</topic><topic>Bacterial Proteins - genetics</topic><topic>Biomedical and Life Sciences</topic><topic>Biotechnology</topic><topic>Calcium</topic><topic>Calcium ions</topic><topic>Chryseobacterium</topic><topic>Emulsification</topic><topic>Foaming</topic><topic>Food industry</topic><topic>Food processing industry</topic><topic>Glutamic acid</topic><topic>Glutaminase</topic><topic>Life Sciences</topic><topic>Magnesium</topic><topic>Microbiology</topic><topic>pH effects</topic><topic>Protease</topic><topic>Proteins</topic><topic>Solubility</topic><topic>Substrates</topic><topic>Trypsin</topic><topic>Zinc</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ouyang, Xiaoying</creatorcontrib><creatorcontrib>Liu, Yingjie</creatorcontrib><creatorcontrib>Qu, Ruidan</creatorcontrib><creatorcontrib>Tian, Min</creatorcontrib><creatorcontrib>Yang, Ting</creatorcontrib><creatorcontrib>Zhu, Rui</creatorcontrib><creatorcontrib>Gao, Hongliang</creatorcontrib><creatorcontrib>Jin, Mingfei</creatorcontrib><creatorcontrib>Huang, Jing</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Health & Medical Collection (Proquest)</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Research Library (Alumni Edition)</collection><collection>ProQuest Central (Alumni)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>ProQuest Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>Research Library Prep</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection (Proquest) (PQ_SDU_P3)</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Biological Sciences</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>ProQuest research library</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Research Library (Corporate)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central Basic</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Current microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ouyang, Xiaoying</au><au>Liu, Yingjie</au><au>Qu, Ruidan</au><au>Tian, Min</au><au>Yang, Ting</au><au>Zhu, Rui</au><au>Gao, Hongliang</au><au>Jin, Mingfei</au><au>Huang, Jing</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Optimizing Protein-Glutaminase Expression in Bacillus subtilis</atitle><jtitle>Current microbiology</jtitle><stitle>Curr Microbiol</stitle><addtitle>Curr Microbiol</addtitle><date>2021-05-01</date><risdate>2021</risdate><volume>78</volume><issue>5</issue><spage>1752</spage><epage>1762</epage><pages>1752-1762</pages><issn>0343-8651</issn><eissn>1432-0991</eissn><abstract>Protein-glutaminase (PG) is a promising protein deaminase. It only hydrolyzes the side chain amido groups of protein-bound to generate ammonia and protein-L-glutamic acid and does not catalyze any other undesirable changes in protein structures. Deamidation of proteins via PG can influence the solubility, emulsification, foaming, and gelation properties of proteins, which are important properties for some food proteins. Therefore, there is great potential for the application of PG in the food industry. PG is derived from
Chryseobacterium proteolyticum
(
C. proteolyticum
); however, wild strains are difficult to industrialize because of their low levels of enzyme production. In this article, we studied different strategies for PG expression in
B. subtilis.
Results showed that PG produced from
C. proteolyticum
could be successfully secreted in
B. subtilis WB800N
, and actively secreted in
B. subtilis 168
(
BS168
) or
DB403
containing a pro-peptide (pro-PG). The secreted PG from
B. subtilis WB800N
was inactive unless digested by exogenous proteases, such as trypsin, alkaline protease, and neutral protease. However, active PG was secreted by the self-processing of
BS168 and DB403.
The specific activity of purified PG reached 20.9 U/mg. PG showed maximum activity at pH 5.5, 55 °C and more than 80% of PG activity was retained within a range of pH 3.5–6.5. When Cbz-Gln-Gly was used as the substrate, PG activity was 31.1
±
0.9 μM min
−1
mg
−1
. Mg
2+
, Ca
2+
, and Zn
2+
stabilized and even activated PG activity. These strategies concerning PG expression in
B. subtilis
and the enzymatic properties of PG provide efficient alternatives for PG research and contribute to the industrial-scale production of PG.</abstract><cop>New York</cop><pub>Springer US</pub><pmid>33740115</pmid><doi>10.1007/s00284-021-02404-0</doi><tpages>11</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0343-8651 |
| ispartof | Current microbiology, 2021-05, Vol.78 (5), p.1752-1762 |
| issn | 0343-8651 1432-0991 |
| language | eng |
| recordid | cdi_proquest_journals_2519567031 |
| source | MEDLINE; Springer journals |
| subjects | Alkaline protease Ammonia Bacillus subtilis - genetics Bacterial Proteins - genetics Biomedical and Life Sciences Biotechnology Calcium Calcium ions Chryseobacterium Emulsification Foaming Food industry Food processing industry Glutamic acid Glutaminase Life Sciences Magnesium Microbiology pH effects Protease Proteins Solubility Substrates Trypsin Zinc |
| title | Optimizing Protein-Glutaminase Expression in Bacillus subtilis |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-07T17%3A28%3A01IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Optimizing%20Protein-Glutaminase%20Expression%20in%20Bacillus%20subtilis&rft.jtitle=Current%20microbiology&rft.au=Ouyang,%20Xiaoying&rft.date=2021-05-01&rft.volume=78&rft.issue=5&rft.spage=1752&rft.epage=1762&rft.pages=1752-1762&rft.issn=0343-8651&rft.eissn=1432-0991&rft_id=info:doi/10.1007/s00284-021-02404-0&rft_dat=%3Cproquest_cross%3E2503445288%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2519567031&rft_id=info:pmid/33740115&rfr_iscdi=true |