Highly efficient expression of Rasamsonia emersonii lipase in Pichia pastoris: characterization and gastrointestinal simulated digestion in vitro
BACKGROUND Acidic lipases with high catalytic activities under acidic conditions have important application values in the food, feed and pharmaceutical industries. However, the availability of acidic lipases is still the main obstacle to their industrial applications. Although a novel acidic lipase...
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| Veröffentlicht in: | Journal of the science of food and agriculture 2024-07, Vol.104 (9), p.5603-5613 |
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| creator | Wang, Buqing Wang, Yasen Zhou, Xiaoman Gao, Xiao‐Dong Fujita, Morihisa Li, Zijie |
| description | BACKGROUND
Acidic lipases with high catalytic activities under acidic conditions have important application values in the food, feed and pharmaceutical industries. However, the availability of acidic lipases is still the main obstacle to their industrial applications. Although a novel acidic lipase Rasamsonia emersonii (LIPR) was heterologously expressed in Escherichia coli, the expression level was unsatisfactory.
RESULTS
To achieve the high‐efficiency expression and secretion of LIPR in Pichia pastoris GS115, the combinatorial optimization strategy was adopted including gene codon preference, signal peptide, molecular chaperone co‐expression and disruption of vacuolar sorting receptor VPS10. The activity of the combinatorial optimization engineered strain in a shake flask reached 1480 U mL−1, which was 8.13 times greater than the P. pastoris GS115 parental strain. After high‐density fermentation in a 5‐L bioreactor, the highest enzyme activity reached as high as 11 820 U mL−1. LIPR showed the highest activity at 40 °C and pH 4.0 in the presence of Ca2+ ion. LIPR exhibited strong tolerance to methanol, indicating its potential application in biodiesel biosynthesis. Moreover, the gastrointestinal digestion simulation results demonstrated that LIPR was tolerant to pepsin and trypsin, but its activity was inhibited by sodium taurodeoxycholate.
CONCLUSION
This study provided an effective approach for the high expression of acidic lipase LIPR. LIPR was more appropriate for lipid digestion in the stomach than in intestine according to the gastrointestinal digestion simulation results. © 2024 Society of Chemical Industry. |
| doi_str_mv | 10.1002/jsfa.13390 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_3153621914</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>3067630957</sourcerecordid><originalsourceid>FETCH-LOGICAL-c3490-7384c4efd58f64baad5cf01592e1defa98a65b175380c3640b1aac4b7a06f6133</originalsourceid><addsrcrecordid>eNqFkctu1DAUhi0EokNhwwMgS2wqpJTjOHFidlVFKagSiMs6OuMcz5xRLoOdAMNb8MY4TGHBAla-_J8-HfsX4rGCcwWQP99Fj-dKawt3xEqBrTIABXfFKoV5VqoiPxEPYtwBgLXG3BcnutZGq9ysxI9r3my7gyTv2TENk6Rv-0Ax8jjI0cv3GLGP48Aoqaew7Fh2vMdIkgf5jt02Rek4jYHjC-m2GNBNFPg7TosDh1ZuUhxGHiaKEw_Yycj93OFErWx5s1wmMNm-cMIeinseu0iPbtdT8enq5cfL6-zm7avXlxc3mdOFhazSdeEK8m1Ze1OsEdvSeVClzUm15NHWaMq1qkpdg9OmgLVCdMW6QjDepN86FWdH7z6Mn-c0RNNzdNR1ONA4x0arUptcWVX8F81tXueFNdom9Olf6G6cQ3pzEoKpjAZbVol6dqRcGGMM5Jt94B7DoVHQLJ02S6fNr04T_ORWOa97av-gv0tMgDoCX7mjwz9UzZsPVxdH6U8WKq7m</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>3067630957</pqid></control><display><type>article</type><title>Highly efficient expression of Rasamsonia emersonii lipase in Pichia pastoris: characterization and gastrointestinal simulated digestion in vitro</title><source>Wiley Online Library Journals Frontfile Complete</source><creator>Wang, Buqing ; Wang, Yasen ; Zhou, Xiaoman ; Gao, Xiao‐Dong ; Fujita, Morihisa ; Li, Zijie</creator><creatorcontrib>Wang, Buqing ; Wang, Yasen ; Zhou, Xiaoman ; Gao, Xiao‐Dong ; Fujita, Morihisa ; Li, Zijie</creatorcontrib><description>BACKGROUND
Acidic lipases with high catalytic activities under acidic conditions have important application values in the food, feed and pharmaceutical industries. However, the availability of acidic lipases is still the main obstacle to their industrial applications. Although a novel acidic lipase Rasamsonia emersonii (LIPR) was heterologously expressed in Escherichia coli, the expression level was unsatisfactory.
RESULTS
To achieve the high‐efficiency expression and secretion of LIPR in Pichia pastoris GS115, the combinatorial optimization strategy was adopted including gene codon preference, signal peptide, molecular chaperone co‐expression and disruption of vacuolar sorting receptor VPS10. The activity of the combinatorial optimization engineered strain in a shake flask reached 1480 U mL−1, which was 8.13 times greater than the P. pastoris GS115 parental strain. After high‐density fermentation in a 5‐L bioreactor, the highest enzyme activity reached as high as 11 820 U mL−1. LIPR showed the highest activity at 40 °C and pH 4.0 in the presence of Ca2+ ion. LIPR exhibited strong tolerance to methanol, indicating its potential application in biodiesel biosynthesis. Moreover, the gastrointestinal digestion simulation results demonstrated that LIPR was tolerant to pepsin and trypsin, but its activity was inhibited by sodium taurodeoxycholate.
CONCLUSION
This study provided an effective approach for the high expression of acidic lipase LIPR. LIPR was more appropriate for lipid digestion in the stomach than in intestine according to the gastrointestinal digestion simulation results. © 2024 Society of Chemical Industry.</description><identifier>ISSN: 0022-5142</identifier><identifier>EISSN: 1097-0010</identifier><identifier>DOI: 10.1002/jsfa.13390</identifier><identifier>PMID: 38363126</identifier><language>eng</language><publisher>Chichester, UK: John Wiley & Sons, Ltd</publisher><subject>acidic lipase ; agriculture ; biodiesel ; Biodiesel fuels ; Biofuels ; Bioreactors ; Biosynthesis ; calcium ; Calcium ions ; carboxylic ester hydrolases ; codon usage ; Combinatorial analysis ; culture flasks ; Digestion ; Digestive system ; E coli ; Enzymatic activity ; Enzyme activity ; Escherichia coli ; Fermentation ; Food industry ; gastrointestinal simulated digestion ; Gastrointestinal tract ; in vitro digestion ; Industrial applications ; Intestine ; intestines ; Komagataella pastoris ; Lipase ; lipid metabolism ; Lipids ; methanol ; molecular chaperone ; molecular chaperones ; Optimization ; Pepsin ; Pharmaceutical industry ; Pichia pastoris ; Rasamsonia ; secretion ; signal peptide ; sodium ; stomach ; Trypsin ; vacuoles</subject><ispartof>Journal of the science of food and agriculture, 2024-07, Vol.104 (9), p.5603-5613</ispartof><rights>2024 Society of Chemical Industry.</rights><rights>This article is protected by copyright. All rights reserved.</rights><rights>2024 Society of Chemical Industry</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c3490-7384c4efd58f64baad5cf01592e1defa98a65b175380c3640b1aac4b7a06f6133</cites><orcidid>0000-0002-5618-1862</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fjsfa.13390$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fjsfa.13390$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27901,27902,45550,45551</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/38363126$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wang, Buqing</creatorcontrib><creatorcontrib>Wang, Yasen</creatorcontrib><creatorcontrib>Zhou, Xiaoman</creatorcontrib><creatorcontrib>Gao, Xiao‐Dong</creatorcontrib><creatorcontrib>Fujita, Morihisa</creatorcontrib><creatorcontrib>Li, Zijie</creatorcontrib><title>Highly efficient expression of Rasamsonia emersonii lipase in Pichia pastoris: characterization and gastrointestinal simulated digestion in vitro</title><title>Journal of the science of food and agriculture</title><addtitle>J Sci Food Agric</addtitle><description>BACKGROUND
Acidic lipases with high catalytic activities under acidic conditions have important application values in the food, feed and pharmaceutical industries. However, the availability of acidic lipases is still the main obstacle to their industrial applications. Although a novel acidic lipase Rasamsonia emersonii (LIPR) was heterologously expressed in Escherichia coli, the expression level was unsatisfactory.
RESULTS
To achieve the high‐efficiency expression and secretion of LIPR in Pichia pastoris GS115, the combinatorial optimization strategy was adopted including gene codon preference, signal peptide, molecular chaperone co‐expression and disruption of vacuolar sorting receptor VPS10. The activity of the combinatorial optimization engineered strain in a shake flask reached 1480 U mL−1, which was 8.13 times greater than the P. pastoris GS115 parental strain. After high‐density fermentation in a 5‐L bioreactor, the highest enzyme activity reached as high as 11 820 U mL−1. LIPR showed the highest activity at 40 °C and pH 4.0 in the presence of Ca2+ ion. LIPR exhibited strong tolerance to methanol, indicating its potential application in biodiesel biosynthesis. Moreover, the gastrointestinal digestion simulation results demonstrated that LIPR was tolerant to pepsin and trypsin, but its activity was inhibited by sodium taurodeoxycholate.
CONCLUSION
This study provided an effective approach for the high expression of acidic lipase LIPR. LIPR was more appropriate for lipid digestion in the stomach than in intestine according to the gastrointestinal digestion simulation results. © 2024 Society of Chemical Industry.</description><subject>acidic lipase</subject><subject>agriculture</subject><subject>biodiesel</subject><subject>Biodiesel fuels</subject><subject>Biofuels</subject><subject>Bioreactors</subject><subject>Biosynthesis</subject><subject>calcium</subject><subject>Calcium ions</subject><subject>carboxylic ester hydrolases</subject><subject>codon usage</subject><subject>Combinatorial analysis</subject><subject>culture flasks</subject><subject>Digestion</subject><subject>Digestive system</subject><subject>E coli</subject><subject>Enzymatic activity</subject><subject>Enzyme activity</subject><subject>Escherichia coli</subject><subject>Fermentation</subject><subject>Food industry</subject><subject>gastrointestinal simulated digestion</subject><subject>Gastrointestinal tract</subject><subject>in vitro digestion</subject><subject>Industrial applications</subject><subject>Intestine</subject><subject>intestines</subject><subject>Komagataella pastoris</subject><subject>Lipase</subject><subject>lipid metabolism</subject><subject>Lipids</subject><subject>methanol</subject><subject>molecular chaperone</subject><subject>molecular chaperones</subject><subject>Optimization</subject><subject>Pepsin</subject><subject>Pharmaceutical industry</subject><subject>Pichia pastoris</subject><subject>Rasamsonia</subject><subject>secretion</subject><subject>signal peptide</subject><subject>sodium</subject><subject>stomach</subject><subject>Trypsin</subject><subject>vacuoles</subject><issn>0022-5142</issn><issn>1097-0010</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2024</creationdate><recordtype>article</recordtype><recordid>eNqFkctu1DAUhi0EokNhwwMgS2wqpJTjOHFidlVFKagSiMs6OuMcz5xRLoOdAMNb8MY4TGHBAla-_J8-HfsX4rGCcwWQP99Fj-dKawt3xEqBrTIABXfFKoV5VqoiPxEPYtwBgLXG3BcnutZGq9ysxI9r3my7gyTv2TENk6Rv-0Ax8jjI0cv3GLGP48Aoqaew7Fh2vMdIkgf5jt02Rek4jYHjC-m2GNBNFPg7TosDh1ZuUhxGHiaKEw_Yycj93OFErWx5s1wmMNm-cMIeinseu0iPbtdT8enq5cfL6-zm7avXlxc3mdOFhazSdeEK8m1Ze1OsEdvSeVClzUm15NHWaMq1qkpdg9OmgLVCdMW6QjDepN86FWdH7z6Mn-c0RNNzdNR1ONA4x0arUptcWVX8F81tXueFNdom9Olf6G6cQ3pzEoKpjAZbVol6dqRcGGMM5Jt94B7DoVHQLJ02S6fNr04T_ORWOa97av-gv0tMgDoCX7mjwz9UzZsPVxdH6U8WKq7m</recordid><startdate>202407</startdate><enddate>202407</enddate><creator>Wang, Buqing</creator><creator>Wang, Yasen</creator><creator>Zhou, Xiaoman</creator><creator>Gao, Xiao‐Dong</creator><creator>Fujita, Morihisa</creator><creator>Li, Zijie</creator><general>John Wiley & Sons, Ltd</general><general>John Wiley and Sons, Limited</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QF</scope><scope>7QL</scope><scope>7QQ</scope><scope>7QR</scope><scope>7SC</scope><scope>7SE</scope><scope>7SN</scope><scope>7SP</scope><scope>7SR</scope><scope>7ST</scope><scope>7T5</scope><scope>7T7</scope><scope>7TA</scope><scope>7TB</scope><scope>7TM</scope><scope>7U5</scope><scope>7U9</scope><scope>8BQ</scope><scope>8FD</scope><scope>C1K</scope><scope>F28</scope><scope>FR3</scope><scope>H8D</scope><scope>H8G</scope><scope>H94</scope><scope>JG9</scope><scope>JQ2</scope><scope>KR7</scope><scope>L7M</scope><scope>L~C</scope><scope>L~D</scope><scope>M7N</scope><scope>P64</scope><scope>SOI</scope><scope>7X8</scope><scope>7S9</scope><scope>L.6</scope><orcidid>https://orcid.org/0000-0002-5618-1862</orcidid></search><sort><creationdate>202407</creationdate><title>Highly efficient expression of Rasamsonia emersonii lipase in Pichia pastoris: characterization and gastrointestinal simulated digestion in vitro</title><author>Wang, Buqing ; Wang, Yasen ; Zhou, Xiaoman ; Gao, Xiao‐Dong ; Fujita, Morihisa ; Li, Zijie</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3490-7384c4efd58f64baad5cf01592e1defa98a65b175380c3640b1aac4b7a06f6133</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2024</creationdate><topic>acidic lipase</topic><topic>agriculture</topic><topic>biodiesel</topic><topic>Biodiesel fuels</topic><topic>Biofuels</topic><topic>Bioreactors</topic><topic>Biosynthesis</topic><topic>calcium</topic><topic>Calcium ions</topic><topic>carboxylic ester hydrolases</topic><topic>codon usage</topic><topic>Combinatorial analysis</topic><topic>culture flasks</topic><topic>Digestion</topic><topic>Digestive system</topic><topic>E coli</topic><topic>Enzymatic activity</topic><topic>Enzyme activity</topic><topic>Escherichia coli</topic><topic>Fermentation</topic><topic>Food industry</topic><topic>gastrointestinal simulated digestion</topic><topic>Gastrointestinal tract</topic><topic>in vitro digestion</topic><topic>Industrial applications</topic><topic>Intestine</topic><topic>intestines</topic><topic>Komagataella pastoris</topic><topic>Lipase</topic><topic>lipid metabolism</topic><topic>Lipids</topic><topic>methanol</topic><topic>molecular chaperone</topic><topic>molecular chaperones</topic><topic>Optimization</topic><topic>Pepsin</topic><topic>Pharmaceutical industry</topic><topic>Pichia pastoris</topic><topic>Rasamsonia</topic><topic>secretion</topic><topic>signal peptide</topic><topic>sodium</topic><topic>stomach</topic><topic>Trypsin</topic><topic>vacuoles</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wang, Buqing</creatorcontrib><creatorcontrib>Wang, Yasen</creatorcontrib><creatorcontrib>Zhou, Xiaoman</creatorcontrib><creatorcontrib>Gao, Xiao‐Dong</creatorcontrib><creatorcontrib>Fujita, Morihisa</creatorcontrib><creatorcontrib>Li, Zijie</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>Aluminium Industry Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Ceramic Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Computer and Information Systems Abstracts</collection><collection>Corrosion Abstracts</collection><collection>Ecology Abstracts</collection><collection>Electronics & Communications Abstracts</collection><collection>Engineered Materials Abstracts</collection><collection>Environment Abstracts</collection><collection>Immunology Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Materials Business File</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ANTE: Abstracts in New Technology & Engineering</collection><collection>Engineering Research Database</collection><collection>Aerospace Database</collection><collection>Copper Technical Reference Library</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Materials Research Database</collection><collection>ProQuest Computer Science Collection</collection><collection>Civil Engineering Abstracts</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>Computer and Information Systems Abstracts Academic</collection><collection>Computer and Information Systems Abstracts Professional</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Environment Abstracts</collection><collection>MEDLINE - Academic</collection><collection>AGRICOLA</collection><collection>AGRICOLA - Academic</collection><jtitle>Journal of the science of food and agriculture</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wang, Buqing</au><au>Wang, Yasen</au><au>Zhou, Xiaoman</au><au>Gao, Xiao‐Dong</au><au>Fujita, Morihisa</au><au>Li, Zijie</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Highly efficient expression of Rasamsonia emersonii lipase in Pichia pastoris: characterization and gastrointestinal simulated digestion in vitro</atitle><jtitle>Journal of the science of food and agriculture</jtitle><addtitle>J Sci Food Agric</addtitle><date>2024-07</date><risdate>2024</risdate><volume>104</volume><issue>9</issue><spage>5603</spage><epage>5613</epage><pages>5603-5613</pages><issn>0022-5142</issn><eissn>1097-0010</eissn><abstract>BACKGROUND
Acidic lipases with high catalytic activities under acidic conditions have important application values in the food, feed and pharmaceutical industries. However, the availability of acidic lipases is still the main obstacle to their industrial applications. Although a novel acidic lipase Rasamsonia emersonii (LIPR) was heterologously expressed in Escherichia coli, the expression level was unsatisfactory.
RESULTS
To achieve the high‐efficiency expression and secretion of LIPR in Pichia pastoris GS115, the combinatorial optimization strategy was adopted including gene codon preference, signal peptide, molecular chaperone co‐expression and disruption of vacuolar sorting receptor VPS10. The activity of the combinatorial optimization engineered strain in a shake flask reached 1480 U mL−1, which was 8.13 times greater than the P. pastoris GS115 parental strain. After high‐density fermentation in a 5‐L bioreactor, the highest enzyme activity reached as high as 11 820 U mL−1. LIPR showed the highest activity at 40 °C and pH 4.0 in the presence of Ca2+ ion. LIPR exhibited strong tolerance to methanol, indicating its potential application in biodiesel biosynthesis. Moreover, the gastrointestinal digestion simulation results demonstrated that LIPR was tolerant to pepsin and trypsin, but its activity was inhibited by sodium taurodeoxycholate.
CONCLUSION
This study provided an effective approach for the high expression of acidic lipase LIPR. LIPR was more appropriate for lipid digestion in the stomach than in intestine according to the gastrointestinal digestion simulation results. © 2024 Society of Chemical Industry.</abstract><cop>Chichester, UK</cop><pub>John Wiley & Sons, Ltd</pub><pmid>38363126</pmid><doi>10.1002/jsfa.13390</doi><tpages>11</tpages><orcidid>https://orcid.org/0000-0002-5618-1862</orcidid></addata></record> |
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| subjects | acidic lipase agriculture biodiesel Biodiesel fuels Biofuels Bioreactors Biosynthesis calcium Calcium ions carboxylic ester hydrolases codon usage Combinatorial analysis culture flasks Digestion Digestive system E coli Enzymatic activity Enzyme activity Escherichia coli Fermentation Food industry gastrointestinal simulated digestion Gastrointestinal tract in vitro digestion Industrial applications Intestine intestines Komagataella pastoris Lipase lipid metabolism Lipids methanol molecular chaperone molecular chaperones Optimization Pepsin Pharmaceutical industry Pichia pastoris Rasamsonia secretion signal peptide sodium stomach Trypsin vacuoles |
| title | Highly efficient expression of Rasamsonia emersonii lipase in Pichia pastoris: characterization and gastrointestinal simulated digestion in vitro |
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