The reliance of glycerol utilization by Cupriavidus necator on CO2 fixation and improved glycerol catabolism
While crude glycerol is a cheap carbon source for industrial-scale cultivation of microorganisms, its application relies on fast growth and conversion. The biopolymer producing Cupriavidus necator H16 (synonym: Ralstonia eutropha H16) grows poorly on glycerol. The heterologous expression of glycerol...
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| Veröffentlicht in: | Applied microbiology and biotechnology 2022-04, Vol.106 (7), p.2541-2555 |
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| creator | Strittmatter, Carl Simon Eggers, Jessica Biesgen, Vanessa Pauels, Inga Becker, Florian Steinbüchel, Alexander |
| description | While crude glycerol is a cheap carbon source for industrial-scale cultivation of microorganisms, its application relies on fast growth and conversion. The biopolymer producing
Cupriavidus necator
H16 (synonym:
Ralstonia eutropha
H16) grows poorly on glycerol. The heterologous expression of glycerol facilitator
glpF
, glycerol kinase
glpK
, and glycerol dehydrogenase
glpD
from
E. coli
accelerated the growth considerably. The naturally occurring glycerol utilization is inhibited by low glycerol kinase activity. A limited heterotrophic growth promotes the dependency on autotrophic growth by carbon dioxide (CO
2
) fixation and refixation. As mixotrophic growth occurs in the wildtype due to low consumption rates of glycerol, CO
2
fixation by the Calvin-Benson-Bassham (CBB) cycle is essential. The deletion of both
cbbX
copies encoding putative RuBisCO-activases (AAA + ATPase) resulted in a sharp slowdown of growth and glycerol consumption. Activase activity is necessary for functioning carboxylation by RuBisCO. Each of the two copies compensates for the loss of the other, as suggested by observed expression levels. The strong tendency towards autotrophy supports previous investigations of glycerol growth and emphasizes the versatility of the metabolism of
C. necator
H16. Mixotrophy with glycerol-utilization and CO
2
fixation with a high dependence on the CBB is automatically occurring unless transportation and degradation of glycerol are optimized. Parallel engineering of CO
2
fixation and glycerol degradation is suggested towards application for value-added production from crude glycerol.
Key points
• Growth on glycerol is highly dependent on efficient carbon fixation via CBB cycle.
• CbbX is essential for the efficiency of RuBisCO in C. necator H16.
• Expression of glycerol degradation pathway enzymes accelerates glycerol utilization.
Graphical abstract |
| doi_str_mv | 10.1007/s00253-022-11842-0 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_2644008647</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2647946596</sourcerecordid><originalsourceid>FETCH-LOGICAL-c282t-c5ab32dd0fc308297afa2bf038ce1d149125d22c20b8a7aedab0ac6c34fc06a73</originalsourceid><addsrcrecordid>eNp9kUFr3DAQhUVpodtt_0BOglx6cToay7J9DEuaBBZySc5iLMuJgtbaSvaS7a-vti4s9JDTDMz3Ho95jF0IuBIA9Y8EgFVZAGIhRCOxgA9sJWSZFyXkR7YCUVdFXbXNZ_YlpVcAgY1SK-YfXyyP1jsajeVh4M_-aGwMns-T8-43TS6MvDvyzbyPjg6unxMfraEpRJ4vmwfkg3tbMBp77nb7GA62PxtllrrgXdp9ZZ8G8sl--zfX7OnnzePmrtg-3N5vrreFwQanwlTUldj3MJgSGmxrGgi7AcrGWNEL2QqsekSD0DVUk-2pAzLKlHIwoKgu1-z74puj_JptmvTOJWO9p9GGOWlUUgI0Sp7Qy__Q1zDHMac7UXUrVdWqTOFCmRhSinbQ-Rk7ikctQJ8K0EsBOheg_xagIYvKRZQyPD7beLZ-R_UH4ayKJA</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2647946596</pqid></control><display><type>article</type><title>The reliance of glycerol utilization by Cupriavidus necator on CO2 fixation and improved glycerol catabolism</title><source>Springer Nature - Complete Springer Journals</source><creator>Strittmatter, Carl Simon ; Eggers, Jessica ; Biesgen, Vanessa ; Pauels, Inga ; Becker, Florian ; Steinbüchel, Alexander</creator><creatorcontrib>Strittmatter, Carl Simon ; Eggers, Jessica ; Biesgen, Vanessa ; Pauels, Inga ; Becker, Florian ; Steinbüchel, Alexander</creatorcontrib><description>While crude glycerol is a cheap carbon source for industrial-scale cultivation of microorganisms, its application relies on fast growth and conversion. The biopolymer producing
Cupriavidus necator
H16 (synonym:
Ralstonia eutropha
H16) grows poorly on glycerol. The heterologous expression of glycerol facilitator
glpF
, glycerol kinase
glpK
, and glycerol dehydrogenase
glpD
from
E. coli
accelerated the growth considerably. The naturally occurring glycerol utilization is inhibited by low glycerol kinase activity. A limited heterotrophic growth promotes the dependency on autotrophic growth by carbon dioxide (CO
2
) fixation and refixation. As mixotrophic growth occurs in the wildtype due to low consumption rates of glycerol, CO
2
fixation by the Calvin-Benson-Bassham (CBB) cycle is essential. The deletion of both
cbbX
copies encoding putative RuBisCO-activases (AAA + ATPase) resulted in a sharp slowdown of growth and glycerol consumption. Activase activity is necessary for functioning carboxylation by RuBisCO. Each of the two copies compensates for the loss of the other, as suggested by observed expression levels. The strong tendency towards autotrophy supports previous investigations of glycerol growth and emphasizes the versatility of the metabolism of
C. necator
H16. Mixotrophy with glycerol-utilization and CO
2
fixation with a high dependence on the CBB is automatically occurring unless transportation and degradation of glycerol are optimized. Parallel engineering of CO
2
fixation and glycerol degradation is suggested towards application for value-added production from crude glycerol.
Key points
• Growth on glycerol is highly dependent on efficient carbon fixation via CBB cycle.
• CbbX is essential for the efficiency of RuBisCO in C. necator H16.
• Expression of glycerol degradation pathway enzymes accelerates glycerol utilization.
Graphical abstract</description><identifier>ISSN: 0175-7598</identifier><identifier>EISSN: 1432-0614</identifier><identifier>DOI: 10.1007/s00253-022-11842-0</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer Berlin Heidelberg</publisher><subject>Adenosine triphosphatase ; Applied Genetics and Molecular Biotechnology ; Autotrophy ; Biodegradation ; Biomedical and Life Sciences ; Biopolymers ; Biotechnology ; Carbon cycle ; Carbon dioxide ; Carbon dioxide fixation ; Carbon fixation ; Carbon sources ; Carboxylation ; Catabolism ; Consumption ; Cupriavidus necator ; Degradation ; E coli ; Fixation ; Glycerol ; Glycerol dehydrogenase ; Glycerol kinase ; Heterotrophic growth ; Kinases ; Life Sciences ; Microbial Genetics and Genomics ; Microbiology ; Microorganisms ; Mixotrophy ; Ribulose-bisphosphate carboxylase ; Soil bacteria ; Utilization</subject><ispartof>Applied microbiology and biotechnology, 2022-04, Vol.106 (7), p.2541-2555</ispartof><rights>The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2022</rights><rights>The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2022.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c282t-c5ab32dd0fc308297afa2bf038ce1d149125d22c20b8a7aedab0ac6c34fc06a73</citedby><cites>FETCH-LOGICAL-c282t-c5ab32dd0fc308297afa2bf038ce1d149125d22c20b8a7aedab0ac6c34fc06a73</cites><orcidid>0000-0001-9979-9905 ; 0000-0002-6189-834X ; 0000-0003-0387-833X</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00253-022-11842-0$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00253-022-11842-0$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,776,780,27903,27904,41467,42536,51297</link.rule.ids></links><search><creatorcontrib>Strittmatter, Carl Simon</creatorcontrib><creatorcontrib>Eggers, Jessica</creatorcontrib><creatorcontrib>Biesgen, Vanessa</creatorcontrib><creatorcontrib>Pauels, Inga</creatorcontrib><creatorcontrib>Becker, Florian</creatorcontrib><creatorcontrib>Steinbüchel, Alexander</creatorcontrib><title>The reliance of glycerol utilization by Cupriavidus necator on CO2 fixation and improved glycerol catabolism</title><title>Applied microbiology and biotechnology</title><addtitle>Appl Microbiol Biotechnol</addtitle><description>While crude glycerol is a cheap carbon source for industrial-scale cultivation of microorganisms, its application relies on fast growth and conversion. The biopolymer producing
Cupriavidus necator
H16 (synonym:
Ralstonia eutropha
H16) grows poorly on glycerol. The heterologous expression of glycerol facilitator
glpF
, glycerol kinase
glpK
, and glycerol dehydrogenase
glpD
from
E. coli
accelerated the growth considerably. The naturally occurring glycerol utilization is inhibited by low glycerol kinase activity. A limited heterotrophic growth promotes the dependency on autotrophic growth by carbon dioxide (CO
2
) fixation and refixation. As mixotrophic growth occurs in the wildtype due to low consumption rates of glycerol, CO
2
fixation by the Calvin-Benson-Bassham (CBB) cycle is essential. The deletion of both
cbbX
copies encoding putative RuBisCO-activases (AAA + ATPase) resulted in a sharp slowdown of growth and glycerol consumption. Activase activity is necessary for functioning carboxylation by RuBisCO. Each of the two copies compensates for the loss of the other, as suggested by observed expression levels. The strong tendency towards autotrophy supports previous investigations of glycerol growth and emphasizes the versatility of the metabolism of
C. necator
H16. Mixotrophy with glycerol-utilization and CO
2
fixation with a high dependence on the CBB is automatically occurring unless transportation and degradation of glycerol are optimized. Parallel engineering of CO
2
fixation and glycerol degradation is suggested towards application for value-added production from crude glycerol.
Key points
• Growth on glycerol is highly dependent on efficient carbon fixation via CBB cycle.
• CbbX is essential for the efficiency of RuBisCO in C. necator H16.
• Expression of glycerol degradation pathway enzymes accelerates glycerol utilization.
Graphical abstract</description><subject>Adenosine triphosphatase</subject><subject>Applied Genetics and Molecular Biotechnology</subject><subject>Autotrophy</subject><subject>Biodegradation</subject><subject>Biomedical and Life Sciences</subject><subject>Biopolymers</subject><subject>Biotechnology</subject><subject>Carbon cycle</subject><subject>Carbon dioxide</subject><subject>Carbon dioxide fixation</subject><subject>Carbon fixation</subject><subject>Carbon sources</subject><subject>Carboxylation</subject><subject>Catabolism</subject><subject>Consumption</subject><subject>Cupriavidus necator</subject><subject>Degradation</subject><subject>E coli</subject><subject>Fixation</subject><subject>Glycerol</subject><subject>Glycerol dehydrogenase</subject><subject>Glycerol kinase</subject><subject>Heterotrophic growth</subject><subject>Kinases</subject><subject>Life Sciences</subject><subject>Microbial Genetics and Genomics</subject><subject>Microbiology</subject><subject>Microorganisms</subject><subject>Mixotrophy</subject><subject>Ribulose-bisphosphate carboxylase</subject><subject>Soil bacteria</subject><subject>Utilization</subject><issn>0175-7598</issn><issn>1432-0614</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2022</creationdate><recordtype>article</recordtype><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp9kUFr3DAQhUVpodtt_0BOglx6cToay7J9DEuaBBZySc5iLMuJgtbaSvaS7a-vti4s9JDTDMz3Ho95jF0IuBIA9Y8EgFVZAGIhRCOxgA9sJWSZFyXkR7YCUVdFXbXNZ_YlpVcAgY1SK-YfXyyP1jsajeVh4M_-aGwMns-T8-43TS6MvDvyzbyPjg6unxMfraEpRJ4vmwfkg3tbMBp77nb7GA62PxtllrrgXdp9ZZ8G8sl--zfX7OnnzePmrtg-3N5vrreFwQanwlTUldj3MJgSGmxrGgi7AcrGWNEL2QqsekSD0DVUk-2pAzLKlHIwoKgu1-z74puj_JptmvTOJWO9p9GGOWlUUgI0Sp7Qy__Q1zDHMac7UXUrVdWqTOFCmRhSinbQ-Rk7ikctQJ8K0EsBOheg_xagIYvKRZQyPD7beLZ-R_UH4ayKJA</recordid><startdate>20220401</startdate><enddate>20220401</enddate><creator>Strittmatter, Carl Simon</creator><creator>Eggers, 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reliance of glycerol utilization by Cupriavidus necator on CO2 fixation and improved glycerol catabolism</title><author>Strittmatter, Carl Simon ; Eggers, Jessica ; Biesgen, Vanessa ; Pauels, Inga ; Becker, Florian ; Steinbüchel, Alexander</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c282t-c5ab32dd0fc308297afa2bf038ce1d149125d22c20b8a7aedab0ac6c34fc06a73</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2022</creationdate><topic>Adenosine triphosphatase</topic><topic>Applied Genetics and Molecular Biotechnology</topic><topic>Autotrophy</topic><topic>Biodegradation</topic><topic>Biomedical and Life Sciences</topic><topic>Biopolymers</topic><topic>Biotechnology</topic><topic>Carbon cycle</topic><topic>Carbon dioxide</topic><topic>Carbon dioxide fixation</topic><topic>Carbon fixation</topic><topic>Carbon sources</topic><topic>Carboxylation</topic><topic>Catabolism</topic><topic>Consumption</topic><topic>Cupriavidus necator</topic><topic>Degradation</topic><topic>E coli</topic><topic>Fixation</topic><topic>Glycerol</topic><topic>Glycerol dehydrogenase</topic><topic>Glycerol kinase</topic><topic>Heterotrophic growth</topic><topic>Kinases</topic><topic>Life Sciences</topic><topic>Microbial Genetics and Genomics</topic><topic>Microbiology</topic><topic>Microorganisms</topic><topic>Mixotrophy</topic><topic>Ribulose-bisphosphate carboxylase</topic><topic>Soil bacteria</topic><topic>Utilization</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Strittmatter, Carl Simon</creatorcontrib><creatorcontrib>Eggers, Jessica</creatorcontrib><creatorcontrib>Biesgen, Vanessa</creatorcontrib><creatorcontrib>Pauels, Inga</creatorcontrib><creatorcontrib>Becker, Florian</creatorcontrib><creatorcontrib>Steinbüchel, 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Biotechnol</stitle><date>2022-04-01</date><risdate>2022</risdate><volume>106</volume><issue>7</issue><spage>2541</spage><epage>2555</epage><pages>2541-2555</pages><issn>0175-7598</issn><eissn>1432-0614</eissn><abstract>While crude glycerol is a cheap carbon source for industrial-scale cultivation of microorganisms, its application relies on fast growth and conversion. The biopolymer producing
Cupriavidus necator
H16 (synonym:
Ralstonia eutropha
H16) grows poorly on glycerol. The heterologous expression of glycerol facilitator
glpF
, glycerol kinase
glpK
, and glycerol dehydrogenase
glpD
from
E. coli
accelerated the growth considerably. The naturally occurring glycerol utilization is inhibited by low glycerol kinase activity. A limited heterotrophic growth promotes the dependency on autotrophic growth by carbon dioxide (CO
2
) fixation and refixation. As mixotrophic growth occurs in the wildtype due to low consumption rates of glycerol, CO
2
fixation by the Calvin-Benson-Bassham (CBB) cycle is essential. The deletion of both
cbbX
copies encoding putative RuBisCO-activases (AAA + ATPase) resulted in a sharp slowdown of growth and glycerol consumption. Activase activity is necessary for functioning carboxylation by RuBisCO. Each of the two copies compensates for the loss of the other, as suggested by observed expression levels. The strong tendency towards autotrophy supports previous investigations of glycerol growth and emphasizes the versatility of the metabolism of
C. necator
H16. Mixotrophy with glycerol-utilization and CO
2
fixation with a high dependence on the CBB is automatically occurring unless transportation and degradation of glycerol are optimized. Parallel engineering of CO
2
fixation and glycerol degradation is suggested towards application for value-added production from crude glycerol.
Key points
• Growth on glycerol is highly dependent on efficient carbon fixation via CBB cycle.
• CbbX is essential for the efficiency of RuBisCO in C. necator H16.
• Expression of glycerol degradation pathway enzymes accelerates glycerol utilization.
Graphical abstract</abstract><cop>Berlin/Heidelberg</cop><pub>Springer Berlin Heidelberg</pub><doi>10.1007/s00253-022-11842-0</doi><tpages>15</tpages><orcidid>https://orcid.org/0000-0001-9979-9905</orcidid><orcidid>https://orcid.org/0000-0002-6189-834X</orcidid><orcidid>https://orcid.org/0000-0003-0387-833X</orcidid></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0175-7598 |
| ispartof | Applied microbiology and biotechnology, 2022-04, Vol.106 (7), p.2541-2555 |
| issn | 0175-7598 1432-0614 |
| language | eng |
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| source | Springer Nature - Complete Springer Journals |
| subjects | Adenosine triphosphatase Applied Genetics and Molecular Biotechnology Autotrophy Biodegradation Biomedical and Life Sciences Biopolymers Biotechnology Carbon cycle Carbon dioxide Carbon dioxide fixation Carbon fixation Carbon sources Carboxylation Catabolism Consumption Cupriavidus necator Degradation E coli Fixation Glycerol Glycerol dehydrogenase Glycerol kinase Heterotrophic growth Kinases Life Sciences Microbial Genetics and Genomics Microbiology Microorganisms Mixotrophy Ribulose-bisphosphate carboxylase Soil bacteria Utilization |
| title | The reliance of glycerol utilization by Cupriavidus necator on CO2 fixation and improved glycerol catabolism |
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