Hyperproduction of poly(4-hydroxybutyrate) from glucose by recombinant <it>Escherichia coli</it>
<p>Abstract</p> <p>Background</p> <p>Poly(4-hydroxybutyrate) [poly(4HB)] is a strong thermoplastic biomaterial with remarkable mechanical properties, biocompatibility and biodegradability. However, it is generally synthesized when 4-hydroxybutyrate (4HB) structurally re...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2012-05-01
|
| Series: | Microbial Cell Factories |
| Subjects: | |
| Online Access: | http://www.microbialcellfactories.com/content/11/1/54 |
| _version_ | 1818963339003297792 |
|---|---|
| author | Zhou Xiao-Yun Yuan Xiao-Xi Shi Zhen-Yu Meng De-Chuang Jiang Wen-Jun Wu Lin-Ping Chen Jin-Chun Chen Guo-Qiang |
| author_facet | Zhou Xiao-Yun Yuan Xiao-Xi Shi Zhen-Yu Meng De-Chuang Jiang Wen-Jun Wu Lin-Ping Chen Jin-Chun Chen Guo-Qiang |
| author_sort | Zhou Xiao-Yun |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Poly(4-hydroxybutyrate) [poly(4HB)] is a strong thermoplastic biomaterial with remarkable mechanical properties, biocompatibility and biodegradability. However, it is generally synthesized when 4-hydroxybutyrate (4HB) structurally related substrates such as γ-butyrolactone, 4-hydroxybutyrate or 1,4-butanediol (1,4-BD) are provided as precursor which are much more expensive than glucose. At present, high production cost is a big obstacle for large scale production of poly(4HB).</p> <p>Results</p> <p>Recombinant <it>Escherichia coli</it> strain was constructed to achieve hyperproduction of poly(4-hydroxybutyrate) [poly(4HB)] using glucose as a sole carbon source. An engineering pathway was established in <it>E. coli</it> containing genes encoding succinate degradation of <it>Clostridium kluyveri</it> and PHB synthase of <it>Ralstonia eutropha.</it> Native succinate semialdehyde dehydrogenase genes <it>sad</it> and <it>gabD</it> in <it>E. coli</it> were both inactivated to enhance the carbon flux to poly(4HB) biosynthesis. Four PHA binding proteins (PhaP or phasins) including PhaP1, PhaP2, PhaP3 and PhaP4 from <it>R. eutropha</it> were heterologously expressed in the recombinant <it>E. coli,</it> respectively, leading to different levels of improvement in poly(4HB) production. Among them PhaP1 exhibited the highest capability for enhanced polymer synthesis. The recombinant <it>E. coli</it> produced 5.5 g L<sup>-1</sup> cell dry weight containing 35.4% poly(4HB) using glucose as a sole carbon source in a 48 h shake flask growth. In a 6-L fermentor study, 11.5 g L<sup>-1</sup> cell dry weight containing 68.2% poly(4HB) was obtained after 52 h of cultivation. This was the highest poly(4HB) yield using glucose as a sole carbon source reported so far. Poly(4HB) was structurally confirmed by gas chromatographic (GC) as well as <sup>1</sup>H and <sup>13</sup>C NMR studies.</p> <p>Conclusions</p> <p>Significant level of poly(4HB) biosynthesis from glucose can be achieved in <it>sad</it> and <it>gabD</it> genes deficient strain of <it>E. coli</it> JM109 harboring an engineering pathway encoding succinate degradation genes and PHB synthase gene, together with expression of four PHA binding proteins PhaP or phasins, respectively. Over 68% poly(4HB) was produced in a fed-batch fermentation process, demonstrating the feasibility for enhanced poly(4HB) production using the recombinant strain for future cost effective commercial development.</p> |
| first_indexed | 2024-12-20T12:43:38Z |
| format | Article |
| id | doaj.art-875b973be89c4ff4884c0adb668b082e |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-12-20T12:43:38Z |
| publishDate | 2012-05-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-875b973be89c4ff4884c0adb668b082e2022-12-21T19:40:23ZengBMCMicrobial Cell Factories1475-28592012-05-011115410.1186/1475-2859-11-54Hyperproduction of poly(4-hydroxybutyrate) from glucose by recombinant <it>Escherichia coli</it>Zhou Xiao-YunYuan Xiao-XiShi Zhen-YuMeng De-ChuangJiang Wen-JunWu Lin-PingChen Jin-ChunChen Guo-Qiang<p>Abstract</p> <p>Background</p> <p>Poly(4-hydroxybutyrate) [poly(4HB)] is a strong thermoplastic biomaterial with remarkable mechanical properties, biocompatibility and biodegradability. However, it is generally synthesized when 4-hydroxybutyrate (4HB) structurally related substrates such as γ-butyrolactone, 4-hydroxybutyrate or 1,4-butanediol (1,4-BD) are provided as precursor which are much more expensive than glucose. At present, high production cost is a big obstacle for large scale production of poly(4HB).</p> <p>Results</p> <p>Recombinant <it>Escherichia coli</it> strain was constructed to achieve hyperproduction of poly(4-hydroxybutyrate) [poly(4HB)] using glucose as a sole carbon source. An engineering pathway was established in <it>E. coli</it> containing genes encoding succinate degradation of <it>Clostridium kluyveri</it> and PHB synthase of <it>Ralstonia eutropha.</it> Native succinate semialdehyde dehydrogenase genes <it>sad</it> and <it>gabD</it> in <it>E. coli</it> were both inactivated to enhance the carbon flux to poly(4HB) biosynthesis. Four PHA binding proteins (PhaP or phasins) including PhaP1, PhaP2, PhaP3 and PhaP4 from <it>R. eutropha</it> were heterologously expressed in the recombinant <it>E. coli,</it> respectively, leading to different levels of improvement in poly(4HB) production. Among them PhaP1 exhibited the highest capability for enhanced polymer synthesis. The recombinant <it>E. coli</it> produced 5.5 g L<sup>-1</sup> cell dry weight containing 35.4% poly(4HB) using glucose as a sole carbon source in a 48 h shake flask growth. In a 6-L fermentor study, 11.5 g L<sup>-1</sup> cell dry weight containing 68.2% poly(4HB) was obtained after 52 h of cultivation. This was the highest poly(4HB) yield using glucose as a sole carbon source reported so far. Poly(4HB) was structurally confirmed by gas chromatographic (GC) as well as <sup>1</sup>H and <sup>13</sup>C NMR studies.</p> <p>Conclusions</p> <p>Significant level of poly(4HB) biosynthesis from glucose can be achieved in <it>sad</it> and <it>gabD</it> genes deficient strain of <it>E. coli</it> JM109 harboring an engineering pathway encoding succinate degradation genes and PHB synthase gene, together with expression of four PHA binding proteins PhaP or phasins, respectively. Over 68% poly(4HB) was produced in a fed-batch fermentation process, demonstrating the feasibility for enhanced poly(4HB) production using the recombinant strain for future cost effective commercial development.</p>http://www.microbialcellfactories.com/content/11/1/54Poly(4HB)PHBPolyhydroxyalkanoatesPhaP4-hydroxybutyrate<it>Escherichia coli</it>Metabolic engineeringSynthetic biology |
| spellingShingle | Zhou Xiao-Yun Yuan Xiao-Xi Shi Zhen-Yu Meng De-Chuang Jiang Wen-Jun Wu Lin-Ping Chen Jin-Chun Chen Guo-Qiang Hyperproduction of poly(4-hydroxybutyrate) from glucose by recombinant <it>Escherichia coli</it> Microbial Cell Factories Poly(4HB) PHB Polyhydroxyalkanoates PhaP 4-hydroxybutyrate <it>Escherichia coli</it> Metabolic engineering Synthetic biology |
| title | Hyperproduction of poly(4-hydroxybutyrate) from glucose by recombinant <it>Escherichia coli</it> |
| title_full | Hyperproduction of poly(4-hydroxybutyrate) from glucose by recombinant <it>Escherichia coli</it> |
| title_fullStr | Hyperproduction of poly(4-hydroxybutyrate) from glucose by recombinant <it>Escherichia coli</it> |
| title_full_unstemmed | Hyperproduction of poly(4-hydroxybutyrate) from glucose by recombinant <it>Escherichia coli</it> |
| title_short | Hyperproduction of poly(4-hydroxybutyrate) from glucose by recombinant <it>Escherichia coli</it> |
| title_sort | hyperproduction of poly 4 hydroxybutyrate from glucose by recombinant it escherichia coli it |
| topic | Poly(4HB) PHB Polyhydroxyalkanoates PhaP 4-hydroxybutyrate <it>Escherichia coli</it> Metabolic engineering Synthetic biology |
| url | http://www.microbialcellfactories.com/content/11/1/54 |
| work_keys_str_mv | AT zhouxiaoyun hyperproductionofpoly4hydroxybutyratefromglucosebyrecombinantitescherichiacoliit AT yuanxiaoxi hyperproductionofpoly4hydroxybutyratefromglucosebyrecombinantitescherichiacoliit AT shizhenyu hyperproductionofpoly4hydroxybutyratefromglucosebyrecombinantitescherichiacoliit AT mengdechuang hyperproductionofpoly4hydroxybutyratefromglucosebyrecombinantitescherichiacoliit AT jiangwenjun hyperproductionofpoly4hydroxybutyratefromglucosebyrecombinantitescherichiacoliit AT wulinping hyperproductionofpoly4hydroxybutyratefromglucosebyrecombinantitescherichiacoliit AT chenjinchun hyperproductionofpoly4hydroxybutyratefromglucosebyrecombinantitescherichiacoliit AT chenguoqiang hyperproductionofpoly4hydroxybutyratefromglucosebyrecombinantitescherichiacoliit |