Economical synthesis of γ-cyclodextrin catalyzed by oriented cyclodextrin glycosyltransferase displayed on bacterial polyhydroxyalkanoate nanogranules
Abstract Background The advantages of γ-cyclodextrin (γ-CD) include its high solubility, ability to form inclusion complexes with various poorly water-soluble molecules, and favorable toxicological profile; thus, γ-CD is an attractive functional excipient widely used in many industrial settings. Unf...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2023-09-01
|
| Series: | Microbial Cell Factories |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12934-023-02191-2 |
| _version_ | 1797450981346115584 |
|---|---|
| author | Menglu Duan Yan Wang Dan Tan Guowu Yang Yuan Deng Ganqiao Ran Jiao Li |
| author_facet | Menglu Duan Yan Wang Dan Tan Guowu Yang Yuan Deng Ganqiao Ran Jiao Li |
| author_sort | Menglu Duan |
| collection | DOAJ |
| description | Abstract Background The advantages of γ-cyclodextrin (γ-CD) include its high solubility, ability to form inclusion complexes with various poorly water-soluble molecules, and favorable toxicological profile; thus, γ-CD is an attractive functional excipient widely used in many industrial settings. Unfortunately, the high cost of γ-CD caused by the low activity and stability of γ-cyclodextrin glycosyltransferase (γ-CGTase) has hampered large-scale production and application. Results This study reports the in vivo one-step production of immobilized γ-CGTase decorated on the surface of polyhydroxyalkanoate (PHA) nanogranules by the N-terminal fusion of γ-CGTase to PHA synthase via a designed linker. The immobilized γ-CGTase-PHA nanogranules showed outstanding cyclization activity of 61.25 ± 3.94 U/mg (γ-CGTase protein) and hydrolysis activity of 36,273.99 ± 1892.49 U/mg, 44.74% and 18.83% higher than that of free γ-CGTase, respectively. The nanogranules also exhibited wider optimal pH (cyclization activity 7.0–9.0, hydrolysis activity 10.0–11.0) and temperature (55–60 °C) ranges and remarkable thermo- and pH-stability, expanding its utility to adapt to wider and more severe reaction conditions than the free enzyme. A high yield of CDs (22.73%) converted from starch and a high ratio (90.86%) of γ-CD in the catalysate were achieved at pH 9.0 and 50 °C for 10 h with 1 mmol/L K+, Ca2+, and Mg2+ added to the reaction system. Moreover, γ-CGTase-PHA beads can be used at least eight times, retaining 82.04% of its initial hydrolysis activity and 75.73% of its initial cyclization activity. Conclusions This study provides a promising nanobiocatalyst for the cost-efficient production of γ-CD, which could greatly facilitate process control and economize the production cost. |
| first_indexed | 2024-03-09T14:48:21Z |
| format | Article |
| id | doaj.art-6d7d34656e2247a1a575294473c63aad |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-03-09T14:48:21Z |
| publishDate | 2023-09-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-6d7d34656e2247a1a575294473c63aad2023-11-26T14:37:55ZengBMCMicrobial Cell Factories1475-28592023-09-0122111710.1186/s12934-023-02191-2Economical synthesis of γ-cyclodextrin catalyzed by oriented cyclodextrin glycosyltransferase displayed on bacterial polyhydroxyalkanoate nanogranulesMenglu Duan0Yan Wang1Dan Tan2Guowu Yang3Yuan Deng4Ganqiao Ran5Jiao Li6Shaanxi Institute of MicrobiologyShaanxi Institute of MicrobiologyKey Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi’an Jiaotong UniversityShaanxi Institute of MicrobiologyShaanxi Institute of MicrobiologyBio-Agriculture Institute of ShaanxiShaanxi Institute of MicrobiologyAbstract Background The advantages of γ-cyclodextrin (γ-CD) include its high solubility, ability to form inclusion complexes with various poorly water-soluble molecules, and favorable toxicological profile; thus, γ-CD is an attractive functional excipient widely used in many industrial settings. Unfortunately, the high cost of γ-CD caused by the low activity and stability of γ-cyclodextrin glycosyltransferase (γ-CGTase) has hampered large-scale production and application. Results This study reports the in vivo one-step production of immobilized γ-CGTase decorated on the surface of polyhydroxyalkanoate (PHA) nanogranules by the N-terminal fusion of γ-CGTase to PHA synthase via a designed linker. The immobilized γ-CGTase-PHA nanogranules showed outstanding cyclization activity of 61.25 ± 3.94 U/mg (γ-CGTase protein) and hydrolysis activity of 36,273.99 ± 1892.49 U/mg, 44.74% and 18.83% higher than that of free γ-CGTase, respectively. The nanogranules also exhibited wider optimal pH (cyclization activity 7.0–9.0, hydrolysis activity 10.0–11.0) and temperature (55–60 °C) ranges and remarkable thermo- and pH-stability, expanding its utility to adapt to wider and more severe reaction conditions than the free enzyme. A high yield of CDs (22.73%) converted from starch and a high ratio (90.86%) of γ-CD in the catalysate were achieved at pH 9.0 and 50 °C for 10 h with 1 mmol/L K+, Ca2+, and Mg2+ added to the reaction system. Moreover, γ-CGTase-PHA beads can be used at least eight times, retaining 82.04% of its initial hydrolysis activity and 75.73% of its initial cyclization activity. Conclusions This study provides a promising nanobiocatalyst for the cost-efficient production of γ-CD, which could greatly facilitate process control and economize the production cost.https://doi.org/10.1186/s12934-023-02191-2γ-CGTaseγ-CDPolyhydroxyalkanoatesNanogranulesImmobilization |
| spellingShingle | Menglu Duan Yan Wang Dan Tan Guowu Yang Yuan Deng Ganqiao Ran Jiao Li Economical synthesis of γ-cyclodextrin catalyzed by oriented cyclodextrin glycosyltransferase displayed on bacterial polyhydroxyalkanoate nanogranules Microbial Cell Factories γ-CGTase γ-CD Polyhydroxyalkanoates Nanogranules Immobilization |
| title | Economical synthesis of γ-cyclodextrin catalyzed by oriented cyclodextrin glycosyltransferase displayed on bacterial polyhydroxyalkanoate nanogranules |
| title_full | Economical synthesis of γ-cyclodextrin catalyzed by oriented cyclodextrin glycosyltransferase displayed on bacterial polyhydroxyalkanoate nanogranules |
| title_fullStr | Economical synthesis of γ-cyclodextrin catalyzed by oriented cyclodextrin glycosyltransferase displayed on bacterial polyhydroxyalkanoate nanogranules |
| title_full_unstemmed | Economical synthesis of γ-cyclodextrin catalyzed by oriented cyclodextrin glycosyltransferase displayed on bacterial polyhydroxyalkanoate nanogranules |
| title_short | Economical synthesis of γ-cyclodextrin catalyzed by oriented cyclodextrin glycosyltransferase displayed on bacterial polyhydroxyalkanoate nanogranules |
| title_sort | economical synthesis of γ cyclodextrin catalyzed by oriented cyclodextrin glycosyltransferase displayed on bacterial polyhydroxyalkanoate nanogranules |
| topic | γ-CGTase γ-CD Polyhydroxyalkanoates Nanogranules Immobilization |
| url | https://doi.org/10.1186/s12934-023-02191-2 |
| work_keys_str_mv | AT mengluduan economicalsynthesisofgcyclodextrincatalyzedbyorientedcyclodextringlycosyltransferasedisplayedonbacterialpolyhydroxyalkanoatenanogranules AT yanwang economicalsynthesisofgcyclodextrincatalyzedbyorientedcyclodextringlycosyltransferasedisplayedonbacterialpolyhydroxyalkanoatenanogranules AT dantan economicalsynthesisofgcyclodextrincatalyzedbyorientedcyclodextringlycosyltransferasedisplayedonbacterialpolyhydroxyalkanoatenanogranules AT guowuyang economicalsynthesisofgcyclodextrincatalyzedbyorientedcyclodextringlycosyltransferasedisplayedonbacterialpolyhydroxyalkanoatenanogranules AT yuandeng economicalsynthesisofgcyclodextrincatalyzedbyorientedcyclodextringlycosyltransferasedisplayedonbacterialpolyhydroxyalkanoatenanogranules AT ganqiaoran economicalsynthesisofgcyclodextrincatalyzedbyorientedcyclodextringlycosyltransferasedisplayedonbacterialpolyhydroxyalkanoatenanogranules AT jiaoli economicalsynthesisofgcyclodextrincatalyzedbyorientedcyclodextringlycosyltransferasedisplayedonbacterialpolyhydroxyalkanoatenanogranules |