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Biosynthesis of High-Molecular-Weight Poly(d-lactate)-Containing Block Copolyesters Using Evolved Sequence-Regulating Polyhydroxyalkanoate Synthase PhaC(AR)
| Title: | Biosynthesis of High-Molecular-Weight Poly(d-lactate)-Containing Block Copolyesters Using Evolved Sequence-Regulating Polyhydroxyalkanoate Synthase PhaC(AR) |
| Authors: | Phan, Hien Thi Browse this author | | Furukawa, Shoko Browse this author | | Imai, Koto Browse this author | | Tomita, Hiroya Browse this author | | Isono, Takuya Browse this author →KAKEN DB | | Satoh, Toshifumi Browse this author →KAKEN DB | | Matsumoto, Ken'ichiro Browse this author →KAKEN DB |
| Keywords: | PHA synthase | | in vitro evolution | | sequenceregulation | | PDLA-like copolymers | | block copolymer |
| Issue Date: | 31-Jul-2023 |
| Publisher: | American Chemical Society |
| Journal Title: | ACS sustainable chemistry & engineering |
| Volume: | 11 |
| Issue: | 30 |
| Start Page: | 11123 |
| End Page: | 11129 |
| Publisher DOI: | 10.1021/acssuschemeng.3c01696 |
| Abstract: | Bacterial polyhydroxyalkanoate (PHA) synthase PhaC(AR) is a unique enzyme that can synthesize block copolymers.In thisstudy, poly(d-lactate) (PDLA)-containing block copolymerswere synthesized using PhaC(AR) and its mutated variants.Recombinant Escherichia coli harboring phaC (AR) and relevant genes were cultivated withsupplementation of the corresponding monomer precursors. Consequently,PhaC(AR) synthesized poly(3-hydroxybutyrate)-b-2 mol % PDLA [P(3HB)-b-PDLA]. The incorporationof the d-lactate (LA) enantiomer was confirmed by chiralgas chromatography. Previously identified beneficial mutations inPhaC(AR), N149D (ND), and F314H (FH), which increased activitytoward a medium-chain-length substrate 3-hydroxyhexanoyl (3HHx)-CoA,improved the incorporation of LA units. The combined pairwise mutationNDFH synergistically increased the LA fraction to 21 mol % in P(3HB)-b-PDLA. Interestingly, a large amount of LA units (51 mol%) was incorporated by copolymerization with 3HHx units, which yieldedP(3HHx)-b-PDLA. The block copolymerization of 3HHxand D-LA units was confirmed by NMR analyses and solvent fractionationof polymers. The PDLA crystal in P(3HHx)-b-PDLA wasdetected using differential scanning calorimetry and wide-angle X-raydiffraction. Its mass-average molecular weight was 8.6 x 10(5). Thus, block copolymerization utilized high-molecular-weightPDLA as a component of PHAs. Newsynthetic method of block copolyester with isotacticPDLA segment from renewable resources using engineered polyester synthase. |
| Rights: | This document is the Accepted Manuscript version of a Published Work that appeared in final form in ACS sustainable chemistry & engineering, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://pubs.acs.org/articlesonrequest/AOR-6UBMEBYNUBPMUPB9PUZD. |
| Type: | article (author version) |
| URI: | http://hdl.handle.net/2115/92777 |
| Appears in Collections: | 工学院・工学研究院 (Graduate School of Engineering / Faculty of Engineering) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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Submitter: 松本 謙一郎
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