Biosynthesis of Random-Homo Block Copolymer Poly[Glycolate--3-Hydroxybutyrate (3HB)]--Poly(3HB) Using Sequence-Regulating Chimeric Polyhydroxyalkanoate Synthase in

Overview

Journal Front Bioeng Biotechnol

Date 2020 Dec 28

PMID 33364233

Citations 3

Authors

Shuzo Arai

Sayaka Sakakibara

Robin Mareschal

Toshihiko Ooi

Manfred Zinn

Kenichiro Matsumoto

Affiliations

Soon will be listed here.

Abstract

Glycolate (GL)-containing polyhydroxyalkanoate (PHA) was synthesized in expressing the engineered chimeric PHA synthase PhaC and coenzyme A transferase. The cells produced poly[GL--3-hydroxybutyrate (3HB)] with the supplementation of GL and 3HB, thus demonstrating that PhaC is the first known class I PHA synthase that is capable of incorporating GL units. The triad sequence analysis using H nuclear magnetic resonance indicated that the obtained polymer was composed of two distinct regions, a P(GL--3HB) random segment and P(3HB) homopolymer segment. The random segment was estimated to contain a 71 mol% GL molar ratio, which was much greater than the value (15 mol%) previously achieved by using PhaC1 STQK. Differential scanning calorimetry analysis of the polymer films supported the presence of random copolymer and homopolymer phases. The solvent fractionation of the polymer indicated the presence of a covalent linkage between these segments. Therefore, it was concluded that PhaC synthesized a novel random-homo block copolymer, P(GL--3HB)--P(3HB).

Citing Articles

Toward the production of block copolymers in microbial cells: achievements and perspectives.

Matsumoto K Appl Microbiol Biotechnol. 2024; 108(1):164.

PMID: 38252290 PMC: 10803391. DOI: 10.1007/s00253-023-12973-8.

(R/S)-lactate/2-hydroxybutyrate dehydrogenases in and biosynthesis of block copolyesters by Ralstonia eutropha.

Ishihara S, Orita I, Matsumoto K, Fukui T Appl Microbiol Biotechnol. 2023; 107(24):7557-7569.

PMID: 37773219 PMC: 10656315. DOI: 10.1007/s00253-023-12797-6.

Versatile aliphatic polyester biosynthesis system for producing random and block copolymers composed of 2-, 3-, 4-, 5-, and 6-hydroxyalkanoates using the sequence-regulating polyhydroxyalkanoate synthase PhaC.

Satoh K, Kawakami T, Isobe N, Pasquier L, Tomita H, Zinn M Microb Cell Fact. 2022; 21(1):84.

PMID: 35568875 PMC: 9107728. DOI: 10.1186/s12934-022-01811-7.

References

Matsumoto K, Iijima M, Hori C, Utsunomia C, Ooi T, Taguchi S . In Vitro Analysis of d-Lactyl-CoA-Polymerizing Polyhydroxyalkanoate Synthase in Polylactate and Poly(lactate- co-3-hydroxybutyrate) Syntheses. Biomacromolecules. 2018; 19(7):2889-2895. DOI: 10.1021/acs.biomac.8b00454. View

Matsumoto K, Kageyama Y . Increased Production and Molecular Weight of Artificial Polyhydroxyalkanoate Poly(2-hydroxybutyrate) Above the Glass Transition Temperature Threshold. Front Bioeng Biotechnol. 2019; 7:177. PMC: 6689969. DOI: 10.3389/fbioe.2019.00177. View

Han X, Satoh Y, Satoh T, Matsumoto K, Kakuchi T, Taguchi S . Chemo-enzymatic synthesis of polyhydroxyalkanoate (PHA) incorporating 2-hydroxybutyrate by wild-type class I PHA synthase from Ralstonia eutropha. Appl Microbiol Biotechnol. 2011; 92(3):509-17. DOI: 10.1007/s00253-011-3362-8. View

Matsusaki H, Abe H, Taguchi K, Fukui T, Doi Y . Biosynthesis of poly(3-hydroxybutyrate-co-3-hydroxyalkanoates) by recombinant bacteria expressing the PHA synthase gene phaC1 from Pseudomonas sp. 61-3. Appl Microbiol Biotechnol. 2000; 53(4):401-9. DOI: 10.1007/s002530051633. View

Tan H, Chek M, Lakshmanan M, Foong C, Hakoshima T, Sudesh K . Evaluation of BP-M-CPF4 polyhydroxyalkanoate (PHA) synthase on the production of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) from plant oil using Cupriavidus necator transformants. Int J Biol Macromol. 2020; 159:250-257. DOI: 10.1016/j.ijbiomac.2020.05.064. View

Matsumoto K, Shiba T, Hiraide Y, Taguchi S . Incorporation of Glycolate Units Promotes Hydrolytic Degradation in Flexible Poly(glycolate--3-hydroxybutyrate) Synthesized by Engineered . ACS Biomater Sci Eng. 2021; 3(12):3058-3063. DOI: 10.1021/acsbiomaterials.6b00194. View

Ivorra-Martinez J, Manuel-Manogil J, Boronat T, Sanchez-Nacher L, Balart R, Quiles-Carrillo L . Development and Characterization of Sustainable Composites from Bacterial Polyester Poly(3-Hydroxybutyrate3-hydroxyhexanoate) and Almond Shell Flour by Reactive Extrusion with Oligomers of Lactic Acid. Polymers (Basel). 2020; 12(5). PMC: 7285348. DOI: 10.3390/polym12051097. View

Morohoshi T, Oi T, Aiso H, Suzuki T, Okura T, Sato S . Biofilm Formation and Degradation of Commercially Available Biodegradable Plastic Films by Bacterial Consortiums in Freshwater Environments. Microbes Environ. 2018; 33(3):332-335. PMC: 6167122. DOI: 10.1264/jsme2.ME18033. View

Saad B, Neuenschwander P, Uhlschmid G, Suter U . New versatile, elastomeric, degradable polymeric materials for medicine. Int J Biol Macromol. 1999; 25(1-3):293-301. DOI: 10.1016/s0141-8130(99)00044-6. View

10.

Chen G, Zhang J . Microbial polyhydroxyalkanoates as medical implant biomaterials. Artif Cells Nanomed Biotechnol. 2017; 46(1):1-18. DOI: 10.1080/21691401.2017.1371185. View

11.

Morohoshi T, Oi T, Suzuki T, Sato S . Identification and characterization of a novel extracellular polyhydroxyalkanoate depolymerase in the complete genome sequence of Undibacterium sp. KW1 and YM2 strains. PLoS One. 2020; 15(5):e0232698. PMC: 7199957. DOI: 10.1371/journal.pone.0232698. View

12.

Basnett P, Marcello E, Lukasiewicz B, Panchal B, Nigmatullin R, Knowles J . Biosynthesis and characterization of a novel, biocompatible medium chain length polyhydroxyalkanoate by Pseudomonas mendocina CH50 using coconut oil as the carbon source. J Mater Sci Mater Med. 2018; 29(12):179. DOI: 10.1007/s10856-018-6183-9. View

13.

Hiroe A, Tsuge K, Nomura C, Itaya M, Tsuge T . Rearrangement of gene order in the phaCAB operon leads to effective production of ultrahigh-molecular-weight poly[(R)-3-hydroxybutyrate] in genetically engineered Escherichia coli. Appl Environ Microbiol. 2012; 78(9):3177-84. PMC: 3346486. DOI: 10.1128/AEM.07715-11. View

14.

Taguchi S, Yamada M, Matsumoto K, Tajima K, Satoh Y, Munekata M . A microbial factory for lactate-based polyesters using a lactate-polymerizing enzyme. Proc Natl Acad Sci U S A. 2008; 105(45):17323-7. PMC: 2582312. DOI: 10.1073/pnas.0805653105. View

15.

Dai S, Xue L, Zinn M, Li Z . Enzyme-catalyzed polycondensation of polyester macrodiols with divinyl adipate: a green method for the preparation of thermoplastic block copolyesters. Biomacromolecules. 2009; 10(12):3176-81. DOI: 10.1021/bm9011634. View

16.

Matsumoto K, Hori C, Fujii R, Takaya M, Ooba T, Ooi T . Dynamic Changes of Intracellular Monomer Levels Regulate Block Sequence of Polyhydroxyalkanoates in Engineered Escherichia coli. Biomacromolecules. 2018; 19(2):662-671. DOI: 10.1021/acs.biomac.7b01768. View

17.

Juengert J, Patterson C, Jendrossek D . Poly(3-Hydroxybutyrate) (PHB) Polymerase PhaC1 and PHB Depolymerase PhaZa1 of Ralstonia eutropha Are Phosphorylated . Appl Environ Microbiol. 2018; 84(13). PMC: 6007124. DOI: 10.1128/AEM.00604-18. View

18.

Zheng Y, Chen J, Ma Y, Chen G . Engineering biosynthesis of polyhydroxyalkanoates (PHA) for diversity and cost reduction. Metab Eng. 2019; 58:82-93. DOI: 10.1016/j.ymben.2019.07.004. View

19.

Matsumoto K, Takase K, Yamamoto Y, Doi Y, Taguchi S . Chimeric enzyme composed of polyhydroxyalkanoate (PHA) synthases from Ralstonia eutropha and Aeromonas caviae enhances production of PHAs in recombinant Escherichia coli. Biomacromolecules. 2009; 10(4):682-5. DOI: 10.1021/bm801386j. View

20.

Matsumoto K, Ishiyama A, Sakai K, Shiba T, Taguchi S . Biosynthesis of glycolate-based polyesters containing medium-chain-length 3-hydroxyalkanoates in recombinant Escherichia coli expressing engineered polyhydroxyalkanoate synthase. J Biotechnol. 2011; 156(3):214-7. DOI: 10.1016/j.jbiotec.2011.07.040. View