Fructose-Based Production of Short-Chain-Length and Medium-Chain-Length Polyhydroxyalkanoate Copolymer by Arctic Pseudomonas sp. B14-6

Polymers - Tập 13 Số 9 - Trang 1398
Taekjib Choi1, Hwang‐Phill Kim1, Hun‐Suk Song1, Sun‐Mi Lee1, Sol Lee Park1, Hye Soo Lee1, Hyun‐Joong Kim1, Shashi Kant Bhatia1, Ranjit Gurav1, Kwon‐Young Choi2, Yoo Kyung Lee3, Yung‐Hun Yang1
1Department of Biological Engineering, College of Engineering, Konkuk University, Hwayang-dong, Gwangjin-gu, Seoul 05029, Korea
2Department of Environmental and Safety Engineering, College of Engineering, Ajou University, Suwon-si 16499, Gyeonggi-do, Korea
3Polar Research Institute, Incheon 21990, Korea

Tóm tắt

Arctic bacteria employ various mechanisms to survive harsh conditions, one of which is to accumulate carbon and energy inside the cell in the form of polyhydroxyalkanoate (PHA). Whole-genome sequencing of a new Arctic soil bacterium Pseudomonas sp. B14-6 revealed two PHA-production-related gene clusters containing four PHA synthase genes (phaC). Pseudomonas sp. B14-6 produced poly(6% 3-hydroxybutyrate-co-94% 3-hydroxyalkanoate) from various carbon sources, containing short-chain-length PHA (scl-PHA) and medium-chain-length PHA (mcl-PHA) composed of various monomers analyzed by GC-MS, such as 3-hydroxybutyrate, 3-hydroxyhexanoate, 3-hydroxyoctanoate, 3-hydroxydecanoate, 3-hydroxydodecenoic acid, 3-hydroxydodecanoic acid, and 3-hydroxytetradecanoic acid. By optimizing the PHA production media, we achieved 34.6% PHA content using 5% fructose, and 23.7% PHA content using 5% fructose syrup. Differential scanning calorimetry of the scl-co-mcl PHA determined a glass transition temperature (Tg) of 15.3 °C, melting temperature of 112.8 °C, crystallization temperature of 86.8 °C, and 3.82% crystallinity. In addition, gel permeation chromatography revealed a number average molecular weight of 3.6 × 104, weight average molecular weight of 9.1 × 104, and polydispersity index value of 2.5. Overall, the novel Pseudomonas sp. B14-6 produced a polymer with high medium-chain-length content, low Tg, and low crystallinity, indicating its potential use in medical applications.

Từ khóa


Tài liệu tham khảo

Harding, 2011, Microbes in High Arctic Snow and Implications for the Cold Biosphere, Appl. Environ. Microbiol., 77, 3234, 10.1128/AEM.02611-10

Ayub, 2007, The polyhydroxyalkanoate genes of a stress resistant Antarctic Pseudomonas are situated within a genomic island, Plasmid, 58, 240, 10.1016/j.plasmid.2007.05.003

Deming, 2002, Psychrophiles and polar regions, Curr. Opin. Microbiol., 5, 301, 10.1016/S1369-5274(02)00329-6

Lee, 2004, Growth temperature-dependent conversion of de novo-synthesized unsaturated fatty acids into polyhydroxyalkanoic acid and membrane cyclopropane fatty acids in the psychrotrophic bacterium Pseudomonas fluorescens BM07, J. Microbiol. Biotechnol., 14, 1217

Verlinden, 2007, Bacterial synthesis of biodegradable polyhydroxyalkanoates, J. Appl. Microbiol., 102, 1437, 10.1111/j.1365-2672.2007.03335.x

Obruca, S., Sedlacek, P., Krzyzanek, V., Mravec, F., Hrubanova, K., Samek, O., Kucera, D., Benesova, P., and Marova, I. (2016). Accumulation of Poly(3-hydroxybutyrate) Helps Bacterial Cells to Survive Freezing. PLoS ONE, 11.

Ayub, 2008, Polyhydroxyalkanoates are essential for maintenance of redox state in the Antarctic bacterium Pseudomonas sp. 14-3 during low temperature adaptation, Extremophiles, 13, 59, 10.1007/s00792-008-0197-z

Obruca, 2018, Involvement of polyhydroxyalkanoates in stress resistance of microbial cells: Biotechnological consequences and applications, Biotechnol. Adv., 36, 856, 10.1016/j.biotechadv.2017.12.006

Bhatia, 2021, Biowaste-to-bioplastic (polyhydroxyalkanoates): Conversion technologies, strategies, challenges, and perspective, Bioresour. Technol., 326, 124733, 10.1016/j.biortech.2021.124733

Luengo, 2003, Bioplastics from microorganisms, Curr. Opin. Microbiol., 6, 251, 10.1016/S1369-5274(03)00040-7

Park, 2012, Advanced bacterial polyhydroxyalkanoates: Towards a versatile and sustainable platform for unnatural tailor-made polyesters, Biotechnol. Adv., 30, 1196, 10.1016/j.biotechadv.2011.11.007

Jiang, G., Hill, D.J., Kowalczuk, M., Johnston, B., Adamus, G., Irorere, V., and Radecka, I. (2016). Carbon Sources for Polyhydroxyalkanoates and an Integrated Biorefinery. Int. J. Mol. Sci., 17.

Danis, 2015, Preparation of poly(3-hydroxybutyrate-co-hydroxyvalerate) films from halophilic archaea and their potential use in drug delivery, Extremophiles, 19, 515, 10.1007/s00792-015-0735-4

Chen, 2015, Engineering Biosynthesis Mechanisms for Diversifying Polyhydroxyalkanoates, Trends Biotechnol., 33, 565, 10.1016/j.tibtech.2015.07.007

Cheng, J., and Charles, T.C. (2016). Functional metagenomics using Pseudomonas putida expands the known diversity of polyhydroxyalkanoate synthases and enables the production of novel polyhydroxyalkanoate copolymers. BioRxiv.

Bhatia, 2019, Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) production from engineered Ralstonia eutropha using synthetic and anaerobically digested food waste derived volatile fatty acids, Int. J. Biol. Macromol., 133, 1, 10.1016/j.ijbiomac.2019.04.083

Kiewisz, 2016, Bacterial polyhydroxyalkanoates: Still fabulous?, Microbiol. Res., 192, 271, 10.1016/j.micres.2016.07.010

Philip, 2007, Polyhydroxyalkanoates: Biodegradable polymers with a range of applications, J. Chem. Technol. Biotechnol., 82, 233, 10.1002/jctb.1667

Yang, 2011, Tailor-made type II Pseudomonas PHA synthases and their use for the biosynthesis of polylactic acid and its copolymer in recombinant Escherichia coli, Appl. Microbiol. Biotechnol., 90, 603, 10.1007/s00253-010-3077-2

Metzler, J.B. (2004). Fatty Acid Biosynthesis and Biologically Significant Acyl Transfer Reactions in Pseudomonads, Springer.

Kessler, 2000, Taxonomic implications of synthesis of poly-beta-hydroxybutyrate and other poly-beta-hydroxyalkanoates by aerobic pseudomonads, Int. J. Syst. Evol. Microbiol., 50, 711, 10.1099/00207713-50-2-711

Solaiman, 2005, Genetic Characterization of the Poly(hydroxyalkanoate) Synthases of Various Pseudomonas oleovorans Strains, Curr. Microbiol., 50, 329, 10.1007/s00284-005-4508-7

Oliveira, 2020, Towards the Production of mcl-PHA with Enriched Dominant Monomer Content: Process Development for the Sugarcane Biorefinery Context, J. Polym. Environ., 28, 844, 10.1007/s10924-019-01637-2

Rodriguez, 2013, Improved production of medium-chain-length Polyhydroxyalkanotes in glucose-based fed-batch cultivations of metabolically engineered Pseudomonas putida strains, J. Microbiol. Biotechnol., 24, 59

Schmauder, 2017, Metabolic engineering to expand the substrate spectrum of Pseudomonas putida toward sucrose, Microbiologyopen, 6, e00473, 10.1002/mbo3.473

Sohn, 2020, Biosynthesis of polyhydroxyalkanoates from sucrose by metabolically engineered Escherichia coli strains, Int. J. Biol. Macromol., 149, 593, 10.1016/j.ijbiomac.2020.01.254

Bhatia, 2018, Engineering of artificial microbial consortia of Ralstonia eutropha and Bacillus subtilis for poly(3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer production from sugarcane sugar without precursor feeding, Bioresour. Technol., 257, 92, 10.1016/j.biortech.2018.02.056

Choi, 2020, Effects of a Δ-9-fatty acid desaturase and a cyclopropane-fatty acid synthase from the novel psychrophile Pseudomonas sp. B14-6 on bacterial membrane properties, J. Ind. Microbiol. Biotechnol., 47, 1045, 10.1007/s10295-020-02333-0

Park, 2020, Fructose based hyper production of poly-3-hydroxybutyrate from Halomonas sp. YLGW01 and impact of carbon sources on bacteria morphologies, Int. J. Biol. Macromol., 154, 929, 10.1016/j.ijbiomac.2020.03.129

Sathiyanarayanan, 2017, Production and characterization of medium-chain-length polyhydroxyalkanoate copolymer from Arctic psychrotrophic bacterium Pseudomonas sp. PAMC 28620, Int. J. Biol. Macromol., 97, 710, 10.1016/j.ijbiomac.2017.01.053

Bhatia, 2019, Bioconversion of barley straw lignin into biodiesel using Rhodococcus sp. YHY01, Bioresour. Technol., 289, 121704, 10.1016/j.biortech.2019.121704

Bhatia, 2017, Microbial biodiesel production from oil palm biomass hydrolysate using marine Rhodococcus sp. YHY01, Bioresour. Technol., 233, 99, 10.1016/j.biortech.2017.02.061

Gumel, A.M., Annuar, M.S.M., and Heidelberg, T. (2012). Biosynthesis and Characterization of Polyhydroxyalkanoates Copolymers Produced by Pseudomonas putida Bet001 Isolated from Palm Oil Mill Effluent. PLoS ONE, 7.

Choi, 2020, Production of low molecular weight P (3HB-co-3HV) by butyrateacetoacetate CoA-transferase (cftAB) in Escherichia coli, Biotechnol. Bioprocess Eng., 25, 279, 10.1007/s12257-019-0366-1

Hokamura, 2015, Characterization and identification of the proteins bound to two types of polyhydroxyalkanoate granules in Pseudomonas sp. 61-3, Biosci. Biotechnol. Biochem., 79, 1369, 10.1080/09168451.2015.1023250

Choi, 2020, Microbial Polyhydroxyalkanoates and Nonnatural Polyesters, Adv. Mater., 32, e1907138, 10.1002/adma.201907138

Pacheco, 2019, Exploiting the natural poly(3-hydroxyalkanoates) production capacity of Antarctic Pseudomonas strains: From unique phenotypes to novel biopolymers, J. Ind. Microbiol. Biotechnol., 46, 1139, 10.1007/s10295-019-02186-2

Kato, 1996, Production of a novel copolyester of 3-hydroxybutyric acid and medium-chain-length 3-hydroxyalkanoic acids by Pseudomonas sp. 61-3 from sugars, Appl. Microbiol. Biotechnol., 45, 363, 10.1007/s002530050697

Matsusaki, 1998, Cloning and Molecular Analysis of the Poly(3-hydroxybutyrate) and Poly(3-hydroxybutyrate-co-3-hydroxyalkanoate) Biosynthesis Genes in Pseudomonas sp. Strain 61-3, J. Bacteriol., 180, 6459, 10.1128/JB.180.24.6459-6467.1998

Pacheco, 2019, In-Depth Genomic and Phenotypic Characterization of the Antarctic Psychrotolerant Strain Pseudomonas sp. MPC6 Reveals Unique Metabolic Features, Plasticity, and Biotechnological Potential, Front. Microbiol., 10, 1154, 10.3389/fmicb.2019.01154

Goh, 2012, Polyhydroxyalkanoate production by antarctic soil bacteria isolated from Casey Station and Signy Island, Microbiol. Res., 167, 211, 10.1016/j.micres.2011.08.002

Li, 2013, Psychrotrophic Pseudomonas mandelii CBS-1 produces high levels of poly-β-hydroxybutyrate, SpringerPlus, 2, 335, 10.1186/2193-1801-2-335

Lee, 2001, Accumulation of Polyhydroxyalkanoic Acid Containing Large Amounts of Unsaturated Monomers in Pseudomonas fluorescens BM07 Utilizing Saccharides and Its Inhibition by 2-Bromooctanoic Acid, Appl. Environ. Microbiol., 67, 4963, 10.1128/AEM.67.11.4963-4974.2001

Choonut, A., Prasertsan, P., Klomklao, S., and Sangkharak, K. (2020). Study on mcl-PHA Production by Novel Thermotolerant Gram-Positive Isolate. J. Polym. Environ., 28.

Wellen, 2015, Melting and crystallization of poly(3-hydroxybutyrate): Effect of heating/cooling rates on phase transformation, Polímeros, 25, 296, 10.1590/0104-1428.1961

Kong, 2002, The measurement of the crystallinity of polymers by DSC, Polymer, 43, 3873, 10.1016/S0032-3861(02)00235-5

Matsusaki, 2000, Biosynthesis and properties of poly(3-hydroxybutyrate-co-3-hydroxyalkanoates) by recombinant strains of Pseudomonas sp. 61-3, Biomacromolecules, 1, 17, 10.1021/bm9900040

Abe, 1994, Synthesis and Characterization of Poly[(R,S)-3-hydroxybutyrate-b-6-hydroxyhexanoate] as a Compatibilizer for a Biodegradable Blend of Poly[(R)-3-hydroxybutyrate] and Poly(6-hydroxyhexanoate), Macromole, 27, 6012, 10.1021/ma00099a012

Brandrup, J., Immergut, E.H., Grulke, E.A., Abe, A., and Bloch, D.R. (1999). Polymer Handbook, Wiley.