Polyhydroxyalkanoates (PHA) are microbial polyesters that have recently come to the forefront of interest due to their biodegradability and production from renewable sources. A potential increase in competitiveness of PHA production process comes with a combination of the use of thermophilic bacteria with the mutual use of waste substrates. In this work, the thermophilic bacterium Tepidimonas taiwanensis LMG 22826 was identified as a promising PHA producer. The ability to produce PHA in T. taiwanensis was studied both on genotype and phenotype levels. The gene encoding the Class I PHA synthase, a crucial enzyme in PHA synthesis, was detected both by genome database search and by PCR. The microbial culture of T. taiwanensis was capable of efficient utilization of glucose and fructose. When cultivated on glucose as the only carbon source at 50 °C, the PHA titers reached up to 3.55 g/L, and PHA content in cell dry mass was 65%. The preference of fructose and glucose opens the possibility to employ T. taiwanensis for PHA production on various food wastes rich in these abundant sugars. In this work, PHA production on grape pomace extracts was successfully tested.

Biology Centre The Czech Academy of Sciences v v i Branisovska 31 370 05 Ceske Budejovice Czech Republic

Department of Food Chemistry and Biotechnology Faculty of Chemistry Brno University of Technology Purkynova 118 612 00 Brno Czech Republic

Faculty of Science Charles University Vinicna 7 128 43 Prague 2 Czech Republic

Institute of Scientific Instruments of the Czech Academy of Sciences v v i Kralovopolska 147 612 64 Brno Czech Republic

Zobrazit více v PubMed

Lim H., Chuah J., Chek M., Tan H., Hakoshima T., Sudesh K. Identification of regions affecting enzyme activity, substrate binding, dimer stabilization and polyhydroxyalkanoate (PHA) granule morphology in the PHA synthase of Aquitalea sp. USM4. Int. J. Biol. Macromol. 2021;186:414–423. doi: 10.1016/j.ijbiomac.2021.07.041. PubMed DOI

Raza Z., Abid S., Banat I. Polyhydroxyalkanoates: Characteristics, production, recent developments and applications. Int. Biodeterior. Biodegrad. 2018;126:45–56. doi: 10.1016/j.ibiod.2017.10.001. DOI

Peña C., Castillo T., García A., Millán M., Segura D. Biotechnological strategies to improve production of microbial poly-(3-hydroxybutyrate): A review of recent research work. Microb. Biotechnol. 2014;7:278–293. doi: 10.1111/1751-7915.12129. PubMed DOI PMC

Yadav B., Talan A., Tyagi R., Drogui P. Concomitant production of value-added products with polyhydroxyalkanoate (PHA) synthesis: A review. Bioresour. Technol. 2021;337:125419. doi: 10.1016/j.biortech.2021.125419. PubMed DOI

Ray S., Kalia V. Biomedical Applications of Polyhydroxyalkanoates. Indian J. Microbiol. 2017;57:261–269. doi: 10.1007/s12088-017-0651-7. PubMed DOI PMC

Chen G., Patel M. Plastics Derived from Biological Sources: Present and Future. Chem. Rev. 2012;112:2082–2099. doi: 10.1021/cr200162d. PubMed DOI

Poltronieri P., Kumar P. Polyhydroxyalcanoates (PHAs) in Industrial Applications. In: Martínez L., Kharissova O., Kharisov B., editors. Handbook of Ecomaterials. Springer International Publishing; Cham, Switzerland: 2017. pp. 1–30.

Crutchik D., Franchi O., Caminos L., Jeison D., Belmonte M., Pedrouso A., Val del Rio A., Mosquera-Corral A., Campos J. Polyhydroxyalkanoates (PHAs) Production: A Feasible Economic Option for the Treatment of Sewage Sludge in Municipal Wastewater Treatment Plants? Water. 2020;12:1118. doi: 10.3390/w12041118. DOI

Rampelotto P. Extremophiles and Extreme Environments. Life. 2013;3:482–485. doi: 10.3390/life3030482. PubMed DOI PMC

Berlemont R., Gerday C. Comprehensive Biotechnology. Elsevier; Amsterdam, The Netherlands: 2011. Extremophiles; pp. 229–242.

Irwin J. Overview of extremophiles and their food and medical applications. Physiol. Biotechnol. Asp. Extrem. 2020:65–87. doi: 10.1016/B978-0-12-818322-9.00006-X. DOI

Dumorne K., Cordova D., Astorga-Elo M., Renganathan P. Extremozymes: A Potential Source for Industrial Applications. J. Microbiol. Biotechnol. 2017;27:649–659. doi: 10.4014/jmb.1611.11006. PubMed DOI

Chen G., Jiang X. Next generation industrial biotechnology based on extremophilic bacteria. Curr. Opin. Biotechnol. 2018;50:94–100. doi: 10.1016/j.copbio.2017.11.016. PubMed DOI

Quillaguamán J., Guzmán H., Van-Thuoc D., Hatti-Kaul R. Synthesis and production of polyhydroxyalkanoates by halophiles: Current potential and future prospects. Appl. Microbiol. Biotechnol. 2010;85:1687–1696. doi: 10.1007/s00253-009-2397-6. PubMed DOI

Sedlacek P., Slaninova E., Koller M., Nebesarova J., Marova I., Krzyzanek V., Obruca S. PHA granules help bacterial cells to preserve cell integrity when exposed to sudden osmotic imbalances. New Biotechnol. 2019;49:129–136. doi: 10.1016/j.nbt.2018.10.005. PubMed DOI

Alsafadi D., Al-Mashaqbeh O. A one-stage cultivation process for the production of poly-3-(hydroxybutyrate-co-hydroxyvalerate) from olive mill wastewater by Haloferax mediterranei. New Biotechnol. 2017;34:47–53. doi: 10.1016/j.nbt.2016.05.003. PubMed DOI

Tan D., Wu Q., Chen J., Chen G. Engineering Halomonas TD01 for the low-cost production of polyhydroxyalkanoates. Metab. Eng. 2014;26:34–47. doi: 10.1016/j.ymben.2014.09.001. PubMed DOI

Kucera D., Pernicová I., Kovalcik A., Koller M., Mullerova L., Sedlacek P., Mravec F., Nebesarova J., Kalina M., Marova I., et al. Characterization of the promising poly(3-hydroxybutyrate) producing halophilic bacterium Halomonas halophila. Bioresour. Technol. 2018;256:552–556. doi: 10.1016/j.biortech.2018.02.062. PubMed DOI

Rivera-Terceros P., Tito-Claros E., Torrico S., Carballo S., Van-Thuoc D., Quillaguamán J. Production of poly(3-hydroxybutyrate) by Halomonas boliviensis in an air-lift reactor. J. Biol. Res.-Thessalon. 2015;22:8. doi: 10.1186/s40709-015-0031-6. PubMed DOI PMC

Kulkarni S., Kanekar P., Nilegaonkar S., Sarnaik S., Jog J. Production and characterization of a biodegradable poly (hydroxybutyrate-co-hydroxyvalerate) (PHB-co-PHV) copolymer by moderately haloalkalitolerant Halomonas campisalis MCM B-1027 isolated from Lonar Lake, India. Bioresour. Technol. 2010;101:9765–9771. doi: 10.1016/j.biortech.2010.07.089. PubMed DOI

Pernicova I., Kucera D., Nebesarova J., Kalina M., Novackova I., Koller M., Obruca S. Production of polyhydroxyalkanoates on waste frying oil employing selected Halomonas strains. Bioresour. Technol. 2019;292:122028. doi: 10.1016/j.biortech.2019.122028. PubMed DOI

Ibrahim M., Willems A., Steinbüchel A. Isolation and characterization of new poly(3HB)-accumulating star-shaped cell-aggregates-forming thermophilic bacteria. J. Appl. Microbiol. 2010;109:1579–1590. doi: 10.1111/j.1365-2672.2010.04786.x. PubMed DOI

Sheu D., Chen W., Yang J., Chang R. Thermophilic bacterium Caldimonas taiwanensis produces poly(3-hydroxybutyrate-co-3-hydroxyvalerate) from starch and valerate as carbon sources. Enzym. Microb. Technol. 2009;44:289–294. doi: 10.1016/j.enzmictec.2009.01.004. DOI

Pernicova I., Novackova I., Sedlacek P., Kourilova X., Kalina M., Kovalcik A., Koller M., Nebesarova J., Krzyzanek V., Hrubanova K., et al. Introducing the Newly Isolated Bacterium Aneurinibacillus sp. H1 as an Auspicious Thermophilic Producer of Various Polyhydroxyalkanoates (PHA) Copolymers–1. Isolation and Characterization of the Bacterium. Polymers. 2020;12:1235. doi: 10.3390/polym12061235. PubMed DOI PMC

Kourilova X., Pernicova I., Sedlar K., Musilova J., Sedlacek P., Kalina M., Koller M., Obruca S. Production of polyhydroxyalkanoates (PHA) by a thermophilic strain of Schlegelella thermodepolymerans from xylose rich substrates. Bioresour. Technol. 2020;315:123885. doi: 10.1016/j.biortech.2020.123885. PubMed DOI

2019 Statistical Report on World Vitiviniculture, 1st ed.; International Organisation of Vine and Wine Intergovernmental Organisation: International Organisation of Vine and Wine Intergovernmental Organisation. 2020. [(accessed on 9 September 2021)]. Available online: http://oiv.int/public/medias/6782/oiv-2019-statistical-report-on-world-vitiviniculture.pdf.

Moreno A., Ballesteros M., Negro M. The Interaction of Food Industry and Environment. Academic Press; Cambridge, MA, USA: 2020. Biorefineries for the valorization of food processing waste; pp. 155–190.

Dávila I., Robles E., Egüés I., Labidi J., Gullón P. Handbook of Grape Processing By-Products. Academic Press; Cambridge, MA, USA: 2017. The Biorefinery Concept for the Industrial Valorization of Grape Processing By-Products; pp. 29–53.

Antonić B., Jančíková S., Dordević D., Tremlová B. Grape Pomace Valorization: A Systematic Review and Meta-Analysis. Foods. 2020;9:1627. doi: 10.3390/foods9111627. PubMed DOI PMC

González-Paramás A., Esteban-Ruano S., Santos-Buelga C., de Pascual-Teresa S., Rivas-Gonzalo J. Flavanol Content and Antioxidant Activity in Winery Byproducts. J. Agric. Food Chem. 2004;52:234–238. doi: 10.1021/jf0348727. PubMed DOI

Manios T. The composting potential of different organic solid wastes: Experience from the island of Crete. Environ. Int. 2004;29:1079–1089. doi: 10.1016/S0160-4120(03)00119-3. PubMed DOI

Sánchez A., Ysunza F., Beltrán-García M., Esqueda M. Biodegradation of Viticulture Wastes by Pleurotus: A Source of Microbial and Human Food and Its Potential Use in Animal Feeding. J. Agric. Food Chem. 2002;50:2537–2542. doi: 10.1021/jf011308s. PubMed DOI

Zacharof M. Grape Winery Waste as Feedstock for Bioconversions: Applying the Biorefinery Concept. Waste Biomass Valorization. 2017;8:1011–1025. doi: 10.1007/s12649-016-9674-2. DOI

Taurino R., Ferretti D., Cattani L., Bozzoli F., Bondioli F. Lightweight clay bricks manufactured by using locally available wine industry waste. J. Build. Eng. 2019;26:100892. doi: 10.1016/j.jobe.2019.100892. DOI

Kovalcik A., Pernicova I., Obruca S., Szotkowski M., Enev V., Kalina M., Marova I. Grape winery waste as a promising feedstock for the production of polyhydroxyalkanoates and other value-added products. Food Bioprod. Process. 2020;124:1–10. doi: 10.1016/j.fbp.2020.08.003. DOI

Follonier S. Pilot-scale Production of Functionalized mcl-PHA from Grape Pomace Supplemented with Fatty Acids. Chem. Biochem. Eng. Q. 2015;29:113–121. doi: 10.15255/CABEQ.2014.2251. DOI

Chen T., Chou Y., Chen W., Arun B., Young C. Tepidimonas taiwanensis sp. nov., a novel alkaline-protease-producing bacterium isolated from a hot spring. Extremophiles. 2006;10:35–40. doi: 10.1007/s00792-005-0469-9. PubMed DOI

Obruca S., Petrik S., Benesova P., Svoboda Z., Eremka L., Marova I. Utilization of oil extracted from spent coffee grounds for sustainable production of polyhydroxyalkanoates. Appl. Microbiol. Biotechnol. 2014;98:5883–5890. doi: 10.1007/s00253-014-5653-3. PubMed DOI

Mravec F., Obruca S., Krzyzanek V., Sedlacek P., Hrubanova K., Samek O., Kucera D., Benesova P., Nebesarova J., Steinbüchel A. Accumulation of PHA granules in Cupriavidus necator as seen by confocal fluorescence microscopy. FEMS Microbiol. Lett. 2016;363:fnw094. doi: 10.1093/femsle/fnw094. PubMed DOI

Kouřilová X., Schwarzerová J., Pernicová I., Sedlář K., Mrázová K., Krzyžánek V., Nebesářová J., Obruča S. The First Insight into Polyhydroxyalkanoates Accumulation in Multi-Extremophilic Rubrobacter xylanophilus and Rubrobacter spartanus. Microorganisms. 2021;9:909. doi: 10.3390/microorganisms9050909. PubMed DOI PMC

Obruca S., Sedlacek P., Krzyzanek V., Mravec F., Hrubanova K., Samek O., Kucera D., Benesova P., Marova I., Chen G.-Q. Accumulation of Poly(3-hydroxybutyrate) Helps Bacterial Cells to Survive Freezing. PLoS ONE. 2016;11:e0157778. doi: 10.1371/journal.pone.0157778. PubMed DOI PMC

Singleton V., Rossi J. Colorimetry of Total Phenolics with Phosphomolybdic-Phosphotungstic Acid Reagents. Am. J. Enol. Eiticulture. 1965;16:144–158.

Tan I., Foong C., Tan H., Lim H., Zain N., Tan Y., Hoh C., Sudesh K. Polyhydroxyalkanoate (PHA) synthase genes and PHA-associated gene clusters in Pseudomonas spp. and Janthinobacterium spp. isolated from Antarctica. J. Biotechnol. 2020;313:18–28. doi: 10.1016/j.jbiotec.2020.03.006. PubMed DOI

Pan W., Perrotta J., Stipanovic A., Nomura C., Nakas J. Production of polyhydroxyalkanoates by Burkholderia cepacia ATCC 17759 using a detoxified sugar maple hemicellulosic hydrolysate. J. Ind. Microbiol. Biotechnol. 2012;39:459–469. doi: 10.1007/s10295-011-1040-6. PubMed DOI

Kourmentza C., Costa J., Azevedo Z., Servin C., Grandfils C., De Freitas V., Reis M. Burkholderia thailandensis as a microbial cell factory for the bioconversion of used cooking oil to polyhydroxyalkanoates and rhamnolipids. Bioresour. Technol. 2018;247:829–837. doi: 10.1016/j.biortech.2017.09.138. PubMed DOI

Ibrahim M., Steinbüchel A. High-Cell-Density Cyclic Fed-Batch Fermentation of a Poly(3-Hydroxybutyrate)-Accumulating Thermophile, Chelatococcus sp. Strain MW10. Appl. Environ. Microbiol. 2010;76:7890–7895. doi: 10.1128/AEM.01488-10. PubMed DOI PMC

Keenan T., Nakas J., Tanenbaum S. Polyhydroxyalkanoate copolymers from forest biomass. J. Ind. Microbiol. Biotechnol. 2006;33:616–626. doi: 10.1007/s10295-006-0131-2. PubMed DOI

Norhafini H., Huong K., Amirul A. High PHA density fed-batch cultivation strategies for 4HB-rich P(3HB-co-4HB) copolymer production by transformant Cupriavidus malaysiensis USMAA1020. Int. J. Biol. Macromol. 2019;125:1024–1032. doi: 10.1016/j.ijbiomac.2018.12.121. PubMed DOI

Venkitasamy C., Zhao L., Zhang R., Pan Z. Integrated Processing Technologies for Food and Agricultural By-Products. Elsevier; Amsterdam, The Netherlands: 2019. Grapes; pp. 133–163.

Chowdhary P., Gupta A., Gnansounou E., Pandey A., Chaturvedi P. Current trends and possibilities for exploitation of Grape pomace as a potential source for value addition. Environ. Pollut. 2021;278:116796. doi: 10.1016/j.envpol.2021.116796. PubMed DOI

Hogan S., Zhang L., Li J., Sun S., Canning C., Zhou K. Antioxidant rich grape pomace extract suppresses postprandial hyperglycemia in diabetic mice by specifically inhibiting alpha-glucosidase. Nutr. Metab. 2010;7:71. doi: 10.1186/1743-7075-7-71. PubMed DOI PMC

Pinto D., Cádiz-Gurrea M., Silva A., Delerue-Matos C., Rodrigues F. Food Waste Recovery. Elsevier; Amsterdam, The Netherlands: 2021. Cosmetics—food waste recovery; pp. 503–528.

Luchian C., Cotea V., Vlase L., Toiu A., Colibaba L., Răschip I., Nadăş G., Gheldiu A., Tuchiluş C., Rotaru L., et al. Antioxidant and antimicrobial effects of grape pomace extracts. BIO Web Conf. 2019;15:04006. doi: 10.1051/bioconf/20191504006. DOI

Shang L., Jiang M., Chang H.N. P oly(3-hydroxybutyrate) synthesis in fed-batch culture of Ralstonia eutropha with phosphate limitation under different glucose concentrations. Biotechnol. Lett. 2003;25:1415–1419. doi: 10.1023/A:1025047410699. PubMed DOI

Polyhydroxyalkanoate involvement in stress-survival of two psychrophilic bacterial strains from the High Arctic

Genetic engineering of low-temperature polyhydroxyalkanoate production by Acidovorax sp. A1169, a psychrophile isolated from a subglacial outflow

Combination of Hypotonic Lysis and Application of Detergent for Isolation of Polyhydroxyalkanoates from Extremophiles

Complete Genome Sequence of the Type Strain Tepidimonas taiwanensis LMG 22826T, a Thermophilic Alkaline Protease and Polyhydroxyalkanoate Producer