Polyhydroxyalkanoates, microbial polyesters produced in vivo starting from renewable resources, are considered the future materials of choice to compete recalcitrant petro-chemical plastic on the polymer market. In order to make polyhydroxyalkanoates market-fit, (techno)economics of their production need to be improved. Among the multifarious factors affecting costs of polyhydroxyalkanoate production, increased volumetric productivity is of utmost importance. Improving microbial growth kinetics and increasing cell density are strategies leading to a high concentration of catalytically active biomass within a short time; after changing cultivation conditions, these cells can accumulate polyhydroxyalkanoates as intracellular products. The resulting increase of volumetric productivity for polyhydroxyalkanoates can be realized by supplying complex nitrogen sources to growing microbial cultures. In the present study, the impact of different expensive and inexpensive complex nitrogen sources, in particular whey retentate, on the growth and specific growth rates of Hydrogenophaga pseudoflava was tested.
Based on a detailed kinetic process analysis, the study demonstrates that especially whole (not hydrolyzed) whey retentate, an amply available surplus material from dairy industry, displays positive effects on cultivations of H. pseudoflava in defined media (increase of concentration of catalytically active biomass after 26.25 h of cultivation by about 50%, increase of specific growth rate μ from 0.28 to 0.41 1/h during exponential growth), while inhibiting effects (inhibition constant Ki = 6.1 g/L) of acidically hydrolyzed whey retentate need to be overcome. Considering the huge amounts of surplus whey accruing especially in Europe, the combined utilization of whey permeate (carbon source) and whey retentate (complex nitrogen source) for biopolyester production can be considered a viable bioeconomic strategy for the next future.
1. Haider TP, Völker C, Kramm J, Landfester K, Wurm FR. Plastics of the future? The impact of biodegradable polymers on the environment and on society. Angew Chem Int Edit 2019; 58: 50-62
2. Koller M. Switching from petro-plastics to microbial polyhydroxyalkanoates (PHA): the biotechnological escape route of choice out of the plastic predicament? The EuroBiotech Journal 2019; 3(1): 32-44.
3. Narodoslawsky M, Shazad K, Kollmann R, Schnitzer H. LCA of PHA production–Identifying the ecological potential of bio-plastic. Chem Biochem Eng Q 2015; 29(2): 299-305.
4. Koller M, Maršálek L, Miranda de Sousa Dias M, Braunegg G. Producing microbial polyhydroxyalkanoate (PHA) biopolyesters in a sustainable manner. New Biotechnol 2017; 37(A): 24-38.
5. Kourmentza C, Plácido J, Venetsaneas N, Burniol-Figols A, Varrone C, Gavala HN, Reis MAM. Recent advances and challenges towards sustainable polyhydroxyalkanoate (PHA) production. Bioengineering 2017; 4(2): 55.
6. Bugnicourt E, Cinelli P, Lazzeri A, Alvarez VA. Polyhydroxyalkanoate (PHA): Review of synthesis, characteristics, processing and potential applications in packaging. eXPRESS Polym Lett 2014; 8(11): 791-808.
7. Khosravi-Darani K, Bucci DZ. Application of poly(hydroxyalkanoate) in food packaging: Improvements by nanotechnology. Chem Biochem Engineering Q 2015; 29(2): 275-285.
8. Koller M. Biodegradable and biocompatible polyhydroxy-alkanoates (PHA): Auspicious microbial macromolecules for pharmaceutical and therapeutic applications. Molecules 2018; 23(2): 362.
9. Drosg B, Fritz I, Gattermayr F, Silvestrini L. Photo-autotrophic production of poly(hydroxyalkanoates) in cyanobacteria. Chem Biochem Engineering Q 2015; 29(2): 145-156.
10. Troschl C, Meixner K, Drosg B. Cyanobacterial PHA production—Review of recent advances and a summary of three years’ working experience running a pilot plant. Bioengineering 2017: 4(2): 26.
11. Koller M. Production of polyhydroxyalkanoate (PHA) biopolyesters by extremophiles. MOJ Polym Sci 2017; 1(2): 1-19.
12. Koller M, Obruca S, Pernicova I, Braunegg G. Physiological, kinetic, and process engineering aspects of polyhydroxyalkanoate biosynthesis by extremophiles. In: Williams H, Kelly P (Eds.) Polyhydroxyalkanoates: Biosynthesis, Chemical Structures and Applications. 2018. ISBN 978-1-53613-439-1; Nova Science Publishers, New York, pp. 1-70.
13. Willems A, Busse J, Goor M, Pot B, Falsen E, Jantzen, E, et al. Hydrogenophaga, a new genus of hydrogen-oxidizing bacteria that includes Hydrogenophaga flava comb. nov.(formerly Pseudomonas flava), Hydrogenophaga palleronii (formerly Pseudomonas palleronii), Hydrogenophaga pseudoflava (formerly Pseudomonas pseudoflava and “Pseudomonas carboxydoflava”), and Hydrogenophaga taeniospiralis (formerly Pseudomonas taeniospiralis). Int J Syst Evol Microbiol1989; 39(3): 319-333.
14. Mahmoudi M, Baei MS, Najafpour GD, Tabandeh F, Eisazadeh H. Kinetic model for polyhydroxybutyrate (PHB) production by Hydrogenophaga pseudoflava and verification of growth conditions. Afr J Biotechnol 2010; 9(21): 3151-3157.
15. Povolo S, Romanelli MG, Basaglia M, Ilieva VI, Corti A, Morelli A, Chiellini E, Casella S. Polyhydroxyalkanoate biosynthesis by Hydrogenophaga pseudoflava DSM1034 from structurally unrelated carbon sources. New Biotechnol 2013; 30(6): 629-634.
16. Koller M, Hesse P, Bona R, Kutschera C, Atlić A, Braunegg G. Potential of various archae-and eubacterial strains as industrial polyhydroxyalkanoate producers from whey. Macromol Biosci 2007; 7(2): 218-226.
17. Koller M, Atlić A, Gonzalez-Garcia Y, Kutschera C, Braunegg G. Polyhydroxyalkanoate (PHA) biosynthesis from whey lactose. Macromol Symp 2008; 272(1): 87-92).
18. Choi MH, Song JJ, Yoon SC. Biosynthesis of copolyesters by Hydrogenophaga pseudoflava from various lactones. Can J Microbiol 1995; 41(13): 60-67.
19. Yoon SC, Choi MH. Local sequence dependence of polyhydroxyalkanoic acid degradation in Hydrogenophaga pseudoflava. J Biol Chem 1999; 274(53): 37800-37808.
20. Koller M, Hesse P, Fasl H, Stelzer F, Braunegg G. Study on the effect of levulinic acid on whey-based biosynthesis of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) by Hydrogenophaga pseudoflava. Appl Food Biotechnol 2017; 4(2): 65-78.
21. Choi MH, Yoon SC, Lenz RW. Production of poly (3-hydroxybutyric acid-co-4-hydroxybutyric acid) and poly(4-hydroxybutyric acid) without subsequent degradation by Hydrogenophaga pseudoflava. Appl Environ Microbiol 1999; 65(4): 1570-1577.
22. Choi MH, Lee HJ, Rho JK, Yoon SC, Nam JD, Lim D, Lenz RW. Biosynthesis and local sequence specific degradation of poly(3-hydroxyvalerate-co-4-hydroxybutyrate) in Hydrogenophaga pseudoflava. Biomacromolecules 2003; 4(1): 38-45.
23. Brigham C, Kehail AA, Palmer JD. Ralstonia eutropha and the production of value added products: metabolic background of the wild-type strain and its role as a diverse, genetically-engineered biocatalyst organism. In: Koller M (Ed.): Recent Advances in Biotechnology Volume 1: Microbial Biopolyester Production, Performance and Processing: Microbiology, Feedstocks, and Metabolism. Potomac, Maryland, USA. Bentham Science Publishers Ltd. 2016. pp. 265-347.
24. Kaur G, Roy I. Strategies for large-scale production of polyhydroxyalkanoates. Chem Biochem Eng Q 2015; 29(2): 157-172.
25. Lillo JG, Rodriguez-Valera F. Effects of culture conditions on poly(β-hydroxybutyric acid) production by Haloferax mediterranei. Appl Environ Microbiol 1990; 56(8): 2517-2521.
26. Page WJ, Cornish A. Growth of Azotobacter vinelandii UWD in fish peptone medium and simplified extraction of poly-β-hydroxybutyrate. Appl Environ Microbiol 1993; 59(12): 4236-4244.
27. Koller M, Bona R, Hermann C, Horvat P, Martinz J, Neto J, Pereira L, Varila P, Braunegg, G. Biotechnological production of poly(3-hydroxybutyrate) with Wautersia eutropha by application of green grass juice and silage juice as additional complex substrates. Biocat Biotrans 2005; 23(5): 329-337.
28. Davis R, Kataria R, Cerrone F, Woods T, Kenny S, O’Donovan A, et al. Conversion of grass biomass into fermentable sugars and its utilization for medium chain length polyhydroxyalkanoate (mcl-PHA) production by Pseudomonas strains. Bioresource Technol 2013; 150: 202-209.
29. Koller M, Sandholzer D, Salerno A, Braunegg G, Narodoslawsky M. Biopolymer from industrial residues: Life cycle assessment of poly(hydroxyalkanoates) from whey. Resour Conserv Recy 2013; 73: 64-71.
30. Obruca S, Benesova P, Oborna J, Marova I. Application of protease-hydrolyzed whey as a complex nitrogen source to increase poly(3-hydroxybutyrate) production from oils by Cupriavidus necator. Biotechnol Lett 2014; 36(4): 775-781.
31. Schmid M, Dallmann K, Bugnicourt E, Cordoni D, Wild F, Lazzeri A, Noller K. Properties of whey-protein-coated films and laminates as novel recyclable food packaging materials with excellent barrier properties. Int J Polym Sci 2012; 2012.
32. Cinelli P, Schmid M, Bugnicourt E, et al. Whey protein layer applied on biodegradable packaging film to improve barrier properties while maintaining biodegradability. Polym Degrad Stabil 2014; 108: 151-7.
33. Koller M, Marsalek L, Braunegg G. PHA Biopolyester Production from Surplus Whey: Microbiological and Engineering Aspects. In: Koller M (Ed.): Recent Advances in Biotechnology Volume 1: Microbial Biopolyester Production, Performance and Processing: Microbiology, Feedstocks, and Metabolism. Potomac, Maryland, USA. Bentham Science Publishers Ltd. 2016. pp. 100-172.
34. Koller M, Braunegg G. Advanced approaches to produce polyhydroxyalkanoate (PHA) biopolyesters in a sustainable and economic fashion. The EuroBiotech Journal 2018; 2(2): 89-103.
35. Koller M, Puppi D, Chiellini F, Braunegg G. Comparing chemical and enzymatic hydrolysis of whey lactose to generate feedstocks for haloarchaeal poly(3-hydroxybutyrate-co-3-hydroxyvalerate) biosynthesis. Int J Pharm Sci Res 2016; 3(1).
36. Braunegg G, Sonnleitner BY, Lafferty RM. A rapid gas chromatographic method for the determination of poly-β-hydroxybutyric acid in microbial biomass. Eur J Appl Microbiol Biotechnol 1978; 6(1): 29-37.
37. Daiber KH. Enzyme inhibition by polyphenols of sorghum grain and malt. J Sci Food Agric 1975; 26(9): 1399-1411.
38. Obruca S, Sedlacek P, Koller M, Kucera D, Pernicova I. Involvement of polyhydroxyalkanoates in stress resistance of microbial cells: Biotechnological consequences and applications. Biotechnol Adv 2018; 36(3): 856-870.
39. Novak M, Koller M, Braunegg M, Horvat P. Mathematical modelling as a tool for optimized PHA production. Chem Biochem Eng Q 2015; 29(2): 183-220.
40. Koller M, Vadlja D, Braunegg G, Atlić A, Horvat P. Formal-and high-structured kinetic process modelling and footprint area analysis of binary imaged cells: Tools to understand and optimize multistage-continuous PHA biosynthesis. The EuroBiotech Journal 2017; 1(3): 203-211.
41. Sindhu R, Silviya N, Binod P, Pandey A. Pentose-rich hydrolysate from acid pretreated rice straw as a carbon source for the production of poly-3-hydroxybutyrate. Biochem Eng J 2013; 78: 67-72.
42. Koller M, Bona R, Chiellini E, Fernandes EG, Horvat P, Kutschera C, Hesse P, Braunegg G. Polyhydroxyalkanoate production from whey by Pseudomonas hydrogenovora. Bioresource Technol 2008; 99(11): 4854-4863.
43. Obruca S, Benesova P, Marsalek L, Marova I. Use of lignocellulosic materials for PHA production. Chem Biochem Eng Q 2015; 29(2): 135-144.
44. Lopes MSG, Gomez JGC, Taciro MK, Mendonça TT, Silva LF. Polyhydroxyalkanoate biosynthesis and simultaneous remotion of organic inhibitors from sugarcane bagasse hydrolysate by Burkholderia sp. J Ind Microbiol Biotechnol 2014; 41(9): 1353-1363.
45. Kucera D, Benesova P, Ladicky P, Pekar M, Sedlacek P, Obruca S. Production of polyhydroxyalkanoates using hydrolyzates of spruce sawdust: Comparison of hydrolyzates detoxification by application of overliming, active carbon, and lignite. Bioengineering 2017: 4(2): 53.