Research on the ability of propionic acid and vitamin B12 biosynthesis by Propionibacterium freudenreichii strain T82

Antonie van Leeuwenhoek, Nov 2017

The purpose of this study was to determine the potential for biosynthesis of propionic acid and vitamin B12 by Propionibacterium freudenreichii T82 in a medium containing various sources of carbon (glucose, fructose, and saccharose). These sugars are present in apple pomaces, which are the waste from the production of apple juice. Using statistical analysis design of experiments (DoE), the results allowed us to determine which sugars (carbon sources) exert the most beneficial influence on the biosynthesis of propionic acid and cobalamin. The highest production of propionic acid by the tested bacterial strain was obtained in a medium in which glucose accounted for at least 50% of the available carbon sources. Depending on the culture medium, the concentration of this metabolite ranged from 23 to 40 g/L. P. freudenreichii T82 produced the smallest amount of acid in medium in which the dominant nutrient source was saccharose. The results obtained indicated an inverse relationship between the amount of acid produced by the bacteria and vitamin B12 biosynthesis. Because of the high efficiency of propionic acid biosynthesis by P. freudenreichii T82, the prospect of using this strain to obtain propionate with the simultaneous disposal of waste materials (such as apple pomaces) which contain glucose and/or fructose is very promising.

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Research on the ability of propionic acid and vitamin B12 biosynthesis by Propionibacterium freudenreichii strain T82

Research on the ability of propionic acid and vitamin B12 biosynthesis by Propionibacterium freudenreichii strain T82 0 K. Piwowarek (&) E. Lipin ́ska E. Hac ́-Szyman ́czuk A. Bzducha-Wro ́bel A. Synowiec Department of Biotechnology , Microbiology and Food Evaluation , Division of Food Biotechnology and Microbiology, Faculty of Food Sciences, Warsaw University of Life Sciences - SGGW (WULS-SGGW) , Nowoursynowska 159c Street, 02-776 Warsaw , Poland The purpose of this study was to determine the potential for biosynthesis of propionic acid and vitamin B12 by Propionibacterium freudenreichii T82 in a medium containing various sources of carbon (glucose, fructose, and saccharose). These sugars are present in apple pomaces, which are the waste from the production of apple juice. Using statistical analysis design of experiments (DoE), the results allowed us to determine which sugars (carbon sources) exert the most beneficial influence on the biosynthesis of propionic acid and cobalamin. The highest production of propionic acid by the tested bacterial strain was obtained in a medium in which glucose accounted for at least 50% of the available carbon sources. Depending on the culture medium, the concentration of this metabolite ranged from 23 to 40 g/L. P. freudenreichii T82 produced the smallest amount of acid in medium in which the dominant nutrient source was saccharose. The results obtained indicated an inverse relationship between the amount of acid produced by the bacteria and vitamin B12 biosynthesis. Because of the high efficiency of propionic acid biosynthesis by P. freudenreichii T82, the prospect of using this strain to obtain propionate with the simultaneous disposal of waste materials (such as apple pomaces) which contain glucose and/or fructose is very promising. Propionic acid; Acetic acid; Vitamin; B12; Propionibacterium; Carbon sources; DoE Introduction Bacteria of genus Propionibacterium have been traditionally divided into two groups: skin (acnes) and classic (dairy). Classical strains include, among others, the species Propionibacterium acidipropionici, Propionibacterium jensenii, Propionibacterium thoenii and Propionibacterium freudenreichii (ssp. shermanii, ssp. freudenreichii) (Meile et al. 1999) , of which the first three have recently been reclassified as members of the genus Acidipropionibacterium (Scholz and Kilian 2016) . Classical Propionibacterium are a source of useful metabolites such as propionic acid and vitamin B12 (Meile et al. 1999; Patrick and McDowell 2015) . Propionic acid is used to inhibit the growth of yeast and molds in prepacked sliced bread, rye bread, breads with reduced calories, and partially baked rolls, pita bread, pastry products and animal feed. Propionic acid also is an essential indirect component in production process of the cellulose fibers, herbicides, perfumes, and pharmaceuticals (Suomalainen and Ma¨yra¨Makinen 1999, Gwiazdowski and Gwiazdowska 2008) . Propionic acid for industrial purposes is currently synthesised only in chemical processes, as this is still more economical than microbial processes using propionic acid bacteria. However, due to the serious environmental damage that can be caused by chemical production of propionic acid, as well as due to the rise in demand for natural and ecological food products, there is an increasing demand for the microbial production of propionic acid, along with the desirability of using waste materials. This should reduce the cost of natural production of propionc acid and make it profitable and should have environmental benefits (Baumann and Westermann 2016). Such waste materials could include apple pomaces, which contain sugars (glucose, fructose, saccharose), proteins, pectins, fiber, vitamins and organic acids, which may affect the efficiency of synthesis of propionic acid or cobalamin by Propionibacterium spp. and relatives. Bacteria of the genus Propionibacterium and relatives seem the most appropriate for the biotechnological production of propionic acid. Due to their wide variety of enzymatic systems, they can use carbon from various sources, for example: glucose (Himmi et al. 2000) , xylose (Carrondo et al. 1988) , lactose (Hsu and Yang 1991) , saccharose (QuesadaChanto et al. 1994), lactic acid (Barbirato et al. 1997) , maltose (Zhu et al. 2012) and whey (Lewis and Yang 1992) . These bacteria can be used in the reprocessing of waste materials including glycerol (Yazdani and Gonzales 2007; Zhu et al. 2010) , hemicellulose hydrolysates (Ramsay et al. 1998), corn flour (Huang et al. 2002) and cane molasses (Feng et al. 2011) . The most favourable bacteria for industrial production of vitamin B12—due to their Generally Recognized as Safe status (GRAS) and ability to synthesise active forms of this metabolite—may be strains of P. freudenreichii. However, currently commercially produced cobalamin uses a genetically modified strain of Pseudomonas denitrificans (without GRAS stat (...truncated)


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Kamil Piwowarek, Edyta Lipińska, Elżbieta Hać-Szymańczuk, Anna Bzducha-Wróbel, Alicja Synowiec. Research on the ability of propionic acid and vitamin B12 biosynthesis by Propionibacterium freudenreichii strain T82, Antonie van Leeuwenhoek, 2017, pp. 1-12, DOI: 10.1007/s10482-017-0991-7