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)