Exobiopolymer production of Ophiocordyceps dipterigena BCC 2073: optimization, production in bioreactor and characterization.
BACKGROUND
Biopolymers have various applications in medicine, food and petroleum industries. The ascomycetous fungus Ophiocordyceps dipterigena BCC 2073 produces an exobiopolymer, a (1-->3)-beta-D-glucan, in low quantity under screening conditions. Optimization of O. dipterigena BCC 2073 exobiopolymer production using experimental designs, a scale-up in 5 liter bioreactor, analysis of molecular weight at different cultivation times, and levels of induction of interleukin-8 synthesis are described in this study.


RESULTS
In order to improve and certify the productivity of this strain, a sequential approach of 4 steps was followed. The first step was the qualitative selection of the most appropriate carbon and nitrogen sources (general factorial design) and the second step was quantitative optimization of 5 physiological factors (fractional factorial design). The best carbon and nitrogen source was glucose and malt extract respectively. From an initial production of 2.53 g x L(-1), over 13 g x L(-1) could be obtained in flasks under the improved conditions (5-fold increase). The third step was cultivation in a 5 L bioreactor, which produced a specific growth rate, biomass yield, exobiopolymer yield and exobiopolymer production rate of 0.014 h-1, 0.32 g x g(-1) glucose, 2.95 g x g biomass(-1) (1.31 g x g(-1) sugar), and 0.65 g x (L x d)(-1), respectively. A maximum yield of 41.2 g x L(-1) was obtained after 377 h, a dramatic improvement in comparison to the initial production. In the last step, the basic characteristics of the biopolymer were determined. The molecular weight of the polymer was in the range of 6.3 x 10(5) - 7.7 x 10(5) Da. The exobiopolymer, at 50 and 100. microg x mL(-1), induced synthesis in normal dermal human fibroblasts of 2227 and 3363 pg x mL(-1) interleukin-8 respectively.


CONCLUSIONS
High exobiopolymer yield produced by O. dipterigena BCC 2073 after optimization by qualitative and quantitative methods is attractive for various applications. It induced high IL-8 production by normal dermal fibroblasts, which makes it promising for application as wound healing material. However, there are still other possible applications for this biopolymer, such as an alternative source of biopolymer substitute for hyaluronic acid, which is costly, as a thickening agent in the cosmetic industry due to its high viscosity property, as a moisturizer, and in encapsulation.
Kocharin
Kanokarn
K
National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, 113 Thailand Science Park, Klong 1, Klong Luang, Pathumthani 12120, Thailand.
Rachathewee
Pranee
P
Sanglier
Jean-Jacques
JJ
Prathumpai
Wai
W
D018533:Biomass; D001704:Biopolymers / Q000096:biosynthesis*; D019149:Bioreactors; D001709:Biotechnology / Q000379:methods; D002478:Cells, Cultured; D005285:Fermentation; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D006999:Hypocreales / Q000378:metabolism*; D016209:Interleukin-8 / Q000096:biosynthesis
D001704:Biopolymers; D016209:Interleukin-8