The effect of 0.3 and 0.7 M NaCl on biomass yield, total nitrogen content, intracellular lipid content, and fatty acid profile of the lipids of the alga Botryococcus braunii IPPAS H-252 in different phases of the culture cycle was studied. The presence of sodium chloride in the medium inhibited the growth of algal cells for the first 3 days of the experiment, causing a decrease in total nitrogen, enhanced synthesis of triacylglycerols, and considerable changes in the lipid fatty acid profile: decreases in polyenoic acid contents (from 68.34% to 29.38% and 12.8%) and proportions of long-chain saturated acids (from 0.53% to 5.3% and 14.13% of the total fatty acids) at 0.3 M NaCl and 0.7 M NaCl, respectively. In later phases of the culture, at 0.3 M NaCl, the content of polyenoic acids rose to the values characteristic of the active growth phase of this alga. At 0.7 M NaCl, the proportion of polyenoic acids grew less significantly, but biomass concentration and total nitrogen increased, similarly to the experiment with 0.3 M NaCl.
Samples of olyhydroxyalkanoates (PHAs), polyhydroxybutyrate (PHB) and copolymers poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) with 4 and 18 mol% hydroxyvalerate, synthesized by the bacteria Ralstonia eutropha B5786, were investigated. PHA films in contact with blood did not activate the hemostasis system at the level of cell response, but they did activate the coagulation system and the complement reaction. To detect biologically-active components in the PHAs, a detailed analysis of the composition of the polymers was conducted. Gas chromatography-mass spectrometry revealed long-chain fatty acids (FAs) in the tested PHAs. Their total concentration in the polymer ranged from tenths of mol% to 2-3 mol%, depending on the purification method. C16:0 constituted the largest proportion, up to 70%. Of the long-chain hydroxy acids, only beta-OH-C14:0 was detected and it did not exceed 0.06 mol%. The analysis of the hemocompatibility properties of the PHAs purified by a specialized procedure, including the quantitative and morphological estimation of platelets adherent to the surface of polymer films, the plasma recalcification time and complement activation studies, indicated that PHB and PHBV can be used in contact with blood. It has been found out that the lipopolysaccharides of bacteria producing PHAs, which contain mostly long-chain hydroxy acids, can be the factor activating the hemostasis systems. Thus, the technology of PHA purification must satisfy rather stringent specific requirements.
INTRODUCTIONThe green alga Botryococcus braunii attracts attention because of its surprising ability to form a considerable amount of liquid hydrocarbons. Their composition is determined by the nature of a respective algal race (A, B, or L), and their content is believed to depend also on growth conditions [1]. Provision of biogenic elements, mainly nitrogen, is one of the main factors affecting algal metabolism. The change in the carbon/nitrogen ratio in a medium is known to result in a change in the directionality of metabolism. In many algae, an increase in this ratio results in an accumulation of neutral lipids, mainly triacylglycerols and/or carbohydrates [2][3][4][5] and is accompanied by considerable rearrangements in FA composition. These rearrangements consist in an increase in the content of saturated acids and a decrease in polyenoic acids [4,6,7]. Moreover, a decrease in the concentration of chlorophyll a accompanied by an increase in the content of carotenoids were noticed under the conditions of nitrogen limitation [8].Unfortunately, studies devoted to investigating the effect of nitrogen concentration on the biochemical composition of B. braunii are few in number and are mainly related to the production of hydrocarbons. For instance, a study showed that an increase in the nitrate content in a growth medium resulted in an increase in the duration of exponential phase of growth [9] and, consequently, in an increase in algal yield. In this case, the relative hydrocarbon content decreased; however, their total yield increased by 25% due to a higher biomass yield. On the contrary, culturing of the alga in a medium with decreased nitrogen content resulted in a decrease in both biomass and hydrocarbon content [10].Abstract -The effect of nitrogen limitation in a medium on the composition of intracellular lipids in the alga Botryococcus braunii Kütz IPPAS H-252 in the course of culture development was investigated. Under the conditions of nitrogen limitation, the alga under investigation accumulated lipids as triacylglycerols, and this process was accompanied by substantial changes in the total fatty acid (FA) composition, which were manifested in a decrease in trienoic acids (from 52.8-57.2 to 19.5-24.7% of total FAs) and an increase in the content of oleic (from 1.1-1.2 to 17.1-24.4%) and saturated (from 23.7-26.0 to 32.9-46.1%) acids. In the control culture, the directionality of FA redistribution was less marked, and these changes were noticed at the later stages of culture development. Under nitrogen limitation, marked changes in the FA composition of polar lipids occurred by the 13th day, and they were characterized by an increase in the content of saturated acids (up to 76.8%) and a dramatic decrease in the content of all polyenoic acids (up to 6.8%). The changes in the FA composition of triacylglycerols were noticed as early as by the 7th day; these changes consisted in an increase in the content of oleic acid, and its high content (28.4-38.4%) was maintained up to the end of culturing. In the cont...
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