Effects of exopolysaccharide derived from Pantoea alhagi NX-11 on drought resistance of rice and its efficient fermentation preparation.

Exopolysaccharide (EPS) plays an important role in plant growth-promoting bacteria (PGPB)-mediated enhancement of plant abiotic stress resistance. In this study, it is found that EPS from Pantoea alhagi NX-11 foliar sprayed at 20, 50, and 100 ppm could significantly enhance drought resistance of rice seedlings. The fresh weight and relative water content of EPS sprayed were increased. In addition, malondialdehyde content reduced while total chlorophyll, proline and soluble sugar content, prominent enhanced. Meanwhile, the antioxidant enzymes, CAT, POD and SOD, were also significantly increased. The drought resistance of rice was most pronounced at the 50 ppm EPS dose. For the sake of commercializing the gram-negative EPS-producing PGPB which were difficult to preserve, it is vital to improve the EPS yield. First, the carbon source, nitrogen source and inorganic salt were optimized. Subsequently, the effect of three oxygen vectors, which could increase the efficiency of oxygen mass transfer, on EPS yield was studied by response surface methodology. The maximum EPS yield (19.27 g/L) was obtained, which is 51.7% higher than the initial yield of 12.7 g/L. Overall, it may provide a new way for the industrialization of PGPB to increase the yield of EPS.

Enhanced growth and ß-galactosidase production on Escherichia coli using oxygen vectors.

The addition of n-dodecane (between 1-3%) to the Escherichia coli fermentation broth in a mechanically agitated and aerated bioreactor revealed improved DO (dissolved oxygen) levels induced during fermentation which lead to an increase in biomass productivity and faster glucose consumption. The maximum values for enzyme activity (increased with 43% compared with the control) and k L a (the volumetric mass transfer coefficient) were obtained for the addition of 2% v/v n-dodecane in the bioreactor, due to the fact that oxygen limitation during the exponential growth phase of the bacterium can repress ß-galactosidase production. The oxygen vector addition increased the available dissolved oxygen and activated a redox-sensitive regulation and an elevated intracellular oxidative metabolism that lead to the enhancement in E. coli biomass accumulation and a more accurate protein folding of ß-galactosidase that would increase its activity. In addition to the experimental analysis, a complex model, developed using an improved version of Bacterial Foraging Algorithm and Artificial Neural Networks, was proposed, with a good average absolute value (6.2% in the training phase and 7.28% in the testing phase) between the process dynamic and the predictions generated by the model.

A new combined approach to improved lipid production using a strictly aerobic and oleaginous yeast.

Microbial lipids have potential applications in energy, and food industry, because most of those lipids are triacylglycerol with long-chain fatty-acids that are comparable to conventional vegetable oils and can be obtained without arable land requirement. Rhodosporidium toruloides is a strictly aerobic strain, where oxygen plays a crucial role in growth, maintenance, and metabolite production, such as lipids and carotenoids. Dissolved oxygen concentration is one of the major factors affecting yeast physiological and biochemical characteristics. In this context, different approaches have been developed to increase available oxygen by the increasing the aeration and the addition of an oxygen-vector. The growth of R. toruloides in 2-L mechanical stirred tank reactor equipped with 1 or 2 porous spargers and a 70 C/N ratio, revealed a lipid content of 0.47 and 0.52 g/g and a lipidic productivity of 0.16 and 0.17 g/L day, respectively. The oxygen-vector addition, increased the lipidic productivity for 0.20 g/L day and a lipid contend of 0.51 g of lipids/g of biomass. The combined approach, combining high aeration (AA), and 1% of n-dodecane addition (DA), produced a significant improvement in the lipid accumulation (62%, w/w), when compared with the DA (51%, w/w) and the AA (52%, w/w) approaches. The increasing of lipids accumulation and smaller culture time are key factors for the success of scale-up and profitability of a bioprocess.

Enhancement of antroquinonol production during batch fermentation using pH control coupled with an oxygen vector.

BACKGROUND: Antroquinonol, a ubiquinone derivative that shows anticancer and anti-inflammatory activities, is produced during solid-state fermentation of Antrodia camphorata; however, it cannot be biosynthesized via conventional submerged fermentation. RESULTS: A method for enhancing the biosynthesis of antroquinonol by controlling pH and adding an oxygen vector in a 7 L bioreactor was studied. In shake-flask experiments, a maximum antroquinonol production of 31.39 ± 0.78 mg L-1 was obtained by fermentation with adding 0.2 g L-1 coenzyme Q0 (CoQ0 ), at the 96th hour. Following kinetic analysis of the fermentation process, pH control strategies were investigated. A maximum antroquinonol production of 86.47 ± 3.65 mg L-1 was achieved when the pH was maintained at 5.0, which exhibited an increase of 348.03% higher than the batch without pH regulation (19.30 ± 0.88 mg L-1 ). The conversion rate of CoQ0 improved from 1.51% to 20.20%. Further research revealed that the addition of n-tetradecane could increase the production of antroquinonol to 115.62 ± 4.87 mg L-1 by increasing the dissolved oxygen in the fermentation broth. CONCLUSION: The results demonstrated that pH played an important role in antroquinonol synthesis in the presence of the effective precursor CoQ0 . It was a very effective strategy to increase the yield of antroquinonol by controlling pH and adding oxygen vector. © 2018 Society of Chemical Industry.

Application of n-dodecane as an oxygen vector to enhance the activity of fumarase in recombinant Escherichia coli: role of intracellular microenvironment

Abstract The effect of the intracellular microenvironment in the presence of an oxygen vector during expression of a fusion protein in Escherichia coli was studied. Three organic solutions at different concentration were chosen as oxygen vectors for fumarase expression. The addition of n-dodecane did not induce a significant change in the expression of fumarase, while the activity of fumarase increased significantly to 124% at 2.5% n-dodecane added after 9 h induction. The concentration of ATP increased sharply during the first 6 h of induction, to a value 7600% higher than that in the absence of an oxygen-vector. NAD/NADH and NADP/NADPH ratios were positively correlated with fumarase activity. n-Dodecane can be used to increase the concentration of ATP and change the energy metabolic pathway, providing sufficient energy for fumarase folding.

Application of n-dodecane as an oxygen vector to enhance the activity of fumarase in recombinant Escherichia coli: role of intracellular microenvironment.

The effect of the intracellular microenvironment in the presence of an oxygen vector during expression of a fusion protein in Escherichia coli was studied. Three organic solutions at different concentration were chosen as oxygen vectors for fumarase expression. The addition of n-dodecane did not induce a significant change in the expression of fumarase, while the activity of fumarase increased significantly to 124% at 2.5% n-dodecane added after 9h induction. The concentration of ATP increased sharply during the first 6h of induction, to a value 7600% higher than that in the absence of an oxygen-vector. NAD/NADH and NADP/NADPH ratios were positively correlated with fumarase activity. n-Dodecane can be used to increase the concentration of ATP and change the energy metabolic pathway, providing sufficient energy for fumarase folding.

Metabolomics analysis of the effect of dissolved oxygen on spinosad production by Saccharopolyspora spinosa.

Spinosad, a universal bio-pesticide, is obtained from the soil actinomycete Saccharopolyspora spinosa. Dissolved oxygen, an important contributing factor in aerobic microbial fermentation, however, is not always available in sufficient amounts. To alleviate oxygen limitation in spinosad production, three different oxygen vectors, namely oleic acid, toluene, and n-dodecane, were added into early fermentation. Results indicated that n-dodecane was the optimal oxygen vector. Spinosad yield was increased by 44.2% compared to that in the control group in the presence of 0.5% n-dodecane, added after 120 h of incubation. Yields of the test group reached 6.52 mg/g dry cell weight (DCW), while that of the control group was limited to 4.52 mg/g DCW. Metabolomics analysis by gas chromatography coupled to mass spectrometry was performed to demonstrate the metabolism mechanism in the presence and absence of oxygen vector. In total, 78 principal intracellular metabolites in S. spinosa were detected and quantified in the presence and absence of n-dodecane. Levels of some metabolites that were related to the tricarboxylic acid cycle and pentose phosphate pathway varied significantly. Aspartic acid and glucose-1-phosphate levels varied significantly and contributed most in the distinction of the fermentation conditions and phases. The above findings give new insights into the improvement and the metabolomic characteristics of industrial spinosad production.

Improvement of AD Biosynthesis Response to Enhanced Oxygen Transfer by Oxygen Vectors in Mycobacterium neoaurum TCCC 11979.

In steroid biotransformation, soybean oil can improve the productivity of steroids by increasing substrate solubility and strengthen the cell membrane permeability. However, little is known of its role as oxygen carrier and its mechanism of promoting the steroid biotransformation. In this work, soybean oil used as oxygen vector for the enhancement of androst-4-ene-3,17-dione (AD) production by Mycobacterium neoaurum TCCC 11979 (MNR) was investigated. Upon the addition of 16% (v/v) soybean oil, the volumetric oxygen transfer coefficient (K L a) value increased by 44%, and the peak molar yield of AD (55.76%) was achieved. Analysis of intracellular cofactor levels showed high NAD+, ATP level, and a low NADH/NAD+ ratio. Meanwhile, the two key enzymes of the tricarboxylic acid (TCA) cycle, namely, isocitrate dehydrogenase and α-ketoglutarate dehydrogenase, were upregulated after incubation with soybean oil. These enhancements induced by the increasing of oxygen supply showed positive effects on phytosterol (PS) bioconversion. Results could contribute to the understanding of effects of soybean oil as oxygen vector on steroid biotransformation and provided a convenient method for enhancing the efficiency of aerobic steroid biocatalysis.

Optimization of the Biosynthesis Conditions of Daptomycin by the Biostatistical Methodology.

Response surface methodology (RSM) was employed to optimize medium components including oxygen vector of n-dodecane of a mutant strain GC-63 of Streptomyces roseosporus NRRL 11379. The two-level Plackett-Burman design (PB factorial design) with fourteen variables including oxygen vector was used to screen the most significant factors affecting antibiotic production. Then, the RSM based on center composite design was used to identify the optimum levels of the significant variables to generate optimal response. Glucose, soybean meal, asparagine and n-dodecane were screened to significantly influence the daptomycin production. The medium composition optimized with response surface methodology was (g/L): glucose, 9.46; soluble starch, 25; dextrin, 12.5; yeast extract, 12.5; soybean meal, 21.34; peptone, 25; casein, 5; asparagine, 2.68; K2SO4, 6; (NH4)2Fe(SO4)2, 2; MgSO4, 1; CaCO3, 5; MnCl2, 0.5; n-dodecane, 7.47 % (v/v). The maximum daptomycin concentration reached 979.36 mg/L which was nearly 2.2-fold higher compared to that in the basal medium, with predicted optimal concentrations in a 7.5-L fermentor.