Response surface optimisation of medium components for naringinase production from staphylococcus xylosus MAK 2

Response surface methodology was used to optimize the fermentation medium for enhancing naringinase production by Staphylococcus xylosus. The first step of this process involved the individual adjustment and optimization of various medium components at shake flask level. Sources of carbon (sucrose) and nitrogen (sodium nitrate), as well as an inducer (naringin) and pH levels were all found to be the important factors significantly affecting naringinase production. In the second step, a 22 full factorial central composite design was applied to determine the optimal levels of each of the significant variables. A second-order polynomial was derived by multiple regression analysis on the experimental data. Using this methodology, the optimum values for the critical components were obtained as follows: sucrose, 10.0%; sodium nitrate, 10.0%; pH 5.6; biomass concentration, 1.58%; and naringin, 0.50% (w/v), respectively. Under optimal conditions, the experimental naringinase production was 8.45 U/mL. The determination coefficients (R ²) were 0.9908 and 0.9950 for naringinase activity and biomass production, respectively, indicating an adequate degree of reliability in the model.

Bead milling for lipid recovery from thraustochytrid cells and selective hydrolysis of Schizochytrium DT3 oil using lipase

Marine microalgae present a renewable alternative source for sustainable production of omega-3 fatty acids, as compared to conventional sources such as krill oil and fish oil. In this study, we optimised a method for lipid extraction from marine thraustochytrids using a bead mill and enzymatic concentration of omega-3 fatty acids from the thraustochytrid oil. The optimised lipid extraction conditions were, bead size 0.4-0.6μm, 4500rpm, 4min of processing time at 5g biomass concentration. The maximum lipid yield (% dry weight basis) achieved at optimum conditions were 40.5% for Schizochytrium sp. S31 (ATCC) and 49.4% for Schizochytrium sp. DT3 (in-house isolate). DT3 oil contained 39.8% docosahexaenoic acid (DHA) as a percentage of lipid, a higher DHA percentage than S31. Partial hydrolysis of DT3 oil using Candida rugosa lipase was performed to enrich omega-3 polyunsaturated fatty acids (PUFAs) in the glyceride portion. Total omega-3 fatty acid content was increased to 88.7%.

Combination of calcium and magnesium ions prevents substrate inhibition and promotes biomass and lipid production in thraustochytrids under higher glycerol concentration

Effect of calcium and magnesium ions was studied in detail in batch mode in shake flask cultures of two fast growing strains of thraustochytrids (Aurantiochytrium sp. DBTIOC-18 and Schizochytrium sp. DBTIOC-1) for biomass and lipid production. These strains were previously isolated from Indian marine biodiversity. Screening of these two strains on different carbon and nitrogen sources revealed the suitability of glycerol over glucose and sodium nitrate over yeast extract for the cultivation of these strains. The presence of higher concentration of glycerol in the medium inhibited the glycerol utilization by the cell thus resulting in lower biomass and lipid production in both the strains. Supplementing media with calcium and magnesium ions promoted glycerol utilization thus resulted in a substantial rise in volumetric production of biomass (55.12 g L-1, 48.12 g L-1), fatty acid for biodiesel (27.14 g L-1, 22.15 g L-1) and docosahexaenoic acid (14.57 g L-1, 10.12 g L-1) with both strains Aurantiochytrium sp. DBTIOC-18 and Schizochytrium sp. DBTIOC-1, respectively. Growth profile study of these two strains showed further improvement in production of biomass, fatty acid for biodiesel and docosahexaenoic acid when cultures were extended up to 7 days. Finding of this work underlines the importance of calcium and magnesium salts in designing new fermentation strategies to prevent substrate inhibition and achieve high cell density culture under high nutrient concentration especially carbon sources.

Can mud (silt and clay) concentration be used to predict soil organic carbon content within seagrass ecosystems?

The emerging field of blue carbon science is seeking cost-effective ways to estimate the organic carbon content of soils that are bound by coastal vegetated ecosystems. Organic carbon (Corg) content in terrestrial soils and marine sediments has been correlated with mud content (i.e. silt and clay), however, empirical tests of this theory are lacking for coastal vegetated ecosystems. Here, we compiled data (n = 1345) on the relationship between Corg and mud (i.e. silt and clay, particle sizes <63 μm) contents in seagrass ecosystems (79 cores) and adjacent bare sediments (21 cores) to address whether mud can be used to predict soil Corg content. We also combined these data with the δ13C signatures of the soil Corg to understand the sources of Corg stores. The results showed that mud is positively correlated with soil Corg content only when the contribution of seagrass-derived Corg to the sedimentary Corg pool is relatively low, such as in small and fast growing meadows of the genera Zostera, Halodule and Halophila, and in bare sediments adjacent to seagrass ecosystems. In large and long-living seagrass meadows of the genera Posidonia and Amphibolis there was a lack of, or poor relationship between mud and soil Corg content, related to a higher contribution of seagrass-derived Corg to the sedimentary Corg pool in these meadows. The relative high soil Corg contents with relatively low mud contents (i.e. mud-Corg saturation) together with significant allochthonous inputs of terrestrial organic matter could overall disrupt the correlation expected between soil Corg and mud contents. This study shows that mud (i.e. silt and clay content) is not a universal proxy for blue carbon content in seagrass ecosystems, and therefore should not be applied generally across all seagrass habitats. Mud content can only be used as a proxy to estimate soil Corg content for scaling up purposes when opportunistic and/or low biomass seagrass species (i.e. Zostera, Halodule and Halophila) are present (explaining 34 to 91% of variability), and in bare sediments (explaining 78% of the variability).