ABSTRACT
Haematococcus pluvialis was modified for higher astaxanthin production compatible with the superiorities of high biomass and high activity by three-stage mutagenesis breeding. UV irradiation mutants named UV11-4 made an increase on cell dry weight, but showed a longer growth circle than the wild type. On the basis of UV mutants, ethyl methane sulphonate (EMS) mutants E2-5 cut down the latent phase, brought forward and extended the logarithmic phase. The inhibitor diphenylamine (DPA) was employed to screen high-yield astaxanthin producer by the color change of colonies from green to red on solid medium. Via the contravariant cultivation, proliferation and transformation, the mutant DPA12-2 possessed an 1.7-fold astaxanthin production compared to the wild type, reaching 47.21±3.30mg/g dry cells.
Subject(s)
Chlorophyta/metabolism , Metabolic Engineering/methods , Mutagenesis/genetics , Biomass , Biotechnology , Chlorophyta/genetics , Xanthophylls/analysis , Xanthophylls/metabolismABSTRACT
The operation of cell disruption is indispensable but cost much in microalgae industry. To be simplified, two different reaction mechanisms await in the cell to respond to moderated or stressed environment. The physical and chemical changes of enzyme and turgor pressure of cell in this conversion play an important role in the enhancement of biomass and metabolites. Repeated turgor pressure (based on the structure and mechanics of cell wall) and converted enzyme system (based on photosynthesis) were used to loosen cell wall and then repeated cultivation of Haematococcus pluvialis for astaxanthin extraction was proposed. There was no significant difference of extraction yield between the broken cell (94.75 ± 3.13%) and non-broken cell (92.32 ± 3.24%) treated by the repeated cultivation. Meanwhile, fed-batch culture according to the relationship among pH and nutrient concentration was used to enhance the biomass of Haematococcus pluvialis with the dry cell weight of 1.63 ± 0.07 g/L.
Subject(s)
Batch Cell Culture Techniques/methods , Chlorophyta/growth & development , Biomass , Cell Wall/physiology , Chlorophyta/chemistry , Hydrogen-Ion Concentration , Plant Cells/metabolism , Xanthophylls/isolation & purificationABSTRACT
Fed-batch culture and the transformation conditions of Haematococcus pluvialis in a 5L photobioreactor were investigated. Methods of feeding model, low temperature at night and proper feeding time were used to increase both cell biomass and cell activity. Dry cell weight of 1.87 g/L which was 2.0-fold of batch culture and the specific growth rate of 0.43 d(-1) suggested the superduper results of these methods to increase the dry cell weight in the short cultivation time. Furthermore, mixed lights of blue and white (ratio of 3:1) at 7000 xl were used to expedite the morphologic changes of H. pluvialis from green cells to red cyst cells with the yield of 91.8±2.53 mg/L.
