Crypthecodinium cohnii and Zymomonas mobilis syntrophy for production of omega 3 fatty acid

One of missions of industrial biotechnology is to produce valuable products from waste. In the current study we aim to establish a bioprocess of syntrophic co-cultivation of two microorganisms (Crypthecodinium cohnii and Zymomonas mobilis) at laboratory prototype level for bioconversion of renewables (molasses, the byproduct of sugar industry, and glycerol, byproduct of biodiesel production) into valuable vegetarian (produced by microorganisms) docosahexaenoic acid (DHA) from omega 3 fatty acid family. Cells of Z.mobilis strain (respiratory-deficient mutant derivative of the wild type strain Z.mobilis ATCC 29191) will be immobilized, following the previously described procedure of calcium alginate gel encapsulation. Mathematical modelling will be used for different components and at different stages of the syntrophic process to assist its improvement. The syntrophic bioprocess will be optimized with respect to (i) the medium composition, (ii) feed rates, (iii) oxygen supply, and (iv) the amounts of added immobilized Z. mobilis cells at particular stages of the fermentation. Genome and/or central carbon metabolism scale stoichiometric modelling and optimization will be performed to increase the utilization of cheap substrates and pre-treated residuals of C.cohnii. Bioprocess dynamics with respect to growth speed of batch fermentations will be modelled to ensure optimal proportions of syntrophic organisms, taking into account Z.mobilis production and immobilization as “slave” process of C.cohnii cultivation. Within several iterative cycles the fermentation data will be used for modelling and computer simulation of the bioprocess, and at the same time, the model-predicted dynamic behaviour of the system will aid further improvement of fermentation performance.publishersversionPeer reviewe

Investigation of a Broad-Bean Based Low-Cost Medium Formulation for <i>Bacillus subtilis</i> MSCL 897 Spore Production

Bacillus subtilis (Bs) is a bacterium that benefits plants and is used in the production of bio-fungicides. The cultivation of Bs is a crucial step in bio-control preparation production, as it greatly impacts the quality and price of the final product. In a series of shake flask experiments, we investigated the economically feasible broth composition for spore production of Bacillus subtilis MSCL 897, a Latvian soil isolate. Our study investigated the impact of utilizing legume-based flours (such as broad bean, grey pea, and soybean) as the primary nitrogen source, along with sugar-beet molasses, sucrose, or glucose as the carbon source, and yeast extract, peptone, and corn-steep liquor as growth factor additives. Additionally, we examined the effect of using (NH4)2HPO4 or urea as supplementary nitrogen sources, as well as previously established media formulations, on spore yield. Our results showed that a culture medium composed of broad bean flour (10 g/L) and molasses (10 g/L) led to spore productivity of 1.35 ± 0.47 × 108 CFU/mL at 48 h. By enriching the culture medium base constituents with a minor (0.5–1.0 g/L) yeast extract or corn-steep liquor additive, a notable increase in spore productivity was observed, with values of 2.00 ± 0.28 × 108 and 2.34 ± 0.18 × 108 CFU/mL at 48 h, respectively, and sporulation efficiency > 80–90%. As a result, we achieved a high spore yield of the Bacillus subtilis MSCL 897 strain, demonstrating the competitiveness of our approach, which relied on a low-cost medium made mainly from locally available and renewable raw materials

The role of magnetic field in the biopharmaceutical production: Current perspectives

Current scientific evidence on the influence of magnetic field on mammalian cell lines used for industrial production of biopharmaceuticals, on human cell lines and on potential cell lines for the biopharmaceutical production is presented in this review. A novel magnetic coupling induced agitation could be the best solution to eliminate sources of contamination in stirred tank bioreactors which is especially important for mammalian cell cultures. Nevertheless, the side effect of magnetically-coupled stirring mechanism is that cells are exposed to the generated magnetic field. The influence of magnetic field on biological systems has been investigated for several decades. The research continues nowadays as well, investigating the influence of various types of magnetic field in a variety of experimental setups. In the context of bioreactors, only the lower frequencies and intensities of the magnetic field are relevant. Keywords: Time-varying magnetic field, Static magnetic field, Magnetic field influence, Mammalian cells, Biopharmaceutical

Crypthecodinium cohnii Growth and Omega Fatty Acid Production in Mediums Supplemented with Extract from Recycled Biomass

Crypthecodinium cohnii is a marine heterotrophic dinoflagellate that can accumulate high amounts of omega-3 polyunsaturated fatty acids (PUFAs), and thus has the potential to replace conventional PUFAs production with eco-friendlier technology. So far, C. cohnii cultivation has been mainly carried out with the use of yeast extract (YE) as a nitrogen source. In the present study, alternative carbon and nitrogen sources were studied: the extraction ethanol (EE), remaining after lipid extraction, as a carbon source, and dinoflagellate extract (DE) from recycled algae biomass C. cohnii as a source of carbon, nitrogen, and vitamins. In mediums with glucose and DE, the highest specific biomass growth rate reached a maximum of 1.012 h&minus;1, while the biomass yield from substrate reached 0.601 g&middot;g&minus;1. EE as the carbon source, in comparison to pure ethanol, showed good results in terms of stimulating the biomass growth rate (an 18.5% increase in specific biomass growth rate was observed). DE supplement to the EE-based mediums promoted both the biomass growth (the specific growth rate reached 0.701 h&minus;1) and yield from the substrate (0.234 g&middot;g&minus;1). The FTIR spectroscopy data showed that mediums supplemented with EE or DE promoted the accumulation of PUFAs/docosahexaenoic acid (DHA), when compared to mediums containing glucose and commercial YE

Pichia pastoris growth—coupled heme biosynthesis analysis using metabolic modelling

Abstract Soy leghemoglobin is one of the most important and key ingredients in plant-based meat substitutes that can imitate the colour and flavour of the meat. To improve the high-yield production of leghemoglobin protein and its main component—heme in the yeast Pichia pastoris, glycerol and methanol cultivation conditions were studied. Additionally, in-silico metabolic modelling analysis of growth-coupled enzyme quantity, suggests metabolic gene up/down-regulation strategies for heme production. First, cultivations and metabolic modelling analysis of P. pastoris were performed on glycerol and methanol in different growth media. Glycerol cultivation uptake and production rates can be increased by 50% according to metabolic modelling results, but methanol cultivation—is near the theoretical maximum. Growth-coupled metabolic optimisation results revealed the best feasible upregulation (33 reactions) (1.47% of total reactions) and 66 downregulation/deletion (2.98% of total) reaction suggestions. Finally, we describe reaction regulation suggestions with the highest potential to increase heme production yields