Laboratory- and Pilot-Scale Cultivation of Tetraselmis striata to Produce Valuable Metabolic Compounds.

Marine microalgae are considered an important feedstock of multiple valuable metabolic compounds of high biotechnological potential. In this work, the marine microalga Tetraselmis striata was cultivated in different scaled photobioreactors (PBRs). Initially, experiments were performed using two different growth substrates (a modified F/2 and the commercial fertilizer Nutri-Leaf (30% TN-10% P-10% K)) to identify the most efficient and low-cost growth medium. These experiments took place in 4 L glass aquariums at the laboratory scale and in a 9 L vertical tubular pilot column. Enhanced biomass productivities (up to 83.2 mg L-1 d-1) and improved biomass composition (up to 41.8% d.w. proteins, 18.7% d.w. carbohydrates, 25.7% d.w. lipids and 4.2% d.w. total chlorophylls) were found when the fertilizer was used. Pilot-scale experiments were then performed using Nutri-Leaf as a growth medium in different PBRs: (a) a paddle wheel, open, raceway pond of 40 L, and (b) a disposable polyethylene (plastic) bag of 280 L working volume. Biomass growth and composition were also monitored at the pilot scale, showing that high-quality biomass can be produced, with important lipids (up to 27.6% d.w.), protein (up to 45.3% d.w.), carbohydrate (up to 15.5% d.w.) and pigment contents (up to 4.2% d.w. total chlorophylls), and high percentages of eicosapentaenoic acid (EPA). The research revealed that the strain successfully escalated in larger volumes and the biochemical composition of its biomass presents high commercial interest and could potentially be used as a feed ingredient.

Adaptive laboratory evolution principles and applications in industrial biotechnology.

Adaptive laboratory evolution (ALE) is an innovative approach for the generation of evolved microbial strains with desired characteristics, by implementing the rules of natural selection as presented in the Darwinian Theory, on the laboratory bench. New as it might be, it has already been used by several researchers for the amelioration of a variety of characteristics of widely used microorganisms in biotechnology. ALE is used as a tool for the deeper understanding of the genetic and/or metabolic pathways of evolution. Another important field targeted by ALE is the manufacturing of products of (high) added value, such as ethanol, butanol and lipids. In the current review, we discuss the basic principles and techniques of ALE, and then we focus on studies where it has been applied to bacteria, fungi and microalgae, aiming to improve their performance to biotechnological procedures and/or inspect the genetic background of evolution. We conclude that ALE is a promising and efficacious method that has already led to the acquisition of useful new microbiological strains in biotechnology and could possibly offer even more interesting results in the future.

Laboratory evolution strategies for improving lipid accumulation in Yarrowia lipolytica.

Oleaginous microorganisms are of high biotechnological interest being considered as alternative sources of oil (single cell oil-SCO). Current research for increasing productivity of oleaginous microorganisms is focused on the overexpression of genes implicated in lipid synthesis, the inactivation of genes implicated in storage lipid turnover, and on the suppression of competitive to lipid biosynthesis pathways. An alternative strategy, described here, relies on evolution of Yarrowia lipolytica under alternating environments that promote growth, encourage storage lipid synthesis, and reward high energy-containing cells. Derived populations were characterized biochemically, especially on their ability to accumulate lipids, and compared with the starting strain. Interestingly, lipid-accumulating ability early in the evolution was decreased compared with the starting strain. Subsequently, oleaginous lineages dominated, leading to populations able to accumulate lipids in high amounts. A population obtained after 77 generations was able to accumulate 44% w/w of lipid, which was 30% higher than that of the starting strain. We conclude that evolution-based strategies can be utilized as a robust tool for improving lipid accumulation capacity in oleaginous microorganisms.

Data on cellular lipids of Yarrowia lipolytica grown on fatty substrates.

Yarrowia lipolytica, which is model oleaginous yeast with high industrial interest, was cultivated on fatty substrates. Data concerning fatty acid composition of both substrate and yeast lipids and comparisons of the experimental data with model predictions presented in "Biomodification of fats and oils and scenarios of adding value on renewable fatty materials through microbial fermentations: Modelling and trials with Yarrowia lipolytica" (Vasiliadou et al., 2018) were provided. Furthermore, the total yeast lipids were fractionated into their main fractions, that is, phospholipids, glucolipids plus sphingolipids and neutral lipids, and the fatty acid composition of each lipid fraction was reported.