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The effect of dissolved oxygen concentration on lipid accumulation in Trichosporon oleaginosus has been investigated. The experiment was performed in 15 L fermenters. The dissolved oxygen concentration varied by adjusting the agitation and aeration. High dissolved oxygen level at 50%–60% enhanced cell growth. Maintaining low dissolved oxygen concentration at 20%–30% during lipogenesis phase led to high final lipid content (51%) in Trichosporon oleaginosus. The consumptions of energy and cost of the process were evaluated. The energy consumption in the dissolved oxygen level optimized process was 41% less than that with dissolved oxygen level at 50%–60%. In addition, the cost was also reduced around one time in the dissolved oxygen level optimized process compared to the one with dissolved oxygen level at 50%–60%. The study provided a feasible way of enhancing lipid accumulation in Trichosporon oleaginosus and reducing the consumption of energy and cost of lipid production from Trichosporon oleaginosus.

In this study, two types of organic waste including sludge and crude glycerol were employed for lipid production from oleaginous yeast Trichosporon oleaginosus. As wastewater sludge contains metals and other substances, which could inhibit growth of the microorganisms, sludge washing was performed to reduce/remove the substance in order to enhance the lipid production in this study. The results revealed that the lipid production by Trichosporon oleaginosus was higher in the medium prepared using washed sludge fortified with glycerol compared to that of the unwashed sludge fortified with glycerol. The maximum biomass and lipid concentrations were 44.48 ± 2.34 g/L and 17.37 ± 1.22 g/L, respectively in the fermentation with washed sludge and glycerol and 39.17 ± 2.06 g/L and 14.26 ± 1.17 g/L in the fermentation with unwashed sludge and glycerol. The study showed that sludge washing had improved lipid production from sludge.

Mitochondrial pyruvate dehydrogenase (PDH) is important in the production of lipids in oleaginous yeast, but other yeast may bypass the mitochondria (PDH bypass), converting pyruvate in the cytosol to acetaldehyde, then acetate and acetyl CoA which is further converted to lipids. Using a metabolic model based on the oleaginous yeast Yarrowia lipolytica, we found that introduction of this bypass to an oleaginous yeast should result in enhanced yield of triacylglycerol (TAG) on substrate. Trichosporon oleaginosus (formerly Cryptococcus curvatus) is an oleaginous yeast which can produce TAGs from both glucose and xylose. Based on the sequenced genome, it lacks at least one of the enzymes needed to complete the PDH bypass, acetaldehyde dehydrogenase (ALD), and may also be deficient in pyruvate decarboxylase and acetyl-CoA synthetase under production conditions. We introduced these genes to T. oleaginosus in various combinations and demonstrated that the yield of TAG on both glucose and xylose was improved, particularly at high C/N ratio. Expression of a phospholipid:diacyltransferase encoding gene in conjunction with the PDH bypass further enhanced lipid production. The yield of TAG on xylose (0.27 g/g) in the engineered strain approached the theoretical maximum yield of 0.289 g/g. Interestingly, TAG production was also enhanced compared to the control in some strains which were given only part of the bypass pathway, suggesting that these genes may contribute to alternative routes to cytoplasmic acetyl CoA. The metabolic model indicated that the improved yield of TAG on substrate in the PDH bypass was dependent on the production of NADPH by ALD. NADPH for lipid synthesis is otherwise primarily supplied by the pentose phosphate pathway (PPP). This would contribute to the greater improvement of TAG production from xylose compared to that observed from glucose when the PDH bypass was introduced, since xylose enters metabolism through the non-oxidative part of the PPP. Yield of TAG from xylose in the engineered strains (0.21-0.27 g/g) was comparable to that obtained from glucose and the highest so far reported for lipid or TAG production from xylose.

In this study, it was found that the optimal pH for the growth of Trichosporon oleaginosus was related to the fermentation medium. A neutral or weak acid pH condition was optimal for the growth of Trichosporon oleaginosus in the extract-peptone-dextrose and wastewater sludge medium. Significant inhibition was observed at neutral pH in the wastewater sludge + crude glycerol medium due to the high soap content of the crude glycerol. By converting the soap to free fatty acid (FFA) at pH 5, the soap inhibition could be prevented. Fed-batch fermentation was employed to produce lipid from Trichosporon oleaginosus at pH 5 controlled by feeding crude glycerol. A remarkably high biomass (65.63 g/L) and lipid (35.79 g/L) concentration were achieved from the pH-based fed-batch fermentation in this study.

Background The oleaginous yeast, Trichosporon oleaginosus, has been extensively studied for its ability to metabolize non-conventional feedstocks. These include phenol-containing waste streams, such as distillery wastewater, or streams consisting of non-conventional sugars, such as hydrolyzed biomass and various bagasse. An initial BLAST search suggests this yeast has putative aromatic metabolizing genes. Given the desirability to valorize underutilized feedstocks such as lignin, we investigated the ability of T. oleaginosus to tolerate and metabolize lignin-derived aromatic compounds. Results Trichosporon oleaginosus can tolerate and metabolize model lignin monoaromatics and associated intermediates within funneling pathways. Growth rates and biomass yield were similar to glucose when grown in 4-hydroxybenzoic acid (pHBA) and resorcinol, but had an increased lag phase when grown in phenol. Oleaginous behavior was observed using resorcinol as a sole carbon source. Fed-batch feeding resulted in lipid accumulation of 69.5% on a dry weight basis. Conclusions Though the exact pathway of aromatic metabolism remains to be determined for T. oleaginosus, the results presented in this work motivate use of this organism for lignin valorization and phenolic wastewater bioremediation. Trichosporon oleaginosus is the first yeast shown to be oleaginous while growing on aromatic substrates, and shows great promise as a model industrial microbe for biochemical and biofuel production from depolymerized lignin. Electronic supplementary material The online version of this article (10.1186/s12934-017-0820-8) contains supplementary material, which is available to authorized users.

The article Engineering Trichosporon oleaginosus for enhanced production of lipid from volatile fatty acids as carbon source, written by Kyungsoo Lee*,‡, Yong Jae Lee**,‡, Ho Nam Chang***, and Ki Jun Jeong*,****,†, was originally published on the publisher’s internet portal (currently SPringerLink) on 08 February 2019 with misprinted DOI number, 10.1007/s11814-018-0229-7, due to the technical error from converting manuscript file from Microsoft Word to PDF. The correct DOI number for the article is 10.1007/s11814-019-0312-0.