scholarly journals The native acyltransferase-coding genes DGA1 and DGA2 affect lipid accumulation in Blastobotrys raffinosifermentans differently when overexpressed

2020 ◽  
Vol 20 (8) ◽  
Author(s):  
Daniel Ruben Akiola Sanya ◽  
Djamila Onesime ◽  
Gotthard Kunze ◽  
Cécile Neuveglise ◽  
Anne-Marie Crutz-Le Coq
Keyword(s):  
Lipid Content ◽  
Lipid Accumulation ◽  
Parental Strain ◽  
Nitrogen Limitation ◽  
Lipid Production ◽  
Strain Analysis ◽  
Ascomycetous Yeast ◽  
Transcript Levels ◽  
Dry Cell Weight

ABSTRACT Blastobotrys raffinosifermentans is an ascomycetous yeast with biotechnological applications, recently shown to be an oleaginous yeast accumulating lipids under nitrogen limitation. Diacylglycerol acyltransferases (DGATs) act in the lipid storage pathway, in the last step of triacylglycerol biosynthesis. Two DGAT families are widespread in eukaryotes. We first checked that B. raffinosifermentans strain LS3 possessed both types of DGAT, and we then overexpressed the native DGAT-encoding genes, DGA1 and DGA2, separately or together. DGA2 (from the DGAT1 family) overexpression was sufficient to increase lipid content significantly in LS3, to up to 26.5% of dry cell weight (DCW), 1.6 times the lipid content of the parental strain (16.90% of DCW) in glucose medium under nitrogen limitation. By contrast, DGA1 (of the DGAT2 type) overexpression led to a large increase (up to 140-fold) in the amount of the corresponding transcript, but had no effect on overall lipid content relative to the parental strain. Analysis of the expression of the native genes over time in the parental strain revealed that DGA2 transcript levels quadrupled between 8 and 24 h in the N-limited lipogenic medium, whereas DGA1 transcript levels remained stable. This survey highlights the predominant role of the DGAT1 family in lipid accumulation and demonstrates the suitability of B. raffinosifermentans for engineering for lipid production.

scholarly journals Fast media optimization for mixotrophic cultivation of Chlorella vulgaris

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Valerie C. A. Ward ◽  
Lars Rehmann
Keyword(s):  
Cell Density ◽  
Lipid Content ◽  
Lipid Accumulation ◽  
Accumulation Rate ◽  
Lipid Production ◽  
Total Lipid Content ◽  
Mixotrophic Cultivation ◽  
Cell Weight ◽  
Dry Cell Weight ◽  
Dry Cell

AbstractMicroalgae can accumulate large proportions of their dry cell weight as storage lipids when grown under appropriate nutrient limiting conditions. While a high ratio of carbon to nitrogen is often cited as the primary mode of triggering lipid accumulation in microalgae, fast optimization strategies to increase lipid production for mixotrophic cultivation have been difficult to developed due to the low cell densities of algal cultures, and consequently the limited amount of biomass available for compositional analysis. Response surface methodologies provide a power tool for assessing complex relationships such as the interaction between the carbon source and nitrogen source. A 15 run Box-Behnken design performed in shaker flasks was effective in studying the effect of carbon, nitrogen, and magnesium on the growth rate, maximum cell density, lipid accumulation rate, and glucose consumption rate. Using end-point dry cell weight and total lipid content as assessed by direct transesterification to FAME, numerical optimization resulted in a significant increase in lipid content from 18.5 ± 0.76% to 37.6 ± 0.12% and a cell density of 5.3 ± 0.1 g/L to 6.1 ± 0.1 g/L between the centre point of the design and the optimized culture conditions. The presented optimization process required less than 2 weeks to complete, was simple, and resulted in an overall lipid productivity of 383 mg/L·d.

scholarly journals A Mathematical Model of Neutral Lipid Content in terms of Initial Nitrogen Concentration and Validation in Coelastrum sp. HA-1 and Application in Chlorella sorokiniana

2017 ◽  
Vol 2017 ◽  
pp. 1-10 ◽  
Author(s):  
Zhenhua Yang ◽  
Yue Zhao ◽  
Zhiyong Liu ◽  
Chenfeng Liu ◽  
Zhipeng Hu ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Lipid Content ◽  
Lipid Accumulation ◽  
Nitrogen Concentration ◽  
Lipid Production ◽  
Lipid Synthesis ◽  
Quantitative Relationship ◽  
Chlorella Sorokiniana ◽  
High Lipid ◽  
Nitrogen Concentrations

Microalgae are considered to be a potential major biomass feedstock for biofuel due to their high lipid content. However, no correlation equations as a function of initial nitrogen concentration for lipid accumulation have been developed for simplicity to predict lipid production and optimize the lipid production process. In this study, a lipid accumulation model was developed with simple parameters based on the assumption protein synthesis shift to lipid synthesis by a linear function of nitrogen quota. The model predictions fitted well for the growth, lipid content, and nitrogen consumption of Coelastrum sp. HA-1 under various initial nitrogen concentrations. Then the model was applied successfully in Chlorella sorokiniana to predict the lipid content with different light intensities. The quantitative relationship between initial nitrogen concentrations and the final lipid content with sensitivity analysis of the model were also discussed. Based on the model results, the conversion efficiency from protein synthesis to lipid synthesis is higher and higher in microalgae metabolism process as nitrogen decreases; however, the carbohydrate composition content remains basically unchanged neither in HA-1 nor in C. sorokiniana.

scholarly journals Molecular Mechanism of Lipid Accumulation and Metabolism of Oleaginous Chlorococcum sphacosum GD from Soil under Salt Stress

2021 ◽  
Vol 22 (3) ◽  
pp. 1304
Author(s):  
Hang Su ◽  
Jia Feng ◽  
Junping Lv ◽  
Qi Liu ◽  
Fangru Nan ◽  
...  
Keyword(s):  
Salt Stress ◽  
Fatty Acid ◽  
Lipid Content ◽  
Salt Concentration ◽  
Lipid Accumulation ◽  
Transcriptome Sequencing ◽  
High Throughput Sequencing ◽  
Lipid Production ◽  
Biodiesel Production ◽  
Oleaginous Microalgae

The oleaginous microalgae species Chlorococcum sphacosum GD is a promising feedstock for biodiesel production from soil. However, its metabolic mechanism of lipid production remains unclear. In this study, the lipid accumulation and metabolism mechanisms of Chlorococcum sphacosum GD were analyzed under salt stress based on transcriptome sequencing. The biomass and lipid content of the alga strain were determined under different NaCl concentrations, and total RNA from fresh cells were isolated and sequenced by HiSeq 2000 high throughput sequencing technology. As the salt concentration increased in culture medium, the algal lipid content increased but the biomass decreased. Following transcriptome sequencing by assembly and splicing, 24,128 unigenes were annotated, with read lengths mostly distributed in the 200–300 bp interval. Statistically significant differentially expressed unigenes were observed in different experimental groups, with 2051 up-regulated genes and 1835 down-regulated genes. The lipid metabolism pathway analysis showed that, under salt stress, gene-related fatty acid biosynthesis (ACCase, KASII, KAR, HAD, FATA) was significantly up-regulated, but some gene-related fatty acid degradation was significantly down-regulated. The comprehensive results showed that salt concentration can affect the lipid accumulation and metabolism of C. sphacosum GD, and the lipid accumulation is closely related to the fatty acid synthesis pathway.

scholarly journals miR-146a modulates TLR1/2 and 4 induced inflammation and links it with proliferation and lipid production via the indirect regulation of GNG7 in human SZ95 sebocytes

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Katalin Dull ◽  
Fruzsina Fazekas ◽  
Dávid Deák ◽  
Dóra Kovács ◽  
Szilárd Póliska ◽  
...  
Keyword(s):  
Gene Expression ◽  
In Situ Hybridization ◽  
Gene Expression Profile ◽  
Lipid Content ◽  
Lipid Production ◽  
Sebaceous Glands ◽  
Down Regulation ◽  
Protein Coding

AbstractActivation of Toll-like receptors (TLR) 1/2 and 4 are central in inducing inflammation in sebocytes by regulating the expression of protein coding mRNAs, however the microRNA (miRNA) profile in response to TLR activation and thus the possible role of miRNAs in modulating sebocyte functions has not been elucidated. In this work we identified miR-146a to have the highest induction in the TLR1/2 and 4 activated SZ95 sebocytes and found that its increased levels led to the down-regulation of IL-8 secretion, decreased the chemoattractant potential and stimulated the proliferation of sebocytes. Assessing the gene expression profile of SZ95 sebocytes treated with a miR-146a inhibitor, the induction of GNG7 was one of the highest, while when cells were treated with a miR-146a mimic, the expression of GNG7 was down-regulated. These findings correlated with our in situ hybridization results, that compared with control, miR-146a showed an increased, while GNG7 a decreased expression in sebaceous glands of acne samples. Further studies revealed, that when inhibiting the levels of GNG7 in SZ95 sebocytes, cells increased their lipid content and decreased their proliferation. Our findings suggest, that miR-146a could be a potential player in acne pathogenesis by regulating inflammation, inducing proliferation and, through the indirect down-regulation of GNG7, promoting the lipid production of sebocytes.

scholarly journals Investigation of the Relationship between Bacteria Growth and Lipid Production Cultivating of Microalgae Chlorella Vulgaris in Seafood Wastewater

Energies ◽  
2019 ◽  
Vol 12 (12) ◽  
pp. 2282 ◽  
Author(s):  
Thi Dong Phuong Nguyen ◽  
Duc Huy Nguyen ◽  
Jun Wei Lim ◽  
Chih-Kai Chang ◽  
Hui Yi Leong ◽  
...  
Keyword(s):  
Chlorella Vulgaris ◽  
Lipid Content ◽  
Lipid Accumulation ◽  
Lipid Production ◽  
Oxygen Demand ◽  
Value Added ◽  
Bacteria Growth ◽  
Microalgae Culture ◽  
The Relationship ◽  
Microalgae Growth

Algae biorefinery is gaining much attention for the sustainable production of value-added products (e.g., biofuels, protein supplements etc.) globally. The current study aimed to investigate the relationship between lipid production and bacteria growth by an initial microalgae Chlorella vulgaris density culture in seafood wastewater effluent (SWE). According to our results, the initial C. vulgaris concentration in SWE influenced lipid accumulation. The concentration ranged from 25–35 mg·L−1 which corresponds to SWE’s chemical oxygen demand concentration of 365.67 ± 3.45 mg·L−1. A higher microalgae growth rate and lipid content of 32.15 ± 1.45% was successfully attained. A higher lipid content, approximately double, was observed when compared to the control (16.8 ± 0.5%). Moreover, this study demonstrates that bacteria inhibited microalgae growth as the initial cell density stepped over 35 mg·L−1, which also affected lipid accumulation. This study shows an optimal lipid accumulation attained at moderate Chlorella vulgaris density culture in SWE. Hence, wastewater treatment incorporating microalgae culture could be greatly developed in the future to achieve a greener environment.

scholarly journals DGA1 (diacylglycerol acyltransferase gene) overexpression and leucine biosynthesis significantly increase lipid accumulation in the Δsnf2 disruptant of Saccharomyces cerevisiae

2007 ◽  
Vol 408 (1) ◽  
pp. 61-68 ◽  
Author(s):  
Yasushi Kamisaka ◽  
Nao Tomita ◽  
Kazuyoshi Kimura ◽  
Kumiko Kainou ◽  
Hiroshi Uemura
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Lipid Content ◽  
Lipid Accumulation ◽  
Lipid Production ◽  
Total Lipid Content ◽  
Diacylglycerol Acyltransferase ◽  
Leucine Biosynthesis ◽  
Gene Encoding ◽  
Acyltransferase Activity ◽  
Isopropylmalate Dehydrogenase

We previously found that SNF2, a gene encoding a transcription factor forming part of the SWI/SNF (switching/sucrose non-fermenting) chromatin-remodelling complex, is involved in lipid accumulation, because the Δsnf2 disruptant of Saccharomyces cerevisiae has a higher lipid content. The present study was conducted to identify other factors that might further increase lipid accumulation in the Δsnf2 disruptant. First, expression of LEU2 (a gene encoding β-isopropylmalate dehydrogenase), which was used to select transformed strains by complementation of the leucine axotroph, unexpectedly increased both growth and lipid accumulation, especially in the Δsnf2 disruptant. The effect of LEU2 expression on growth and lipid accumulation could be reproduced by adding large amounts of leucine to the culture medium, indicating that the effect was not due to Leu2p (β-isopropylmalate dehydrogenase) itself, but rather to leucine biosynthesis. To increase lipid accumulation further, genes encoding the triacylglycerol biosynthetic enzymes diacylglycerol acyltransferase (DGA1) and phospholipid:diacylglycerol acyltransferase (LRO1) were overexpressed in the Δsnf2 disruptant. Overexpression of DGA1 significantly increased lipid accumulation, especially in the Δsnf2 disruptant, whereas LRO1 overexpression decreased lipid accumulation in the Δsnf2 disruptant. Furthermore, the effect of overexpression of acyl-CoA synthase genes (FAA1, FAA2, FAA3 and FAA4), which each supply a substrate for Dga1p (diacylglycerol acyltransferase), was investigated. Overexpression of FAA3, together with that of DGA1, did not further increase lipid accumulation in the Δsnf2 disruptant, but did enhance lipid accumulation in the presence of exogenous fatty acids. Lastly, the total lipid content in the Δsnf2 disruptant transformed with DGA1 and FAA3 overexpression vectors reached approx. 30%, of which triacylglycerol was the most abundant lipid. Diacylglycerol acyltransferase activity was significantly increased in the Δsnf2 disruptant strain overexpressing DGA1 as compared with the wild-type strain overexpressing DGA1; this higher activity may account for the prominent increase in lipid accumulation in the Δsnf2 disruptant with DGA1 overexpression. The strains obtained have a lipid content that is high enough to act as a model of oleaginous yeast and they may be useful for the metabolic engineering of lipid production in yeast.

scholarly journals Role of Snf-β in lipid accumulation in the high lipid‐producing fungus Mucor circinelloides WJ11

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Shaista Nosheen ◽  
Tahira Naz ◽  
Junhuan Yang ◽  
Syed Ammar Hussain ◽  
Abu Bakr Ahmad Fazili ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Lipid Metabolism ◽  
Lipid Content ◽  
Lipid Accumulation ◽  
Dry Weight ◽  
Mucor Circinelloides ◽  
Control Strain ◽  
High Lipid ◽  
Overexpressing Strain

Abstract Background Mucor circinelloides WJ11 is a high-lipid producing strain and an excellent producer of γ-linolenic acid (GLA) which is crucial for human health. We have previously identified genes that encode for AMP-activated protein kinase (AMPK) complex in M. circinelloides which is an important regulator for lipid accumulation. Comparative transcriptional analysis between the high and low lipid-producing strains of M. circinelloides showed a direct correlation in the transcriptional level of AMPK genes with lipid metabolism. Thus, the role of Snf-β, which encodes for β subunit of AMPK complex, in lipid accumulation of the WJ11 strain was evaluated in the present study. Results The results showed that lipid content of cell dry weight in Snf-β knockout strain was increased by 32 % (from 19 to 25 %). However, in Snf-β overexpressing strain, lipid content of cell dry weight was decreased about 25 % (from 19 to 14.2 %) compared to the control strain. Total fatty acid analysis revealed that the expression of the Snf-β gene did not significantly affect the fatty acid composition of the strains. However, GLA content in biomass was increased from 2.5 % in control strain to 3.3 % in Snf-β knockout strain due to increased lipid accumulation and decreased to 1.83 % in Snf-β overexpressing strain. AMPK is known to inactivate acetyl-CoA carboxylase (ACC) which catalyzes the rate-limiting step in lipid synthesis. Snf-β manipulation also altered the expression level of the ACC1 gene which may indicate that Snf-β control lipid metabolism by regulating ACC1 gene. Conclusions Our results suggested that Snf-β gene plays an important role in regulating lipid accumulation in M. circinelloides WJ11. Moreover, it will be interesting to evaluate the potential of other key subunits of AMPK related to lipid metabolism. Better insight can show us the way to manipulate these subunits effectively for upscaling the lipid production. Up to our knowledge, it is the first study to investigate the role of Snf-β in lipid accumulation in M. circinelloides.

scholarly journals Transcriptional profiling reveals differentially expressed genes involved in lipid biosynthesis during cacao seed development

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Fupeng Li ◽  
Baoduo Wu ◽  
Lin Yan ◽  
Chaoyun Hao ◽  
Xiaowei Qin ◽  
...  
Keyword(s):  
Differentially Expressed Genes ◽  
Lipid Content ◽  
Lipid Accumulation ◽  
Unsaturated Fatty Acids ◽  
Transcriptional Profiling ◽  
Lipid Production ◽  
Economic Value ◽  
Lipid Synthesis ◽  
Differentially Expressed ◽  
Seed Lipid

AbstractTheobroma cacao is a plant of economic value due to the use of its seed lipid for chocolate, confectionery, and cosmetic industries. The seed lipid contains a stable ratio of saturated and unsaturated fatty acids, which determines its unique melting temperature. However, little is known about the molecular mechanism determining the fatty acid ratio and lipid content in cacao. To gain insight into the unique properties of lipid synthesis in cacao, biochemical and transcriptomic approaches were used to compare the lipid accumulation between high and low lipid content cacao accessions. Lipid accumulation rates and lipid content were different between the two accessions. Moreover, differentially expressed genes were detected between high and low lipid content cacao accessions. The data allowed the identification of distinct candidate genes and furthered our understanding of lipid accumulation, potentially explaining the differences in lipid content between various cacao accessions. The results might be used to develop molecular tools and engineer alternative pathways for cacao breeding with improved lipid production potentials.

scholarly journals Statistical study of growth kinetics and lipid content of microalgae grown in brackish waters for bioenergetic purposes

2021 ◽  
Vol 16 (3) ◽  
pp. 1
Author(s):  
Bruna Da Silveira Guimarães ◽  
Kepler Borges França
Keyword(s):  
Biomass Production ◽  
Growth Kinetics ◽  
Lipid Content ◽  
Lipid Accumulation ◽  
Lipid Production ◽  
Maximum Growth ◽  
Growth Speed ◽  
Saline Stress ◽  
Dry Biomass ◽  
Scenedesmus Acuminatus

This study aimed to statistically prove the influence of salinity on growth kinetics and intracellular lipid accumulation in microalgae. The species Chlorella sp., Scenedesmus acuminatus, Nannochloropsis sp., Monoraphidium contortum and Pediastrum tetras were studied, which were isolated in the Brazilian semi-arid region and cultivated in synthetic media with the addition of NaCl. From the crops, growth kinetics, dry biomass production and lipid content were analyzed, using the experimental planning for one factor as a tool, with the NaCl concentration as an independent variable. The kinetic parameters maximum growth speed (µmax) and generation time (tg), as well as lipid concentration were the response variables studied. The results showed that, in the species Scenedesmus acuminatus, Nannochloropsis sp. and Pediastrum tetras, salt stress contributed to the increase in µmax and the consequent decrease in tg. The highest levels of lipid accumulation were obtained in Nannochloropsis sp. (62.04%), in the medium with the highest salinity and Pediastrum tetras (54.04%) in the lowest. Dry biomass production was higher in Scenedesmus acuminatus (1,941.37 mg.L-1) and Nannochloropsis sp. (1237.05 mg.L-1), both at a concentration of 4.0 g.L-1. Therefore, saline stress acted directly on the cell growth and lipid content of the species, which can be used as a device to enhance lipid production for the purpose of producing biofuels.

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