Overexpression of an Inulinase Gene in an Oleaginous Yeast, Aureobasidium melanogenum P10, for Efficient Lipid Production from Inulin

2018 ◽  
Vol 28 (4) ◽  
pp. 190-200 ◽  
Author(s):  
Yan-Feng Li ◽  
Hong Jiang ◽  
Zhong Hu ◽  
Guang-Lei Liu ◽  
Zhen-Ming Chi ◽  
...  
Keyword(s):  
Lipid Production ◽  
Dry Weight ◽  
Genetically Engineered ◽  
Biodiesel Production ◽  
Yeast Cells ◽  
Gene Encoding ◽  
Direct Production ◽  
Inulinase Gene ◽  
Inulinase Activity ◽  
Engineered Yeast

In this study, in order to directly and efficiently convert inulin into a single-cell oil (SCO), an <i>INU1</i> gene encoding inulinase from<b><i></i></b> <i>Kluyveromyces marxianus</i> was integrated into the genomic DNA and actively expressed in an SCO producer <i>Aureobasidium</i> <i>melanogenum</i> P10. The transformant API41 obtained produced 28.5 U/mL of inulinase and its wild-type strain P10 yielded only 8.62 U/mL. Most (97.5%) of the inulinase produced by the transformant API41 was secreted into the culture. During a 10-L fermentation, 66.2% (w/w) lipid in the yeast cells of the transformant API41 and 14.38 g/L of cell dry weight were attained from inulin of 80.0 g/L within 120 h, high inulinase activity (23.7 U/mL) was also produced within 72 h, and the added inulin was actively hydrolyzed. This confirmed that the genetically engineered yeast of <i>A. melanogenum</i> P10 is suitable for direct production of lipids from inulin. The lipids produced could be used as feedstocks for biodiesel production.

scholarly journals Consortium Growth of Filamentous Fungi and Microalgae: Evaluation of Different Cultivation Strategies to Optimize Cell Harvesting and Lipid Accumulation

Energies ◽  
2020 ◽  
Vol 13 (14) ◽  
pp. 3648
Author(s):  
Savienne M. F. E. Zorn ◽  
Cristiano E. R. Reis ◽  
Messias B. Silva ◽  
Bo Hu ◽  
Heizir F. De Castro
Keyword(s):  
Lipid Accumulation ◽  
Unsaturated Fatty Acids ◽  
Lipid Production ◽  
Synthetic Medium ◽  
Fungal Spores ◽  
Dry Weight ◽  
Biodiesel Production ◽  
Biomass Composition ◽  
Fungal Mycelium ◽  
Fourfold Increase

This study aims to evaluate the potential of consortium biomass formation between Mucor circinelloides, an oleaginous filamentous fungal species, and Chlorella vulgaris, in order to promote a straightforward approach to harvest microalgal cells and to evaluate the lipid production in the consortium system. A synthetic medium with glucose (2 g·L−1) and mineral nutrients essential for both fungi and algae was selected. Four different inoculation strategies were assessed, considering the effect of simultaneous vs. separate development of fungal spores and algae cells, and the presence of a supporting matrix aiming at the higher recovery of algae cell rates. The results were evaluated in terms of consortium biomass composition, demonstrating that the strategy using a mature fungal mycelium with a higher algae count may provide biomass samples with up to 79% of their dry weight as algae, still promoting recovery rates greater than 97%. The findings demonstrate a synergistic effect on the lipid accumulation by the fungal strain, at around a fourfold increase when compared to the axenic control, with values in the range of 23% of dry biomass weight. Furthermore, the fatty acid profile from the samples presents a balance between saturated and unsaturated fatty acids that is likely to present an adequate balance for applications such as biodiesel production.

Novo human insulin - the same but different

1990 ◽  
Vol 28 (6) ◽  
pp. 24-24
Keyword(s):  
Human Insulin ◽  
Manufacturing Process ◽  
Genetically Engineered ◽  
Yeast Cells ◽  
Enzymatic Modification ◽  
Engineered Yeast ◽  
Direct Biosynthesis

Unannounced to prescribers or patients, Novo Nordisk have changed the manufacturing process of their human insulins from enzymatic modification of pig insulin (emp) to direct biosynthesis of genetically engineered yeast cells (pyr). The preservative for Human Actrapid insulin has also been changed from phenol to meta-cresol. No warning of this change was sent to pharmacists, doctors, diabetes specialists or patients.

Detection of hormone active chemicals using genetically engineered yeast cells and microfluidic devices with interdigitated array electrodes

2009 ◽  
Vol 30 (19) ◽  
pp. 3406-3412 ◽  
Author(s):  
Kosuke Ino ◽  
Yusuke Kitagawa ◽  
Tsuyoshi Watanabe ◽  
Hitoshi Shiku ◽  
Masahiro Koide ◽  
...  
Keyword(s):  
Microfluidic Devices ◽  
Genetically Engineered ◽  
Yeast Cells ◽  
Interdigitated Array Electrodes ◽  
Engineered Yeast ◽  
Interdigitated Array

Genetically engineered yeast cells and their applications

1995 ◽  
Vol 82-83 ◽  
pp. 815-822 ◽  
Author(s):  
Denis Pompon ◽  
Alain Perret ◽  
Aouatef Bellamine ◽  
Romuald Laine ◽  
Jean-Charles Gautier ◽  
...  
Keyword(s):  
Genetically Engineered ◽  
Yeast Cells ◽  
Engineered Yeast

scholarly journals Sugarcane Distillery Spent Wash, a New Resource for Third-Generation Biodiesel Production

Water ◽  
2018 ◽  
Vol 10 (11) ◽  
pp. 1623 ◽  
Author(s):  
Julien Hoarau ◽  
Isabelle Grondin ◽  
Yanis Caro ◽  
Thomas Petit
Keyword(s):  
Low Cost ◽  
Lipid Production ◽  
Liquid Waste ◽  
Fungal Growth ◽  
Dry Weight ◽  
Biodiesel Production ◽  
Aspergillus Awamori ◽  
Cryptococcus Curvatus ◽  
Spent Wash ◽  
Distillery Spent Wash

Industrial production of biodiesel from microbial catalysts requires large volume of low-cost feedstock for lipid production. Vinasse, also known as distillery spent wash (DSW), is a liquid waste produced in large amounts by ethanol distilleries. This effluent is particularly rich in organic matter, and may be considered as a potential resource for the production of fungal lipids. The present study aimed at evaluating the potential of vinasse from a distillery located in Reunion Island for yeast and fungal growth, lipid production, and suitability for biodiesel requirements. Among the 28 different strains tested, we found that Aspergillus niger grown on pure vinasse allowed biomass production of up to 24.05 g/L (dry weight), whereas Aspergillus awamori produced the maximum amount of lipid, at 2.27 g/L. Nutrient removal and vinasse remediation were found to be the best for A. niger and Cryptococcus curvatus, reaching a maximum of 50% for nitrogen, and A. awamori showed 50% carbon removal. Lipids produced were principally composed of C16:0, C18:1 (n-9), and C18:2 (n-6), thus resembling the vegetal oil used in the biodiesel production. This work has shown that vinasse can support production of biomass and lipids from fungi and yeast suitable for energetic use and that its polluting charge can be significantly reduced through this process.

scholarly journals Utilization of Clarified Butter Sediment Waste as a Feedstock for Cost-Effective Production of Biodiesel

Foods ◽  
2019 ◽  
Vol 8 (7) ◽  
pp. 234 ◽  
Author(s):  
Alok Patel ◽  
Km Sartaj ◽  
Parul A. Pruthi ◽  
Vikas Pruthi ◽  
Leonidas Matsakas
Keyword(s):  
Fossil Fuels ◽  
Neutral Lipids ◽  
Renewable Energy Sources ◽  
Lipid Production ◽  
Cost Effective ◽  
Dry Weight ◽  
International Standards ◽  
Biodiesel Production ◽  
Gas Chromatography Mass Spectrometry ◽  
High Lipid

The rising demand and cost of fossil fuels (diesel and gasoline), together with the need for sustainable, alternative, and renewable energy sources have increased the interest for biomass-based fuels such as biodiesel. Among renewable sources of biofuels, biodiesel is particularly attractive as it can be used in conventional diesel engines without any modification. Oleaginous yeasts are excellent oil producers that can grow easily on various types of hydrophilic and hydrophobic waste streams that are used as feedstock for single cell oils and subsequently biodiesel production. In this study, cultivation of Rhodosporidium kratochvilovae on a hydrophobic waste (clarified butter sediment waste medium (CBM)) resulted in considerably high lipid accumulation (70.74% w/w). Maximum cell dry weight and total lipid production were 15.52 g/L and 10.98 g/L, respectively, following cultivation in CBM for 144 h. Neutral lipids were found to accumulate in the lipid bodies of cells, as visualized by BODIPY staining and fluorescence microscopy. Cells grown in CBM showed large and dispersed lipid droplets in the intracellular compartment. The fatty acid profile of biodiesel obtained after transesterification was analyzed by gas chromatography-mass spectrometry (GC–MS), while its quality was determined to comply with ASTM 6751 and EN 14214 international standards. Hence, clarified sediment waste can be exploited as a cost-effective renewable feedstock for biodiesel production.

Effects of an Autoregulatory Senescence-inhibitor Gene Construct on Nicotiana alata Link and Otto.

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 519d-519 ◽  
Author(s):  
Kenneth R. Schroeder ◽  
Dennis P. Stimart
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Plant Height ◽  
Dry Weight ◽  
Nicotiana Alata ◽  
Wild Type ◽  
Gene Encoding ◽  
Delayed Senescence ◽  
Gene Construct ◽  
Shoot Dry Weight ◽  
Inhibitor System

Nicotiana alata Link and Otto. was transformed via Agrobacterium tumefaciens encoding a senescence-specific promoter SAG12 cloned from Arabidopsis thaliana fused to a Agrobacterium tumefaciens gene encoding isopentenyl transferase (IPT) that catalyzes cytokinin synthesis. This was considered an autoregulatory senescence-inhibitor system. In 1996, we reported delayed senescence of intact flowers by 2 to 6 d and delayed leaf senescence of transgenic vs. wild-type N. alata. Further evaluations in 1997 revealed several other interesting effects of the SAG12-IPT gene construct. Measurement of chlorophyll content of mature leaves showed higher levels of both chlorophyll a and b in transgenic material under normal fertilization and truncated fertilization regimes. At 4 to 5 months of age transgenic plants expressed differences in plant height, branching, and dry weight. Plant height was reduced by 3 to 13 cm; branch counts increased 2 to 3 fold; and shoot dry weight increased up to 11 g over wild-type N. alata. These observations indicate the system is not tightly autoregulated and may prove useful to the floriculture industry for producing compact and more floriferous plants.

scholarly journals Bicarbonate for microalgae cultivation: a case study in a chlorophyte, Tetradesmus wisconsinensis isolated from a Norwegian lake

2021 ◽  
Author(s):  
Ikumi Umetani ◽  
Eshetu Janka ◽  
Michal Sposób ◽  
Chris J. Hulatt ◽  
Synne Kleiven ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Lipid Production ◽  
Cetane Number ◽  
Fluorescence Analysis ◽  
Dry Weight ◽  
Maximum Growth ◽  
Biodiesel Fuel ◽  
Fatty Acid Production ◽  
Total Fatty Acids

AbstractBicarbonate was evaluated as an alternative carbon source for a green microalga, Tetradesmus wisconsinensis, isolated from Lake Norsjø in Norway. Photosynthesis, growth, and lipid production were studied using four inorganic carbon regimes: (1) aeration only, (2) 20 mM NaHCO3, (3) 5% (v/v) CO2 gas, and (4) combination of 20 mM NaHCO3 and 5% CO2. Variable chlorophyll a fluorescence analysis revealed that the bicarbonate treatment supported effective photosynthesis, while the CO2 treatment led to inefficient photosynthetic activity with a PSII maximum quantum yield as low as 0.31. Conversely, bicarbonate and CO2 treatments gave similar biomass and fatty acid production. The maximum growth rate, the final cell dry weight, and total fatty acids under the bicarbonate-only treatment were 0.33 (± 0.06) day−1, 673 (± 124) mg L−1 and 75 (± 5) mg g−1 dry biomass, respectively. The most abundant fatty acid components were α-linolenic acid and polyunsaturated fatty acids constituting 69% of the total fatty acids. The fatty acid profile eventuated in unsuitable biodiesel fuel properties such as high degree of unsaturation and low cetane number; however, it would be relevant for food and feed applications. We concluded that bicarbonate could give healthy growth and comparative product yields as CO2.

scholarly journals Mutation in the ntrR Gene, a Member of the vap Gene Family, Increases the Symbiotic Efficiency of Sinorhizobium meliloti

2001 ◽  
Vol 14 (7) ◽  
pp. 887-894 ◽  
Author(s):  
Boglárka Oláh ◽  
Erno Kiss ◽  
Zoltán Györgypál ◽  
Judit Borzi ◽  
Gyöngyi Cinege ◽  
...  
Keyword(s):  
Nitrogen Fixation ◽  
Pathogenic Bacteria ◽  
Sinorhizobium Meliloti ◽  
Root Nodules ◽  
Nifh Gene ◽  
Dry Weight ◽  
Nodule Occupancy ◽  
Gene Encoding ◽  
Symbiotic Efficiency ◽  
Host Interactions

In specific plant organs, namely the root nodules of alfalfa, fixed nitrogen (ammonia) produced by the symbiotic partner Sinorhizobium meliloti supports the growth of the host plant in nitrogen-depleted environment. Here, we report that a derivative of S. meliloti carrying a mutation in the chromosomal ntrR gene induced nodules with enhanced nitrogen fixation capacity, resulting in an increased dry weight and nitrogen content of alfalfa. The efficient nitrogen fixation is a result of the higher expression level of the nifH gene, encoding one of the subunits of the nitrogenase enzyme, and nifA, the transcriptional regulator of the nif operon. The ntrR gene, controlled negatively by its own product and positively by the symbiotic regulator syrM, is expressed in the same zone of nodules as the nif genes. As a result of the nitrogen-tolerant phenotype of the strain, the beneficial effect of the mutation on efficiency is not abolished in the presence of the exogenous nitrogen source. The ntrR mutant is highly competitive in nodule occupancy compared with the wild-type strain. Sequence analysis of the mutant region revealed a new cluster of genes, termed the “ntrPR operon,” which is highly homologous to a group of vap-related genes of various pathogenic bacteria that are presumably implicated in bacterium-host interactions. On the basis of its favorable properties, the strain is a good candidate for future agricultural utilization.

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