scholarly journals Study of Metabolic Adaptation of Red Yeasts to Waste Animal Fat Substrate

2019 ◽  
Vol 7 (11) ◽  
pp. 578 ◽  
Author(s):  
Martin Szotkowski ◽  
Dana Byrtusova ◽  
Andrea Haronikova ◽  
Marie Vysoka ◽  
Marek Rapta ◽  
...  

Carotenogenic yeasts are non-conventional oleaginous microorganisms capable of utilizing various waste substrates. In this work, four red yeast strains (Rhodotorula, Cystofilobasidium, and Sporobolomyces sp.) were cultivated in media containing crude, emulsified, and enzymatically hydrolyzed animal waste fat, compared with glucose and glycerol, as single C-sources. Cell morphology (cryo-SEM (cryo-scanning electron microscopy), TEM (transmission electron microscopy)), production of biomass, lipase, biosurfactants, lipids (gas chromatography/flame ionization detection, GC/FID) carotenoids, ubiquinone, and ergosterol (high performance liquid chromatography, HPLC/PDA) in yeast cells was studied depending on the medium composition, the C source, and the carbon/nitrogen (C/N) ratio. All studied strains are able to utilize solid and processed fat. Biomass production at C/N = 13 was higher on emulsified/hydrolyzed fat than on glucose/glycerol. The production of lipids and lipidic metabolites was enhanced for several times on fat; the highest yields of carotenoids (24.8 mg/L) and lipids (54.5%/CDW (cell dry weight)) were found in S. pararoseus. Simultaneous induction of lipase and biosurfactants was observed on crude fat substrate. An increased C/N ratio (13–100) led to higher biomass production in fat media. The production of total lipids increased in all strains to C/N = 50. Oppositely, the production of carotenoids, ubiquinone, and ergosterol dramatically decreased with increased C/N in all strains. Compounds accumulated in stressed red yeasts have a great application potential and can be produced efficiently during the valorization of animal waste fat under the biorefinery concept.

2010 ◽  
Vol 42 (6) ◽  
pp. 739-749 ◽  
Author(s):  
Annika I. SMEDS ◽  
Minna-Maarit KYTÖVIITA

AbstractThe ecologically important lichen Cladonia stellaris forms thick carpets in boreal forest floors. In addition to affecting temperature and water conditions in the soil underneath, the secondary metabolites formed by the lichen layer are of ecological interest. In this paper, we investigated the distribution of lichen acids in C. stellaris collected at different latitudes in Finland and developed methods to quantify the two optical enantiomers of usnic acid separately. The lichen extracts were analysed by high-performance liquid chromatography (HPLC) with UV and mass spectrometric (MS) detection and by gas chromatography with flame ionization (GC-FID) and MS detection. Usnic acid and perlatolic acid were quantified using GC-FID. The concentration of usnic acid in the top 20 mm of the lichen thallus ranged from 0·48–3·08% of dry weight, and that of perlatolic acid from 0·08–0·54%. The enantiomeric composition of usnic acid was determined using a chiral HPLC column coupled to an electrospray ionization-tandem mass spectrometer. (−)-Usnic acid was found to be the predominating enantiomer in all extracts; the proportion of (+)-usnic acid ranged from 0·4%–10·0%. Olivetoric acid methyl ester, diphenylmethanol, and 5-pentylresorcinol were identified, and several other olivetoric acid-type compounds were tentatively identified in the extracts.


2007 ◽  
Vol 73 (13) ◽  
pp. 4342-4350 ◽  
Author(s):  
René Verwaal ◽  
Jing Wang ◽  
Jean-Paul Meijnen ◽  
Hans Visser ◽  
Gerhard Sandmann ◽  
...  

ABSTRACT To determine whether Saccharomyces cerevisiae can serve as a host for efficient carotenoid and especially β-carotene production, carotenogenic genes from the carotenoid-producing yeast Xanthophyllomyces dendrorhous were introduced and overexpressed in S. cerevisiae. Because overexpression of these genes from an episomal expression vector resulted in unstable strains, the genes were integrated into genomic DNA to yield stable, carotenoid-producing S. cerevisiae cells. Furthermore, carotenoid production levels were higher in strains containing integrated carotenogenic genes. Overexpression of crtYB (which encodes a bifunctional phytoene synthase and lycopene cyclase) and crtI (phytoene desaturase) from X. dendrorhous was sufficient to enable carotenoid production. Carotenoid production levels were increased by additional overexpression of a homologous geranylgeranyl diphosphate (GGPP) synthase from S. cerevisiae that is encoded by BTS1. Combined overexpression of crtE (heterologous GGPP synthase) from X. dendrorhous with crtYB and crtI and introduction of an additional copy of a truncated 3-hydroxy-3-methylglutaryl-coenzyme A reductase gene (tHMG1) into carotenoid-producing cells resulted in a successive increase in carotenoid production levels. The strains mentioned produced high levels of intermediates of the carotenogenic pathway and comparable low levels of the preferred end product β-carotene, as determined by high-performance liquid chromatography. We finally succeeded in constructing an S. cerevisiae strain capable of producing high levels of β-carotene, up to 5.9 mg/g (dry weight), which was accomplished by the introduction of an additional copy of crtI and tHMG1 into carotenoid-producing yeast cells. This transformant is promising for further development toward the biotechnological production of β-carotene by S. cerevisiae.


Author(s):  
M. T. Postek ◽  
A. E. Vladar

One of the major advancements applied to scanning electron microscopy (SEM) during the past 10 years has been the development and application of digital imaging technology. Advancements in technology, notably the availability of less expensive, high-density memory chips and the development of high speed analog-to-digital converters, mass storage and high performance central processing units have fostered this revolution. Today, most modern SEM instruments have digital electronics as a standard feature. These instruments, generally have 8 bit or 256 gray levels with, at least, 512 × 512 pixel density operating at TV rate. In addition, current slow-scan commercial frame-grabber cards, directly applicable to the SEM, can have upwards of 12-14 bit lateral resolution permitting image acquisition at 4096 × 4096 resolution or greater. The two major categories of SEM systems to which digital technology have been applied are:In the analog SEM system the scan generator is normally operated in an analog manner and the image is displayed in an analog or "slow scan" mode.


Toxins ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 434
Author(s):  
Pascaline Bahati ◽  
Xuejun Zeng ◽  
Ferdinand Uzizerimana ◽  
Ariunsaikhan Tsoggerel ◽  
Muhammad Awais ◽  
...  

In the food industry, microbiological safety is a major concern. Mycotoxin patulin represents a potential health hazard, as it is heat-resistant and may develop at any stage during the food chain, especially in apple-based products, leading to severe effects on human health, poor quality products, and profit reductions. The target of the study was to identify and characterize an excellent adsorbent to remove patulin from apple juice efficiently and to assess its adsorption mechanism. To prevent juice fermentation and/or contamination, autoclaving was involved to inactivate bacteria before the adsorption process. The HPLC (high-performance liquid chromatography) outcome proved that all isolated strains from kefir grains could reduce patulin from apple juice. A high removal of 93% was found for juice having a 4.6 pH, 15° Brix, and patulin concentration of 100 μg/L by Lactobacillus kefiranofacien, named JKSP109, which was morphologically the smoothest and biggest of all isolates in terms of cell wall volume and surface area characterized by SEM (Scanning electron microscopy) and TEM (transmission electron microscopy). C=O, OH, C–H, and N–O were the main functional groups engaged in patulin adsorption indicated by FTIR (Fourier transform–infrared). E-nose (electronic nose) was performed to evaluate the aroma quality of the juices. PCA (Principal component analysis) results showed that no significant changes occurred between control and treated juice.


2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Madhavi Latha Gandla ◽  
Niklas Mähler ◽  
Sacha Escamez ◽  
Tomas Skotare ◽  
Ogonna Obudulu ◽  
...  

Abstract Background Bioconversion of wood into bioproducts and biofuels is hindered by the recalcitrance of woody raw material to bioprocesses such as enzymatic saccharification. Targeted modification of the chemical composition of the feedstock can improve saccharification but this gain is often abrogated by concomitant reduction in tree growth. Results In this study, we report on transgenic hybrid aspen (Populus tremula × tremuloides) lines that showed potential to increase biomass production both in the greenhouse and after 5 years of growth in the field. The transgenic lines carried an overexpression construct for Populus tremula × tremuloides vesicle-associated membrane protein (VAMP)-associated protein PttVAP27-17 that was selected from a gene-mining program for novel regulators of wood formation. Analytical-scale enzymatic saccharification without any pretreatment revealed for all greenhouse-grown transgenic lines, compared to the wild type, a 20–44% increase in the glucose yield per dry weight after enzymatic saccharification, even though it was statistically significant only for one line. The glucose yield after enzymatic saccharification with a prior hydrothermal pretreatment step with sulfuric acid was not increased in the greenhouse-grown transgenic trees on a dry-weight basis, but increased by 26–50% when calculated on a whole biomass basis in comparison to the wild-type control. Tendencies to increased glucose yields by up to 24% were present on a whole tree biomass basis after acidic pretreatment and enzymatic saccharification also in the transgenic trees grown for 5 years on the field when compared to the wild-type control. Conclusions The results demonstrate the usefulness of gene-mining programs to identify novel genes with the potential to improve biofuel production in tree biotechnology programs. Furthermore, multi-omic analyses, including transcriptomic, proteomic and metabolomic analyses, performed here provide a toolbox for future studies on the function of VAP27 proteins in plants.


2020 ◽  
Vol 11 (1) ◽  
pp. 174
Author(s):  
Konstantinos P. Papadopoulos ◽  
Christina N. Economou ◽  
Athanasia G. Tekerlekopoulou ◽  
Dimitris V. Vayenas

Algal/cyanobacterial biofilm photobioreactors provide an alternative technology to conventional photosynthetic systems for wastewater treatment based on high biomass production and easy biomass harvesting at low cost. This study introduces a novel cyanobacteria-based biofilm photobioreactor and assesses its performance in post-treatment of brewery wastewater and biomass production. Two different supporting materials (glass/polyurethane) were tested to investigate the effect of surface hydrophobicity on biomass attachment and overall reactor performance. The reactor exhibited high removal efficiency (over 65%) of the wastewater’s pollutants (chemical oxygen demand, nitrate, nitrite, ammonium, orthophosphate, and total Kjeldahl nitrogen), while biomass per reactor surface reached 13.1 and 12.8 g·m−2 corresponding to 406 and 392 mg·L−1 for glass and polyurethane, respectively, after 15 days of cultivation. The hydrophilic glass surface favored initial biomass adhesion, although eventually both materials yielded complete biomass attachment, highlighting that cell-to-cell interactions are the dominant adhesion mechanism in mature biofilms. It was also found that the biofilm accumulated up to 61% of its dry weight in carbohydrates at the end of cultivation, thus making the produced biomass a suitable feedstock for bioethanol production.


2017 ◽  
Vol 12 (3) ◽  
pp. 1934578X1701200 ◽  
Author(s):  
Katarzyna Sułkowska-Ziaja ◽  
Anna Maślanka ◽  
Agnieszka Szewczyk ◽  
Bożena Muszyńska

The content of two groups of compounds with biological activity (non-hallucinogenic indole compounds and free phenolic acids) were analyzed in extracts of fruiting bodies of four species of Phellinus: P. igniarius, P. pini, P. pomaceus and P. robustus. The presence of indole compounds in methanolic extracts was analyzed by high-performance liquid chromatography and thin-layer chromatography coupled with densitometric detection. Three metabolites (serotonin, tryptamine, and L-tryptophan) were identified. The contents of individual indole compounds ranged from 1.70 (tryptamine in P. robustus) to 8.32 mg x 100 g1 dry weight (L-tryptophan in P. robustus). Four free phenolic acids were detected in methanolic extracts by the HPLC method. The total content ranged from 9.9 mg x 100 g1 DW (P. igniarius) to 32.5 mg x 100 g1 DW (P. robustus).


1989 ◽  
Vol 35 (12) ◽  
pp. 1081-1086 ◽  
Author(s):  
Byron F. Johnson ◽  
L. C. Sowden ◽  
Teena Walker ◽  
Bong Y. Yoo ◽  
Gode B. Calleja

The surfaces of flocculent and nonflocculent yeast cells have been examined by electron microscopy. Nonextractive preparative procedures for scanning electron microscopy allow comparison in which sharp or softened images of surface details (scars, etc.) are the criteria for relative abundance of flocculum material. Asexually flocculent budding-yeast cells cannot be distinguished from nonflocculent budding-yeast cells in scanning electron micrographs because the scar details of both are well resolved, being hard and sharp. On the other hand, flocculent fission-yeast cells are readily distinguished from nonflocculent cells because fission scars are mostly soft or obscured on flocculent cells, but sharp on nonflocculent cells. Sexually and asexually flocculent fission-yeast cells cannot be distinguished from one another as both are heavily clad in "mucilaginous" or "hairy" coverings. Examination of lightly extracted and heavily extracted flocculent fission-yeast cells by transmission electron microscopy provides micrographs consistent with the scanning electron micrographs.Key words: flocculation, budding yeast, fission yeast, scanning, transmission.


1990 ◽  
Vol 68 (7) ◽  
pp. 1578-1588 ◽  
Author(s):  
Brian T. Luck ◽  
Colette Breuil ◽  
David L. Brown

An enzyme-linked immunosorbent assay (ELISA) was used to detect a sap-staining fungus, Ophiostoma piceae, and a biological-control agent, Gliocladium roseum, grown in liquid culture and in wood. A polyclonal serum prepared against whole cell fragments from broken mycelia of O. piceae detected O. piceae in liquid culture at 0.25 μg dry weight/mL; however, there was moderate cross-reactivity with G. roseum. Antiserum adsorbed on G. roseum had almost no reactivity with G. roseum but still reacted strongly with O. piceae. The specificity of these sera was verified, and the antigenic sites were localized, by immunogold labeling and electron microscopy. These studies confirmed that the adsorbed serum could differentiate between G. roseum and O. piceae and showed that the cell wall was the most reactive cellular component. These results are discussed in relation to the development of immunological probes for the detection of sap-staining and biological control fungi. Key words: polyclonal serum, enzyme-linked immunosorbent assay, immunogold labeling, sap-staining and biological control fungi, electron microscopy.


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