Recombinant Saccharomyces cerevisiae Expressing P450 in Artificial Digestive Systems: a Model for Biodetoxication in the Human Digestive Environment

ABSTRACT The use of genetically engineered microorganisms such as bacteria or yeasts as live vehicles to carry out bioconversion directly in the digestive environment is an important challenge for the development of innovative biodrugs. A system that mimics the human gastrointestinal tract was combined with a computer simulation to evaluate the survival rate and cinnamate 4-hydroxylase activity of a recombinant model of Saccharomyces cerevisiae expressing the plant P450 73A1. The yeasts showed a high level of resistance to gastric and small intestinal secretions (survival rate after 4 h of digestion, 95.6% ± 10.1% [n = 4]) but were more sensitive to the colonic conditions (survival rate after 4 h of incubation, 35.9% ± 2.7% [n = 3]). For the first time, the ability of recombinant S. cerevisiae to carry out a bioconversion reaction has been demonstrated throughout the gastrointestinal tract. In the gastric-small intestinal system, 41.0% ± 5.8% (n = 3) of the ingested trans-cinnamic acid was converted into p-coumaric acid after 4 h of digestion, as well as 8.9% ± 1.6% (n = 3) in the stomach, 13.8% ± 3.3% (n = 3) in the duodenum, 11.8% ± 3.4% (n = 3) in the jejunum, and 6.5% ± 1.0% (n = 3) in the ileum. In the large intestinal system, cinnamate 4-hydroxylase activity was detected but was too weak to be quantified. These results suggest that S. cerevisiae may afford a useful host for the development of biodrugs and may provide an innovative system for the prevention or treatment of diseases that escape classical drug action. In particular, yeasts may provide a suitable vector for biodetoxication in the digestive environment.

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Effect of microencapsulation on Saccharomyces cerevisiae var. boulardii viability in the gastrointestinal tract and level of some blood biochemical factors in wistar rats

Iranian Journal of Microbiology ◽

10.18502/ijm.v11i2.1082 ◽

2019 ◽

Author(s):

Hassan Ghorbani-Choboghlo ◽

Donya Nikaein ◽

Ali-Reza Khosravi ◽

Reza Rahmani ◽

Zohreh Farahnejad

Keyword(s):

Saccharomyces Cerevisiae ◽

Survival Rate ◽

Gastrointestinal Tract ◽

Wistar Rats ◽

Health Benefit ◽

Control Group ◽

Blood Biochemical ◽

Probiotic Treatment ◽

Male Wistar Rats ◽

The Impact

ABSTRACT Background and Objectives: Probiotics are live microorganisms that, when administered in an adequate amount, confer a health benefit on the host through the gut. Saccharomyces cerevisiae is a widespread yeast found in nature. This microor- ganism has been used as a probiotic agent in recent years. In this study, the effect of microencapsulation on survival rate of S. cerevisiae var. boulardii in the simulated gastrointestinal tract medium and the impact of microencapsulated S. cerevisiae var. boulardii on some serum biochemical factors in a rat model was evaluated. Materials and Methods: 30 male wistar rats were divided into three groups (control, rats receiving microencapsulated S. cerevisiae var. boulardii, and rats receiving S. cerevisiae var. boulardii alone). The probiotic was gavaged at a dosage of 2 gr/ kg BW for 8 weeks. Blood was collected from rats at the end of the treatment period and biochemical factors were measured using Mancompany kits. Results: The results showed a significant increase in viability of microencapsulated S. cerevisiae var. boulardii in compar- ison with free S. cerevisiae var. boulardii (p<0.05). Weight of rats in probiotic treated groups was significantly higher in comparison with the control group (p<0.05). Moreover, probiotic treatment reduced mean levels of triglycerides, cholesterol, free blood sugar and liver enzymes in rats. Conclusion: Microencapsulation could increase the survival rate of yeast probiotics in the gastrointestinal tract; however, more studies are needed for better understanding of the exact effect of microencapsulation on probiotics’ function.

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Evidence for two main domestication trajectories in Saccharomyces cerevisiae linked to distinct bread-making processes

10.1101/2020.05.28.120584 ◽

2020 ◽

Author(s):

Frédéric Bigey ◽

Diego Segond ◽

Anne Friedrich ◽

Stephane Guezenec ◽

Aurélie Bourgais ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Copy Number ◽

Population Divergence ◽

Bread Making ◽

Number Of Genes ◽

Maltose Utilization ◽

Selection For ◽

Bakery Yeast ◽

High Level ◽

First Time

SummaryDespite bread being one of the most historically and culturally important fermented products, its history and influence on the evolution of associated microbial species remains largely unknown. The first evidence of leavened bread dates to the second millenium BCE in Egypt and since, the art of bread-making developed and spread worldwide. Nowadays, leavened bread is made either by using a pure commercial culture of the yeast Saccharomyces cerevisiae or by propagating a sourdough, which is a mix of flour and water spontaneously fermented by yeast and bacteria. We studied the domestication of S. cerevisiae populations originating from industry and sourdough and tested whether these different bread-making processes led to population divergence. We found that the origin of S. cerevisiae bakery strains is polyphyletic with 67 % of strains clustering in two main clades: most commercial strains were tetraploid and clustered with strains having diverse origins, including beer. By contrast, most sourdough strains were diploids and found in a second clade of strains having mosaic genomes and diverse origins including fruits, or clinical and wild environments. When compared to the others, sourdough strains harboured in average a higher copy number of genes involved in maltose utilization, a common sugar produced from dough starch. Overall, a high level of gene flow from multiple contributors was detected. Phenotyping of bakery and non bakery strains further showed that sourdough and industrial bakery populations have undergone human selection for rapidly starting fermentations and for high CO2 production. Interestingly, sourdough strains also showed a better adaptation to a sourdough mimicking environment, suggesting that natural selection occurred as well. In summary, our results revealed that the domestication of bakery yeast populations has been accompanied by dispersion, hybridization and divergent selection through industrial and artisanal bakery processes. In addition, they unveiled for the first time a case of fungus domestication where species divergence occurred through autotetraploidisation.

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Alkaloids of Crinum x powellii “Album” (Amaryllidaceae) and their Topoisomerase Inhibitory Activity

Zeitschrift für Naturforschung C ◽

10.1515/znc-2007-3-411 ◽

2007 ◽

Vol 62 (3-4) ◽

pp. 223-226 ◽

Cited By ~ 11

Author(s):

Jaime Niño ◽

Gina M. Hincapié ◽

Yaned M. Correa ◽

Oscar M. Mosquera

Keyword(s):

Saccharomyces Cerevisiae ◽

Inhibitory Activity ◽

Topoisomerase I ◽

Soluble Fraction ◽

Genetically Engineered ◽

Yeast Saccharomyces Cerevisiae ◽

Spectroscopic Analyses ◽

Nmr Techniques ◽

First Time

Abstract The alkaloids lycorine, 1-O-acetyllycorine and ismine were isolated from the basic dichloromethane- soluble fraction of Crinum x powellii “Album” bulbs. The alkaloid structures were established by physical and spectroscopic analyses, including 1D NMR techniques and GCMS analysis. The three alkaloids are reported for the first time for this hybrid. Additionally, the three alkaloids isolated were tested against a mechanism-based bioassay utilizing genetically engineered mutants of the yeast Saccharomyces cerevisiae strains RAD+, RAD52Y and RS321 where lycorine was the only alkaloid that displayed moderate topoisomerase I inhibitory activity.

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A Confrontation between Two Doctrines: The Birth of Struggle for Hegemony in Hebrew Children's Literature during the 1930s and 1940s

International Research in Children s Literature ◽

10.3366/ircl.2008.0003 ◽

2008 ◽

Vol 1 (2) ◽

pp. 139-155 ◽

Cited By ~ 2

Author(s):

YAEL DARR

Keyword(s):

Children's Literature ◽

Educational System ◽

Single Channel ◽

Children’S Literature ◽

The Political ◽

Literary Creation ◽

History Of ◽

High Level ◽

First Time ◽

Literary Quality

This article describes a crucial and fundamental stage in the transformation of Hebrew children's literature, during the late 1930s and 1940s, from a single channel of expression to a multi-layered polyphony of models and voices. It claims that for the first time in the history of Hebrew children's literature there took place a doctrinal confrontation between two groups of taste-makers. The article outlines the pedagogical and ideological designs of traditionalist Zionist educators, and suggests how these were challenged by a group of prominent writers of adult poetry, members of the Modernist movement. These writers, it is argued, advocated autonomous literary creation, and insisted on a high level of literary quality. Their intervention not only dramatically changed the repertoire of Hebrew children's literature, but also the rules of literary discourse. The article suggests that, through the Modernists’ polemical efforts, Hebrew children's literature was able to free itself from its position as an apparatus controlled by the political-educational system and to become a dynamic and multi-layered field.

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ANTICYTOKINE ACTIVITY OF THE PROTOZOA BLASTOCYSTIS SPP

Bulletin Biomedicine and sociology ◽

10.26787/nydha-2618-8783-2020-5-4-44-48 ◽

2020 ◽

pp. 44-48

Author(s):

Bugero N.V. ◽

Ilyina N.A. ◽

Aleksandrova S.M.

Keyword(s):

Gastrointestinal Tract ◽

Gastrointestinal Diseases ◽

Control Group ◽

Ulcer Disease ◽

Cytokine Balance ◽

High Prevalence ◽

Blastocystis Spp ◽

High Level ◽

Eukaryotic Organisms ◽

High Degree

In addition to the classical pathogens, which are well understood and well identified, new pathogens with the potential to spread epidemiologically are being identified. Some of these little-known organisms are the simplest Blastocystis spp. blastocystostosis. The clinical significance of Blastocystis spp. and its pathogenicity are still under discussion. This parasite belongs to a group of single-celled eukaryotic organisms living in the colon of the human intestine. Blastocystis spp. is known to be found both in people with reduced immune status and in individuals without any clinical manifestation. It has been established that a sufficiently high degree of invasiveness is observed in persons with gastrointestinal tract diseases, dermatosis, allergic reactions, in patients with carriers of the human immunodeficiency virus, etc. Possessing persistence factors, protozoa blastocysts contribute to the inactivation of host defensive mechanisms, providing a stable anthogonistic effect. In recent years, many works have been devoted to the characteristics of the persistent properties of Blastocystis spr., however, individual properties of blastocysts, in particular, anticytokine activity (ACA), have not yet been studied. In this regard, the work studied the anticytokine activity of microorganisms isolated from healthy subjects and patients with gastrointestinal tract diseases. A high prevalence of the studied characteristic in the subjects was shown. The expression of anticytokine activity in the obtained isolates of blastocysts was the highest in the group of persons with gastric ulcer disease, which decreased in the order of duodenal ulcer, chronic cholecystitis, chronic gastritis, etc. The data obtained in this work on the high level of ACA expression in blastocyst isolates obtained from individuals with gastrointestinal diseases as compared with the control group enables to conclude that their exometabolites may influence the local cytokine balance [1], which supports the inflammatory process.

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Bacterial cellulose spheroids as building blocks for 3D and patterned living materials and for regeneration

Nature Communications ◽

10.1038/s41467-021-25350-8 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Joaquin Caro-Astorga ◽

Kenneth T. Walker ◽

Natalia Herrera ◽

Koon-Yang Lee ◽

Tom Ellis

Keyword(s):

Bacterial Cellulose ◽

Building Blocks ◽

Genetically Engineered ◽

High Yield ◽

Synthetic Materials ◽

Chemical Inputs ◽

First Time ◽

3D Shapes ◽

Promising Avenue ◽

Living Materials

AbstractEngineered living materials (ELMs) based on bacterial cellulose (BC) offer a promising avenue for cheap-to-produce materials that can be programmed with genetically encoded functionalities. Here we explore how ELMs can be fabricated in a modular fashion from millimetre-scale biofilm spheroids grown from shaking cultures of Komagataeibacter rhaeticus. Here we define a reproducible protocol to produce BC spheroids with the high yield bacterial cellulose producer K. rhaeticus and demonstrate for the first time their potential for their use as building blocks to grow ELMs in 3D shapes. Using genetically engineered K. rhaeticus, we produce functionalized BC spheroids and use these to make and grow patterned BC-based ELMs that signal within a material and can sense and report on chemical inputs. We also investigate the use of BC spheroids as a method to regenerate damaged BC materials and as a way to fuse together smaller material sections of cellulose and synthetic materials into a larger piece. This work improves our understanding of BC spheroid formation and showcases their great potential for fabricating, patterning and repairing ELMs based on the promising biomaterial of bacterial cellulose.

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Tempo and Mode of Ty Element Evolution in Saccharomyces cerevisiae

Genetics ◽

10.1093/genetics/151.4.1341 ◽

1999 ◽

Vol 151 (4) ◽

pp. 1341-1351 ◽

Cited By ~ 2

Author(s):

I King Jordan ◽

John F McDonald

Keyword(s):

Saccharomyces Cerevisiae ◽

Negative Selection ◽

Sequence Variation ◽

Size Variation ◽

Sequence Comparisons ◽

Ty Elements ◽

Ty Element ◽

Amino Acid Sequence Variation ◽

Element Evolution ◽

High Level

Abstract The Saccharomyces cerevisiae genome contains five families of long terminal repeat (LTR) retrotransposons, Ty1–Ty5. The sequencing of the S. cerevisiae genome provides an unprecedented opportunity to examine the patterns of molecular variation existing among the entire genomic complement of Ty retrotransposons. We report the results of an analysis of the nucleotide and amino acid sequence variation within and between the five Ty element families of the S. cerevisiae genome. Our results indicate that individual Ty element families tend to be highly homogenous in both sequence and size variation. Comparisons of within-element 5′ and 3′ LTR sequences indicate that the vast majority of Ty elements have recently transposed. Furthermore, intrafamily Ty sequence comparisons reveal the action of negative selection on Ty element coding sequences. These results taken together suggest that there is a high level of genomic turnover of S. cerevisiae Ty elements, which is presumably in response to selective pressure to escape host-mediated repression and elimination mechanisms.

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Transcriptome Analysis Reveals a Promotion of Carotenoid Production by Copper Ions in Recombinant Saccharomyces cerevisiae

Microorganisms ◽

10.3390/microorganisms9020233 ◽

2021 ◽

Vol 9 (2) ◽

pp. 233

Author(s):

Buli Su ◽

Anzhang Li ◽

Ming-Rong Deng ◽

Honghui Zhu

Keyword(s):

Saccharomyces Cerevisiae ◽

Transcriptome Analysis ◽

Gene Expression Analysis ◽

Target Genes ◽

Copper Ions ◽

Carotenoid Production ◽

Carotenoid Accumulation ◽

Nutritional Components ◽

Differential Gene ◽

First Time

We previously constructed a Saccharomyces cerevisiae carotenoid producer BL03-D-4 which produced much more carotenoid in YPM (modified YPD) media than YPD media. In this study, the impacts of nutritional components on carotenoid accumulation of BL03-D-4 were investigated. When using YPM media, the carotenoid yield was increased 10-fold compared to using the YPD media. To elucidate the hidden mechanism, a transcriptome analysis was performed and showed that 464 genes changed significantly in YPM media. Furthermore, inspired by the differential gene expression analysis which indicated that ADY2, HES1, and CUP1 showed the most remarkable changes, we found that the improvement of carotenoid accumulation in YPM media was mainly due to the copper ions, since supplementation of 0.08 mM CuSO4 in YPD media could increase carotenoid yield 9.2-fold. Reverse engineering of target genes was performed and carotenoid yield could be increased 6.4-fold in YPD media through overexpression of ACE1. The present study revealed for the first time the prominent promotion of carotenoid yield by copper ions in engineered S. cerevisiae and provided a new target ACE1 for genetic engineering of S. cerevisiae for the bioproduction of carotenoids.

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Aberrant Expression of TLR2, TLR7, TLR9, Splicing Variants of TLR4 and MYD88 in Chronic Lymphocytic Leukemia Patients

Journal of Clinical Medicine ◽

10.3390/jcm10040867 ◽

2021 ◽

Vol 10 (4) ◽

pp. 867

Author(s):

Katarzyna Skorka ◽

Paulina Wlasiuk ◽

Agnieszka Karczmarczyk ◽

Krzysztof Giannopoulos

Keyword(s):

Bone Marrow ◽

Chronic Lymphocytic Leukemia ◽

Cytotoxic T Cells ◽

Lymphocytic Leukemia ◽

Aberrant Expression ◽

Downstream Signaling ◽

Splicing Variants ◽

High Level ◽

Time To First Treatment ◽

First Time

Functional toll-like receptors (TLRs) could modulate anti-tumor effects by activating inflammatory cytokines and the cytotoxic T-cells response. However, excessive TLR expression could promote tumor progression, since TLR-induced inflammation might stimulate cancer cells expansion into the microenvironment. Myd88 is involved in activation NF-κB through TLRs downstream signaling, hence in the current study we provided, for the first time, a complex characterization of expression of TLR2, TLR4, TLR7, TLR9, and MYD88 as well as their splicing forms in two distinct compartments of the microenvironment of chronic lymphocytic leukemia (CLL): peripheral blood and bone marrow. We found correlations between MYD88 and TLRs expressions in both compartments, indicating their relevant cooperation in CLL. The MYD88 expression was higher in CLL patients compared to healthy volunteers (HVs) (0.1780 vs. 0.128, p < 0.0001). The TLRs expression was aberrant in CLL compared to HVs. Analysis of survival curves revealed a shorter time to first treatment in the group of patients with low level of TLR4(3) expression compared to high level of TLR4(3) expression in bone marrow (13 months vs. 48 months, p = 0.0207). We suggest that TLRs expression is differentially regulated in CLL but is similarly shared between two distinct compartments of the microenvironment.

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Designing P. aeruginosa synthetic phages with reduced genomes

Scientific Reports ◽

10.1038/s41598-021-81580-2 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Diana P. Pires ◽

Rodrigo Monteiro ◽

Dalila Mil-Homens ◽

Arsénio Fialho ◽

Timothy K. Lu ◽

...

Keyword(s):

Galleria Mellonella ◽

Antibacterial Properties ◽

Genetically Engineered ◽

In Vivo Studies ◽

Infection Model ◽

Promising Therapeutic Approach ◽

Genes Encoding ◽

First Time

AbstractIn the era where antibiotic resistance is considered one of the major worldwide concerns, bacteriophages have emerged as a promising therapeutic approach to deal with this problem. Genetically engineered bacteriophages can enable enhanced anti-bacterial functionalities, but require cloning additional genes into the phage genomes, which might be challenging due to the DNA encapsulation capacity of a phage. To tackle this issue, we designed and assembled for the first time synthetic phages with smaller genomes by knocking out up to 48% of the genes encoding hypothetical proteins from the genome of the newly isolated Pseudomonas aeruginosa phage vB_PaeP_PE3. The antibacterial efficacy of the wild-type and the synthetic phages was assessed in vitro as well as in vivo using a Galleria mellonella infection model. Overall, both in vitro and in vivo studies revealed that the knock-outs made in phage genome do not impair the antibacterial properties of the synthetic phages, indicating that this could be a good strategy to clear space from phage genomes in order to enable the introduction of other genes of interest that can potentiate the future treatment of P. aeruginosa infections.

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