scholarly journals Influence of prefoldin subunit 4 on the tolerance of Kluyveromyces marxianus to lignocellulosic biomass-derived inhibitors

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Nini Zhang ◽  
Yingying Shang ◽  
Feier Wang ◽  
Dongmei Wang ◽  
Jiong Hong

Abstract Background Kluyveromyces marxianus is a potentially excellent host for microbial cell factories using lignocellulosic biomass, due to its thermotolerance, high growth rate, and wide substrate spectrum. However, its tolerance to inhibitors derived from lignocellulosic biomass pretreatment needs to be improved. The prefoldin complex assists the folding of cytoskeleton which relates to the stress tolerance, moreover, several subunits of prefoldin have been verified to be involved in gene expression regulation. With the presence of inhibitors, the expression of a gene coding the subunit 4 of prefoldin (KmPFD4), a possible transcription factor, was significantly changed. Therefore, KmPFD4 was selected to evaluate its functions in inhibitors tolerance. Results In this study, the disruption of the prefoldin subunit 4 gene (KmPFD4) led to increased concentration of intracellular reactive oxygen species (ROS) and disturbed the assembly of actin and tubulin in the presence of inhibitors, resulting in reduced inhibitor tolerance. Nuclear localization of KmPFD4 indicated that it could regulate gene expression. Transcriptomic analysis showed that upregulated gene expression related to ROS elimination, ATP production, and NAD+ synthesis, which is a response to the presence of inhibitors, disappeared in KmPFD4-disrupted cells. Thus, KmPFD4 impacts inhibitor tolerance by maintaining integration of the cytoskeleton and directly or indirectly affecting the expression of genes in response to inhibitors. Finally, overexpression of KmPFD4 enhanced ethanol fermentation with a 46.27% improvement in productivity in presence of the inhibitors. Conclusion This study demonstrated that KmPFD4 plays a positive role in the inhibitor tolerance and can be applied for the development of inhibitor-tolerant platform strains.

PLoS Genetics ◽  
2021 ◽  
Vol 17 (10) ◽  
pp. e1009826
Author(s):  
Gert Vanmarcke ◽  
Quinten Deparis ◽  
Ward Vanthienen ◽  
Arne Peetermans ◽  
Maria R. Foulquié-Moreno ◽  
...  

Development of cell factories for conversion of lignocellulosic biomass hydrolysates into biofuels or bio-based chemicals faces major challenges, including the presence of inhibitory chemicals derived from biomass hydrolysis or pretreatment. Extensive screening of 2526 Saccharomyces cerevisiae strains and 17 non-conventional yeast species identified a Candida glabrata strain as the most 5-hydroxymethylfurfural (HMF) tolerant. Whole-genome (WG) transformation of the second-generation industrial S. cerevisiae strain MD4 with genomic DNA from C. glabrata, but not from non-tolerant strains, allowed selection of stable transformants in the presence of HMF. Transformant GVM0 showed the highest HMF tolerance for growth on plates and in small-scale fermentations. Comparison of the WG sequence of MD4 and GVM1, a diploid segregant of GVM0 with similarly high HMF tolerance, surprisingly revealed only nine non-synonymous SNPs, of which none were present in the C. glabrata genome. Reciprocal hemizygosity analysis in diploid strain GVM1 revealed AST2N406I as the only causative mutation. This novel SNP improved tolerance to HMF, furfural and other inhibitors, when introduced in different yeast genetic backgrounds and both in synthetic media and lignocellulose hydrolysates. It stimulated disappearance of HMF and furfural from the medium and enhanced in vitro furfural NADH-dependent reducing activity. The corresponding mutation present in AST1 (i.e. AST1D405I) the paralog gene of AST2, also improved inhibitor tolerance but only in combination with AST2N406I and in presence of high inhibitor concentrations. Our work provides a powerful genetic tool to improve yeast inhibitor tolerance in lignocellulosic biomass hydrolysates and other inhibitor-rich industrial media, and it has revealed for the first time a clear function for Ast2 and Ast1 in inhibitor tolerance.


2011 ◽  
Vol 22 (17) ◽  
pp. 3127-3138 ◽  
Author(s):  
Amit Sadeh ◽  
Natalia Movshovich ◽  
Misha Volokh ◽  
Larisa Gheber ◽  
Amir Aharoni

The Msn2 and Msn4 transcription factors play major roles in the yeast general stress response by mediating the transcription of hundreds of genes. Despite extensive information on Msn2/4–mediated gene expression profiles, much less is known regarding the network of proteins that regulate its activity. Here we describe a systematic approach designed to examine the roles of 35 Msn2/4 partners in regulating Msn2/4 transcriptional activity in the face of four different environmental conditions. Our analysis indicates that single deletions of 26 Msn2/4 partners significantly affect Msn2/4 transcription activity under four different conditions. The low functional redundancy of the Msn2 regulatory network indicates that Msn2/4 activity is finely tuned by many of Msn2/4 partners to provide an optimized stress response through differential activation, nuclear localization, degradation, and chromatin remodeling. Our specific analysis of Msn2 activity showed that a relatively large number of partners act to suppress Msn2 activity under nonstress conditions through independent mechanisms, including cytoplasmic retention, proteosome-mediated Msn2 degradation, and chromatin remodeling. Such negative regulation is crucial to minimize the cost of uncontrolled stress response gene expression and ensures a high growth rate in the absence of stress.


2014 ◽  
Vol 84 (3-4) ◽  
pp. 0183-0195 ◽  
Author(s):  
Takashi Nakamura ◽  
Tomoya Takeda ◽  
Yoshihiko Tokuji

The common water-soluble organic germanium compound poly-trans-[(2-carboxyethyl) germasesquioxane] (Ge-132) exhibits activities related to immune responses and antioxidant induction. In this study, we evaluated the antioxidative effect of dietary Ge-132 in the plasma of mice. Male ICR mice (seven mice per group) received an AIN-76 diet with 0.05 % Ge-132; three groups received the Ge-132-containing diet for 0, 1 or 4 days. The plasma alpha-tocopherol (α-tocopherol) concentration increased from 6.85 to 9.60 μg/ml after 4 days of Ge-132 intake (p < 0.05). We evaluated the changes in hepatic gene expression related to antioxidative activity as well as in the entire expression profile after one day of Ge-132 intake, using DNA microarray technology. We identified 1,220 genes with altered expression levels greater than 1.5-fold (increased or decreased) as a result of Ge-132 intake, and α-tocopherol transfer protein (Ttpa) gene expression was increased 1.62-fold. Immune activation was identified as the category with the most changes (containing 60 Gene Ontology (GO) term biological processes (BPs), 41 genes) via functional clustering analysis of altered gene expression. Ge-132 affected genes in clusters related to ATP production (22 GO term BPs, 21 genes), lipid metabolism (4 GO term BPs, 38 genes) and apoptosis (5 GO term BPs). Many GO term BPs containing these categories were significantly affected by the Ge-132 intake. Oral Ge-132 intake may therefore have increased plasma α-tocopherol levels by up-regulating α-tocopherol transfer protein (Ttpa) gene expression.


2019 ◽  
Vol 118 (8) ◽  
pp. 236-240
Author(s):  
Dr.R. Murugesan ◽  
M. Leelavathi ◽  
Dr. K. Ravindran

towards jumping from the category of developing economy to developed economy there is one big factor that stops and poses a hindrance in its path of advancement and that obstacle is termed as Poverty. The Indian economic policy focuses on a high growth rate along with a equal participation of the poor so that they avail the opportunities available in the market economy. And in order to ensure the participation of the poor it has become important for the country to create a platform where the poor can easily access the various financial products. Microfinance is one such strategy for inclusive growth. Microfinance can change the life of the poor though not completely but a reasonable change can be ensured. In different phases of life women play a crucial role despite the discrimination that is faced by them. But equality can be endowed to women by enhancing the entrepreneurial skills in them. This is possible through Self Help Groups (SHGs). In India women produce around 30% of the total food consumed but she gets only 10% of the property or wealth of the country. Development of women is inevitable for the development and growth of any economy. SHGs happen to be a positive step in this direction. Along with these mediums there should be a cheap and easy source of credit for them and Microfinance fulfills the requirement. This study aims to find the role of this strong medium of Microfinance in the advancement of SHGs in India


2013 ◽  
Vol 740-742 ◽  
pp. 323-326
Author(s):  
Kassem Alassaad ◽  
François Cauwet ◽  
Davy Carole ◽  
Véronique Soulière ◽  
Gabriel Ferro

Abstract. In this paper, conditions for obtaining high growth rate during epitaxial growth of SiC by vapor-liquid-solid mechanism are investigated. The alloys studied were Ge-Si, Al-Si and Al-Ge-Si with various compositions. Temperature was varied between 1100 and 1300°C and the carbon precursor was either propane or methane. The variation of layers thickness was studied at low and high precursor partial pressure. It was found that growth rates obtained with both methane and propane are rather similar at low precursor partial pressures. However, when using Ge based melts, the use of high propane flux leads to the formation of a SiC crust on top of the liquid, which limits the growth by VLS. But when methane is used, even at extremely high flux (up to 100 sccm), no crust could be detected on top of the liquid while the deposit thickness was still rather small (between 1.12 μm and 1.30 μm). When using Al-Si alloys, no crust was also observed under 100 sccm methane but the thickness was as high as 11.5 µm after 30 min growth. It is proposed that the upper limitation of VLS growth rate depends mainly on C solubility of the liquid phase.


2013 ◽  
Vol 687 ◽  
pp. 255-261 ◽  
Author(s):  
Sandra Cunha ◽  
José Barroso Aguiar ◽  
Victor Ferreira ◽  
António Tadeu

Increasingly in a society with a high growth rate and standards of comfort, the need to minimize the currently high energy consumption by taking advantage of renewable energy sources arises. The mortars with incorporation of phase change materials (PCM) have the ability to regulate the temperature inside buildings, contributing for an increase in the level of thermal comfort and reduction of the use of heating, ventilation and air conditioning (HVAC) equipment, using only the energy supplied by the sun. However, the incorporation of PCM in mortars modifies some of its characteristics. Therefore, the main objective of this study was the characterization of mortars doped with two different phase change materials. Specific properties of different PCM, such as particle size, shape and enthalpy were studied, as well as the properties of the fresh and hardened state of these mortars. Nine different compositions were developed which were initially doped with microcapsules of PCM A and subsequently doped with microcapsules of PCM B. It was possible to observe that the incorporation of phase change materials in mortars causes differences in properties such as compressive strength, flexural strength and shrinkage. After the study of the behaviour of these mortars with the incorporation of two different phase change materials, it was possible to select the composition with a better compromise between its aesthetic appearance, physical and mechanical characteristics.


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