scholarly journals Database mining and transcriptional analysis of genes encoding inulin-modifying enzymes of Aspergillus niger

Microbiology ◽  
2006 ◽  
Vol 152 (10) ◽  
pp. 3061-3073 ◽  
Author(s):  
Xiao-Lian Yuan ◽  
Coenie Goosen ◽  
Harrie Kools ◽  
Marc J. E. C. van der Maarel ◽  
Cees A. M. J. J van den Hondel ◽  
...  
Keyword(s):  
Aspergillus Niger ◽  
Carbon Sources ◽  
Soil Fungus ◽  
Invertase Activity ◽  
Transcriptional Analysis ◽  
Database Mining ◽  
Inulinase Activity ◽  
Genes Encoding ◽  
Catabolite Repressor ◽  
Plant Storage

As a soil fungus, Aspergillus niger can metabolize a wide variety of carbon sources, employing sets of enzymes able to degrade plant-derived polysaccharides. In this study the genome sequence of A. niger strain CBS 513.88 was surveyed, to analyse the gene/enzyme network involved in utilization of the plant storage polymer inulin, and of sucrose, the substrate for inulin synthesis in plants. In addition to three known activities, encoded by the genes suc1 (invertase activity; designated sucA), inuE (exo-inulinase activity) and inuA/inuB (endo-inulinase activity), two new putative invertase-like proteins were identified. These two putative proteins lack N-terminal signal sequences and therefore are expected to be intracellular enzymes. One of these two genes, designated sucB, is expressed at a low level, and its expression is up-regulated when A. niger is grown on sucrose- or inulin-containing media. Transcriptional analysis of the genes encoding the sucrose- (sucA) and inulin-hydrolysing enzymes (inuA and inuE) indicated that they are similarly regulated and all strongly induced on sucrose and inulin. Analysis of a ΔcreA mutant strain of A. niger revealed that expression of the extracellular inulinolytic enzymes is under control of the catabolite repressor CreA. Expression of the inulinolytic enzymes was not induced by fructose, not even in the ΔcreA background, indicating that fructose did not act as an inducer. Evidence is provided that sucrose, or a sucrose-derived intermediate, but not fructose, acts as an inducer for the expression of inulinolytic genes in A. niger.

scholarly journals Purification and characterization of β-Fructosidase with inulinase activity from Aspergillus niger - 245

1998 ◽  
Vol 41 (3) ◽  
pp. 288-295
Author(s):  
Vinícius D'Arcadia Cruz ◽  
Juliana Gisele Belote ◽  
Claudia Dorta ◽  
Luíza Helena Oliveira dos Santos ◽  
Cláudia Regina Andriolo ◽  
...  
Keyword(s):  
Aspergillus Niger ◽  
Invertase Activity ◽  
Inulinase Activity ◽  
Km Value ◽  
Good Potential ◽  
Chicory Inulin ◽  
Different Origins ◽  
Hydrolysis Of ◽  
Assay Conditions

Aspergillus niger - 245, a strain isolated from soil samples showed good β-fructosidase activity when inoculated in medium formulated with dahlia extract tubers. The enzyme was purified by precipitation in ammonium sulphate and percolated in DEAE-Sephadex A-50 and CM-cellulose columns, witch showed a single peack in all the purification steps, maintaining the I/S ratio between 0.32 to, 0.39. Optimum pH for inulinase activity (I) was between 4.0 - 4.5 and for invertase activity (S) between 2.5 and 5.0. The optimum temperature was 60O.C for both activities and no loss in activity was observed when it was maintained at this temperature for 30 min. The Km value was 1.44 and 5.0, respectively, for I and S and Vm value 10.48 and 30.55, respectively. The I activity was strongly inhibited by Hg2+ and Ag+ and 2 x 10-3 M of glucose, but not by fructose at the same concentration. The enzyme showed an exo-action mechanism, acting on the inulin of different origins. In assay conditions total hydrolysis of all the frutans was obtained, although it has shown larger activity on the chicory inulin than that one from artichoke Jerusalem and dahlia, in the first 30 min. The obtained results suggested that the enzyme presented good potential for industrial application in the preparing the fructose syrups

Long-Living Budding Yeast Cell Subpopulation Induced by Ethanol/Acetate and Respiration

2019 ◽  
Vol 75 (8) ◽  
pp. 1448-1456 ◽  
Author(s):  
Young-Yon Kwon ◽  
Seung-Soo Kim ◽  
Han-Jun Lee ◽  
Seo-Hyeong Sheen ◽  
Kyoung Heon Kim ◽  
...  
Keyword(s):  
Differentially Expressed Genes ◽  
Budding Yeast ◽  
Gradient Centrifugation ◽  
Carbon Sources ◽  
Cell Types ◽  
Glucose Sensing ◽  
Differentially Expressed ◽  
Cell Subpopulation ◽  
Genes Encoding ◽  
Cell Transcriptome

Abstract Budding yeast generate heterogeneous cells that can be separated into two distinctive cell types: short-living low-density and long-living high-density (HD) cells by density gradient centrifugation. We found that ethanol and acetate induce formation of HD cells, and mitochondrial respiration is required. From their transcriptomes and metabolomes, we found upregulated differentially expressed genes in HD cells involved in the RGT2/RGT1 glucose sensing pathway and its downstream genes encoding hexose transporters. For HD cells, we determined an abundance of various carbon sources including glucose, lactate, pyruvate, trehalose, mannitol, mannose, and galactose. Other upregulated differentially expressed genes in HD cells were involved in the TORC1–SCH9 signaling pathway and its downstream genes involved in cytoplasmic translation. We also measured an abundance of free amino acids in HD cells including valine, proline, isoleucine, and glutamine. These characteristics of the HD cell transcriptome and metabolome may be important conditions for maintaining a long-living phenotype.

scholarly journals Transcriptional analysis of genes encoding proteins significantly increased in Kitasatospora setae KM-6054T under submerged culture

2014 ◽  
Vol 60 (6) ◽  
pp. 276-280
Author(s):  
Hiromi Miura ◽  
Yasufumi Yagisawa ◽  
Yasuki Kato ◽  
Kenji Hayashi ◽  
Nobuyuki Fujita ◽  
...  
Keyword(s):  
Submerged Culture ◽  
Transcriptional Analysis ◽  
Genes Encoding

scholarly journals Analysis of Cytadherence-Deficient, GapA-NegativeMycoplasma gallisepticum Strain R

2000 ◽  
Vol 68 (12) ◽  
pp. 6643-6649 ◽  
Author(s):  
L. Papazisi ◽  
K. E. Troy ◽  
T. S. Gorton ◽  
X. Liao ◽  
S. J. Geary
Keyword(s):  
Shuttle Vector ◽  
Phenotypic Expression ◽  
Mycoplasma Genitalium ◽  
Transcriptional Analysis ◽  
Start Codon ◽  
Wild Type ◽  
Gene Encoding ◽  
Genes Encoding ◽  
Low Passage ◽  
Translational Start

ABSTRACT Comparison of the phenotypic expression of Mycoplasma gallisepticum strain R low (passage 15) to that of strain R high (passage 164) revealed that three proteins, i.e., the cytadhesin molecule GapA, a 116-kDa protein (p116), and a 45-kDa protein (p45), are missing in strain R high. Sequence analysis confirmed that the insertion of an adenine 105 bp downstream of the gapAtranslational start codon resulted in premature termination of translation in R high. A second adenine insertion had also occurred at position 907. Restoration of expression of wild-type gapAin R high (clone designated GT5) allowed us to evaluate the extent to which the diminished cytadherence capacity could be attributed to GapA alone. The results indicated that GT5 attached to the same limited extent as the parental R high, from which it was derived. The cytadherence capability of the parental R high was not restored solely by gapA complementation alone, indicating that either p116 or p45 or both may play a role in the overall cytadherence process. The gene encoding p116 was found to be immediately downstream ofgapA in the same operon and was designatedcrmA. This gene exhibited striking homology to genes encoding molecules with cytadhesin-related functions in bothMycoplasma pneumoniae and Mycoplasma genitalium. Transcriptional analysis revealed thatcrmA is not transcribed in R high. We are currently constructing a shuttle vector containing both the wild-typegapA and crmA for transformation into R high to assess the role of CrmA in the cytadherence process.

Sugarcane Bagasse Hydrothermal Pretreatment Liquors as Suitable Carbon Sources for Hemicellulase Production by Aspergillus niger

2018 ◽  
Vol 11 (2) ◽  
pp. 316-329 ◽  
Author(s):  
Caio de Oliveira Gorgulho Silva ◽  
José Antonio de Aquino Ribeiro ◽  
Augusto Lopes Souto ◽  
Patrícia Verardi Abdelnur ◽  
Luís Roberto Batista ◽  
...  
Keyword(s):  
Aspergillus Niger ◽  
Sugarcane Bagasse ◽  
Carbon Sources ◽  
Hydrothermal Pretreatment

Organization of the regulatory region of the yeast CYC7 gene: multiple factors are involved in regulation

1987 ◽  
Vol 7 (9) ◽  
pp. 3252-3259
Author(s):  
T Prezant ◽  
K Pfeifer ◽  
L Guarente
Keyword(s):  
Cytochrome C ◽  
Regulatory Region ◽  
Carbon Sources ◽  
Binding Studies ◽  
Multiple Factors ◽  
Vitro Binding ◽  
Genes Encoding ◽  
Nonfermentable Carbon Source

Regulation of the CYC7 gene of Saccharomyces cerevisiae, encoding iso-2-cytochrome c, was studied. Expression was induced about 20-fold by heme and derepressed 4- to 8-fold by a shift from glucose medium to one containing a nonfermentable carbon source. Deletion analysis showed that induction by heme depends upon sequences between -250 and -228 (from the coding sequence) and upon the HAP1 activator gene, previously shown to be required for CYC1 expression (L. Guarente et al., Cell 36:503-511, 1984). Thus, HAP1 coordinates expression of CYC7 and CYC1, the two genes encoding isologs of cytochrome c in S. cerevisiae. HAP1-18, a mutant allele of HAP1, which increased CYC7 expression more than 10-fold, also acted through sequences between -250 and -228. In vitro binding studies showed that the HAP1 product binds to these sequences (see also K. Pfeifer, T. Prezant, and L. Guarente, Cell 49:19-28, 1987) and an additional factor binds to distal sequences that lie between -201 and -165. This latter site augmented CYC7 expression in vivo. Derepression of CYC7 expression in a medium containing nonfermentable carbon sources depended upon sequences between -354 and -295. The interplay of these multiple sites and the factors that bind to them are discussed.

scholarly journals The Tolerance of Salinity in Rice Requires the Presence of a Functional Copy of FLN2

Biomolecules ◽  
2019 ◽  
Vol 10 (1) ◽  
pp. 17 ◽  
Author(s):  
Guang Chen ◽  
Jiang Hu ◽  
Liuliu Dong ◽  
Dali Zeng ◽  
Longbiao Guo ◽  
...  
Keyword(s):  
Salinity Stress ◽  
Salinity Tolerance ◽  
Sugar Metabolism ◽  
Invertase Activity ◽  
Leaf Sheath ◽  
Japonica Rice ◽  
Wild Type ◽  
Bisphosphate Carboxylase ◽  
Genes Encoding ◽  
Knockout Line

A panel of ethane-methyl-sulfonate-mutagenized japonica rice lines was grown in the presence of salinity in order to identify genes required for the expression of salinity tolerance. A highly nontolerant selection proved to harbor a mutation in FLN2, a gene which encodes fructokinase-like protein2. Exposure of wild-type rice to salinity up-regulated FLN2, while a CRISPR/Cas9-generated FLN2 knockout line was hypersensitive to the stress. Both ribulose 1,5-bisphosphate carboxylase/oxygenase activity and the abundance of the transcript generated by a number of genes encoding components of sucrose synthesis were lower in the knockout line than in wild-type plants’ leaves, while the sucrose contents of the leaf and root were, respectively, markedly increased and decreased. That sugar partitioning to the roots was impaired in FLN2 knockout plants was confirmed by the observation that several genes involved in carbon transport were down-regulated in both the leaf and in the leaf sheath. The levels of sucrose synthase, acid invertase, and neutral invertase activity were distinctly lower in the knockout plants’ roots than in those of wild-type plants, particularly when the plants were exposed to salinity stress. The compromised salinity tolerance exhibited by the FLN2 knockout plants was likely a consequence of an inadequate supply of the assimilate required to support growth, a problem which was rectifiable by providing an exogenous supply of sucrose. The conclusion was that FLN2, on account of its influence over sugar metabolism, is important in the context of seedling growth and the rice plant’s response to salinity stress.

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