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Amylase Gene (amylase + gene)
Selected AbstractsCLONING AND SEQUENCING OF THE ,-AMYLASE GENE FROM BACILLUS SUBTILIS US116 STRAIN ENCODING AN ENZYME CLOSELY IDENTICAL TO THAT FROM BACILLUS AMYLOLIQUEFACIENS BUT DISTINCT IN THERMAL STABILITYJOURNAL OF FOOD BIOCHEMISTRY, Issue 2 2010EZZEDINE BEN MESSAOUD ABSTRACT The gene encoding for the ,-amylase AMYUS116 was cloned and sequenced. The amino acid sequence of AMYUS116 exhibited an almost perfect homology with the ,-amylase BACAAM, excluding the residues N205 and N217 of AMYUS116 that were changed to H205 and I217 into BACAAM. Three mutant derivatives from AMYUS116 (N205H, N217I and N205H/N217I) were created by site-directed mutagenesis and their physicochemical and kinetic properties were compared with those of the wild-type enzymes. Therefore, the undertaken amylases mainly generated maltohexaose from starch and had radically the same kinetic parameters and optimum pH and temperature. They, however, were significantly distinct in thermal stability; AMYUS116 was more thermosensible as its half-life time at 80C was 13 min, while those of BACAAM and the double mutant were likewise 38 min. The single-mutant amylases exhibited an identically intermediate thermal stability as their half-life times at 80C were roughly 22 min. PRACTICAL APPLICATIONS Of particular interest to the current search is that the different thermal stability between AMYUS116 and BACAAM can lead to novel findings pertaining to protein stability, which can bring about new strategies for protein engineering. Basically, the comparative study of closely related amylases and the protein engineering of already existing ones are certainly important because they offer opportunities to understand the structure,function relationships of these biocatalysts. [source] Expression of 2 Lipomyces kononenkoae,-Amylase Genes in Selected Whisky Yeast StrainsJOURNAL OF FOOD SCIENCE, Issue 7 2004K. la Grange-Nel ABSTRACT: Nineteen whisky yeasts were evaluated according to their fermentation performance and ability to produce a palatable spirit. Four of these strains were selected and, together with a commercial wine yeast strain (control), were transformed with integration plasmids containing the LKA1 and LKA2 , -amylase genes from the yeast Lipomyces kononenkoae. Fermentation trials with starch-containing media indicated that the transformants produced between 47% and 66% of the theoretical ethanol yield. This study has resulted in progress toward the development of whisky yeast that could ultimately be used in a process during which production of amylases, hydrolysis of starch, and fermentation of resulting sugars to grain whisky occur in a single step. [source] Subcellular localization of proteins labeled with GFP in Xanthomonas citri ssp. citri: targeting the division septumFEMS MICROBIOLOGY LETTERS, Issue 1 2010Paula M.M. Martins Abstract Xanthomonas citri ssp. citri (Xac) is the causal agent of citrus canker, an economically important disease that affects citrus worldwide. To initiate the characterization of essential biological processes of Xac, we constructed integrative plasmids for the ectopic expression of green fluorescent protein (GFP)-labeled proteins within this bacterium. Here, we show that the disruption of the ,-amylase gene (amy), the site of plasmid integration into the bacterial chromosome, does not alter its pathogenesis while abolishing completely the ability of Xac to degrade starch. Furthermore, our GFP expression system was used to characterize ORF XAC3408, a hypothetical protein encoded by Xac that shares significant homology to the FtsZ-stabilizing factor ZapA from Bacillus subtilis (ZapABsu). GFP-XAC3408 expressed in Xac exhibited a septal localization pattern typical of GFP-ZapABsu, which indicates that XAC3408 is the Xac orthologue of the cell division protein ZapABsu. The results demonstrate the potential of GFP labeling for protein functional characterizations in Xac, and, in addition, the Xac mutant strain labeled at the septum constitutes a biological model for the exploration of antibacterial compounds able to inhibit cell division in this plant pathogen. [source] Expression and secretion of an ,-amylase gene from a native strain of Bacillus licheniformis in Escherichia coli by T7 promoter and putative signal peptide of the geneJOURNAL OF APPLIED MICROBIOLOGY, Issue 6 2003M. Shahhoseini Abstract The gene encoding a hyperthermostable , -amylase from a Bacillus licheniformis native strain was cloned in pET24d transcription vector containing T7 promoter, and expressed in Escherichia coli BL21(DE3) cells. Having confirmed the , -amylase activity through activity staining method on SDS,PAGE gel, the yields of production were determined in two separated intra and inter-cellular phases and compared using enzymatic assay methods. Extracellular production of the active recombinant enzyme implies the recognition of the putative signal peptide of this Bacillus sp. by E. coli secretory system. This may be because of the amino acid sequence of this signal peptide which covers all the structural parameters of a standard signal peptide processed by Lep B, the major signal peptidase in E. coli secretory system. This study recommends the use of this signal peptide for extracellular production of other foreign proteins in E. coli. [source] An ABA-responsive bZIP protein, OsBZ8, mediates sugar repression of , -amylase gene expressionPHYSIOLOGIA PLANTARUM, Issue 1 2003Yi-Ching Lee Expression of some , -amylase genes in cereals is suppressed by sugars and activated by sugar starvation. A 100-bp sugar response sequence (SRS) identified in the promoter of a rice , -amylase gene, ,Amy3, contains three essential motifs: the GC box, the G box, and the TATCCA element. To study the mechanism of sugar regulation of ,Amy3 transcription, an ABA-responsive bZIP protein, OsBZ8, which binds specifically to the G box in ,Amy3 SRS was characterized and function analysed. In sucrose-starved rice suspension cells and embryos, decline in OsBZ8 mRNA levels coincided with the induction of ,Amy3 mRNA accumulation. In vivo gain- and loss-of-function studies by transient expression assays in rice embryos revealed that OsBZ8 suppresses SRS activity through the G box and overrides the activity of an activator, OsMYBS1, which binds to the TATCCA element. Gel mobility shift assays revealed that OsBZ8 binds specifically to the G box in vitro. These studies suggest that OsBZ8 is a suppressor responsible for sugar repression of ,Amy3 expression, and OsMYBS1 is responsible for sugar starvation induced expression of ,Amy3. [source] Ovine alpha-amylase genes: isolation, linkage mapping and association analysis with milk traitsANIMAL GENETICS, Issue 4 2004J. H. Calvo Summary On the basis of comparisons between cattle and sheep genome mapping information the ovine , - amylase gene was examined as a possible genetic marker for milk traits in sheep. The objective of the present study was to isolate, map and determine whether this gene is a candidate gene for milk traits. DNA fragments (832 and 2360 bp) corresponding to two different AMY genes were isolated, and one SNP in intron 3 and one GTG deletion in exon 3 of the 2360 bp DNA fragment were found. The 2360 bp ovine AMY DNA fragment was located on chromosome 1 by linkage mapping using the International Mapping Flock. No association was found between estimated breeding values for milk yield, protein and fat contents and AMY genotypes in a daughter design comprising 13 Manchega families with an average of 29 daughters (12,62) per sire. [source] A three-way contact zone between forms of Patella rustica (Mollusca: Patellidae) in the central Mediterranean SeaBIOLOGICAL JOURNAL OF THE LINNEAN SOCIETY, Issue 1 2010ALEXANDRA SÁ-PINTO Previous studies have reported the occurrence of three differentiated mtDNA lineages within Patella rustica in the Mediterranean Sea. Two hypotheses have been proposed to explain these observations: (1) the maintenance of ancestral polymorphism within a single species; (2) the occurrence of cryptic species not identified previously. To distinguish between these hypotheses, we screened the genetic variability at nine allozyme loci, an intron from the ,-amylase gene and a mitochondrial gene for 187 individuals of P. rustica sampled from seven Mediterranean localities. Eight additional localities were screened for the last two markers to place the differentiated lineages in a clear geographic context. Our results demonstrate that the three mtDNA lineages correspond to three distinct nuclear genotype clusters and provide further details on their distribution: the cluster corresponding to the mtDNA lineage from the Atlantic and western Mediterranean extends as far as the south coast of Italy, whereas the remaining two clusters occur in sympatry in the eastern Mediterranean. One of the eastern Mediterranean clusters is highly differentiated and seems to be reproductively isolated from the codistributed form; we therefore suggest that it corresponds to a new species. The remaining two clusters are less differentiated and form a contact zone across south Italian shores. This three-way contact zone constitutes an interesting model for the study of speciation in the marine realm. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 100, 154,169. [source] Expression of 2 Lipomyces kononenkoae,-Amylase Genes in Selected Whisky Yeast StrainsJOURNAL OF FOOD SCIENCE, Issue 7 2004K. la Grange-Nel ABSTRACT: Nineteen whisky yeasts were evaluated according to their fermentation performance and ability to produce a palatable spirit. Four of these strains were selected and, together with a commercial wine yeast strain (control), were transformed with integration plasmids containing the LKA1 and LKA2 , -amylase genes from the yeast Lipomyces kononenkoae. Fermentation trials with starch-containing media indicated that the transformants produced between 47% and 66% of the theoretical ethanol yield. This study has resulted in progress toward the development of whisky yeast that could ultimately be used in a process during which production of amylases, hydrolysis of starch, and fermentation of resulting sugars to grain whisky occur in a single step. [source] An ABA-responsive bZIP protein, OsBZ8, mediates sugar repression of , -amylase gene expressionPHYSIOLOGIA PLANTARUM, Issue 1 2003Yi-Ching Lee Expression of some , -amylase genes in cereals is suppressed by sugars and activated by sugar starvation. A 100-bp sugar response sequence (SRS) identified in the promoter of a rice , -amylase gene, ,Amy3, contains three essential motifs: the GC box, the G box, and the TATCCA element. To study the mechanism of sugar regulation of ,Amy3 transcription, an ABA-responsive bZIP protein, OsBZ8, which binds specifically to the G box in ,Amy3 SRS was characterized and function analysed. In sucrose-starved rice suspension cells and embryos, decline in OsBZ8 mRNA levels coincided with the induction of ,Amy3 mRNA accumulation. In vivo gain- and loss-of-function studies by transient expression assays in rice embryos revealed that OsBZ8 suppresses SRS activity through the G box and overrides the activity of an activator, OsMYBS1, which binds to the TATCCA element. Gel mobility shift assays revealed that OsBZ8 binds specifically to the G box in vitro. These studies suggest that OsBZ8 is a suppressor responsible for sugar repression of ,Amy3 expression, and OsMYBS1 is responsible for sugar starvation induced expression of ,Amy3. [source] An , -amylase (At4g25000) in Arabidopsis leaves is secreted and induced by biotic and abiotic stressPLANT CELL & ENVIRONMENT, Issue 4 2007ELIZABETH A. DOYLE ABSTRACT Leaves are reported to contain a secreted , -amylase that accumulates during senescence or after biotic or abiotic stress; however, a gene encoding this enzyme has not been described. Because a secreted amylase is isolated from plastidic starch, the function of this enzyme is difficult to predict, but circumstantial evidence suggests that it may degrade starch after cell death. The Arabidopsis thaliana genome contains three , -amylase genes, one of which, AMY1 (At4g25000), has a putative signal sequence suggesting that the protein may be secreted. Two independent T-DNA insertion mutants in AMY1 lacked an amylase band on starch zymograms, which was previously named ,A1'. Washed leaf protoplasts contained reduced A1 activity suggesting that the enzyme is secreted. Native AMY1, fused to a weakly fluorescent form of GFP, was sensitive to proteinase K infiltrated into leaf apoplastic spaces, while a cytosolic form of GFP was unaffected until cell breakage, confirming that the AMY1 protein is secreted. Amylase A1 was transcriptionally induced in senescing leaves and in leaves exposed to heat stress, treated with abscisic acid or infected with Pseudomonas syringae pv. tomato expressing avrRpm1. The A1 amylase was also extremely heat resistant and its expression was up-regulated in cpr5-2, an activated defence response mutant. [source] | |||||||||||||