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1.
Arch Microbiol ; 202(5): 1077-1084, 2020 Jul.
Article in English | MEDLINE | ID: mdl-32030461

ABSTRACT

Plant material falling into the ultra-basic (pH 11.5-11.9) springs within The Cedars, an actively serpentinizing site in Sonoma County, California, is subject to conditions that mimic the industrial pretreatment of lignocellulosic biomass for biofuel production. We sought to obtain hemicellulolytic/cellulolytic bacteria from The Cedars springs that are capable of withstanding the extreme alkaline conditions wherein calcium hydroxide-rich water removes lignin, making cell wall polysaccharides more accessible to microorganisms and their enzymes. We enriched for such bacteria by adding plant debris from the springs into a synthetic alkaline medium with ground tissue of the biofuel crop switchgrass (Panicum virgatum L.) as the sole source of carbon. From the enrichment culture we isolated the facultative anaerobic bacterium Cellulomonas sp. strain FA1 (NBRC 114238), which tolerates high pH and catabolizes the major plant cell wall-associated polysaccharides cellulose, pectin, and hemicellulose. Strain FA1 in monoculture colonized the plant material and degraded switchgrass at a faster rate than the community from which it was derived. Cells of strain FA1 could be acclimated through subculturing to grow at a maximal concentration of 13.4% ethanol. A strain FA1-encoded ß-1, 4-endoxylanase expressed in E. coli was active at a broad pH range, displaying near maximal activity at pH 6-9. Discovery of this bacterium illustrates the value of extreme alkaline springs in the search for microorganisms with potential for consolidated bioprocessing of plant biomass to biofuels and other valuable bio-inspired products.


Subject(s)
Biofuels/microbiology , Cellulomonas/isolation & purification , Cellulomonas/metabolism , Endo-1,4-beta Xylanases/metabolism , Lignin/metabolism , Base Composition/genetics , Biomass , Cellulose/metabolism , Endo-1,4-beta Xylanases/genetics , Escherichia coli/genetics , Escherichia coli/metabolism , Ethanol/metabolism , Panicum/chemistry , Panicum/genetics , Panicum/metabolism , Pectins/metabolism , Phylogeny , Plants/metabolism , Polysaccharides/metabolism , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA
2.
Mar Genomics ; 48: 100698, 2019 Dec.
Article in English | MEDLINE | ID: mdl-31307923

ABSTRACT

Despite the lack of an inducible heat shock response (HSR), the Antarctic notothenioid fish, Trematomus bernacchii, has retained a level of physiological plasticity that can at least partially compensate for the effects of acute heat stress. Over the last decade, both physiological and transcriptomic studies have signaled these fish can mitigate the effects of acute heat stress by employing other aspects of the cellular stress response (CSR) that help confer thermotolerance as well as drive homeostatic mechanisms during long-term thermal acclimations. However, the regulatory mechanisms that determine temperature-induced changes in gene expression remain largely unexplored in this species. Therefore, this study utilized next generation sequencing coupled with an in silico approach to explore the regulatory role of microRNAs in governing the transcriptomic level response observed in this Antarctic notothenioid with respect to the CSR. Using RNAseq, we characterized the expression of 125 distinct miRNA orthologues in T. bernacchii gill tissue. Additionally, we identified 12 miRNAs that appear to be thermally responsive based on differential expression (DE) analyses performed between fish acclimated to control (-1.5 °C) and an acute heat stress (+4 °C). We further characterized the functional role of these DE miRNAs using bioinformatics pipelines to identify putative gene targets of the DE miRNAs and subsequent gene set enrichment analyses, which together suggest these miRNAs are involved in regulating diverse aspects of the CSR in T. bernacchii.


Subject(s)
Forkhead Box Protein O1/genetics , Heat-Shock Response , MicroRNAs/genetics , Perciformes/genetics , Signal Transduction , Acclimatization , Animals , Gills , Hot Temperature , Nucleic Acid Conformation , Transcriptome
3.
Org Lett ; 7(19): 4145-8, 2005 Sep 15.
Article in English | MEDLINE | ID: mdl-16146373

ABSTRACT

[reaction: see text] The reaction outcome of 2-azidoethanol and aliphatic aldehyde is found to be dependent on the catalyst and the structure of the azido alcohol. Under the catalysis of Cu(II) triflate, the corresponding acetal is obtained. A similar reaction between 2-aryl-2-azidoethanol and aldehyde catalyzed by BF3 yields a mixture of 3-oxazoline and 2-oxazoline. The latter reaction has been used for the preparation of 3-oxazolines in good enantioselectivity.

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