scholarly journals Biochemical Characterization of a Mycobacteriophage Derived DnaB Ortholog Reveals New Insight into the Evolutionary Origin of DnaB Helicases

PLoS ONE ◽  
2015 ◽  
Vol 10 (8) ◽  
pp. e0134762 ◽  
Author(s):  
Priyanka Bhowmik ◽  
Sujoy K. Das Gupta
2016 ◽  
Vol 11s1 ◽  
pp. ACI.S40292
Author(s):  
Tom Manczak ◽  
Henrik Toft Simonsen

A fast and reproducible protocol was established for enzymatic characterization of plant sesquiterpene synthases that can incorporate radioactivity in their products. The method utilizes the 96-well format in conjunction with cluster tubes and enables processing of >200 samples a day. Along with reduced reagent usage, it allows further reduction in the use of radioactive isotopes and flammable organic solvents. The sesquiterpene synthases previously characterized were expressed in yeast, and the plant-derived Thapsia garganica kunzeaol synthase TgTPS2 was tested in this method. KM for TgTPS2 was found to be 0.55 μM; the turnover number, kcat, was found to be 0.29 s−1, kcat for TgTPS2 is in agreement with that of terpene synthases of other plants, and kcat/ KM was found to be 0.53 s−1 μM−1 for TgTPS2. The kinetic parameters were in agreement with previously published data.


2011 ◽  
Vol 2011 ◽  
pp. 1-7 ◽  
Author(s):  
José M. Eirín-López ◽  
Juan Ausió

The evolution of sex remains a hotly debated topic in evolutionary biology. In particular, studying the origins of the molecular mechanisms underlying sexual reproduction and gametogenesis (its fundamental component) in multicellular eukaryotes has been difficult due to the rapid divergence of many reproductive proteins, pleiotropy, and by the fact that only a very small number of reproductive proteins specifically involved in reproduction are conserved across lineages. Consequently, during the last decade, many efforts have been put into answering the following question: did gametogenesis evolve independently in different animal lineages or does it share a common evolutionary origin in a single ancestral prototype? Among the various approaches carried out in order to solve this question, the characterization of the evolution of the DAZ gene family holds much promise because these genes encode reproductive proteins that are conserved across a wide range of animal phyla. Within this family, BOULE is of special interest because it represents the most ancestral member of this gene family (the “grandfather” of DAZ). Furthermore, BOULE has attracted most of the attention since it represents an ancient male gametogenic factor with an essential reproductive-exclusive requirement in urbilaterians, constituting a core component of the reproductive prototype. Within this context, the aim of the present work is to provide an up-to-date insight into the studies that lead to the characterization of the DAZ family members and the implications in helping decipher the evolutionary origin of gametogenesis in metazoan animals.


2008 ◽  
Vol 3 (12) ◽  
pp. 1934578X0800301
Author(s):  
Salvador Máñez

The discovery that various drugs (e.g., phenobarbital) stimulate their own metabolism through a mechanism coined as enzymatic induction opened up a fascinating road that eventually led to the accurate biochemical characterization of the pregnane X receptor. After numerous studies, researchers have concluded that this receptor is activated by different endogenous steroids and a number of foreign lipophile ligands. Once activated, it induces the synthesis of oxygenases and conjugating enzymes. The activating ligands identified to date include many synthetic drugs, along with a number of natural products. The present review summarizes the data relating to the origin, chemistry, and pharmacological activity of the newest natural products that have been found to interact with the pregnane X receptor.


1998 ◽  
Vol 64 (10) ◽  
pp. 3939-3947 ◽  
Author(s):  
Yaowei Kang ◽  
Russell Carlson ◽  
Wendy Tharpe ◽  
Mark A. Schell

ABSTRACT Genetic manipulation of fluorescent pseudomonads has provided major insight into their production of antifungal molecules and their role in biological control of plant disease. Burkholderia cepaciaalso produces antifungal activities, but its biological control activity is much less well characterized, in part due to difficulties in applying genetic tools. Here we report genetic and biochemical characterization of a soil isolate of B. cepacia relating to its production of an unusual antibiotic that is very active against a variety of soil fungi. Purification and preliminary structural analyses suggest that this antibiotic (called AFC-BC11) is a novel lipopeptide associated largely with the cell membrane. Analysis of conditions for optimal production of AFC-BC11 indicated stringent environmental regulation of its synthesis. Furthermore, we show that production of AFC-BC11 is largely responsible for the ability ofB. cepacia BC11 to effectively control the damping-off of cotton caused by the fungal pathogen Rhizoctonia solani in a gnotobiotic system. Using Tn5 mutagenesis, we identified, cloned, and characterized a region of the genome of strain BC11 that is required for production of this antifungal metabolite. DNA sequence analysis suggested that this region encodes proteins directly involved in the production of a nonribosomally synthesized lipopeptide.


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