Aspergillus westerdijkiae polyketide synthase gene “aoks1” is involved in the biosynthesis of ochratoxin A

OchratoxinA (OTA) is a potential nephrotoxic, teratogenic, immunogenic, hepatotoxic and carcinogenic mycotoxin, produced by Aspergillus westerdijkiae NRRL 3174. Herein we describe the characterization of a putative OTA-polyketide synthasegene “aoks1”, cloned by using gene walking approach. The predicted amino acid sequence of the 2 kb clone display 34–60% similarities to different polyketide synthasegenes including lovastatine biosynthesis gene “lovb” in A. terreus, compactin biosynthesis gene “mlcA” in Penicillium citrinum and OTA biosynthesis gene “otapksPN” in P. nordicum. Based on the reverse transcription PCR and kinetic secondary metabolites production studies, aoks1 expression was found to be associated with OTA biosynthesis. Further a mutant, in which the aoks1gene was inactivated by Escherichia coli hygromycin B phosphotransferase gene, lost the capacity to produce OTA, but still producing mellein. To our knowledge this report describes for the first time characterization of a gene involved in OTA biosynthesis, with the information about mellein which was proposed in the literature to be an intermediate OTA. This study also suggests that aoks1 may be the second polyketide synthase gene required for OTA biosynthesis in A. westerdijkiae NRRL 3174

Differential regulation of Ota and Otb, two primary glycine betaine transporters in the methanogenic archaeon Methanosarcina mazei go1

Methanogenic archaea accumulate glycine betaine in response to hypersalinity, but the regulation of proteins involved, their mechanism of activation and regulation of the corresponding genes are largely unknown. Methanosarcina mazei differs from most other methanoarchaea in having two gene clusters both encoding a potential glycine betaine transporter, Ota and Otb. Western blot as well as quantitative real-time PCR revealed that Otb is not regulated by osmolarity. On the other hand, cellular levels of Ota increased with increasing salt concentrations. A maximum was reached at 300-500 m M NaCl. Ota concentrations reached a maximum 4 h after an osmotic upshock. Hyperosmolarity also caused an increase in cellular Ota concentrations. In addition to osmolarity Ota expression was regulated by the growth phase. Expression of Ota as well as transport of betaine was downregulated in the presence of glycine betaine. Copyright (c) 2007 S. Karger AG, Basel

A CMOS class-AB transconductance amplifier for switched-capacitor applications

A CMOS operational transconductance amplifier (OTA) using a fully differential single-stage core OTA as the input stage and a differential to single current converter as the output stage, each biased at a separate current level, is presented. A large gain-bandwidth product (2.7 MHz) and a high slew-rate (5 V/μs) can be obtained by applying a large bias current to the core OTA. Due to the class-AB operation of the output stage, a high output impedance can be obtained by applying a small bias current to the output stage, resulting in a high DC-gain (61.6 dB). When the performance of this class-AB OTA is compared with that of basic single-stage OTAs it is found that the output impedance of the class-AB OTA is increased without limiting the bandwidth or slew-rat

Nonlinear time-domain macromodeling of OTA circuits

The authors present an accurate nonlinear macromodel of the operational transconductance amplifier (OTA) which is suitable for the transient simulation of OTA-based CMOS analog integrated circuits. As compared to device-level OTA models, the proposed macromodel is advantageous in terms of CPU time. Also, in circuits with many OTAs, it does not have the problems of convergence that the device-level MODEL has. All the macromodel parameters can be calculated from measurements made at the OTA terminals. Experimental results from a 3-μm CMOS OTA prototype as well as simulation results from device-level models are included and compared to simulation results from the macromodel