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The codon sequence optimized bovine prochymosin B gene was cloned under the control of the alcohol oxidase 1 promoter (AOX1) in the vector pPIC9K and integrated into the genome of the methylotrophic yeast Pichia (Komagataella) pastoris (P. pastoris) strain GS115. A transformant clone that showed resistance to over 4 mg G418/ml and displayed the highest(More)
A clone of the methylotrophic yeast Pichia pastoris strain GS115 transformed with the bovine prochymosin B gene was used to optimize the production and downstream of recombinant bovine chymosin expressed under the methanol-inducible AOXI promoter. Cell growth and recombinant chymosin production were analyzed in flask cultures containing basal salts medium(More)
Endo-polygalacturonase (endo-PGase) activity determinations generally rely on viscosity changes or reducing sugar ends produced by this activity over polygalacturonic acid. Torres and coworkers [Enzyme Microb. Technol. 48 (2011) 123-128] showed that ruthenium red (RR) is useful for endo-PGase determination. In this article, we present a high-throughput(More)
The application of cellulases in saccharification processes is restricted by its production cost. Consequently, new fungal strains able to elaborate higher cellulases titers and with special activity profiles are required to make the process economical. The aim of this investigation was to find a promising wild-type Trichoderma strain for cellulases(More)
DNA breakage detection-fluorescence in situ hybridization (DBD-FISH) is a new procedure for detecting and quantifying DNA breaks and alkali-labile sites in single cells. Cells trapped within an agarose matrix are deproteinized and treated with an alkaline unwinding solution that transforms DNA breaks and alkali-labile sites into single-strand DNA (ssDNA)(More)
Sex steroids are known to be involved in gonadal differentiation in fish, but whether androgens are early mediators of testis differentiation remains unclear. We studied the sex-related developmental variations in the gene expression of two key enzymes involved in steroids and androgen synthesis (cyp11a1 and cyp11b1) in trunks and isolated gonads of(More)
The Alexandrite laser system has proven to be an effective and safe method of ureteral lithotripsy. Some authors have recently reported the risk of interspersion of fiber splinters into tissue during lithotripsy, when short pulses and high power densities are employed. In vitro lithotripsy on renal calculi artificially placed in human ureters was realized(More)
The alexandrite laser system has proven to be an effective and safe method of treating ureteral stones. When the electromagnetic energy of a laser light pulse is selectively absorbed by the stone, a plasma forms at the surface. This plasma, which is composed of ions and electrons, continues to absorb laser energy, reaching very high pressure and generating(More)
Extraction of oil from almond fruits using supercritical carbon dioxide (SC-CO(2)) was carried out at 50 degrees C and 330 bar on three sets of almonds: raw almond seeds, raw almond kernels, and toasted almond seeds. Three different oil extraction percentages were applied on each set ranging from approximately 15 to 16%, from approximately 27 to 33%, and(More)
A simple and novel HPLC-MS method for the simultaneous quantification of testosterone, 11-ketotestosterone, and 11beta-hydroxyandrostenedione in fish serum was developed and validated. Separation was achieved on a C-18 column using a water-acetonitrile mobile-phase with a cycle time of 12 min. Ion detection was performed using ESI positive SIM at [M+H] (m/z(More)