Ultra-cold atoms in an optical cavity: two-mode laser locking to the cavity avoiding radiation pressure

The combination of ultra-cold atomic clouds with the light fields of optical cavities provides a powerful model system for the development of new types of laser cooling and for studying cooperative phenomena. These experiments critically depend on the precise tuning of an incident pump laser with respect to a cavity resonance. Here, we present a simple and reliable experimental tuning scheme based on a two-mode laser spectrometer. The scheme uses a first laser for probing higher-order transversal modes of the cavity having an intensity minimum near the cavity's optical axis, where the atoms are confined by a magnetic trap. In this way the cavity resonance is observed without exposing the atoms to unwanted radiation pressure. A second laser, which is phase-locked to the first one and tuned close to a fundamental cavity mode drives the coherent atom-field dynamics.Comment: 7 pages, 7 figure

Ethanol production from xylose by pichia stipitis NRRL Y-7124 in a stirred tank bioreactor

The ethanol production by Pichia stipitis was evaluated in a stirred tank bioreactor using semidefined medium containing xylose (90.0 g/l) as the main carbon source. Experimental assays were performed according to a 22 full factorial design to evaluate the influence of aeration (0.25 to 0.75 vvm) and agitation (150 to 250 rpm) conditions on ethanol production. In the studied range of values, the agitation increase and aeration decrease favored ethanol production, which was maximum (26.7 g/l) using 250 rpm and 0.25 vvm, conditions that gave a volumetric oxygen transfer coefficient (kLa value) of 4.9 h-1. Under these conditions, the ethanol yield factor, ethanol productivity, and the process efficiency were 0.32 g/g, 0.32 g/l.h, and 63%, respectively. These results are promising and contribute to the development of a suitable process for ethanol production from xylose by Pichia stipitis.The authors gratefully acknowledge Santander, Fapesp, Capes, and CNPq (Brazil)

The influence of initial xylose concentration, agitation, and aeration on ethanol production by Pichia stipitis from rice straw hemicellulosic hydrolysate

Rice straw hemicellulosic hydrolysate was used as fermentation medium for ethanol production by Pichia stipitis NRRL Y-7124. Shaking bath experiments were initially performed aiming to establish the best initial xylose concentration to be used in this bioconversion process. In the sequence, assays were carried out under different agitation (100 to 200 rpm) and aeration (V flask/V medium ratio varying from 2.5 to 5.0) conditions, and the influence of these variables on the fermentative parameters values (ethanol yield factor, Y P/S; cell yield factor, Y X/S; and ethanol volumetric productivity, Q P) was investigated through a 22 full-factorial design. Initial xylose concentration of about 50 g/l was the most suitable for the development of this process, since the yeast was able to convert substrate in product with high efficiency. The factorial design assays showed a strong influence of both process variables in all the evaluated responses. The agitation and aeration increase caused a deviation in the yeast metabolism from ethanol to biomass production. The best results (Y P/S = 0.37 g/g and Q P = 0.39 g/l.h) were found when the lowest aeration (2.5 V flask/V medium ratio) and highest agitation (200 rpm) levels were employed. Under this condition, a process efficiency of 72.5% was achieved. These results demonstrated that the establishment of adequate conditions of aeration is of great relevance to improve the ethanol production from xylose by Pichia stipitis, using rice straw hemicellulosic hydrolysate as fermentation medium.The financial support from Fapesp (Brazil) is gratefully acknowledged

Random UV-C mutagenesis of Scheffersomyces (formerly Pichia) stipitis NRRL Y-7124 to improve anaerobic growth on lignocellulosic sugars

Scheffersomyces (formerly Pichia) stipitis NRRL Y-7124 was mutagenized using UV-C irradiation to produce yeast strains for anaerobic conversion of lignocellulosic sugars to ethanol. UV-C irradiation potentially produces large numbers of random mutations broadly and uniformly over the whole genome to generate unique strains. Wild-type cultures of S. stipitis NRRL Y-7124 were subjected to UV-C (234 nm) irradiation targeted at approximately 40% cell survival. When surviving cells were selected in sufficient numbers via automated plating strategies and cultured anaerobically on xylose medium for 5 months at 28°C, five novel mutagenized S. stipitis strains were obtained. Variable number tandem repeat analysis revealed that mutations had occurred in the genome, which may have produced genes that allowed the anaerobic utilization of xylose. The mutagenized strains were capable of growing anaerobically on xylose/glucose substrate with higher ethanol production during 250- to 500-h growth than a Saccharomyces cerevisiae yeast strain that is the standard for industrial fuel ethanol production. The S. stipitis strains resulting from this intense multigene mutagenesis strategy have potential application in industrial fuel ethanol production from lignocellulosic hydrolysates

Deletion of the Pichia pastoris KU70 Homologue Facilitates Platform Strain Generation for Gene Expression and Synthetic Biology

Targeted gene replacement to generate knock-outs and knock-ins is a commonly used method to study the function of unknown genes. In the methylotrophic yeast Pichia pastoris, the importance of specific gene targeting has increased since the genome sequencing projects of the most commonly used strains have been accomplished, but rapid progress in the field has been impeded by inefficient mechanisms for accurate integration. To improve gene targeting efficiency in P. pastoris, we identified and deleted the P. pastoris KU70 homologue. We observed a substantial increase in the targeting efficiency using the two commonly known and used integration loci HIS4 and ADE1, reaching over 90% targeting efficiencies with only 250-bp flanking homologous DNA. Although the ku70 deletion strain was noted to be more sensitive to UV rays than the corresponding wild-type strain, no lethality, severe growth retardation or loss of gene copy numbers could be detected during repetitive rounds of cultivation and induction of heterologous protein production. Furthermore, we demonstrated the use of the ku70 deletion strain for fast and simple screening of genes in the search of new auxotrophic markers by targeting dihydroxyacetone synthase and glycerol kinase genes. Precise knock-out strains for the well-known P. pastoris AOX1, ARG4 and HIS4 genes and a whole series of expression vectors were generated based on the wild-type platform strain, providing a broad spectrum of precise tools for both intracellular and secreted production of heterologous proteins utilizing various selection markers and integration strategies for targeted or random integration of single and multiple genes. The simplicity of targeted integration in the ku70 deletion strain will further support protein production strain generation and synthetic biology using P. pastoris strains as platform hosts

Effect of cell immobilization and pH on Scheffersomyces stipitis growth and fermentation capacity in rich and inhibitory media

Background A wide range of value-added products can potentially be produced by bioprocessing hardwood spent sulfite liquors (HSSLs) that are by-products of pulp and paper industry with a high pentose sugar content. However, besides sugars, HSSLs contain considerable amounts of sulfonated lignin derivatives and acetic acid that inhibit the metabolic activity of most microorganisms. Scheffersomyces stipitis is a yeast with high capacity to ferment the pentose sugar xylose under appropriate microaerophilic conditions but it has limited tolerance to HSSL inhibitors. In the present study, cultivations of suspended and immobilized S. stipitis were compared in terms of growth capacity and by-product formation using rich medium and HSSL to investigate whether the immobilization of cells in calcium alginate beads could be a protection against inhibitors while favoring the presence of microaerophilic conditions. Results Whereas cell immobilization clearly favored the fermentative metabolism in rich medium, pH control was found to play a more important role than cell immobilization on the ethanol production efficiency from bio-detoxified HSSL (bdHSSL), leading to an improvement of 1.3-fold on the maximum ethanol productivity than using suspended cells. When immobilization and pH control were applied simultaneously, the ethanol yield improved by 1.3-fold with unchanged productivity, reaching 0.26 g ethanol.(g glucose\ +\ xylose)\−1. Analysis of the immobilized beads inside revealed that the cells had grown in the opposite direction of the cortex. Conclusions Immobilization and pH control at 5.5, when applied simultaneously, have a positive impact on the fermentative metabolism of S. stipitis, improving the ethanol production efficiency. For the first time light microscopic analysis of the beads suggested that the nutrient and mass transfer limitations played a more important role in the fermentation than a possible protective role against inhibitors. Keywords Scheffersomyces stipitis Hardwood spent sulfite liquor Cell immobilization Light microscopy Ca alginate beads Xylose fermentation Stress toleranc

Identification of a mitotic recombination hotspot on chromosome III of the asexual fungus Aspergillus niger and its possible correlation elevated basal transcription

Genetic recombination is an important tool in strain breeding in many organisms. We studied the possibilities of mitotic recombination in strain breeding of the asexual fungus Aspergillus niger. By identifying genes that complemented mapped auxotrophic mutations, the physical map was compared to the genetic map of chromosome III using the genome sequence. In a program to construct a chromosome III-specific marker strain by selecting mitotic crossing-over in diploids, a mitotic recombination hotspot was identified. Analysis of the mitotic recombination hotspot revealed some physical features, elevated basal transcription and a possible correlation with purine stretches

Biological Earth observation with animal sensors.

Space-based tracking technology using low-cost miniature tags is now delivering data on fine-scale animal movement at near-global scale. Linked with remotely sensed environmental data, this offers a biological lens on habitat integrity and connectivity for conservation and human health; a global network of animal sentinels of environmen-tal change