Mixing power, external convection, and effectiveness in bioreactors

The phenomena of mixing and mass transfer of substrates to microorganisms greatly affect the biochemical reactions which take place in fermentation processes. The effect that agitation power has on the observable reaction kinetics involved in beer fermentation has been studied in different types of bioreactors, from laboratory to industrial scale. With this aim in mind, an effectiveness factor, eta, is introduced which is defined as the relation between the existing rate of reaction, whichever bioreactor is considered, and the reaction rate in the well-mixed, and therefore presumably homogeneous, bioreactor with no diffusional limits. The limitation to homogeneously supplying nutrient material to the cells produces a decrease in this effectiveness factor, which has been correlated to the energy dissipation rate with a similar slope to that which appears in an existing correlation in the literature between this energy and the mass transfer coefficient. Additionally, a dimensionless reaction-convection number, N(RC), which is a function of the power input per unit volume, is proposed, which has been appropriately employed in correlating the effectiveness factor for the types of processes in which convection may be the key resistance factor. (c) 1996 John Wiley & Sons, Inc.     

Xanthan production in a plunging jet reactor

Summary A plunging jet reactor (0.04–0.08 m³) was used for the production of the exopolysaccharide xanthan with Xanthomonas campestris. The microorganism was not affected by the pump shear force. Similar specific growth rates and xanthan space-time yields to those in other reactor types were achieved at much lower specific power input. The better oxygen sorption efficiency in the jet reactor overcompensated for the effect of poor mixing in the wall region at high xanthan concentrations. Offprint requests to: A. Schumpe     

Kinetic and metabolic effects of nitrogen, magnesium and sulphur restriction in Xanthomonas campestris batch cultures

By reducing the concentration of nitrogen (from 5.0 to 2.5 mmol 1^-1), batch cultures of Xanthomonas campestris induced the enzyme UDP-glucose dehydrogenase and stimulated the Entner-Doudoroff pathway enzyme glucose-6-P dehydrogenase. The surplus energy generation was directed to xanthan biosynthesis resulting in a 10% polysaccharide increase. The nitrogen restriction led to a higher consumption of nitrogen (93%) whereas glucose consumption did not surpass 75% utilization. Low concentrations of both magnesium and sulphur exerted a negative effect on xanthan formation. Both restrictions reduced the phosphomannose isomerase enzyme activity by 10-fold turning the mannose transference presumably into the rate-limiting step for xanthan biosynthesis. Conversely, the rate of synthesis of glucuronic acid residues did not affect the rate of xanthan biosynthesis. Polysaccharide synthesis in magnesium and sulphur cultures was negatively affected in comparison with cell formation as the cell volumetric production rate increased from 0.037 to 0.091 g 1^-1 h^-1 and the xanthan volumetric production rate dropped from 0.133 g 1^-1 h^-1 to the minimum obtained at 0.083 g 1^-1 h^-1. The efficiency of the carbon substrate conversion was also greatly changed.     

华北平原种养一体规模化农场氮素流动特征及利用效率——以河北津龙循环农业园区为例

种养一体规模化、集约化是华北平原农业发展的必然趋势,而氮素是连接种植养殖的主要养分资源,以河北津龙循环农业园区为例,采用文献资料、实地调查方法分析农场水平氮素流动特征及利用率,并通过情景分析方法提出农场氮素管理措施,为实现农场水平氮养分资源高效利用、提高农场生产系统生产力和改善华北平原循环农业模式提供技术支撑和科学依据.结果表明: 在农场水平下,化肥和有机肥输入氮量674.6 kg·hm^-2·a^-1,占总输入氮量的88.3%,氮利用率为41.5%,种植系统氮盈余量190.7 kg·hm^-2·a^-1,施氮量过多是造成种植系统氮利用率低和氮素盈余量高的主要原因.养殖系统中外购饲料提供氮量占饲料总输入氮量的83.2%,粪尿排氮量为776.6 t·a^-1,而还田比例仅为36.3%,氮利用率19.7%.农场水平氮总利用率为40.7%.情景分析表明,农田减少化肥施氮量50%(情景1)、增加来自农场内部玉米籽粒产量(情景2)措施,可分别使种植系统氮利用率提高34.6%和15.6%,同时农场水平氮总利用率分别提高18.7%和9.8%;另外,优化养殖系统饲料结构(情景3),可使氮总利用率提高19.1%.因此,减少化肥氮施用量、调整作物种植结构、优化饲料结构等,是提高农场氮生产力和实现环境友好双赢效果的措施和途径.     

Expression of the gum operon directing xanthan biosynthesis in Xanthomonas campestris and its regulation in planta

The gum gene cluster of Xanthomonas campestris pv. campestris comprises 12 genes whose products are involved in the biosynthesis of the extracellular polysaccharide xanthan. These genes are expressed primarily as an operon from a promoter upstream of the first gene, gumB. Although the regulation of xanthan synthesis in vitro has been well studied, nothing is known of its regulation in planta. A reporter plasmid was constructed in which the promoter region of the gum operon was fused to gusA. In liquid cultures, the expression of the gumgusA reporter was correlated closely with the production of xanthan, although a low basal level of beta-glucuronidase activity was seen in the absence of added carbon sources when xanthan production was very low. The expression of the gumgusA fusion also was subject to positive regulation by rpfF, which is responsible for the synthesis of the diffusible signal factor (DSF). The expression of the gumgusA fusion in bacteria recovered from inoculated turnip leaves was maximal at the later phases of growth and was subject to regulation by rpfF. These results provide indirect support for the operation of the DSF regulatory system in bacteria in planta.     

Power consumption of three impeller combinations in mixing Xanthan fermentation broths

Xanthan gum fermentation represents a good model for the study of the mixing of rheologically complex culture broths. Most of the previous work on power consumption dealt with ‘standard’, single impellers and used model fluids to simulate xanthan broths. This work describes the characterization of three dual-impeller combinations (D/T = 0·53) for the mixing of dehydrated—reconstituted fermentation broths of Xanthomonas campestris that had matched rheology to the actual broths. The bottom impeller was a Rushton turbine (RT) and the top impeller was another RT, a 45° pitched blade turbine (PT) or an A-310 Lightnin mixer (A310). The experiments were carried out in a tank of 0·0094 m³ working volume equipped with an air bearing dynamometer. The power was measured in a wide range of xanthan concentrations (5–40 kg m⁻³) in aerated (0·25, 0·5 and 1·0 vvm) and unaerated conditions. Unaerated power number (Po) vs. Reynolds number (Re) curves showed similar trends for the three combinations. Exponents close to −1 were obtained in the laminar region. A minimum in Po (Po_min) occurred at Re = 30–40, then increasing to a plateau value which was evident at Re> 200. In the transition region Po_min values were 4·3 (RT and RT), 3·6 (RT and PT) and 2·4 (RT and A310). The aerated power data for (RT and PT) and (RT and A-310) showed higher torque instabilities than the dual RT combinations at higher xanthan concentrations. The higher the xanthan concentrations, the higher the drop in power and the less important the effect of the aeration rate. Among the combinations tested, when using Rushton turbines, the well-mixed ‘cavern’ reached the tank wall (i.e., fluid motion was observed) at the lowest volumetric power input. High     

Effects of melanin on the accumulation of exopolysaccharides by Aureobasidium pullulans grown on nitrate

Aureobasidium pullulans produced pullulan and melanin when grown in medium containing low nitrate levels. With high nitrate concentrations, however, this fungus produced a mixture of exopolysaccharides (EPS) without melanin synthesis. At 0.78 g l(-1) N as nitrate, where no melanin synthesis occurred, maximum EPS yields reached 6.92 g l(-1) and then decreased to the final yield of 2.36 g l(-1). Following melanin addition (0.1 g l(-1)), yields reached 7.02 g l(-1) at 48 h and fell to a final yield of 5.21 g l(-1). The EPS produced in high nitrate medium contained both pullulan and (1-->3)-beta-glucan, but only pullulan was produced with melanin-supplementation. With melanin addition a doubling of (1-->3)-beta-glucanase activity was observed in high nitrate medium compared to that without supplementation. On the other hand amylolytic activities disappeared in medium with melanin production or addition. Culture filtrates sustained a higher reducing capacity (RC) when melanin was present. Low RC appeared to reduce (1-->3)-beta-glucanase activity and increase amylolytic activities. Thus, higher RC appears to inhibit production/activity of amylose-degrading enzymes capable of degrading pullulan, and stimulates (1-->3)-beta-glucanase synthesis/activity, leading to a preferential accumulation of pullulan.     

Productivity, 15N dynamics and water use efficiency in low‐ and high‐input switchgrass systems

Sustainable and environmentally benign switchgrass production systems need to be developed for switchgrass to become a large‐scale dedicated energy crop. An experiment was conducted in California from 2009 to 2011 to determine the sustainability of low‐ and high‐input irrigated switchgrass systems as a function of yield, irrigation requirement, crop N removal, N translocation from aboveground (AG) to belowground (BG) biomass during senescence, and fertilizer ¹⁵N recovery (FNR) in the AG and BG biomass (0–300 cm), and soil (0–300 cm). The low‐input system consisted of a single‐harvest (mid‐fall) irrigated until flowering (early summer), while the high‐input system consisted of a two‐harvest system (early summer and mid‐fall) irrigated throughout the growing season. Three N fertilization rates (0, 100, and 200 kg N ha^?1 yr^?1) were applied as subtreatments in a single application in the spring of each year. A single pulse of ¹⁵N enriched fertilizer was applied in the first year of the study to micro‐plots within the 100 kg N ha^?1 subplots. Average yields across years under optimal N rates (100 and 200 kg ha^?1 yr^?1 for low‐ and high‐input systems, respectively) were 20.7 and 24.8 Mg ha^?1. However, the low input (372 ha mm) required 47% less irrigation than the high‐input system (705 ha mm) and achieved higher irrigation use efficiency. In addition, the low‐input system had 46% lower crop N removal, 53% higher N stored in BG biomass, and a positive N balance, presumably due to 49% of ¹⁵N translocation from AG to BG biomass during senescence. Furthermore, at the end of 3 years, the low‐input system had lower fertilizer ¹⁵N removed by harvest (26%) and higher FNR remaining in the system in BG biomass plus soil (31%) than the high‐input system (45% and 21%, respectively). Based on these findings, low‐input systems are more sustainable than high‐input systems in irrigated Mediterranean climates.     

Effect of corn steep liquor on xanthan production by Xanthomonas campestris

Summary The addition of corn steep liquor (CSL) to batch cultures of Xanthomonas campestris using sucrose as carbon source stimulated cell growth rate, viscosity and xanthan production as compared to non-supplemented cultures. The addition of CSL to a basal medium at a dose of 1 g/l, increased xanthan production and viscosity by 22% and 44% respectively. CSL also shortened the cultivation time and promoted a more efficient sucrose utilization for polymer synthesis. After 72 h of incubation the xanthan yield per sucrose consumed in the CSL-amended culture was 0.63 g/g, this is, 15% higher than without CSL addition. At higher doses of CSL cell growth rate was also increased but not polymer production.     

Xanthan production from olive-mill wastewaters

Olive mill wastewaters (OMW) are a by-product from olive oil manufacture that cause environmental pollution. These wastes have been used as substrate for the production of the extracellular polysaccharide xanthan by Xanthomonas campestris NRRL B1459-S4L41. Growth and xanthan production on dilute OMW as a sole source of nutrients were obtained at OMW concentrations below 60%, yielding a maximal xanthan production of 4.4gl⁻¹ at 30–40% OMW concentration. Addition of nitrogen and/or salts led to significantly increased xanthan yields with a maximum of 7.7gl⁻¹. The N/salts supplements also allowed an increase in the optimal OMW concentration. Inocula pre-grown on OMW can be used. Results suggest that an improved xanthan yield could be obtained with adequate balance between waste concentration and nitrogen or salt supplementation. OMW is proposed as a low-cost substrate for xanthan production with the additional environmental benefit of this use.     

Effects of nitrogen mineralization on paddy rice yield under low nitrogen input conditions in irrigated rice-based multiple cropping with intensive cropping of vegetables in southwest China

The farm household responsibility system (FHRS) was adopted in Chinese rural areas during the economic reform in the early 1980s. Since then, many farm households have increased cropping intensity by using large quantities of nitrogen (N) fertilizers in their responsible fields to increase agricultural income. However, intensive cropping systems with low N input are still common in remote places of the southwestern region of China. Maintenance and improvement of soil quality in intensive cropping systems is critical for sustaining agricultural productivity and environmental quality for future generations. The effects of intensive cropping of vegetables on paddy rice (Oryza sativa L.) yield using small quantities of N fertilizers through N mineralization of paddy soil in irrigated rice-based multiple cropping systems were studied in 15 paddy fields in Sichuan Province, China for 3 years. Intensification of vegetable cropping with negative N balance and removal of vegetable crop residues has greatly decreased total N (TN) contents in paddy soil leading to low levels of effective cumulated soil temperature and thickness of plow layer. As a result, the N mineralization in paddy field during paddy rice growing period was decreased. In addition to the low levels of chemical fertilizer N input and residual mineral N input, the lower level of N mineralization in paddy fields and low N recovery efficiency decreased the amount of N accumulated in aboveground biomass of paddy rice at maturity, resulting in limited rice yields. The amount of mineralized N only correlated with TN (partial correlation analysis). Therefore, in paddy fields with very low N input, the N mineralization in paddy soil during the paddy rice-growing period was the major limiting factor affecting the total yield increases. In addition, a decline in soil fertility can be determined using TN as an indicator. To improve paddy rice yield and avoid soil deterioration, the development and adoption of rational soil management programs are needed. These include input of plant residues, conscientious soil tillage for the maintenance of soil temperature and thickness of the plow layer, and the split application of fertilizer for the improvement of N recovery efficiency.     

Modelling Xanthomonas campestris batch fermentations in a bubble column

Rate and yield expressions relating to biomass and xanthan formation and to nitrogen, glucose, and oxygen consumption were established for Xanthomonas campestris batch fermentations in a bubble column. Microbial growth was described by the logistic rate equation, characterized by a maximum specific growth rate mu(M) = 0.5 h(-1) and a maximum attainable cell concentration provided by nitrogenous compounds. With regard to carbon metabolism, the decrease with time in experimental yields and in the experimental specific rates of xanthan production and glucose assimilation demonstrated the inadequacy of the Luedeking-Piret model. These decreases were connected to the simultaneous drop in dissolved-oxygen tension observed during xanthan synthesis. The knowledge of metabolic pathways and energetic balance were used to establish the relationships between substrate utilization, ATP generation, and xanthan production. The model was structured by assuming the oxygen limitation of both the respiration rate and the efficiency of the oxidative phosphorylation mechanism (P/O ratio). Consequently, the specific rates and yield expressions became dependent on the dissolved-oxygen tension, i.e., of the volumetric oxygen transfer in the fermentor.     

Strain-Dependent Differences in the Efficiency of Transgenic Mouse Production

Transgenic mouse production via pronuclear microinjection is a complex process consisting of a number of sequential steps. Many different factors contribute to the effectiveness of each step and thus influence the overall efficiency of transgenic mouse production. The response of egg donor females to superovulation, the fertilization rate, egg survival after injection, ability of manipulated embryos to implant and develop to term, and concentration and purity of the injected DNA all contribute to transgenic production efficiency. We evaluated and compared the efficiency of transgenic mouse production using four different egg donor mouse strains: B6D2/F1 hybrids, Swiss Webster (SW) outbred, and inbred FVB/N and C57BL/6. The data included experiments involving 350 DNA transgene constructs performed by a high capacity core transgenic mouse facility. Significant influences of particular genetic backgrounds on the efficiency of different steps of the production process were found. Except for egg production, FVB/N mice consistently produced the highest efficiency of transgenic mouse production at each step of the process. B6D2/F2 hybrid eggs are also quite efficient, but lyze more frequently than FVB/N eggs after DNA microinjection. SW eggs on the other hand block at the 1-cell stage more often than eggs from the other strains. Finally, using C57BL/6 eggs the main limiting factor is that the fetuses derived from injected eggs do not develop to term as often as the other strains. Based on our studies, the procedure for transgenic mouse production can be modified for each egg donor strain in order to overcome any deficiencies, and thus to increase the overall efficiency of transgenic mouse production.     

Genetic and physical analyses of a cluster of genes essential for xanthan gum biosynthesis in Xanthomonas campestris.

Xanthomonas campestris produces copious amounts of a complex exopolysaccharide, xanthan gum. Nonmucoid mutants, defective in synthesis of xanthan polysaccharide, were isolated after nitrosoguanidine mutagenesis. To isolate genes essential for xanthan polysaccharide synthesis (xps), a genomic library of X. campestris DNA, partially digested with SalI and ligated into the broad-host-range cloning vector pRK293, was constructed in Escherichia coli. The pooled clone bank was conjugated en masse from E. coli into three nonmucoid mutants by using pRK2013, which provides plasmid transfer functions. Kanamycin-resistant exconjugants were then screened for the ability to form mucoid colonies. Analysis of plasmids from several mucoid exconjugants indicated that overlapping segments of DNA had been cloned. These plasmids were tested for complementation of eight additional nonmucoid mutants. A 22-kilobase (kb) region of DNA was defined physically by restriction enzyme analysis and genetically by ability to restore mucoid phenotype to 10 of the 11 nonmucoid mutants tested. This region was further defined by subcloning and by transposon mutagenesis with mini-Mu(Tetr), with subsequent analysis of genetic complementation of nonmucoid mutants. A region of 13.5 kb of DNA was determined to contain at least five complementation groups. The effect of plasmids containing cloned xps genes on xanthan gum synthesis was evaluated. One plasmid, pCHC3, containing a 12.4-kb insert and at least four linked xanthan biosynthetic genes, increased the production of xanthan gum by 10% and increased the extent of pyruvylation of the xanthan side chains by about 45%. This indicates that a gene affecting pyruvylation of xanthan gum is linked to this cluster of xps genes.     

Harmful algal blooms mitigation using clay/soil/sand modified with xanthan and calcium hydroxide

A method was studied for marine harmful algal blooms (HABs) mitigation using clay, soil, or sand modified with xanthan and calcium hydroxide. Results showed that xanthan could trap and wrap Amphidinium carterae cells via bridging and netting interactions due to its superior salt compatibility in seawater. The maximum cell removal efficiency was 55% when xanthan was used alone. The removal effect of xanthan was enhanced by the addition of appropriate calcium hydroxide that decreased the repulsive interaction between anionic xanthan and negatively charged algal cells. Three kinds of minerals (clays, soils, and sands) were ineffective in removing algal cells before treatment. When xanthan and calcium hydroxide were used together as modifiers, the removal efficiency increased to 83–89% within 30?min using 300?mg?L^?1 clays, soils, or sands modified with 20?mg?L^?1 xanthan and 100?mg?L^?1 calcium hydroxide. After several hours, 95–98% cell removal was achieved and there was no significant difference in the removal efficiencies among clays, soils, and sands after being modified with xanthan and calcium hydroxide. Thus, the method would provide an alternative modification approach to suppress and mitigate HABs using local soils/sands and polymers in marine systems.     

Kinetics of xanthan production when NH3-N limits biomass synthesis and glucose limits polysaccharide synthesis

The bacterium Xanthomonas campestris, which synthesizes the commercially important polysaccharide xanthan, was grown aseptically in 1.2 L fermenters using semicontinuous cell culture technique (d' = 0.0035 h-1). The effects of carbon-substrate concentration on xanthan production were investigated at three initial glucose concentrations (Go = 15, 20, 25 g/L). Cell biomass synthesis was nitrogen-limited by use of a chemically defined medium that contained NH3-N as the sole nitrogen source at a concentration where it was exhausted before glucose. A linear relationship between biomass synthesis and NH3-N depletion was observed. This relationship remained valid only until NH3-N exhaustion, after which biomass concentration slowly rose another 20 percent before declining. Another linear relationship was found between xanthan synthesis and glucose uptake. This relationship was unaffected by the disappearance of NH3-N and held through glucose exhaustion. The quasi-stoichiometric yield coefficients obtained for each linear relationship were not affected by G0-. Biomass synthesis kinetics showed no variation with G0 before NH3-N exhaustion; afterwards, cell biomass decline was delayed by increasing G0. Xanthan synthesis kinetics displayed no detectable response to depletion of NH3-N and plateauing of biomass concentration; however, there was a marked slow down in the net rate of xanthan synthesis and a drop in xanthan yield after cell biomass decline became noticeable.     

京郊典型集约化"农田-畜牧"生产系统氮素流动特征

城郊畜牧业的集约化发展在满足人们日益增长的肉蛋奶等动物性产品需求的同时,也带来了巨大的环境压力。本文利用养分流动和模型分析的方法,分析北京市郊区某村三种不同类型"农田-畜牧"生产系统(大型集约化种猪场、种养结合小规模生态养殖园和集约化单一种植区)的氮素流动特征。结果表明:饲料是集约化种猪场和生态养殖园氮素输入的主要来源,饲料投入氮量分别为12469.0和9268.5 kg.hm-.2a-1);集约化种猪场农牧体系间生产脱节,致使农田氮素输入主要依赖于化肥投入,化肥氮输入量(435.0 kg.hm-.2a-1)占农田氮素输入量的82.7%。相反,生态养殖园农牧体系结合紧密,猪粪尿氮还田比例达28.6%,这使得园区化肥氮输入量仅为135.0 kg.hm-.2a-1,因此畜禽粪尿的合理循环利用可作为减少化肥投入的有效途径;集约化种猪场、生态养殖园和单一种植区农牧生产系统氮素利用效率分别为18.8%、20.6%和17.3%,均处于较低水平;集约化种猪场猪粪尿在猪舍和储藏处理过程的氮素损失率为15.8%和25.4%,分别低于小规模生态养殖园相应损失率约8.7和4.8个百分点;生态养殖园粪肥氮还田量高,导致农田氮素盈余量高达1962.8 kg.hm-.2a-1,未能实现生态型养殖的理想效果,优化氮素管理、确定合理的消纳畜禽粪尿的农田面积和调整畜禽养殖密度是解决该问题的关键。     

Exposure Assessment in Millimeter-Wave Reverberation Chamber Using Murine Phantoms

This study deals with the design and calibration of the first mode-stirred reverberation chamber (RC) in the 60-GHz-band adapted for in vivo bioelectromagnetic studies. In addition to the interface for electromagnetic and thermal dosimetry, the interfaces for lighting and ventilation were integrated into the RC walls while preserving acceptable shielding. The RC with mechanical and electronic steering capabilities is characterized in the 55–65 GHz range. To this end, murine skin-equivalent phantoms of realistic shape were designed and fabricated. Their complex permittivity is within ±12% of the target value of murine skin (6.19–j5.81 at 60 GHz). The quality factor of the RC loaded with an animal cage, bedding litter, and five murine phantoms was found to be 1.2 × 10⁴. The losses inside the RC were analyzed, and it was demonstrated that the main sources of the power dissipation were the phantoms and mice cage. The input power required to reach the average incident power density of 1 and 5 mW/cm² was found to be 0.23 and 1.14 W, respectively. Surface heating of the mice models was measured in the infrared (IR) range using a specifically designed interface, transparent at IR and opaque at millimeter waves (mmW). Experimental results were compared with an analytical solution of the heat transfer equation and to full-wave computations. Analytical and numerical results were in very good agreement with measurements (the relative deviation after 90 min of exposure was within 4.2%). Finally, a parametric study was performed to assess the impact of the thermophysical parameters on the resulting heating. Bioelectromagnetics. 2020;41:121–135. © 2020 Bioelectromagnetics Society.     

Input-output budget of nitrogen and the effect of experimentally changed deposition in the forest ecosystems in central Japan

To evaluate the current nitrogen (N) status in Japanese forests, field measurements of rainfall, throughfall, litter layer percolation, and soil solution percolation were conducted in a red pine stand (Kannondai) and a deciduous stand (Yasato) located in central Japan. N input via throughfall was 31 and 14 kg ha(-1) year(-1) and output below rooting zone was 9.6 and 5.5 kg ha1 year(-1) in Kannondai and in Yasato, respectively. Two thirds of input N were retained in plant-soil systems. Manipulation of N input was carried out. Ionic constituents were removed from throughfall with ion exchange resin at removal sites and ammonium nitrate containing twice the N of the throughfall was applied at N addition sites periodically. SO4(2-) output below 20-cm soil layer changed depending on the input, while NO3- output was regulated mainly by the internal cycle and effect of manipulation was undetected. These Japanese stands were generally considered to have a larger capacity to assimilate N than NITREX sites in Europe. However, N output fluxes had large spatial variability and some sites in Kannondai showed high N leaching below rooting zone almost balanced with the input via throughfall.