He-Ne激光诱变高活力苯酚降解菌热带假丝酵母CT43的研究

运用He—Ne激光辐照热带假丝酵母(Candida tropicalis)对激光诱变参数进行优化,考察了突变株的遗传稳定性和降酚特性。结果表明：经过激光辐照处理的热带假丝酵母CT43降解2000mg／L的苯酚仅需要54．5h,降解效率显著提高。表明He-Ne激光诱变育种技术可以在环境微生物领域中应用。     

热带假丝酵母及其重组菌产十三碳二元酸的补糖发酵实验

利用热带假丝酵母原始菌F10-1及已构建的肉毒碱乙酰转移酶基因单拷贝缺失重组菌CZ-15,在5 L发酵罐中进行了补糖发酵批式培养,生产十三碳二元酸(DCA13). 结果表明,肉毒碱乙酰转移酶基因单拷贝缺失后,CZ-15菌株的最佳补糖时间和补糖浓度都与原始菌F10-1不同. 采用补糖工艺后,F10-1与CZ-15的十三碳二元酸的产量分别提高了25.0%和23.4%,达到99.5和108.1 g/L,烷烃转化率分别提高了35.5%和28.6%.     

Studies of L-phenylalanine production by immobilised aminoacylase in stabilized calcium alginate beads

Aminoacylase I (EC 3.5. 1.14) was immobilised by entrapment in uncoated calcium alginate beads and calcium alginate beads coated with chitosan, polyethyleneimine and polyethyleneimine-glutaraldehyde for the production of L-phenylalanine by the hydrolysis of a racemic mixture of N-acetyl-DL-phenylalanine. The operational stability, thermal stability, effects of pH and temperature and kinetic constants, K_m and V_max values of free and immobilised enzymes were studied. Scanning electron micrographs revealed differences in the structures of the surfaces of coated and uncoated beads. Chitosan-coated calcium alginate beads was found to be the best among the immobilised systems studied. The activity and the optimum temperature of immobilised aminoacylase were higher than those of the free enzyme. In addition, the beads showed stable activity under operational conditions. The immobilised aminoacylase lost about 20% of its initial activity after ten cycles of reuse. Polyethyleneimine and polyethyleneimine-glutaraldehyde treatments were also found to enhance the operational stability of the enzyme but its activity was greatly reduced.     

CAT基因敲除对热带假丝酵母产DCA13代谢网络的影响

肉毒碱乙酰转移酶在脂肪酸β-氧化过程中起着重要作用.利用同源重组技术,构建了热带假丝酵母CAT基因单拷贝缺失菌CZ-15和双拷贝缺失菌CKC-11后,发现CZ-15的β-氧化过程受到抑制,与原始菌F10-1相比,十三碳二元酸的产量和烷烃转化率都得到了明显提高,菌体生长情况与F10-1基本一致;CKC-11则无法代谢烷烃,菌体生长也受到了较大的影响.采用代谢动力学模型对发酵数据进行了计算与分析,实验结果与模型相符.根据实验和模型计算结果完善了热带假丝酵母代谢葡萄糖和十三碳二元酸的途径.     

Modeling for local dynamic behaviors of phenol biodegradation in bubble columns

A coupled three-dimensional (3-D) model, combining hydrodynamics with biochemical reactions, was developed to simulate the local transient flow patterns and the dynamic behaviors of cell growth and phenol biodegradation by yeast Candida tropicalis in the bubble-column bioreactor, using the computational fluid dynamic (CFD) method. In order to validate this proposed model effectively, the validation of the local hydrodynamic characteristics of the gas-mineral salt solution (gas-liquid) two-phase system, with the phenol concentration of 1200 mg/L, and with the absence of cells, was performed in a square-sectioned bubble column bioreactor using the LDA system and conductivity probe. Furthermore, the validation of phenol biodegradation behaviors by yeast Candida tropicalis at different initial concentrations of phenol and cell was also carried out in the above bubble-column bioreactor. The results indicated that the model simulations had a satisfying agreement with the experimental data. Finally, the local instantaneous flow and phenol biodegradation features including gas holdup, gas velocity, liquid velocity, cell concentration and phenol concentration inside the bioreactor were successfully predicted in different-scale bubble columns by the proposed model. © 2006 American Institute of Chemical Engineers AIChE J, 2006     

Xylitol production by Candida tropicalis from corn cob hemicellulose hydrolysate in a two‐stage fed‐batch fermentation process

BACKGROUND: Xylitol, a sugar alcohol widely used in food and pharmaceutical industries, can be produced through biological reduction of xylose present in hemicellulose hydrolysates by Candida tropicalis. However, the aeration rate and by‐products originating from hemicellulose hydrolysis strongly inhibit the production of xylitol in a fermentation process. A two‐stage fed‐batch fermentation system was developed to reduce these inhibitory effects and to improve xylitol production from corn cob hemicellulose hydrolysates by C. tropicalis. RESULTS: Results of batch fermentations indicated that high xylitol production could be obtained from C. tropicalis at an initial xylose concentration of 80 g L^?1 in corn cob hydrolysate medium at an aeration rate of 0.4 vvm at the micro‐aeration stage. In the two‐stage fed‐batch fermentation process, 96.5 g L^?1 xylitol was obtained after 120 h, giving a yield of 0.83 g g^?1 and a productivity of 1.01 g L^?1 h^?1, which were 12.16% and 65.57% higher than those in a batch fermentation. CONCLUSION: High xylitol production can be achieved in a two‐stage fed‐batch fermentation process, in which the negative effects of aeration rate and inhibitory compounds on xylitol formation can be considerably reduced. Copyright © 2011 Society of Chemical Industry     

Studies on biodegradation of phenol using response surface methodology

Biodegradation of phenol was studied using Pseudomonas pictorum (NCIM 2077) immobilized on alginate and activated carbon – alginate beads. Control experiments were also performed using free cells and non‐inoculated activated carbon – alginate beads. The entrapped alginate and activated carbon – alginate beads suffer from a concentration gradient for oxygen in the interior of the beads and hence free cells showed better degradation at lower concentrations of phenol. The results on entrapped alginate beads were modeled using response surface methodology which determines the dependency of the maximum percentage of phenol degraded as a function of the independent variables, namely initial phenol concentration, initial pH, temperature, and diameter of the immobilized beads. The predicted values are in close agreement with the experimental values with the coefficient of correlation equal to 0.9999 and 0.9993 for both P pictorum – alginate beads and activated carbon – P pictorum – alginate beads respectively. © 2002 Society of Chemical Industry     

一株高效苯酚降解菌的分离鉴定及动力学研究

从活性污泥中分离出了热带假丝酵母。该菌株可利用苯酚做碳源和能源,降酚能力达到2 000 mg/L。通过对该菌株在不同温度、pH值以及不同苯酚组成下的生长和降解情况的研究,确定了该菌株最适宜的降解工艺条件;获得了细胞生长动力学模型。     

Anaerobic biodegradation of phenol in sulfide‐rich media

In the refinery industry, the washing processes of middle‐distillates using caustic solutions generate phenol‐ and sulfide‐containing waste streams. The spent caustic liquors generated contain phenols at concentrations higher than 60 g dm^?3(638.3 mmol dm^?3). For sulfur compounds, the average sulfide concentration was 48 g dm^?3(1500 mmol dm^?3) in these streams. The goal of this study was to evaluate the specific impact of phenol and sulfide concentrations towards the phenol‐biodegradation activity of a phenol‐acclimated anaerobic granular sludge. An inhibition model was used to calculate the phenol and sulfide inhibitory concentrations that completely stopped the phenol‐biodegradation activity (IC100). A maximum phenol‐biodegradation activity of 83 µmol g^?1 VSS h^?1 was assessed and the IC100 values were 21.8 mmol dm^?3 and 13.4 mmol dm^?3 for phenol and sulfide respectively. The limitation of the phenol biodegradation flow by phenol inhibition seemed to be related to the more important sensitivity of phenol‐degrading bacteria. The up‐flow anaerobic sludge bed reactor operating in a non‐phenol‐dependent inhibition condition did not present any sensitivity to sulfide concentrations below 9.6 mmol dm^?3. At this residual concentration, the pH and bisulfide ions' concentration might be responsible for the general collapsing of the reactor activity. Copyright © 2004 Society of Chemical Industry     

高压静电法制备单分散性的海藻酸钙微胶珠

In this paper, the effect of operation parameters on the monodispersity of calcium-alginate gel beads prepared in high-voltage electrostatic field were analyzed. The parameters included the applied potential （U）, the distribution of electrostatic field, the frequency and the flow rate of solution through the capillary. Furthermore, the process is analyzed with the theory of jet break-up in electrostatic field. The result showed that the modified symmetrical electrostatic field was more advantageous in preparing the monodispersed gel beads than the traditional method. Meanwhile, gel beads with good monodispersity （CV〈20%） and stability without aggregation were produced under a range of applied potential （U）, frequency and flow rate of solution through the capillary.     

载药蒙脱石/海藻酸钠凝胶球的制备和释放性能

以离子交换方式将硫酸氢氯吡格雷插入蒙脱石层间,再用海藻酸钠包衣得到凝胶球。利用XRD、FTIR进行表征以及体外释放实验检测缓释效果。结果表明,硫酸氢氯吡格雷成功地插入蒙脱石层间,蒙脱石的层间距由1.25 nm增大到1.48 nm;体外释放实验显示在1 h内载药凝胶球的累积释放量为12.9%,表明载药蒙脱石/海藻酸钠凝胶球能明显的减少药物突释,因此可将它作为硫酸氢氯吡格雷的缓释载体。     

海藻酸凝胶性质对蛋白质扩散的影响

通过观察冷冻干燥海藻酸凝胶的断面扫描电镜(SEM)照片与卵清蛋白从海藻酸凝胶中的释放试验,分析了海藻酸凝胶性质对蛋白质在凝胶中扩散的影响。凝胶的SEM照片可见,海藻酸钙的冷冻干燥颗粒内为较大的圆孔,海藻酸锌凝胶内为较小的长孔,表明海藻酸锌高分子链在凝胶颗粒中的体积分率相对较大同时刚性较强;卵清蛋白从海藻酸凝胶颗粒中释放的试验结果表明,由于上述海藻酸锌凝胶的特性,导致海藻酸锌对卵清蛋白扩散阻滞作用相对较强;根据试验数据计算得卵清蛋白在海藻酸钙、海藻酸锌凝胶颗粒中的扩散系数分别为1.19×10-7、0.07×10-7cm2/s,利用阻滞模型计算得海藻酸锌高分子链在凝胶颗粒中体积分率约为海藻酸钙高分子链在凝胶颗粒中体积分率的1.7倍。     

多通道微胶囊制备系统规模化制备海藻酸钙微胶珠

利用自制多通道强制电场微胶囊制备系统制备海藻酸钙微胶珠,分析了微胶囊制备系统中电极电场分布规律,考察了进料气压、脉冲电压、脉冲频率、脉冲宽度、辅助电压等系统调节参数对微胶珠制备过程的影响,发现制备过程中微胶珠粒径大小与产率主要由进料气压决定,而微胶珠的分散性则是由多个因素共同作用的结果。通过优化条件制备的微胶珠分散系数较小（CV<20%）,球形度良好,表面光洁,并且产率是传统静电法的20~40倍,使海藻酸钙微胶珠大规模制备成为可能。     

Characterization of the structure and diffusion behavior of calcium alginate gel beads

A simple and novel method using gel shrinkage to indirectly characterize the structure of calcium alginate gel (CAG) beads during the calcium alginate gelation process was presented in this study. The effect of preparation process parameters (gelling cations, bead diameter, and alginate M _w and concentration) on the structure of the CAG bead formation process was thoroughly investigated. It was found that (a) the concentration of the Na⁺ and Ca²⁺ ion in gel bath was found to be the determining factor in the gel structure formation process by regulating the dissociation of alginate and the complexation of the calcium; (b) Na⁺ acts as a competitor with calcium and a screen in the electrostatic repulsion; (c) the effect of beads size below 700 μm on the structure of CAG beads can be neglected; and (d) the sodium alginate concentration has no significant effect on the gel formation process. Furthermore, the diffusion of bovine serum albumin (BSA) was controlled by the density of CAG bead. Consequently, a faster diffusion rate of BSA within the looser structure of beads can be observed. These results are keys to understanding the behavior and performance of beads in their utilization medium. © 2020 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2020 , 137, 48923.     

Ethanol production by alginate immobilised yeast in a fluidised bed bioreactor

Yeast, immobilised in alginate beads of known standard size and mechanical strength, has been utilised in a novel design of fluidised bed bioreactor which avoids problems of particle flotation and gas logging. Circulating substrate simultaneously entered the top and bottom of the bed. The bioreactor operated reliably for periods of up to 20 days. Increasing alginate concentration in the range 1–5% (w/w) had little effect on the performance of the immobilised yeast in converting ethanol to glucose but reduced the tendency of beads to split. Increasing bead diameter in the range 1–5 mm increased the tendency to split and reduced overall conversion of glucose. A model was developed to describe the consumption of glucose within beads based on Michaelis–Menten kinetics and the diffusion of glucose into beads. Application of the model to experimental results showed maximum reaction velocity to be independent of bead diameter and alginate concentration. The model confirmed that the observed reduction in ethanol yield compared with free yeast cells was caused by the lower substrate concentration towards the centre of the bead as opposed to any change in the metabolic rate of the immobilised cells.     

十三碳二元酸发酵与肉毒碱乙酰基转移酶酶活动力学研究

在利用热带假丝酵母发酵生产十三碳二元酸(DCA13)的过程中,建立了用荧光法测定酵母体内肉毒碱乙酰基转移酶(CAT)酶活的方法,方法简单,重现性高,测量精确。研究了热带假丝酵母中CAT酶活动力学与产酸速率的关系,结果表明:代谢前期细胞以生长为主,CAT酶活较高可提供更多的生长动力,而代谢后期细胞进入产酸期,CAT酶活与产酸速率的变化趋势成反比,即产酸速率高时,CAT的酶活较低。     

Modeling of batch phenol biodegradation in internal loop airlift bioreactor with gas recirculation by Candida tropicalis

A mathematical model for simulating the dynamic behaviors of batch phenol biodegradation processes in internal loop airlift bioreactor (ILAB) with gas recirculation using free cells of the yeast Candida tropicalis was established by coupling the fluid dynamic model and the mass balance model. Based on the coupling arithmetic, the program for evaluating batch phenol biodegradation processes was achieved. The predicted results of linear liquid velocities, gas holdups, linear gas velocities, cell and phenol concentration profiles in the riser and the downcomer of the ILAB with gas recirculation agreed very well with the corresponding experimental data, and the applicability and reliability of this proposed model were validated.     

假丝酵母菌对高浓度油脂废水的降解性能

对本实验室分离筛选的优良油脂分解菌株——假丝酵母WZFF．G-284的各种微生物学和生化分析试验结果表明．该菌株对于高含油废水具有很强的适应能力,在以高浓度食用色拉油作为唯一碳源、添加适当无机盐的实际工业废水开放处理体系中,该菌株生长繁殖力强,能迅速形成优势菌群,分解油脂。该菌株除菌体细胞具有高强乳化作用能力外。还可合成乳化性能强的生物乳化剂达21．5g／L,同时生产68．3U／mL的高稳定性胞外脂肪酶。该微生物净化处理技术可以在高负荷下持续稳定运转,对油脂、COD和BOD的降解速度快,去除率均在95％以上,分解效果显著。