Continuous Asymmetric Reduction Of 4-Oxoisophorone By Thermophilic Bacteria Using A Hollow Fiber Reactor

Continuous asymmetric reduction of 4-oxoisophorone by the thermophilic bacterium Thermomonospora curvata JTS321 was examined using three reactor systems: packed bed, fluidized bed and hollow fiber. T. curvata was immobilized in polyacrylamide-hydrazide gels when used in the packed and fluidized bed reactors. Of the three reactor systems, the highest productivity (964 mg.1^-1.h^-1) was observed in the fluidized bed reactor. However, many cells grew outside of the gel matrix, causing product contamination. The productivity of the hollow fiber reactor was 504 mg.1^-1.h^-1; the problem of cell contamination of the product was avoided, as the molecular cut-off of the hollow fibers (400 000) was of an appropriate size to prevent cell leakage to the product stream. We therefore consider that the hollow fiber reactor is most suitable for continuous microbial conversions.     

Sorbitol and Gluconate Production in a Hollow Fibre Membrane Reactor by Immobilized Zymomonas Mobilis

This study describes the results of a hollow fibre membrane reactor with immobilized treated cells of Zymomonas mobilis which produced sorbitol and gluconic acid continuously from fructose and glucose respectively. A productivity of 10-20 g sorbitol · L^-1 · h^-1 and 10-20 gluconate · L^-1 · h^-1 (based on total bioreactor volume) from a feed of 100 g · L^-1 each of glucose and fructose was possible at high dilution rates. Kinetic parameters describing the reaction rate of treated cells in batch reactors were used to analyse the performance of the hollow fibre membrane reactor employing significant convective mass transfer. No significant mass transfer limitation was apparent.     

虎斑乌贼的胚胎耗氧率

为了探究虎斑乌贼胚胎不同发育时期的耗氧率变化和几种生态因子对胚胎发育过程耗氧率的影响,试验采用封闭静水装置,对不同发育时期(12期)的耗氧率进行测定,并研究不同盐度(21、24、27、30、33)、温度(18、21、24、27、30 ℃)和pH(7.0、7.5、8.0、8.5、9.0)对胚胎4个主要发育时期(受精卵期、原肠胚期、器官形成期和内骨骼形成期)耗氧率的影响.结果表明: 胚胎各个发育时期耗氧率不同,随着发育的进程而增大,受精卵期为0.082 mg·(100 eggs)^-1·h^-1,而到原肠胚期的耗氧率显著升高,为0.279 mg·(100 eggs)^-1·h^-1,到孵化期时,耗氧率达到1.367 mg·(100 eggs)^-1·h^-1;盐度对器官形成期和内骨骼形成期的耗氧率均有显著影响(P<0.05),对受精卵期和原肠胚期影响不显著(P>0.05),当盐度为30时,4个发育时期耗氧率均达到最大值,分别为0.082、0.200、0.768和1.301 mg·(100 eggs)^-1·h^-1;温度对原肠胚期、器官形成期和内骨骼形成期的耗氧率有显著影响(P<0.05),对受精卵期无显著性影响(P>0.05),在27 ℃时,胚胎4个发育时期均达到最大值,分别为0.082、0.286、0.806和1.338 mg·(100 eggs)^-1·h^-1;而pH对4个发育时期的耗氧率均无显著性影响(P>0.05),受精卵期在pH 8.0时达到最大值,为0.116 mg·(100 eggs)^-1·h^-1,原肠胚期、器官形成期、内骨骼形成期在pH 8.5时达到最大值,分别为0.281 、0.799和1.130 mg·(100 eggs)^-1·h^-1.     

虎斑乌贼的胚胎耗氧率

为了探究虎斑乌贼胚胎不同发育时期的耗氧率变化和几种生态因子对胚胎发育过程耗氧率的影响,试验采用封闭静水装置,对不同发育时期(12期)的耗氧率进行测定,并研究不同盐度(21、24、27、30、33)、温度(18、21、24、27、30 ℃)和pH(7.0、7.5、8.0、8.5、9.0)对胚胎4个主要发育时期(受精卵期、原肠胚期、器官形成期和内骨骼形成期)耗氧率的影响.结果表明: 胚胎各个发育时期耗氧率不同,随着发育的进程而增大,受精卵期为0.082 mg·(100 eggs)^-1·h^-1,而到原肠胚期的耗氧率显著升高,为0.279 mg·(100 eggs)^-1·h^-1,到孵化期时,耗氧率达到1.367 mg·(100 eggs)^-1·h^-1;盐度对器官形成期和内骨骼形成期的耗氧率均有显著影响(P<0.05),对受精卵期和原肠胚期影响不显著(P>0.05),当盐度为30时,4个发育时期耗氧率均达到最大值,分别为0.082、0.200、0.768和1.301 mg·(100 eggs)^-1·h^-1;温度对原肠胚期、器官形成期和内骨骼形成期的耗氧率有显著影响(P<0.05),对受精卵期无显著性影响(P>0.05),在27 ℃时,胚胎4个发育时期均达到最大值,分别为0.082、0.286、0.806和1.338 mg·(100 eggs)^-1·h^-1;而pH对4个发育时期的耗氧率均无显著性影响(P>0.05),受精卵期在pH 8.0时达到最大值,为0.116 mg·(100 eggs)^-1·h^-1,原肠胚期、器官形成期、内骨骼形成期在pH 8.5时达到最大值,分别为0.281 、0.799和1.130 mg·(100 eggs)^-1·h^-1.     

Continuous ethanol production by immobilized yeast in a fluidized reactor

Summary In order to minimize the adverse effect of CO₂ gas in a packed bed immobilized yeast reactor, a fluidized bed reactor was used for the continuous production of ethanol from glucose. Immobilized yeast was prepared by entrapping whole cells of Saccharomyces cerevisiae within a Caalginate matrix. It was found that the efficiency of the ethanol production in a fluidized bed reactor was 100% better than that for a packed bed reactor system. The alcohol productivity obtained was 21 g/l/hr in a fluidized bed reactor at 94% of conversion level.     

Enzymatic synthesis of new aromatic and aliphatic esters of flavonoids using Candida antarctica lipase as biocatalyst

Enzymatic acylation of rutin and esculin with aromatic, aliphatic and aryl-aliphatic acids using Candida antarctica lipase in tert amyl alcohol as solvent was investigated under low water content. Whatever the acyl donor used, the conversion yields and initial rates for esculin were higher than for rutin. For a given flavonoid, the performance of these reactions depended on the acyl donor structures. For aliphatic acids, conversion yields and initial rates of both flavonoids were respectively in the ranges of 68-90% and of 9.5×10^-2-72×10^-2 mmol l^-1 h^-1. For aromatic acids, the reaction occurred only with the aryl subgroup (cinnamic, hydrocinnamic, 3,4-dihydroxyhydrocinnamic and 4-hydroxyphenyl acetic acids) and was drastically influenced by the presence of side chain and substitution patterns of the aromatic ring. Except for hydrocinnamic acid (75%, 23.4×10^-2 mmol l^-1 h^-1), with these acids the conversion yields and initial rates were lower and in the range of 10-45% and of 0.7×10^-2 to 12.1×10^-2 mmol l^-1 h^-1. Unsaturation of the side chain of the hydrocinnamic acid decreased the esculin conversion rate from 75 to 13% and initial rate from 23.4 to 1.76×10^-2 mmol l^-1 h^-1. The presence of hydroxyl or nitro-groups on the aromatic ring of the aryl aliphatic acid also reduced conversion yields and initial rates. Even without a spacer, the non-phenolic ring acid (quinic acid) was reactive and lead to conversion yields of about 20 and 23% respectively for rutin and esculin.     

Application of fluidized carrier to bacterial sulphate-reduction in industrial wastewaters purification

Summary Comparative laboratory investigation of two types bioreactors with iron as a carrier of biofilm was made. One was a packed bed reactor and the another was fluidized bed. The results showed that maximum productivity of carrier in fluidized bed bioreactor {7,97 g/m²·d} is two times higher than productivity of carrier in packed bed one {3,45 g/m²·d}.     

Immobilized Cyclodextrin Glycosyl Transferase for the Continuous Production of Cyclodextrins

Cyclodextrin glycosyl transferase (E.C: 2.4.1.19) from Bacillus, macerans and from a Bacillus sp. isolate was immobilized by two methods, viz. to epoxy-activated Sepharose and to alkylamine silica treated with glutaraldehyde. Because of the ready availability, low cost ($0.01/g), good surface area (30 M²/g) and ease of operation of a continuous cylindrical reactor, the high silica fabric was chosen. The immobilized enzyme had a pH optimum shifted to the alkaline side (from 6.5 to 7.5) and had a reduced temperature optimum (from 60°C to 50-55°C). Reuse efficiency showed 65% reduction in the overall activity of the immobilized enzyme after 10 cycles of 48 h each. Continuous operation at 55°C of a cylindrical reactor of 141 ml capacity, using the immobilized enzyme (80 g of high - silica fabric containing 114 mg of purified enzyme) gave a maximum productivity of 10.2 g of cyclodextrins L^-1 h,^-1, at a dilution rate of 0.32 h^-1 and a substrate concentration of 20 g L^-1. The half life of the biocatalyst was found to be 22 days, which could be further improved by using a lower operating temperature. Over the useful life time of the immobilized biocatalyst (22 days), the total Cyclodextrin produced was of the order of 88 Kg.     

Continuous adsorption and recovery of Cr(VI) in different types of reactors

This study reports the results of experiments on continuous adsorption and desorption of Cr(VI) ions by a chemically modified and polysulfone-immobilized biomass of the fungus Rhizopus nigricans. A fixed quantity of polymer-entrapped biomass beads corresponding to 2 g of dry biomass powder was employed in packed bed, fluidized bed, and stirred tank reactor for monitoring the continuous removal and recovery of Cr(VI) ions from aqueous solution and synthetic chrome plating effluent. Parameters such as flow rate (5, 10 and 15 mL/min), inlet concentration of Cr(VI) ions (50, 100, 150 and 250 mg/L) and the depth of biosorbent packing (22.8, 11.2 and 4.9 cm) were evaluated for the packed bed reactor. The breakthrough time and the adsorption rates in the packed bed column were found to decrease with increasing flow rate and higher Cr inlet concentrations and to increase with higher depths of sorbent packing. To have a comparative analysis of Cr adsorption efficiency in different types of reactors, the fluidized bed reactor and stirred tank reactor were operated using the same quantities of biosorbent material. For the fluidized bed reactor, Cr(VI) solution of 100 mg/L was pumped at 5 mL/min and fluidized by compressed air at a flow rate of 0.5 kg/cm.(2) The stirred tank reactor had a working volume of 200 mL capacity and the inlet/outlet flow rate was 5 mL/min. The maximum removal efficiency (mg Cr/g biomass) was obtained for the stirred tank reactor (159.26), followed by the fluidized reactor (153.04) and packed bed reactor (123.33). In comparison to the adsorption rate from pure chromate solution, approximately 16% reduction was monitored for synthetic chrome plating effluent in the packed bed. Continuous desorption of bound Cr ions from the reactors was effective with 0.01 N Na(2)CO(3) and nearly 80-94% recoveries have been obtained for all the reactors.     

Kinetics of Pyruvate Decarboxylase Deactivation by Benzaldehyde

Based on experimental data, a kinetic model for the deactivation of partially purified pyruvate decarboxylase (PDC) by benzaldehyde (0-200 mM) in MOPS buffer (2.5 M) has been developed. An initial lag period prior to deactivation was found to occur. With first order dependencies of PDC deactivation on exposure time and on benzaldehyde concentration, a reaction time deactivation constant of 2.64×10^-3 h^-1 and a benzaldehyde deactivation coefficient of 1.98×10^-4 mM^-1 h^-1 were determined for benzaldehyde concentrations up to 200 mM. The PDC deactivation kinetic equations established in this study are an essential component in an overall model being developed to describe the enzymatic biotransformation of benzaldehyde and pyruvate to produce the pharmaceutical intermediate (R)-phenylacetylcarbinol (R-PAC).     

Continuous biocatalytic synthesis of epoxypropane using a biofilm reactor

Mixed culture methanotrophic attached biofilms immobilized on diatomite particles in a three-phase fluidized bed reaction system were developed. Methane monooxygenase (MMO) activity on diatomite particles increased as soon as the lag phase ended. More than 90% of the MMO activity in the fluidized bed was attached. A biofilm concentration of 3.3c3.7mg dry weight cell (dwc) per g dry solid (DS) was observed. Batch experiments were performed to explore the possibility of producing epoxypropane by a propene–methane co-oxidation process. The effect of methane on the epoxidation of propene and the effect of propene on the growth of methanotroph was also studied. In continuous experiments, optimum mixed gas containing 35 methane, 20 propene and 45% oxygen were continuously circulated through the fluidized bed reactor to deliver substrates and extract product. Initial epoxypropane productivity was 110–150 μmol/day. The bioreactor operated continuously for 53 days without obvious loss of epoxypropane productivity.     

Effect of light-path length in outdoor flat plate reactors on output rate of cell mass and of EPA in Nannochloropsis sp.

The effect of light-path length (i.e. reactor width or thickness) of flat plate glass reactors on outdoor production of eicosapentaenoic acid (EPA) and cell mass of Nannochloropsis sp. was tested, using a range of light-paths from 1.3 to 17.0 cm. Volumetric productivity of cell mass and optimal, as well as maximal cell density which represents the highest sustainable cell density under the experimental conditions, decreased with increase in light-path. Daily areal output rate (g dry weight m⁻² day⁻¹) increased with increased light-path, in contrast with results obtained in similar reactors with Spirulina cultures, in which areal output rates increased when the light-path was reduced. Maximal areal productivity of Nannochloropsis sp. (12.8 and 22.4 g ash-free dry weight per day per m² of irradiated reactor surfaces, in winter and summer, respectively), reflecting maximal efficiency in light utilization, was obtained with the long light-paths, i.e. 10.4 and 17.0 cm. Increasing the light-path from 1.3 to 17.0 cm resulted in an increase in areal EPA productivity, from 66.7 to 278.2 mg m⁻² day⁻¹ in winter and from 232.1 to 515.7 mg m⁻² day⁻¹ in summer. This enhancement in areal productivity of EPA stems from increased productivity of cell mass which was associated with the increase in light-path. We concluded that the optimal light-path, which must be defined for each algal species, represents an important parameter which determines optimal culture density (i.e. resulting in the highest output rate of cell mass per irradiated reactor surface), as well as productivity of cell mass and cell products. Under our conditions the optimal light-path for culturing Nannochloropsis in vertical reactors was ca 10 cm.     

Development of a Bioprocess for Murine Dimeric IgA Production

Monoclonal IgA was produced under serum free conditions by a murine hybridoma cell line (ZAC3) in a hollow fibre, a continuous stirred tank and a fluidized bed reactor. Differences in the antibody adsorption to DEAE chromatography matrices, an essential step in downstream processing, were related to the production systems. Chromatography on hydroxyapatite was used to separate monomeric, dimeric and polymeric IgA. This method was successfully applied to IgA produced by all three reactor configurations. The binding of dimeric and polymeric IgA to antigen was tested by ELISA. Using 2-dimensional SDS-PAGE analysis, dimeric IgA from the three sources was shown to be identical with respect to isoelectric points of alpha, kappa and J-chain but showed marked differences in purity. Finally the efficiency of the three bioprocesses was assessed by comparing the product yields after purification. This included the reactor specific production rates, media and time requirements and time consumption for the production of 1 g purified dimeric IgA.     

甲烷氧化菌吸附膜反应器中环氧丙烷的连续生物转化

以流化床作为固定化体系 ,在硅藻土颗粒表面构建了混合培养的甲烷氧化细菌的吸附膜。研究发现延迟期后固定化细胞的甲烷单加氧酶活性明显增加。流化床中 90 %以上的甲烷氧化细菌以吸附形式存在。吸附膜浓度为 3.3～3.7 mgdryweightcell gDS。通过批式反应考察了丙烯 甲烷共氧化过程合成环氧丙烷的可能性。研究了甲烷对丙烯环氧化以及丙烯对甲烷氧化细菌生长的影响。通过最佳配比的混合反应气体 (methane :35 % ;propene :20% ;oxygen :45 % )连续循环通入流化床反应器中抽提产物环氧丙烷 ,克服了产物抑制。该生物反应器最初产生环氧丙烷的日产量为 110～ 150μmol d ,连续操作25d ,未观察到环氧丙烷生产能力的明显减小.     

Effects of Oxygenation on Hydrocarbon Biodegradation in a Hypoxic Environment

In 1984, an underground storage tank leaked approximately 41,000 L of gasoline into the ground water at the Naval Construction Battalion Command in Port Hueneme, CA (USA). Benzene, toluene, ethylbenzene, and xylenes (BTEX) contamination stimulated remedial action. In 1995, a ground water circulation well (GCW) and network of surrounding monitoring wells were installed. After year of operation, dissolved oxygen and nitrate concentrations remained low in all monitoring wells. Benzene utilization (the sum of respiration, uptake, and conversion to polar compounds) ranged from 0.03 to 4.6 µg L^-1 h^-1, and toluene utilization ranged from 0.01 to 5.2 µg L^-1 h^-1. Heterotrophic bacterial productivity (total carbon assimilation) increased dramatically in the GCW, although benzene and toluene utilization decreased markedly relative to surrounding wells. Benzene and toluene uptake accounted for a significant proportion (mean=22%) of the heterotrophic bacterial productivity except within the GCW, indicating other fuel contaminant or indigenous organic carbon and not BTEX compounds served as primary carbon source. The GCW effectively air-stripped BTEX compounds, but failed to stimulate benzene and toluene biodegradation and thus would not be effective for stimulating BTEX bioremediation under current deployment parameters. Air stripping was three orders of magnitude more effective than biodegradation for removing benzene and toluene in the GCW.     

金黄花滇百合植株再生与离体快繁技术体系的建立

以金黄花滇百合(Lilium bakerianum var. aureum)的鳞片为外植体, 探讨不同部位外植体和不同配比的植物生长调节剂对愈伤组织诱导和植株再生的影响。研究结果表明, 鳞片诱导愈伤组织和不定芽的最适培养基为MS+1.0 mg∙L ^-16-BA+0.1 mg∙L ^-1NAA+30 g∙L ^-1蔗糖; 不定芽增殖的最适培养基为MS+0.5 mg∙L ^-16-BA+0.1 mg∙L ^-1NAA+30 g∙L ^-1蔗糖; 不定芽诱导小鳞茎的最适培养基为MS+1.0 mg∙L ^-1NAA+30 g∙L ^-1蔗糖; 小鳞茎膨大生根的最适培养基为1/2MS+0.01 mg∙L ^-1NAA+60 g∙L ^-1蔗糖。该研究为金黄花滇百合种质资源保护和快速繁殖提供了技术支撑。     

金黄花滇百合植株再生与离体快繁技术体系的建立

以金黄花滇百合(Lilium bakerianum var. aureum)的鳞片为外植体, 探讨不同部位外植体和不同配比的植物生长调节剂对愈伤组织诱导和植株再生的影响。研究结果表明, 鳞片诱导愈伤组织和不定芽的最适培养基为MS+1.0 mg?L ^-16-BA+0.1 mg?L ^-1NAA+30 g?L ^-1蔗糖; 不定芽增殖的最适培养基为MS+0.5 mg?L ^-16-BA+0.1 mg?L ^-1NAA+30 g?L ^-1蔗糖; 不定芽诱导小鳞茎的最适培养基为MS+1.0 mg?L ^-1NAA+30 g?L ^-1蔗糖; 小鳞茎膨大生根的最适培养基为1/2MS+0.01 mg?L ^-1NAA+60 g?L ^-1蔗糖。该研究为金黄花滇百合种质资源保护和快速繁殖提供了技术支撑。     

宁南山区典型草本植物茎叶分解对土壤酶活性及微生物多样性的影响

采用凋落物分解袋法,以宁南山区典型草本植物长芒草、铁杆蒿、百里香为研究对象,分析了3种植物茎叶分解过程中土壤酶活性变化特征和分解后期微生物多样性特征,以及土壤酶活性与初始土壤化学性质的关系.结果表明: 植物茎叶分解480 d后,各处理土壤酶活性均有不同程度的增加,且长芒草处理土壤蔗糖酶活性和碱性磷酸酶活性最高,分别为32.40和1.99 mg·g^-1·24 h^-1,百里香处理土壤脲酶活性最高(2.66 mg·g^-1·24 h^-1),铁杆蒿处理土壤纤维素酶活性最高(1.42 mg·g^-1·72 h^-1).分解末期土壤纤维素酶活性与土壤初始微生物生物量碳呈显著正相关;分解末期土壤纤维素酶活性与土壤初始硝态氮含量呈显著负相关.添加植物茎叶处理土壤细菌和真菌的Ace指数、Chao指数和Shannon指数均显著大于对照,Simpson指数显著小于对照.植物茎叶的分解显著提高了土壤细菌和真菌的丰度及多样性,进而提高了植物茎叶的分解速率,促进了生态系统营养物质的循环与转化.     

库布齐沙漠东部不同生物结皮发育阶段土壤温室气体通量

以流动沙地为对照,采用时空替代法分析库布齐沙漠东部固定沙地上不同发育阶段生物结皮藻类结皮和地衣结皮土壤温室气体通量特征及其与环境因子之间的关系,研究生物结皮发育对荒漠土壤温室气体通量的影响.结果表明: 荒漠土壤CO₂排放通量大小为地衣结皮(128.5 mg·m^-2·h^-1)>藻结皮(70.2 mg·m^-2·h^-1)>流动沙地(48.2 mg·m^-2·h^-1),CH₄吸收通量大小为地衣结皮(30.4 μg·m^-2·h^-1)>藻结皮(21.2 μg·m^-2·h^-1)>流动沙地(18.2 μg·m^-2·h^-1),N₂O排放通量大小为地衣结皮(6.6 μg·m^-2·h^-1)>藻结皮(5.4 μg·m^-2·h^-1)>流动沙地(2.5 μg·m^-2·h^-1).CO₂排放具有明显的季节变化,生长季显著大于非生长季;CH₄和N₂O季节变化差异不显著,前者生长季吸收大于非生长季,后者非生长季排放大于生长季.土壤有机碳和全氮含量、土壤微生物数量均是影响温室气体通量的重要因素,环境水热因子是影响土壤CO₂排放的关键因子,但CH₄和N₂O通量对水热因子的变化不敏感.随着植被恢复和生物结皮发育,荒漠土壤温室气体累积通量的不断增大导致其百年尺度的全球增温潜势亦显著提高,依次为地衣结皮(1135.7 g CO₂-e·m^-2·a^-1)>藻结皮(626.5 g CO₂-e·m^-2·a^-1) >流动沙地(422.7 g CO₂-e·m^-2·a^-1).     

Reactor design for the ABE fermentation using cells of Clostridium acetobutylicum immobilized by adsorption onto bonechar

Cells of Clostridium acetobutylicum were immobilized by adsorption onto bonechar. and used for the production of solvents (ABE fermentation) from whey permeate. When the process was performed in packed bed reactors operated in a vertical or inclined mode, solvent productivities approximating 6 kg/(m³h) were obtained. However, the systems suffered from blockage due to excess biomass production and gas hold-up. These problems were less apparent when a partially-packed bed reactor was operated in the horizontal mode. A fluidized bed reactor proved to be the most stable of the systems investigated, and a productivity of 4.8 kg/(m³h) was maintained over a period of 2000 h of operation. The results demonstrate that this type of reactor may have a useful future role in the ABE fermentation.