光照混菌培养应用研究进展

光合细菌与其他微生物在光照条件下混合培养是近年来的研究热点。综述了光照混菌培养的特点和目前光照混菌培养在水体净化、生物制氢和高价值物质生产方面的应用,并对影响混合菌株生长代谢与繁殖的因素做了总结。分析表明菌株之间存在的相互协同共生作用能促进微生物的生长繁殖,使底物被充分利用,提高物质产率。光照混菌培养工艺简单、成本较低,在水体净化、生物制氢、高价值物质生产方面的应用具有相当好的效果。在影响因素中对混合培养影响最大的因素是菌株接种量、接种比和培养基pH。在总结光照混菌培养应用现存不足的基础上,对其发展前景作出展望。     

Aniline oxidation by microorganisms in chemostat cultivation

Under the conditions of chemostat cultivation, a mixed culture of microorganisms oxidized aniline at a gradually increasing concentration (up to 2.5 g/litre) as a sole source of carbon and nitrogen. The specific rate of aniline oxidation was as high as 160 mg per 1 g of dry biomass when aniline concentration in the growth medium was 2.5 g/litre. As aniline concentration in the growth medium was gradually raised, selection of the microorganisms took place and the number of the strains decreased from ten to two. The growth rate of the microorganisms fell down abruptly when phosphorus concentration in the medium was below 35 mg per 1 g of aniline.     

Dialysis fermentation. II. Kinetic analysis of salicylic acid production via cell growth and maintenance

A kinetic analysis was made of the relationship between salicylate production from naphthalene and growth of Pseudomonas fluorescens in semicontinuous dialysis culture. The specific rates both of product formation and growth initially were increased by the diffusional withdrawal of salicylate, but subsequently were reduced to low levels despite continued salicylate removal. Productivity and growth were correlated by the Luedeking-Piret equation in an initial nondialysis period and in the early stages of dialysis fermentation, when specific growth rates exceeded. 005 hr^?1. Below this level of growth at later stages of dialysis fermentation, the specific production rate was correlated only with total cell mass by a proportionality constant of .035 hr^?1, which was attributed to maintenance metabolism. Maintenance accounted for about 84% of the total salicylate produced, while growth-associated metabolism accounted for the remainder.     

Effects of varied pH, growth rate and temperature using controlled fermentation and batch culture on matrix assisted laser desorption/ionization whole cell protein fingerprints

Rapid identification of microorganisms using matrix assisted laser desorption/ionization (MALDI) is a rapidly growing area of research due to the minimal sample preparation, speed of analysis and broad applicability of the technique. This approach relies on expressed biochemical markers, often proteins, to identify microorganisms. Therefore, variations in culture conditions that affect protein expression may limit the ability of MALDI-MS to correctly identify an organism. We have expanded our efforts to investigate the effects of culture conditions on MALDI-MS signatures to specifically examine the effects of pH, growth rate and temperature. Continuous cultures maintained in bioreactors were used to maintain specific growth rates and pH for E. coli HB 101. Despite measurable morphological differences between growth conditions, the MALDI-MS data associated each culture with the appropriate library entry (E. coli HB 101 generated using batch culture on a LB media), independent of pH or growth rate. The lone exception was for a biofilm sample collected from one of the reactors which had no appreciable degree of association with the correct library entry. Within the data set for planktonic organisms, variations in growth rate created the largest variation between fingerprints. The effect of varying growth temperature on Y. enterocolitica was also examined. While the anticipated effects on phenotype were observed, the MALDI-MS technique provided the proper identification.     

Protein-mediated adhesion of Lactobacillus fermentum strain 737 to mouse stomach squamous epithelium

The mechanism of adhesion of Lactobacillus fermentum strain 737 to mouse stomach squamous epithelium was investigated. Adhesion inhibition tests involving chelators, monosaccharides, periodate and concanavalin A and the use of bacteria grown in the presence of tunicamycin failed to clarify the adhesive mechanism. Washed bacterial cells had reduced adhesive capacity, except in the presence of spent broth culture supernatant fraction or cell washings. Spent culture supernatant fractions of erythrosine-supplemented broth did not enhance adhesion of washed cells. The adhesion-promoting factor(s) in the spent broth culture supernatant fractions and cell washings bound to both bacterial and epithelial cell surfaces, but did not promote adhesion of two other Lactobacillus strains which were not of mouse origin, thereby indicating host specificity for the adhesion-promoting activity. Chemical characteristics of the adhesion-promoting factor were determined by pretreatment of the dialysis retentate of spent broth culture supernatant fractions with proteolytic enzymes, concanavalin A-Sepharose or periodate before the adhesion assay. The adhesin was non-dialysable, pronase-sensitive, heat sensitive at 100 degrees C, had no affinity for concanavalin A-Sepharose and contained no carbohydrate groups active in the adhesion process. The protein profiles of dialysis retentates of spent broth culture supernatant fractions after bacterial growth in the absence and presence of erythrosine were determined by 2-dimensional SDS-PAGE. Gel filtration by HPLC was used for purification of an adhesion-promoting fraction. The host-specific adhesion of L. fermentum strain 737 was mediated by a protein, with an Mr of 12-13000, that was not detectable in cells grown in the presence of erythrosine. A model for the mode of binding of the adhesin to host epithelia and bacterial surfaces is proposed.     

The half-saturation coefficient for dissolved oxygen: A dynamic method for its determination and its effect on dual species competition

A simple approach was developed to determine the half-saturation coefficient for dissolved oxygen (K(DO)) for three bacteria by maintaining a constant oxygen concentration in continuous culture, and employing a dynamic method to obtain the specific growth rate (mu) for each species. Measurement of mu at selected dissolved oxygen concentrations (DO) resulted in a typical Monod curve for a plot of mu vs. DO. Values for K(DO) and mu(max) were obtained from the Lineweaver-Burk reciprocal plot. The bacteria studied included representative strains of three microorganisms isolated in pure culture from poorly settling activated sludge: two filamentous microorganisms, Sphaerotilus natans and a second Sphaerotilus sp., and an unidentified floc-forming microorganism. The K(DO) values obtained for Sphaerotilus sp., S. natans, and the floc former were 0.014, 0.033, and 0.073 mg/L, respectively. Dual species competition experiments were conducted in continuous culture under low and high DO conditions. Successful growth competition by these microorganisms under DO-limiting conditions was consistent with experimentally determined K(DO) values. The finding of lower K(DO) values for the two Sphaerotilus species, compared to the floc former, confirmed the hypothesis that these filamentous microorganisms can outgrow floc-forming microorganisms in activated sludge when DO in the aeration basin is low.     

The influence of high substrate concentrations on microbial kinetics

High substrate concentrations inhibit growth and may distort the metabolism of microorganisms. Mechanisms causing substrate inhibition are discussed and used to derive several mathematical models representative of the entire concentration range, including stimulation of growth by low substrate concentrations. These kinetic models are tested with a variety of batch culture measurements of specific growth rate and respiration rate at widely-ranging substrate concentrations. Using one of the kinetic models, equations are developed for batch, continuous, and exponential-feed reactors. Comparison of results obtained in continuous culture with results from exponential-feed culture systems is shown to offer a novel experimental method for evaluating the effect of the cell age distribution on the properties and metabolic activity of a culture.     

Method for studying the adaptation of Escherichia coli to the acidification of the medium

Apparatus permitting to regulate the growth rate of turbidostat culture of microorganisms has been worked out. The process of adaptation of culture microorganisms to the medium acidity has been investigated by the stabilization of growth rate method. The correlation coefficients between adaptation time and changes of acid concentration in medium have been determined. The effect of aeration and density of microorganisms on the parameters of adaptation process has been considered. The conclusion concerning prospects of the growth rate stabilization method application for studying the regularities of culture microorganisms adaptation to the action of the stress has been made.     

On relationships between various distribution functions in balanced unicellular growth

The relationships between various size distributions in balanced exponential growth of a batch culture of microorganisms are presented. Starting from the partial differential integral equations (Eakmanet al., 1966; Fredricksonet al., 1967) derived for the growth of a microbial culture expressions are obtained for the growth rate of organisms of specific size and size range. These expressions were first obtained by Collins and Richmond (1962) by an entirely different method. Also derived are equations which link probability functions, which are basic to the growth of a microbial culture, with other size distributions that can be estimated experimentally.     

Measurement of Light Absorption and Determination of the Specific Rate of Light Uptake in Cultures of Phototrophic Microorganisms

This article describes a novel method for the measurement of light absorption by cultures of phototrophic microorganisms. The rate of light absorption is calculated as the difference between the rate of light output from a culture containing cells and the rate of light output from a culture containing only growth medium. The specific rate of light uptake is calculated by dividing the rate of light absorption by the total biomass present in the culture. Application of the method to several case studies shows that light output from a culture varies widely depending on the absorption and scattering characteristics of the suspension.     

Substrate concentration control by dialysis in fermentations of Escherichia coli

A substrate control system based upon dialysis culture techniques has been developed. In fermentations of Escherichia coli where phosphate or sulfate concentrations were controlled, the relation between the apparent specific growth rate, mu, and the phosphate concentration followed Monod kinetics, while the relation between mu and sulfate showed a sharp optimun. The pH of fermentations of E. coli was controlled by dialysis culture techniques without the fluctuations in pH associated with control by direct addition of acid or base.     

干扰素基因工程菌产物表达数学模型的研究

根据阻遏蛋白,辅阻遏因子与启动基因之间的相互作用,建立了采用色氨酸启动子的基因工程重组徽生物的产物表达数学模型。提出了mRNA最大转录速率常数和产物表达速率常数与比生长速率呈线性关系的假设,用干扰素基因工程菌培养过程中的实验数据估计了mR—NA转录速率常数,干扰素表达速率常数和干扰素降解速率常数。推导出了发酵过程中干扰素表达量和细胞内干扰素比活计算公式。运用这些公式可以通过检测色氨酸浓度和细胞比生长速率来计算预测干扰紊表达量。确定最佳的终止培养时间。     

Inhibition of adhesion of Escherichia coli K88 to piglet ileal mucus by Lactobacillus spp.

Enteropathogenic Escherichia coli K88 colonizing the piglet ileum adhere to the mucosa by K88 fimbrial appendages. A recent study in our laboratory has implicated indigenous lactobacilli in the suppression of the colonization potential of enteropathogenic E. coli as measured by adhesion to ileal mucus. The aim of this study was to investigate the effect of Lactobacillus spp. of porcine origin on the adhesion of K88 fimbriae of E. coli. With an in vitro assay, the adhesion of E. coli K88ab strain G1108E and E. coli K88ac strain 1107 to 35-day-old piglet ileal mucus was studied in the presence of spent culture fluid of Lactobacillus spp. Detailed studies focused specifically on culture fluid of Lactobacillus fermentum 104R. Subsequently, the ileal mucus was exposed to the retentate of the spent culture fluid after dialysis and fractionation. Adhesion was confirmed to be attributable to K88 fimbriae when K88-specific monoclonal antibodies and isogenic mutants of E. coli K-12 with and without the plasmid containing the K88 gene were used. The active component was characterized by pretreatment of dialysis retentate with heat, periodate, pronase, and centrifugation, as well as by growth of the lactobacillus in various media and by assays at both 0 and 37 degrees C. All three lactobacilli of porcine origin reduced adhesion of E. coli K88 by approximately 50%. Inhibition occurred when mucus was pretreated with either spent culture dialysis retentate or the void volume (fraction of > 250,000 molecular weight) after gel filtration. The activity of the dialysis retentate was sensitive to pronase, but there was still activity at 0 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS)     

Pentachlorophenol degradation: a pure bacterial culture and an epilithic microbial consortium.

The steady-state growth of a Flavobacterium strain known to utilize pentachlorophenol (PCP) was examined when cellobiose and PCP simultaneously limited its growth rate in continuous culture. A concentration of 600 mg of PCP per liter in influent medium could be continuously degraded without affecting steady-state growth. We measured specific rates of PCP carbon degradation as high as 0.15 +/- 0.01 g (dry weight) of C per h at a growth rate of 0.045 h-1. Comparable specific rates of PCP degradation were obtained and maintained by PCP-adapted, natural consortia of epilithic microorganisms. The consortium results suggest that a fixed-film bioreactor containing a PCP-adapted natural microbial population could be used to treat PCP-contaminated water.     

Removal of inhibitors of bacterial growth by dialysis culture.

Dialysis culture was used to investigate the extent to which growth inhibition in bacterial cultures may be caused by accumulation of metabolites. Escherichia coli B was grown in a glucose/salts medium. A concentrated nutrient solution was pumped at a constant rate into the growing culture to ensure that growth was not limited by exhaustion of nutrients. In this way the only difference between growth conditions in dialysis and non-dialysis cultures was the transfer of dialysable metabolites from the culture vessel to the reservoir in the dialysis culture system. By adjusting the glucose concentration in the feed and maintaining a constant rate of feeding, glucose-limited growth could be achieved. Under these conditions, with oxygen in excess, bacterial yields of 140 to 150 g dry wt l-1 were obtained in dialysis culture compared with 30 to 40 g l-1 in non-dialysis culture. The high yields in dialysis culture depended on the removal of end-products of glucose metabolism. Growth inhibition was demonstrated to be the result of the combined influence of acetate, lactate, pyruvate, succinate, propionate and isobutyrate in concentrations found at the end of growth in non-dialysis cultures of Escherichia coli B.     

Pentachlorophenol degradation: a pure bacterial culture and an epilithic microbial consortium

The steady-state growth of a Flavobacterium strain known to utilize pentachlorophenol (PCP) was examined when cellobiose and PCP simultaneously limited its growth rate in continuous culture. A concentration of 600 mg of PCP per liter in influent medium could be continuously degraded without affecting steady-state growth. We measured specific rates of PCP carbon degradation as high as 0.15 +/- 0.01 g (dry weight) of C per h at a growth rate of 0.045 h-1. Comparable specific rates of PCP degradation were obtained and maintained by PCP-adapted, natural consortia of epilithic microorganisms. The consortium results suggest that a fixed-film bioreactor containing a PCP-adapted natural microbial population could be used to treat PCP-contaminated water.     

Monoclonal antibody productivity and the metabolic pattern of perfusion cultures under varying oxygen tensions

The metabolic pattern and cell culture kinetics of high-cell-density perfusion cultures were compared under two different oxygen transfer conditions: oxygen limiting and not limiting. When oxygen was a limiting factor during perfusion culture, both specific glucose uptake and lactate production rates increased, compared to non-oxygen-limited condition, by about 60% and 30%, respectively. The specific glutamine uptake rate under oxygen-limited conditions was almost 4.0 times higher than that under non-oxygen-limited conditions. The activity of lactate dehydrogenase (LDH) released into the medium by the dead cells can be used as an indicator for the metabolic and physiological conditions related to oxygen limitation. There was a 3.2 times higher specific rate of LDH activity released by dead cells in oxygen-limited cultures than those in non-oxygen-limited cultures. The specific production rate of monoclonal antibody was not significantly affected by the oxygen transfer conditions during the rapid cell growth period, but it rapidly increased toward the end of perfusion cultures. The higher perfusion rate may have limited further cell growth during high-cell-density perfusion culture, because cell damage was caused by the hydrodynamic shear within a hollow fiber microfiltration cartridge installed to withdraw the spent medium and the waste metabolites. (c) 1993 John Wiley & Sons, Inc.     

Inferential control of the specific growth rate in fed-batch cultivation processes

Inferential control algorithms and systems for set-point control of the specific growth rate in fed-batch cultivation processes are presented. Realization of the proposed control systems does not require mathematical model and a priori knowledge of the culture of microorganisms under control. Feed-back signal in PID control loops of the investigated systems is based on measurements of the O₂ concentration in exhaust gas and the air supply rate, however, measurements of the other technological parameters related to the culture growth dynamics can also be applied.     

Effect of surface-active agents (tween-21) on indices of energy metabolism in oleandomycin producers

The specific growth rate of Streptomyces antibioticus, a producer of oleandomycin, and the specific rate of the antibiotic accumulation in the culture medium during fermentation were investigated. On the basis of the results obtained the fermentation period was divided into 7 phases of development. The culture treated with the surfactant (Tween-21) is characterized by a higher specific growth rate during the whole fermentation and a higher specific rate of the antibiotic accumulation at the stage of the highest production as compared to the control. The ATP content, the value of the adenylate energy charge and the contents of high-molecular weight polyphosphates in the mycelium were examined. In the phase of the intensive growth St. antibioticus was characterized by a higher ATP level and a higher energy charge. More active accumulation of polyphosphates was observed in the late intensive growth phase. It was also found that after the treatment of the culture with Tween-21 it utilized polyphosphates more actively during the antibiotic biosynthesis.