Adaptations in bacterial catabolic enzyme activity and community structure in membrane-coupled bioreactors fed simple synthetic wastewater

Membrane-coupled bioreactors (MBRs) offer substantial benefits compared to conventional reactor designs for biological wastewater treatment. MBR treatment efficiency, however, has not been optimized because the effects of the MBR on process microbiology are poorly understood. In this study, the structure and function of the microbial communities growing in MBRs fed simple synthetic wastewater were investigated. In four starch-fed MBRs, the bacterial community substantially increased its alpha-glucosidase affinity (>1000-fold), while the leucine aminopeptidase and heptanoate esterase affinities increased slightly (<40-fold) or remained relatively constant. Concomitant to these physiological adaptations, shifts in the bacterial community structure in two of the starch-fed MBRs were detected by PCR-DGGE. Four of the bacterial populations detected by PCR-DGGE were isolated and exhibited specific growth rates in batch culture ranging from 0.009 to 0.22 h(-1). Our results suggest that bacterial communities growing under increasingly stringent nutrient limitation adapt their enzyme activities primarily for the nutrients provided, but that there is also a more subtle response not linked to the substrates included in the feed medium. Our research also demonstrates that MBRs can support relatively complex bacterial communities even on simple feed media.     

Aerobic Biological Treatment of Low-Strength Synthetic Wastewater in Membrane-Coupled Bioreactors: The Structure and Function of Bacterial Enrichment Cultures as the Net Growth Rate Approaches Zero

The goal of the current research was to determine if the stringent nutrient limitation imposed by membrane-coupled bioreactors (MBRs) could be used to force mixed bacterial communities to exhibit a zero net growth rate over an extended time period. Mechanistically, this zero net growth rate could be achieved when the amount of energy available for growth is balanced by the maintenance requirements of the bacterial community. Bench-scale MBRs were fed synthetic feed medium containing gelatin as the major organic substrate. Biomass concentrations initially increased rapidly, but subsequently declined until an asymptote was reached. Leucine aminopeptidase activities concomitantly increased by at least 10-fold, suggesting that bacterial catabolic activity remained high even while growth rates became negligible. In contrast, α-glucosidase and heptanoate esterase activities decreased, indicating that the bacterial community specifically adapted to the carbon source in the feed medium. Bacterial community analysis by denaturing gradient gel electrophoresis of PCR-amplified 16S rRNA gene fragments (PCR-DGGE) suggested that the bacterial community structure completely changed from the beginning to the end of each MBR. Excision and nucleotide sequence analysis of prominent PCR-DGGE bands suggested that many of the dominant populations were similar to novel bacterial strains that were previously uncultivated or recently cultivated during studies specifically targeting these novel populations. This research demonstrates that MBRs have substantial practical applications for biological wastewater treatment; in addition, MBRs are a useful tool to study the ecology of slow-growing bacteria.     

Removal of carbonaceous and nitrogenous pollutants from a synthetic wastewater using a membrane-coupled bioreactor

Two modified Ludzack-Ettinger (MLE)-type membrane-coupled bioreactors (MBRs) were investigated in this study for the purpose of removing both nitrogenous and carbonaceous pollutants from a synthetic wastewater. During the first MBR experiment, removal efficiencies were high (>90%) for chemical oxygen demand (COD) and ammonia, but total nitrogenous pollutant removal efficiency was poor (~25%). Bacterial community analysis of ammonia oxidizing bacteria (AOB) by a nested PCR-DGGE approach detected two Nitrosomonas-like populations and one Nitrosospira-like population. During the initial portion of the second MBR experiment, COD and ammonia removal efficiencies were similar to the first MBR experiment until the COD of the influent wastewater was increased to provide additional electron donors to support denitrification. Total nitrogen removal efficiencies eventually exceeded 90%, with a hydraulic residence time (HRT) of 24 h and a recirculation ratio of 8. When the HRT of the MBR experiment was decreased to 12 h, however, ammonia removal efficiency was adversely affected. A subsequent increase in the HRT to 18 h helped improve removal efficiencies for both ammonia (>85%) and total nitrogenous compounds (~70%). Our research demonstrates that MBRs can be effectively designed to remove both carbonaceous and nitrogenous pollutants. The ability of the microbial community to switch between anoxic (denitrifying) and oxic (nitrifying) conditions, however, represents a critical process constraint for the application of MLE-type MBR systems, such that little benefit is gained compared to conventional designs.     

Novel application of oxygen-transferring membranes to improve anaerobic wastewater treatment

Anaerobic biological wastewater treatment has numerous advantages over conventional aerobic processes; anaerobic biotechnologies, however, still have a reputation for low-quality effluents and operational instabilities. In this study, anaerobic bioreactors were augmented with an oxygen-transferring membrane to improve treatment performance. Two anaerobic bioreactors were fed a synthetic high-strength wastewater (chemical oxygen demand, or COD, of 11,000 mg l(-1)) and concurrently operated until biomass concentrations and effluent quality stabilized. Membrane aeration was then initiated in one of these bioreactors, leading to substantially improved COD removal efficiency (> 95%) compared to the unaerated control bioreactor (approximately 65%). The membrane-augmented anaerobic bioreactor required substantially less base addition to maintain circumneutral pH and exhibited 75% lower volatile fatty acid concentrations compared to the unaerated control bioreactor. The membrane-aerated bioreactor, however, failed to improve nitrogenous removal efficiency and produced 80% less biogas than the control bioreactor. A third membrane-augmented anaerobic bioreactor was operated to investigate the impact of start-up procedure on nitrogenous pollutant removal. In this bioreactor, excellent COD (>90%) and nitrogenous (>95%) pollutant removal efficiencies were observed at an intermediate COD concentration (5,500 mg l(-1)). Once the organic content of the influent wastewater was increased to full strength (COD = 11,000 mg l(-1)), however, nitrogenous pollutant removal stopped. This research demonstrates that partial aeration of anaerobic bioreactors using oxygen-transferring membranes is a novel approach to improve treatment performance. Additional research, however, is needed to optimize membrane surface area versus the organic loading rate to achieve the desired effluent quality.     

Functional and Structural Adaptations of Bacterial Communities Growing on Particulate Substrates under Stringent Nutrient Limitation

Biomass recycle reactors (BRRs) were used as a model system to study the functional and structural adaptations of mixed bacterial communities in response to the imposition of increasingly severe nutrient limitation. BRRs were fed synthetic media containing either spinach homogenate or autoclaved yeast cells to simulate the complex mixtures of particulate carbon sources that are often present in nature. In the BRRs fed spinach homogenate, the biomass (measured as particulate protein) exhibited a physiological response similar to previous studies as detected by 40–80% reductions in respiratory potential and by relatively stable catabolic ectoenzyme activities. Concomitant adaptations in bacterial community structure were detected by PCR-DGGE and RT-PCR-DGGE of 16S rDNA and 16S rRNA fragments, respectively. The microbial community structure was dynamic even after the biomass had reached a quasi-steady state with respect to physiological measurements. In the BRRs fed yeast cells, respiratory potentials increased 2- to 5-fold during the initial portion of the BRR run and a-glucosidase and b-glucosidase activities increased 2- to 4-fold. Substantial bacterial community shifts were also detected in both the rDNA and rRNA profiles, indicating that this community was also structurally dynamic. These experiments suggest that phylogenetically different bacteria sustained the functional activities in these ecosystems in response to increasingly stringent nutrient limitation.     

Digestion in the soil predatory mite Pergamasus longicornis (Berlese) (Acari: Mesostigmata: Parasitidae)--detectable hydrolases

Nineteen hydrolytic enzymes were detected in individual adult Pergamasus longicornis (Berlese) mites--amylase, hide protease, alkali phosphatase, esterase (C4), esterase lipase (C8), lipase (C14), leucine arylamidase, valine arylamidase, cystine arylamidase, acid phosphatase, phosphoamidase, alpha-galactosidase, beta-galactosidase, beta-glucuronidase, alpha-glucosidase, beta-glucosidase, N-acetyl-beta-glucosaminidase, alpha-mannosidase, and alpha-fucosidase. All but the phosphatases were detected for the first time. Tryptic and chymotryptic activity were consistently not demonstrable. Comparisons are made with saprophagous mites. No clear enzymic specialization for predation was found.     

基于好氧反硝化及反硝化聚磷菌强化的低温低碳氮比生活污水生物处理中试研究

【背景】低碳氮比生活污水很难达标处理,多级A/O工艺、生物强化技术及生物膜技术的有机结合可有效解决这一问题。【目的】开发出一种泥膜共生多级A/O工艺并进行中试研究,驯化出高效脱氮除磷菌剂并对系统进行生物强化。【方法】通过测定中试设备出水及污水处理厂出水化学需氧量(Chemical oxygen demand,COD)、氨氮(NH_4~+-N)、硝氮(NO_3~--N)、总氮(Total nitrogen,TN)、总磷(Total phosphorus,TP)对比分析两种工艺的污染物去除效能,利用高通量测序技术对比生物强化技术对系统微生物群落结构的影响。【结果】中试设备对COD、NH_4~+-N、NO_3~--N、TN、TP的去除效果均优于污水处理厂的处理工艺;驯化的低温好氧反硝化菌TN去除率最大值可达84.21%,驯化的低温反硝化聚磷菌群对磷的去除率最高可达85.75%;利用驯化菌群对中试设备进行生物强化后较好地改善了系统NH_4~+-N、NO_3~--N、TN、TP的去除效果;经生物强化后,具有好氧反硝化和反硝化聚磷功能的Pseudomonas菌群明显增多。【结论】泥膜共生多级A/O工艺对于低碳氮比生活污水的处理具有很好的效果,利用生物强化技术可有效提高低温条件下系统污染物去除效能。     

Human plasma carboxyl esterase-catalyzed triolein hydrolysis. Existence of promoting factor in serum

The possibility that some factor in serum changes the substrate specificity of purified human plasma carboxyl esterase, which hydrolyzes the short chain fatty acid ester, tributyrin, was investigated. The purified carboxyl esterase from human plasma hydrolyzed 48 mmol of tributyrin/mg of protein/h, monoolein at 1560 mumol of released fatty acids/mg of protein/h, diolein at 133 mumol of released fatty acids/mg of protein/h, and triolein at less than 10 mumol of released fatty acids/mg of protein/h. When human serum was applied to phenyl-Sepharose, a triolein hydrolysis-promoting factor (THPF) for purified carboxyl esterase was bound to the gel and was eluted with water. This partially purified human serum THPF enhanced carboxyl esterase-catalyzed triolein hydrolysis about 30-fold, diolein hydrolysis 2-fold, and monoolein hydrolysis 1.5-fold. Hydrolysis of triolein in very low density lipoproteins (d less than 1.006) and intermediate lipoproteins (1.006 less than d less than 1.019) by carboxyl esterase was also enhanced by addition of THPF. THPF activity was reduced by treatment of delipidation, but resistant to trypsin treatment or heating at 50 degrees C. These results indicated that serum carboxyl esterase can hydrolyze the long chain fatty acid ester, triolein, in the presence of triolein hydrolysis-promoting factor in serum.     

Purification and characterization of lysosomal alpha-glucosidase secreted by eukaryote Tetrahymena

A large amount of lysosomal acid hydrolases was released into the medium by Tetrahymena pyriformis strain W during growth. An extracellular lysosomal acid alpha-glucosidase has been purified 500-fold with a 41% yield to homogeneity, as judged by polyacrylamide gel electrophoresis. It was found to be a glycoprotein and to consist of a single 110,000-dalton polypeptide chain. The carbohydrate content of the alpha-glucosidase was equivalent to 2.8% of the total protein content, and the oligosaccharide moiety was composed of mannose and N-acetylglucosamine in a molar ratio of 6.7:2. The optimal pHs for hydrolysis of maltose and p-nitrophenyl-alpha-glucopyranoside, maltose, isomaltose, and glycogen were 1.1 mM, 2.5 mM, 33.0 mM, and 18.5 mg/ml, respectively. This purified enzyme appears to have alpha-1,6-glucosidase as well as alpha-1,4-glucosidase activity. Turanose has a noncompetitive inhibitory effect on the hydrolysis of maltose. The antibody raised against Tetrahymena acid alpha-glucosidase inhibited the hydrolysis of all substrates tested. These properties of Tetrahymena acid alpha-glucosidase were found to be similar to those of the human liver lysosomal alpha-glucosidase.     

The microbial community in a moving bed biotrickling filter operated to remove hydrogen sulfide from gas streams

The information available on the microbial communities responsible for pollutant degradation is increasingly accessible. Its use to optimize process design and operation is an important challenge in the field of effluent treatment research. Therefore, a prototype of a moving bed biotrickling filter (MBBTF) reactor was designed and, for the first time, operated at full-scale for the removal of sulfides desorbing from tannery industrial wastewater. The bacterial community operating in this innovative reactor was studied, and its composition and response to different operating conditions were characterized. A stable biomass, dominated by sulfur-oxidizing bacteria of the genus Acidithiobacillus was selected from inside the MBBTF reactor, and temperature, pH and bed rotation were shown to be the main factors driving the community structure. Moreover, data from different approaches indicated an uneven spatial distribution of biofilm inside the studied reactor, due to the combined effect of fluid dynamics and substrate gradients within the bed volume. Despite the high removal efficiency achieved by this innovative prototype (80% on average), the data suggested that the result could be improved by adopting solutions for a more stable and even biofilm distribution. It was shown that short frequent bed rotations, rather than long scattered rotations, ensured biomass stability. Furthermore, diversifying biofilm support media as a function of expected local pollutant concentrations should be considered. Data obtained from the bacterial community can therefore provide indications for possible further improvement of MBBTF reactor design and performance.     

Microbial population analysis of nutrient removal-related organisms in membrane bioreactors

Membrane bioreactors (MBR) are an important and increasingly implemented wastewater treatment technology, which are operated at low food to microorganism ratios (F/M) and retain slow-growing organisms. Enhanced biological phosphorus removal (EBPR)-related organisms grow slower than ordinary heterotrophs, but have never been studied in detail in MBRs. This study presents a comprehensive analysis of the microorganisms involved in EBPR in pilot- and full-scale MBRs, using fluorescence in situ hybridization (FISH), as well as an overall assessment of other relevant microbial groups. The results showed that polyphosphate accumulating organisms (PAOs) were present at similar levels in all studied MBRs (10% ± 6%), even those without a defined anaerobic zone. Glycogen accumulating organisms were also detected, although rarely. The FISH results correlated well with the observed P removal performance of each plant. The results from this study suggest that a defined anaerobic zone is not necessarily required for putative PAO growth in MBRs, since polyphosphate storage may provide a selective advantage in fulfilling cell maintenance requirements in substrate-limited conditions (low F/M).     

塔拉种子多糖脱蛋白的正交优化及其抗氧化性研究

以塔拉（Caesalpinia spinosa）种子为原料,研究了塔拉种子多糖的脱蛋白工艺及塔拉多糖的抗氧化性质。以多糖损失率和蛋白脱除率为评价指标,比较Sevage法、三氯乙酸法和木瓜蛋白酶法对塔拉多糖的脱蛋白效果。利用正交优化组合实验设计原理,采用四因素三水平的正交分析法,对木瓜蛋白酶法脱蛋白进行正交优化。结果表明：塔拉多糖最佳脱蛋白工艺条件为酶添加量0.15mL、酶解时间90min、酶解温度60℃、酶解pH=6,蛋白脱除率95.19%,多糖保留率75.02%。通过对塔拉多糖抗氧化性的研究,发现塔拉多糖总抗氧化性较好,对DPPH自由基有较强的清除作用。     

Thermophilic aerobic treatment of a synthetic wastewater in a membrane-coupled bioreactor

Synthetic wastewater containing -lactose and gelatin was treated in a thermophilic membrane-coupled bioreactor (MBR). Thermophilic (>45°C) treatment represents a potentially advantageous process for high-temperature as well as high-strength industrial wastewaters susceptible to reactor autoheating. Thermophilic systems, however, generally support a nonflocculating biomass that resists conventional methods of cell separation from the treated wastewater. MBRs were applied to thermophilic treatment systems because bacterial cells can be retained regardless of cell aggregation. Thermophilic aerobic MBRs were successfully operated at high levels of biocatalyst and produced a better effluent quality than analogous thermophilic bioreactors without cell recycle. At a hydraulic residence time (HRT) of 13.1 h, the chemical oxygen demand (COD) of the membrane eluate improved from 760 mg l⁻¹ (without cell recycle) to 160 mg l⁻¹ (with cell recycle). Bacterial community shifts were detected by denaturing gradient gel electrophoresis (DGGE) of polymerase chain reaction (PCR) -amplified 16S rRNA gene fragments — 6 of 13 bands disappeared within 2 days of MBR operation. A concomitant 40–50% reduction in physiological indicators of cell reactivity (RNA:protein; ATP:protein) was also observed. The specific activity of β-galactosidase and aminopeptidase, however, increased by 10–25%, indicating that there is a definite advantage to MBR operation at the highest biomass level possible. Nucleotide sequence analysis of 16S rDNA clones identified phylotypes from the low-G+C Gram-positive division and the β- and γ-subdivisions of Proteobacteria. Journal of Industrial Microbiology & Biotechnology (2001) 26, 203–209. Received 18 March 2000/ Accepted in revised form 26 January 2001     

The substrate specificity of acid α-glucosidase from rabbit muscle

1. Acid alpha-glucosidase was purified 3500-fold from rabbit muscle. 2. The enzyme was activated by cations, the degree of activation varying with the substrate. Enzyme action on glycogen was most strongly activated and activation was apparently of a non-competitive type. With rabbit liver glycogen as substrate, the relative V(max.) increased 15-fold, accompanied by an increase in K(m) from 8.3 to 68.6mm-chain end over the cation range 2-200mm-Na(+) at pH4.5. Action on maltose was only moderately activated (1.3-fold, non-competitively) and action on maltotriose was marginally and competitively inhibited. 3. The pH optimum at 2mm-Na(+) was 4.5 (maltose) and 5.1 (glycogen). Cation activation of enzyme action on glycogen was markedly pH-dependent. At 200mm-Na(+), the pH optimum was 4.8 and activity was maximally stimulated in the range pH4.5-3.3. 4. Glucosidase action on maltosaccharides was associated with pronounced substrate inhibition at concentrations exceeding 5mm. Of the maltosaccharides tested, the enzyme showed a preference for p-nitrophenyl alpha-maltoside (K(m) 1.2mm) and maltotriose (K(m) 1.8mm). The extrapolated K(m) for enzyme action on maltose was 3.7mm. 5. The macromolecular polysaccharide substrate glycogen differed from linear maltosaccharide substrates in the kinetics of its interaction with the enzyme. Activity was markedly dependent on pH, cation concentration and polysaccharide structure. There was no substrate inhibition. 6. The enzyme exhibited constitutive alpha-1,6-glucanohydrolase activity. The K(m) for panose was 20mm. 7. The enzyme catalysed the total conversion of glycogen into glucose. The hydrolysis of alpha-1,6-linkages was apparently rate-limiting during the hydrolysis of glycogen. 8. Enzyme action on glycogen and maltose released the alpha-anomer of d-glucose. 9. The results are discussed in terms of the physiological role of acid alpha-glucosidase in lysosomal glycogen catabolism.     

Assay for determination of α-glucosidase and peptidase activity and location in a nitrifying trickling filter

Enzymatic alpha-glucosidase and peptidase activity in a nitrifying trickling filter (NTF) at the Rya wastewater treatment plant, G?teborg, Sweden, was investigated to evaluate whether these activities can be used as indicators of heterotrophic activity and polymer degradation. Samples of the biofilm were taken from the NTF and incubated in sterile filtered effluent water from the NTF with the addition of soluble starch, peptone, and ammonium chloride. In order to determine the distribution of enzyme activities, the alpha-glucosidase and peptidase activities were measured in the biofilm samples, in the filtered effluent water from the NTF and in the water phase in which the biofilm was incubated. Activities of both enzymes were found both in the effluent water from the NTF and in the biofilm. The enzyme activities were elevated in the samples when starch and peptone were present. In addition, there was a significant inhibition of ammonium oxidation in samples incubated with starch and peptone. Thus, the presence of starch, peptone and ammonium resulted in increased activity of heterotrophs, which lead to an inhibition of the nitrifiers, probably via competition for available oxygen.     

Improvement in post-treatment of digested swine wastewater

The performance of sequencing batch reactor (SBR) during post-treatment of digested effluent of swine wastewater was investigated. While operating SBR to treat the digested effluent directly, the performance was very poor with COD removal rate about 10%, and NH4+-N removal rate nearly 50%, with a scarce removal of total phosphorus. The performance apparently improved after adding raw swine wastewater or alkali to digested effluent. Although similar results for NH4+-N removal were achieved adopting both measures, the addition of raw wastewater proved superior in removing total nitrogen and total phosphorus. The addition of raw wastewater obtained effluent COD around 300 mg/L which was lower than that after alkali addition i.e. around 550 mg/L. Judged from the investment, oxygen demand, sludge yield, biogas production and running cost, the traditional combined anaerobic-SBR process is unfeasible to treat swine wastewater, while the combined anaerobic-SBR process with addition of raw swine wastewater can be a suitable biotechnology.     

Effect of sulfadiazine and trimethoprim on activated sludge performance and microbial community dynamics in laboratory-scale membrane bioreactors and sequencing batch reactors at 8°C

The effect of antibiotics sulfadiazine and trimethoprim on activated sludge operated at 8°C was investigated. Performance and microbial communities of sequencing batch reactors (SBRs) and Membrane Bioreactors (MBRs) were compared before and after the exposure of antibiotics to the synthetic wastewater. The results revealed irreversible negative effect of these antibiotics in environmentally relevant concentrations on nitrifying microbial community of SBR activated sludge. In opposite, MBR sludge demonstrated fast adaptation and more stable performance during the antibiotics exposure. Dynamics of microbial community was greatly affected by presence of antibiotics. Bacteria from classes Betaproteobacteria and Bacteroidetes demonstrated the potential to develop antibiotic resistance in both wastewater treatment systems while Actinobacteria disappeared from all of the reactors after 60 days of antibiotics exposure. Altogether, results showed that operational parameters such as sludge retention time (SRT) and reactor configuration had great effect on microbial community composition of activated sludge and its vulnerability to antibiotics. Operation at long SRT allowed archaea, including ammonium oxidizing species (AOA) such as Nitrososphaera viennensis to grow in MBRs. AOA could have an important role in stable nitrification performance of MBR-activated sludge as a result of tolerance of archaea to antibiotics. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2708, 2019     

Substrate-Specific Development of Thermophilic Bacterial Consortia by Using Chemically Pretreated Switchgrass

Microbial communities that deconstruct plant biomass have broad relevance in biofuel production and global carbon cycling. Biomass pretreatments reduce plant biomass recalcitrance for increased efficiency of enzymatic hydrolysis. We exploited these chemical pretreatments to study how thermophilic bacterial consortia adapt to deconstruct switchgrass (SG) biomass of various compositions. Microbial communities were adapted to untreated, ammonium fiber expansion (AFEX)-pretreated, and ionic-liquid (IL)-pretreated SG under aerobic, thermophilic conditions using green waste compost as the inoculum to study biomass deconstruction by microbial consortia. After microbial cultivation, gravimetric analysis of the residual biomass demonstrated that both AFEX and IL pretreatment enhanced the deconstruction of the SG biomass approximately 2-fold. Two-dimensional nuclear magnetic resonance (2D-NMR) experiments and acetyl bromide-reactive-lignin analysis indicated that polysaccharide hydrolysis was the dominant process occurring during microbial biomass deconstruction, and lignin remaining in the residual biomass was largely unmodified. Small-subunit (SSU) rRNA gene amplicon libraries revealed that although the dominant taxa across these chemical pretreatments were consistently represented by members of the Firmicutes, the Bacteroidetes, and Deinococcus-Thermus, the abundance of selected operational taxonomic units (OTUs) varied, suggesting adaptations to the different substrates. Combining the observations of differences in the community structure and the chemical and physical structure of the biomass, we hypothesize specific roles for individual community members in biomass deconstruction.     

Novel polysaccharide–protein-based amphipathic formulations

Previous results showed that the cell-surface esterase from Acinetobacter venetianus RAG-1 enhances the emulsification properties of the polymeric bioemulsifier emulsan and its deproteinated derivative apoemulsan (Bach H, Berdichevsky Y, Gutnick D (2003) An exocellular protein from the oil-degrading microbe Acinetobacter venetianus RAG-1 enhances the emulsifying activity of the polymeric bioemulsifier emulsan. Appl Environ Microbiol 69:2608–2615). Here we show that in the presence of the his-tagged recombinant esterase from RAG-1, 18 different polysaccharides from microbial, plant, insect and synthetic sources formed hexadecane-in-water emulsions. Emulsifying activities were distributed over a 13-fold range from over 4800 U/mg protein/mg polysaccharide in the case of apoemulsan to 370 U/mg protein/mg polysaccharide in the case of alginic acid. The stability of the emulsions ranged between 95 and 58%. Emulsions formed in the presence of seven of the polysaccharides exhibited stabilities of over 80%. The esterase from A. calcoaceticus BD4, which shows sequence homology to the RAG-1 esterase, was inactive in emulsification enhancement. The sequence of the RAG-1 esterase was shown to contain two conserved peptide sequences previously shown to be implicated in carbohydrate/polysaccharide binding. A hypothetical model illustrating a possible mode of interaction between the esterase, the apoemulsan and the oil droplet is presented. The complex is presumed to generate a series of “coated” oil droplets which are restricted in their ability to coalesce resulting in a relatively stable emulsion.     

Biomass production from glutamate fermentation wastewater by the co-culture of Candida halophila and Rhodotorula glutinis

In this study, the biomass production and pollutant removal from high-strength glutamate fermentation wastewater (GFW) using yeast isolates was investigated. Following enrichment culture, two species of yeasts, Candida halophila and Rhodotorula glutinis, were isolated from raw GFW with chemical oxygen demand (COD) and ammonia-nitrogen levels of 40 and 16 g l(-1), respectively. The binary mixed yeast culture was cultivated batchwise in 2.5-fold diluted GFW from which 85% of COD and 96% of reducing sugar were removed. The resulting yeast biomass contained 56% crude protein, 36.0% carbohydrate and 0.4% crude lipid. The amino acid composition of mixed yeast cells was balanced and was comparable with that of C. utilis and soybean.