Occurrence, removal, and prioritization of organic micropollutants in four full-scale wastewater treatment plants in Korea.

This study investigated the occurrence, removal rate, and potential risks of 43 organic micropollutants (OMPs) in four municipal wastewater treatment plants (WWTPs) in Korea. Results from two-year intensive monitoring confirmed the presence of various OMPs in the influents, including pharmaceuticals such as acetaminophen (pain relief), caffeine (stimulants), cimetidine (H2-blockers), ibuprofen (non-steroidal anti-inflammatory drugs- NSAIDs), metformin (antidiabetics), and naproxen (NSAIDs) with median concentrations of >1 µg/L. Some pharmaceuticals (carbamazepine-anticonvulsants, diclofenac-NSAIDs, propranolol-ß-blockers), corrosion inhibitors (1H-benzotriazole-BTR, 4-methyl-1H-benzotriazole-4-TTR), and perfluorinated compounds (PFCs) were negligibly removed during WWTP treatment. The OMP concentrations in the influents and effluents were mostly lower in August than those of other months (p-value < 0.05) possibly due to wastewater dilution by high precipitation or enhanced biodegradation under high-temperature conditions. The anaerobic-anoxic-oxic process (A2O) with a membrane bioreactor exhibited higher OMP removal than other processes, such as A2O with sedimentation or the conventional activated sludge process (p-value < 0.05). Pesticides (DEET and atrazine), corrosion inhibitors (4-TTR and BTR), and metformin were selected as priority OMPs in toxicity-driven prioritization, whereas PFCs were determined as priority OMPs given their persistence and bioaccumulation properties. Overall, our results contribute to an important database on the occurrence, removal, and potential risks of OMPs in Korean WWTPs.

Predicting micropollutant fate during wastewater treatment using refined classical kinetic model based on quantitative monitoring in multi-metropolitan regions of South Korea.

This study aimed to implement an extensive prediction model for the fate of micropollutants (MPs) in wastewater treatment plants (WWTPs). Five WWTPs equipped with seven different biological treatment processes were monitored from 2020 to 2022 with three to four sampling events in each year, and 27 datasets for 20 MPs were collected. Among these datasets, 12 were used to investigate the behavior and fate of MPs in WWTPs in South Korea. Metformin, acetaminophen, caffeine, naproxen, and ibuprofen were the MPs with the highest influent concentrations (ranging from 3,933.3-187,637.0 ng L-1) at all WWTPs. More than 90% of MPs were removed by biological treatment processes in all WWTPs. The Kruskal-Wallis test verified that their efficacy did not differ statistically (p-value > 0.05). Meanwhile, to refine the performance of the prediction model, this study optimized the biodegradation rate constants (kbio) of each MP according to the variation of seasonal water temperature. As a result, compared to the original prediction model, the mean difference between the actual data and predicted results (MEAN) decreased by 6.77%, while the Nash-Sutcliffe efficiency (NSE) increased by 0.226. The final MEAN and NSE for the refined prediction model were calculated to be 5.09% and 0.964, respectively. The prediction model made accurate predictions, even for MPs exhibiting behaviors different from other cases, such as estriol and atrazine. Consequently, the optimization strategy proposed in this study was determined to be effective because the overall removal efficiencies of MPs were successfully predicted even with limited reference datasets.

Efficiency of ozonation and O3/H2O2 as enhanced wastewater treatment processes for micropollutant abatement and disinfection with minimized byproduct formation.

Ozonation, a viable option for improving wastewater effluent quality, requires process optimization to ensure the organic micropollutants (OMPs) elimination and disinfection under minimized byproduct formation. This study assessed and compared the efficiencies of ozonation (O3) and ozone with hydrogen peroxide (O3/H2O2) for 70 OMPs elimination, inactivation of three bacteria and three viruses, and formation of bromate and biodegradable organics during the bench-scale O3 and O3/H2O2 treatment of municipal wastewater effluent. 39 OMPs were fully eliminated, and 22 OMPs were considerably eliminated (54 ± 14%) at an ozone dosage of 0.5 gO3/gDOC for their high reactivity to ozone or •OH. The chemical kinetics approach accurately predicted the OMP elimination levels based on the rate constants and exposures of ozone and •OH, where the quantum chemical calculation and group contribution method successfully predicted the ozone and •OH rate constants, respectively. Microbial inactivation levels increased with increasing ozone dosage up to ∼3.1 (bacteria) and ∼2.6 (virus) log10 reductions at 0.7 gO3/gDOC. O3/H2O2 minimized bromate formation but significantly decreased bacteria/virus inactivation, whereas its impact on OMP elimination was insignificant. Ozonation produced biodegradable organics that were removed by a post-biodegradation treatment, achieving up to 24% DOM mineralization. These results can be useful for optimizing O3 and O3/H2O2 processes for enhanced wastewater treatment.

In vitro bioanalytical assessment of the occurrence and removal of bioactive chemicals in municipal wastewater treatment plants in Korea.

Effluents of wastewater treatment plants (WWTPs) contain various organic micropollutants, some of which can exert negative effects on the quality of receiving waters or drinking water sources. This study monitored two full-scale WWTPs in Korea for the occurrence and removal of bioactive chemicals for a one-year period using a battery of in vitro bioassays as a complementary approach to chemical analysis. Bioassays covering different endpoints were employed, such as hormone receptor activation (AR and ERα), xenobiotic metabolism (PAH and PXR), oxidative stress response (Nrf2), and cytotoxicity. The WWTP influents showed AR, ERα, and PAH activities at ng/L - µg/L and PXR and Nrf2 activities at µg/L - mg/L as bioanalytical equivalent concentrations of a reference compound for each bioassay. These bioactivities decreased along with the WWTP treatment train, with significant removals achieved by the secondary biological treatment processes. Cytotoxicity was observed only for some municipal wastewater (M-WWTP) influents but was below the limit of quantification for most cases. The influent and effluent bioactivities observed in this study were mostly comparable to those reported in other WWTPs in the literature. Comparison of the bioactivities with the effect-based trigger (EBT) values indicates that the impact of WWTP effluents on receiving water quality was low for most endpoints. For Nrf2, however, further investigation is required to evaluate the observed high bioactivities compared with the current EBT. The observed ERα activity could partly be explained by the presence of some steroid estrogens. Overall, our results contribute to an important database for the concentrations and removal efficiencies of bioactive chemicals in WWTPs and demonstrate bioassays as a useful tool for urban water quality monitoring.

Spatial and temporal effects of fish feed on antibiotic resistance in coastal aquaculture farms.

For the first time, both spatial and temporal effects of fish feed on changes in abundance of antibiotic resistance genes (ARGs) were investigated in South Korea via quantifying ARGs and analyzing physicochemical parameters in the influent (IN) and effluent before (BF) and 30 min after (AF) the fish feeding time of sixteen flow-through fish farms. The absolute abundance of ARGs in AF samples was 5 times higher than in BF and 12 times higher than in IN samples. Values of physicochemical parameters such as ammonia, total nitrogen, suspended solids and turbidity in the effluent significantly increased by 21.6, 4.2, 2.6 and 1.65 times, respectively, after fish feeding. Spatially, the fish farms on Jeju Island exhibited higher relative abundance (3.02 × 10-4 - 6.1 × 10-2) of ARGs compared to the farms in nearby Jeollanam-do (3.4 × 10-5 - 8.3 × 10-3). Seasonally, samples in summer and autumn showed a higher abundance of ARGs than in winter and spring. To assess risk to the food chain as well as public health, further studies are warranted to explore the pathogenic potential of these ARGs.

Removal efficiency of organic micropollutants in successive wastewater treatment steps in a full-scale wastewater treatment plant: Bench-scale application of tertiary treatment processes to improve removal of organic micropollutants persisting after secondary treatment.

The goal of this study was to investigate the occurrence and removal of 52 organic micropollutants (OMPs) during each wastewater treatment step in a full-scale wastewater treatment plant (WWTP). Pharmaceuticals such as metformin, acetaminophen, caffeine, ibuprofen, cimetidine and naproxen were found with high average concentrations in the influent. Most OMPs were not affected by the primary treatment (removal <10%), while secondary biological treatment contributed the most to overall removal of the OMPs. Among the three lanes of the secondary treatment of the WWTP, a combined anaerobic-anoxic-oxic process followed by a membrane bioreactor (A2O-MBR) process effectively facilitated removal of the OMPs (96%) using a different redox: a conventional activated sludge (CAS) process exhibited removal of approximately 85% of total concentrations of the OMPs, while a Modified Ludzack-Ettinger (MLE) process achieved approximately 92.1% removal. Removal of more than 50% of the concentrations of 13 targeted OMPs was observed in the secondary effluent (except for metformin showing only 3.8% removal) via adsorption with powdered activated carbon (PAC) as additional tertiary treatment. Metformin, inadequately removed by additional tertiary treatments, was effectively removed by a biological activated carbon (BAC) process, reaching a removal efficiency of 90.5%. To increase the removal of the amounts and types of OMPs with various physico-chemical properties, hybrid processes through a combination of diverse advanced treatment should be tailored to WWTPs.

Reduction of antibiotic resistome in influent of a wastewater treatment plant (WWTP) via a chemically enhanced primary treatment (CEPT) process.

Chemically enhanced primary treatment (CEPT) has been considered for maximizing wastewater energy recovery by enhancing the carbon captured through the primary treatment. However, evaluating the potential of CEPT as a primary treatment process for removing antibiotic resistance genes (ARGs) in the influent from a wastewater treatment plant (WWTP) has seldom been investigated. In this study, CEPT was conducted to assess simultaneous reduction of 13 major targeted ARGs and common pollutants in wastewater compared with primary sedimentation alone (non-CEPT). CEPT processes using three types of coagulants (PACl, FeCl3 and alum) effectively reduced absolute abundance of ARGs and intI1 in the influent from municipal WWTP. Average log-removal of absolute abundance of ARGs was achieved up to 1.77 ± 0.41 along with 90% turbidity reduction compared to non-CEPT. Through the simultaneous reduction of ARGs and intI1 genes during a CEPT process, ARGs proliferation may be limited directly through reduction of antibiotic resistant bacteria or indirectly through decreasing the possibility of horizontal gene transfer by intI1 removal. Reduction of ARGs and intI1 was improved by increasing coagulants' doses: abundances of residual ARGs under optimal dose conditions were similar, regardless of the different characteristics of coagulant types. The strongly positive correlation between reduction of turbidity/total phosphorus (T-P) and ARGs was explored, identifying that turbidity or T-P might be suitable indicators linked with variations in the abundance of ARGs during CEPT. As a result, CEPT may prove promising in efforts to control ARGs flowing into a WWTP.

Identification of primary effecters of N2O emissions from full-scale biological nitrogen removal systems using random forest approach.

Wastewater treatment plants (WWTPs) have long been recognized as point sources of N2O, a potent greenhouse gas and ozone-depleting agent. Multiple mechanisms, both biotic and abiotic, have been suggested to be responsible for N2O production from WWTPs, with basis on extrapolation from laboratory results and statistical analyses of metadata collected from operational full-scale plants. In this study, random forest (RF) analysis, a machine-learning approach for feature selection from highly multivariate datasets, was adopted to investigate N2O production mechanism in activated sludge tanks of WWTPs from a novel perspective. Standardized measurements of N2O effluxes coupled with exhaustive metadata collection were performed at activated sludge tanks of three biological nitrogen removal WWTPs at different times of the year. The multivariate datasets were used as inputs for RF analyses. Computation of the permutation variable importance measures returned biomass-normalized dissolved inorganic carbon concentration (DIC·VSS-1) and specific ammonia oxidation activity (sOURAOB) as the most influential parameters determining N2O emissions from the aerated zones (or phases) of activated sludge bioreactors. For the anoxic tanks, dissolved-organic-carbon-to-NO2-/NO3- ratio (DOC·(NO2--N + NO3--N)-1) was singled out as the most influential. These data analysis results clearly indicate disparate mechanisms for N2O generation in the oxic and anoxic activated sludge bioreactors, and provide evidences against significant contributions of N2O carryover across different zones or phases or niche-specific microbial reactions, with aerobic NH3/NH4+ oxidation to NO2- and anoxic denitrification predominantly responsible from aerated and anoxic zones or phases of activated sludge bioreactors, respectively.

Antibiotics in coastal aquaculture waters: Occurrence and elimination efficiency in oxidative water treatment processes.

The influents and effluents of coastal flow-through aquacultures in Korea were monitored for four selected antibiotics (amoxicillin-AMX, florfenicol-FLO, oxolinic acid-OXO, and oxytetracycline-OTC). A number of 177 samples were obtained from 16 aquaculture facilities for a monitoring period of two years. OTC was detected in 93 samples with a median concentration of 116 ng/L. OXO, FLO, and AMX were also detected in 36, 34, and 22 samples with median concentrations of 90, 44, and 63 ng/L, respectively. After antibiotics were applied to fish tanks, the aquaculture effluents were found to contain antibiotics up to several hundred µg/L, indicating that some control measures are required. Bench-scale experiments showed that chlorine and ozone fully eliminated AMX and OTC but not FLO at ≤2 mg/L of oxidant dosage. Reactive halogen species formed in the marine water matrix enhanced the antibiotic degradation. UV254 most effectively eliminated FLO, achieving 60-70 % elimination at 1000 mJ/cm2 of UV fluence. Sequential use of chlorine followed by UV254 demonstrated significant elimination of all four selected antibiotics. The obtained kinetic information for the reactions of these oxidants and UV with the antibiotics and marine aquaculture water constituents could be useful for designing and optimizing the aquaculture water treatment processes.

Additional reduction of antibiotic resistance genes and human bacterial pathogens via thermophilic aerobic digestion of anaerobically digested sludge.

Thermophilic aerobic digestion (TAD) was applied to further reduce ARGs and heavy metal resistance genes (HMRGs) as well as class 1 integrons (intI1) in sludge from anaerobic digestion (AnD). Unlike after AnD, there was no enrichment of ARGs, HMRGs and intI1 after TAD. Residual gene fractions of intI1 and total ARGs (sum of targeted ARGs) were 0.03 and 0.08, respectively. Two kinetic models (Collins-Selleck and first-order) described the decay patterns of targeted genes, revealing rapid removal of intI1 during TAD. After TAD, the relative abundance of human bacterial pathogens (HBPs) and the numbers of HBPs species decreased to approximately 68% and 64% compared to anaerobically digested sludge, respectively. Thus, TAD, subsequent to AnD, may possess high potential for reducing biological risks resulting from ARGs, HMRGs, intI1 and HBPs in sewage sludge.

Response of antibiotic and heavy metal resistance genes to two different temperature sequences in anaerobic digestion of waste activated sludge.

Response of antibiotic resistance genes (ARGs) and heavy metal resistance genes (HMRGs) to two different temperature sequences (i.e., mesophilic-thermophilic and thermophilic-mesophilic) were investigated. Higher removal of total ARGs (twenty-one targeted subtypes) and HMRGs (three targeted subtypes) was achieved by the mesophilic-thermophilic sequence than by the thermophilic-mesophilic sequence. The sequence of mesophilic-thermophilic showed the highest removal of total ARGs, but the sequence of thermophilic-mesophilic proved more suitable for removal of class 1 integrons (intI1). Correlation analysis indicated that intI1 correlated significantly with tetG, tetQ, tetX, sul2, aac(6')-lb-cr, blaTEM, ermB and floR. High-throughput sequencing revealed that the mesophilic-thermophilic sequence TPAD removed more human bacterial pathogens (HBPs) than did the thermophilic-mesophilic sequence. Also, significantly positive correlation was observed between ARGs and HBPs. For instance, Mycoplasma pneumonia showed significantly positive correlation with several ARGs including tetE, tetQ, tetX, tetZ, sul1, sul2, aac(6')-lb-cr and floR.

Prevalence of antibiotic resistance genes from effluent of coastal aquaculture, South Korea.

The wide use of antibiotics in aquaculture for prophylactic and therapeutic purposes can potentially lead to the prevalence of antibiotic resistance genes (ARGs). This study reports for the first time the profile of ARGs from effluents of coastal aquaculture located in South Jeolla province and Jeju Island, South Korea. Using quantitative PCR (qPCR), twenty-two ARGs encoding tetracycline resistance (tetA, tetB, tetD, tetE, tetG, tetH, tetM, tetQ, tetX, tetZ, tetBP), sulfonamide resistance (sul1, sul2), quinolone resistance (qnrD, qnrS, aac(6')-Ib-cr), ß-lactams resistance (blaTEM, blaCTX, blaSHV), macrolide resistance (ermC), florfenicol resistance (floR) and multidrug resistance (oqxA) and a class 1 integrons-integrase gene (intI1) were quantified. In addition, Illumina Miseq sequencing was applied to investigate microbial community differences across fish farm effluents. Results from qPCR showed that the total number of detected ARGs ranged from 4.24 × 10-3 to 1.46 × 10-2 copies/16S rRNA gene. Among them, tetB and tetD were predominant, accounting for 74.8%-98.0% of the total ARGs. Furthermore, intI1 gene showed positive correlation with tetB, tetD, tetE, tetH, tetX, tetZ tetQ and sul1. Microbial community analysis revealed potential host bacteria for ARGs and intI1. Two genera, Vibrio and Marinomonas belonging to Gammaproteobacteria, showed significant correlation with tetB and tetD, the most dominant ARGs in all samples. Also, operational taxonomic units (OTUs)-based network analysis revealed that ten OTUs, classified into the phyla Proteobacteria, Cyanobacteria/Chloroplast, Bacteroidetes, Verrucomicrobia and an unclassified phylum, were potential hosts of tetracycline resistance genes (i.e., tetA, tetG, tetH, tetM, tetQ and tetZ). Further systematic monitoring of ARGs is warranted for risk assessment and management of antibacterial resistance from fish farm effluents.

Use of a filtering process to remove solid waste and antibiotic resistance genes from effluent of a flow-through fish farm.

The objective of this study was to investigate reduction in antibiotic resistance genes (ARGs) via targeting solid waste in effluent from a flow-through aquaculture in South Korea. The level of suspended solids in the filtrates was approximately 12.5±2.3mg/L, corresponding to a removal efficiency of 68.8±5.7% irrespective of variations in the size of the filter pores. The total number of particles in the effluent was reduced to the lowest numbers of particles using a filter pore size of 25µm, corresponding to a removal efficiency of 40.3%. Among the 23 ARGs conferring resistance to tetracyclines, beta-lactam antibiotics, sulfonamides, quinolones, macrolides, florfenicol and multidrug, tetracycline resistance genes were the most prevalent with a relative abundance of 67.5%. Of eleven tetracycline resistance genes (tetA, tetB, tetD, tetE, tetG, tetH, tetM, tetQ, tetX, tetZ, tetB/P) analyzed, the relative abundance of tetG was the highest in the effluent. The removal efficiency of the total number of particles showed similar patterns to the removal efficiency of ARGs depending on the size of the filter pores. Levels of ARGs in the filtrates were reduced to approximately 60.5% of those of the ARGs in the effluents. With a filter pore size of 25µm, a maximum removal efficiency of 66.0% was achieved. In particular, the relative abundance of detected tetracycline resistance genes decreased only after passing through the filters, perhaps reflecting the presence of high quantities of tetracycline resistance genes in particles from the fish farm. Using Illumina sequencing based on a 16S rRNA gene, the dominant phyla were found to be Bacteroidetes, Proteobacteria, Planctomycetes and Verrucomicrobia in the effluent. Although the overall composition of the bacterial communities was not significantly changed via filtering tests, only the relative abundance of Bacteroidetes and Proteobacteria was changed. These results demonstrate that a filtering process in aquaculture facilities can be used to reduce solid waste as well as ARGs from aquaculture farms.

Fate of antibiotic resistance genes in mesophilic and thermophilic anaerobic digestion of chemically enhanced primary treatment (CEPT) sludge.

Anaerobic digestion (AD) of chemically enhanced primary treatment (CEPT) sludge and non-CEPT (conventional sedimentation) sludge were comparatively operated under mesophilic and thermophilic conditions. The highest methane yield (692.46±0.46mL CH4/g VSremoved in CEPT sludge) was observed in mesophilic AD of CEPT sludge. Meanwhile, thermophilic conditions were more favorable for the removal of total antibiotic resistance genes (ARGs). In this study, no measurable difference in the fates and removal of ARGs and class 1 integrin-integrase gene (intI1) was observed between treated non-CEPT and CEPT sludge. However, redundancy analysis indicated that shifts in bacterial community were primarily accountable for the variations in ARGs and intI1. Network analysis further revealed potential host bacteria for ARGs and intI1.

Biomethanation of Sewage Sludge with Food Waste Leachate Via Co-Digestion.

Anaerobic mono- and co-digestion of sewage sludge and food waste leachate (FWL) were performed by assessing methane production and characterizing microbial communities. Anaerobic digestion (AD) of waste activated sludge (WAS) alone produced the lowest methane (281 ml CH4), but an approximately 80% increase in methane production was achieved via co-digestion of WAS and FWL (506 ml CH4). There were less differences in the diversity of bacterial communities in anaerobic digesters, while archaeal (ARC) and bacterial (BAC) amounts reflected AD performance. Compared with the total ARC and BAC amounts in the mono-digestion of WAS, the ARC and BAC amounts increased two and three times, respectively, during co-digestion of FWL and WAS. In characterized archaeal communities, the dominant ratio of hydrogenotrophic methanogens in the mono-digestion of WAS approached nearly a 1:1 ratio of the two acetoclastic and hydrogenotrophic methanogens in the co-digestion of FWL and WAS. The ARC/BAC ratio in the digesters varied in the range of 5.9% to 9.1%, indicating a positive correlation with the methane production of AD.

Hybrid selective noncatalytic reduction (SNCR)/selective catalytic reduction (SCR) for NOx removal using low-temperature SCR with Mn-V2O5/TiO2 catalyst.

UNLABELLED: A hybrid selective noncatalytic reduction/selective catalytic reduction (SNCR/SCR) system that uses two types of technology, low-temperature SCR process and SNCR process, was designed to develop nitrogen oxide (NOx) reduction technology. SCR was conducted with space velocity (SV)=2400 hr(-1) and hybrid SNCR/SCR with SV=6000 hr(-1), since the study focused on reducing the amount of catalyst and both achieved 98% NOx reduction efficiency. Characteristics of NOx reduction by NH3 were studied for low-temperature SCR system at 150 °C using Mn-V2O5/TiO2 catalyst. Mn-added V2O5/TiO2 catalyst was produced, and selective catalyst reduction of NOx by NH3 was experimented. NOx reduction rate according to added Mn content in Mn-V2O5/TiO2 catalyst was studied with varying conditions of reaction temperature, normalized stoichiometric ratio (NSR), SV, and O2 concentration. In the catalyst experiment according to V2O5 concentration, 1 wt.% V2O5 catalyst showed the highest NOx reduction rate: 98% reduction at temperature window of 200~250 °C. As a promoter of the V2O5 catalyst, 5 wt.% Mn was added, and the catalyst showed 47~90% higher efficiency even with low temperatures, 100~200 °C. Mn-V2O5/TiO2 catalyst, prepared by adding 5 wt.% Mn in V2O5/TiO2 catalyst, showed increments of catalyst activation at 150 °C as well as NOx reduction. Mn-V2O5/TiO2 catalyst showed 8% higher rate for NOx reduction compared with V2O5/TiO2 catalyst in 150 °C SCR. Thus, (5 wt.%)Mn-(1 wt.%)V2O5/TiO2 catalyst was applied in SCR of hybrid SNCR/SCR system of low temperature at 150 °C. Low-temperature SCR hybrid SNCR/SCR (150 °C) system and hybrid SNCR/SCR (350 °C) showed 91~95% total reduction rate with conditions of SV=2400~6000 hr(-1) SCR and 850~1050 °C SNCR, NSR=1.5~2.0, and 5% O2. Hybrid SNCR/SCR (150 °C) system proved to be more effective than the hybrid SNCR/SCR (350 °C) system at low temperature. IMPLICATIONS: NOx control is very important, since they are the part of greenhouse gases as well as the cause of acid rain and ozone hole. A technology, so-called hybrid SNCR/SCR process, was tested using Mn-V2O5/TiO2 monolithic catalyst for NOx reduction, and the method is promising. The results of this study would provide some ideas to parties such as policy makers, environmental engineers, and so on.

Cloning of the xylose reductase gene of Candida milleri.

The entire nucleotide sequence of the xylose reductase (XR) gene in Candida milleri CBS8195 sourdough yeast was determined by degenerate polymerase chain reaction (PCR) and genome walking. The sequence analysis revealed an open-reading frame of 981 bp that encoded 326 amino acids with a predicted molecular mass of 36.7 kDa. The deduced amino acid sequence of XR of C. milleri was 64.7% homologous to that of Kluyveromyces lactis. The cloned XR gene was expressed in Saccharomyces cerevisiae, and the resulting recombinant S. cerevisiae strain produced xylitol from xylose, indicating that the C. milleri XR introduced into S. cerevisiae is functional. An enzymatic activity assay and semiquantitative reverse transcription-PCR revealed that the expression of CmXR was induced by xylose.

Production, purification, and characterization of a novel thermostable serine protease from soil isolate, Streptomyces tendae.

An isolate of Streptomyces tendae produced a extracellular protease which was purified to apparent homogeneity giving a single band on SDS-PAGE with a molecular mass of 21 kDa. Optimum activity was at 70 degrees C and pH 6. It was stable at 55 degrees C for 30 min and between pH 4 and 9. It was resistant to neutral detergents and organic solvents such as Triton X-100, Tween 80, methanol, ethanol, acetone, and 2-propanol at 5% (v/v). The enzyme was completely inhibited by 5 mM PMSF, indicating it to be a serine protease. N-terminal amino acid sequence did not show any homology with other known proteolytic enzymes. The protease may therefore be a novel neutral serine protease, which is stable at high temperature and over a broad range of pH.