[Impacts of F/M Ratio on Microbial Networks in Activated Sludge].

The performance of wastewater treatment plants (WWTPs) mainly relies on the microbial community in activated sludge (AS). The food to microorganisms (F/M) ratio is an important operational parameter, the recommended value of which is between 0.2 and 0.5 kg·(kg·d)-1. The F/M directly influences microbial growth and metabolism, but how the F/M ratio affects AS microbial networks and the related mechanisms remain unclear. In this study, 63 AS samples with different F/M ratios were analyzed by utilizing a random-matrix-theory based network pipeline. Firstly, all 63 samples were divided into three groups based on their F/M ratios (lower F/M group, normal F/M group, and higher F/M group). The results indicated that the lower F/M decreased microbial diversity and evenness significantly, but the higher F/M had no significant effects on the diversity of the microbial community. All three constructed networks were scale-free, small world, and modular, but network size and complexity were decreased in the lower and higher F/M groups. The network of the normal F/M ratio group was detected with the most nodes, the highest average clustering coefficient, and the shortest geodesic distance. The proportion of positive links was 76.8% when the F/M was within the normal range. On the contrary, 71.1% and 60.0% of positive links were identified when the F/M was below 0.2 and above 0.5 kg·(kg·d)-1. Moreover, 24 keystones were detected in the normal F/M network, whereas only 4 and 7 keystones were detected in the lower and higher F/M networks. Overall, our results provided clear evidence that the AS microbial community could be more stable and resilient to environmental disturbance when the F/M ratio was between 0.2 and 0.5 kg·(kg·d)-1.

[Potential Source Environments for Microbial Communities in Wastewater Treatment Plants (WWTPs) in China].

To better understanding the microbial composition of activated sludge (AS), unique OTUs and their indicators were examined and SourceTracker was used to identify possible source environments for AS communities to thrive in 15 cities across China, based on 16S rRNA gene sequencing. This may help reveal the underlying mechanisms of AS community composition formations in various areas. The results indicated that the dominant phyla in AS in each city were similar; Proteobacteria, Bacteroidetes, and Chloroflexi made up 94.53%-97.04% of the whole microbial communities. Geographic location and environmental conditions were identified as having an effect on microbial compositions. Cities located toward to the edge of China or along the coast tended to harbor more unique OTUs. Moreover, the composition of indicators in wastewater treatment plants (WWTPs) near the sea were more similar, while no indicators were detected in WWTPs in Chengdu and Chongqing. Source environments for AS microbial communities also depended on location and environmental conditions. Overall, microbes from 'plant' and 'soil' sources accounted for a large proportion of the AS microbial communities in each city, while bacteria from 'marine' sources were only detected in coastal AS communities. Cities with more rivers and lakes tended to have more bacteria originating from 'fresh water' sources. Since most of the WWTPs in China play a role in treating industrial wastewater, industrial wastewater was also an important source environment. However, few bacteria originating from the human digestive system were identified in the AS communities. Moreover, a large proportion of bacteria (36%-70%) originated from unknown sources not included in the source database, such as sewage infrastructure, indicating that the source database will need to be expanded in the future.

[Comparison of Polycaprolactone and Poly-3-hydroxybutyrate-co-3-hydroxyvalerate for Nitrogen Removal].

Two types of biodegradable polymers, polycaprolactone (PCL) and poly-3-hydroxybutyrate-co-3-hydroxyvalerate (PHBV), were used as a denitrification slow-release carbon source and a microbial carrier. By comprehensively comparing their performances in denitrification, carbon release, surface morphology, and material composition as well as their microbial community characteristics, the PHBV was determined as the better performer. It had a shorter denitrification start time, a higher denitrification rate, a lower residual organic matter concentration, and a more stable and sustained denitrification performance than PCL. This is because its surface was rough and contained large amounts of hydrophilic groups such as C-O and C=O, which is easily attached and degraded by microorganisms. As a result, the microorganisms on its surface were diverse. The dominant ones were identified with heterotrophic denitrification potentials, such as Thiothrix, Pseudomonas, Zoogloea, Flavobacterium, and Dechloromonas. Therefore, PHBV is suitable as a carbon source medium for tertiary nitrogen removal.

[Characteristics and Regional Heterogeneity of Eukaryotic Microbial Community in Wastewater Treatment Plants].

Wastewater treatment plants (WWTPs) are important facilities to control water pollution and ensure the sustainable development of cities and humans. As an indispensable part of the activated sludge (AS) system, eukaryotic microbes play important roles in indicating the properties of AS, predicting the quality of the effluent, enhancing the purification effect, and ensuring a stable performance of the system in WWTPs. In this study, 61 AS samples from 14 full-scale WWTPs of Beijing, Shenzhen, and Wuxi were collected. Characteristics and regional heterogeneity of eukaryotic microbial community were elucidated via high-throughput sequencing (HTS) of the 18S rDNA and multi ecological and statistical methods. Results showed that eukaryotic microbial communities in different scales shared similar main members, which were mainly composed of fungi, ciliophora, and metazoa in division level with their total relative abundance up to 86.22%-89.40%. Diversity of eukaryotic microbial community in WWTPs of different cites varied. Richness and Shannon Wiener index of eukaryotic microbial communities in the AS system of Wuxi were the highest, while that of Beijing was the lowest. Diversity of eukaryotic microbes from HTS in this study was higher than that of conventional methods, but lower than the diversity of bacteria in AS systems. Regional heterogeneity of eukaryotic microbial community structure was uncovered by nonmetric multidimensional scaling based on Bray-Curtis distance and dissimilarity analysis. Results of partial mantel test and multiple regression matrix analysis showed that the eukaryotic microbial community was significantly correlated with the temperature of the aeration tank mixture and the total nitrogen concentration of the effluent of the AS system. These results help deepen the understanding of eukaryotic microbes in WWTPs.

[Dynamics of quickly absorption of the carbon source in wastewater by activated sludge].

In this paper, absorption characteristics of organic matter in municipal wastewater by three kinds of activated sludge (carbon-enriching, nitrification and denitrification sludge) were studied, and the absorption kinetic data was checked using three kinds of absorption kinetic equations based on Ritchie rate equation. The objectives of this study were to investigate the absorption mechanism of activated sludge to organic matter in municipal wastewater, and to identify the possibility of reclaiming organic matter by activated sludge. Results indicated that in the early 30 min, absorption process of organic matter by activated sludge was found to be mainly physical adsorption, which could be expressed by the Lagergren single-layer adsorption model. The carbon-enriching sludge had the highest adsorption capacity (COD/SS) which was 60 mg/g but the adsorption rate was lower than that of denitrification sludge. While nitrification sludge had the lowest adsorption rate and higher adsorption capacity compared with denitrification sludge, which was about 35 mg/g. The rates of the fitting index theta(0) of carbon-enriching, nitrification and denitrification sludge were 0.284, 0.777 and 0.923, respectively, which indicated that the sorbed organic matter on the surface of carbon-enriching sludge was the easiest fraction to be washed away. That is, the combination intensity of carbon-enriching sludge and organic matter was the feeblest, which was convenient for carbon-enriching sludge to release sorbed carbon. Furthermore, by fitting with Langmuir model, concentration of organic matter was found to be the key parameter influencing the adsorption capacity of activated sludge, while the influence of temperature was not obvious. The kinetic law of organic matter absorption by activated sludge was developed, which introduces a way to kinetically analyze the removing mechanism of pollutant by activated sludge and provides theoretical base for the reclaiming of nutriments in wastewater by the absorption of activated sludge.

[Determination of kinetic parameters of activated sludge in an MBR wastewater treatment plant].

One of the main problems in application of MBR technology is lack of reliable kinetic parameters for process design. The activated sludge in the aerobic tank of an MBR municipal wastewater treatment plant was investigated therefore. Using oxygen utilization rate method, following kinetic parameters were measured: heterotrophic yield coefficient YH = 0.693, autotrophic yield coefficient Y(A) = 0.263, heterotrophic decay coefficient KdH, = 0.108 d(-1), autotrophic yield coefficient KdA = 0.089 d(-1), specific maximum COD removal rate v(mS), = 1.94 mg x (mg x d)(-1), half saturation constant for COD removal K(S) = 34.6 mg x L(-1), specific maximum ammonia removal rate vmN = 0.18 mg x (mg x d)(-1), half saturation constant for ammonia removal K(N) = 1.06 mg x L(-1). Compared with the normal reported data of conventional activated sludge process (CAS), Y(H), KdH obtained are higher and vmS, vmN are lower. The high sludge concentration condition of MBR process may account for those differences.

[Harvest of the carbon source in wastewater by the adsorption and desorption of activated sludge].

The carbon source in municipal wastewater was adsorbed by activated sludge and then harvested through the hydrolysis of activated sludge. Results indicated that activated sludge had high absorbing ability towards organic carbon and phosphorus under continuous operation mode, and the average COD and TP absorption rate reached as high as 63% and 76%, respectively. Moreover, about 50% of the soluble carbon source was outside of the sludge cell and could be released under mild hydrolysis condition. Whereas the absorbed amount of nitrogen was relatively low, and the removal rate of ammonia was only 13% . Furthermore, the releases of organic carbon, nitrogen and phosphorus from the sludge absorbing pollutants in the wastewater were studied. By comparing different hydrolysis conditions of normal (pH 7.5, 20 degrees C), heating (pH 7.5, 60 degrees C) and the alkaline heating (pH 11, 60 degrees C), the last one presented the optimum hydrolysis efficiency. Under which, the release rate of COD could reach 320 mg/g after 24 hours, whereas nitrogen and phosphorus just obtained low release rates of 18 mg/g and 2 mg/g, respectively. Results indicate that the carbon source in wastewater could be harvested by the adsorption and desorption of activated sludge, and the concentrations of nitrogen and phosphorus are low and would not influence the reuse of the harvested carbon source.

[Quickly enrichment of carbon in wastewater by activated sludge].

Pilot tests were carried out to investigate the absorption characteristics of the carbon source in urban wastewater by activated sludge and to analyze the carbon release from the carbon absorbed activated sludge in the settling process. The results indicated that carbon in wastewater could be quickly enriched by activated sludge. The absorption process of indissolvable organic matter could be finished as shortly as less than 10 min, while the absorption process of the dissolved organic matter was relatively slow and should consume up about 30 min. Moreover, carbon release was observed in the settling process of enriched sludge. In the period of 30-100 min, the release amount of total COD (TCOD) was 11.44 mg x g(-1), while in the period of 60-150 min, the release amount of dissolved COD (SCOD) was 6.24 mg x g(-1). Furthermore, based on the results of the bench-scale tests, a pilot-scale plant was built to investigate the absorption of carbon, nitrogen and phosphorus by activated sludge and the settleability of enriched sludge. The results indicated that under continuously operation mode, 60% of COD, 75% of TP and 10% of TN in the wastewater could be removed by the absorption of activated sludge, and the enriched sludge with SVI of 34.2 mL x g(-1) presented good settleability. Carbon enrichment by activated sludge could not only reclaim the carbon source in wastewater, but also reduce the loading of organic matter and give low C/N for the following nitrification unit and improving the nitrification efficiency.

[Bacterial community dynamics in wastewater treatment plants based on T-RFLP profiles].

Bacterial communities in two full-scale wastewater treatment systems (system A and system B) with functional stability were monitored over 105 days using terminal restriction fragment length polymorphism (T-RFLP) based on 16S rRNA genes. During the study period, system A and system B were functionally stable. The effluent BOD concentration of two systems was always around 8 mg/L, and at the same time the effluent TN concentrations of two systems were always below 25 mg/L and 23 mg/L respectively. However, the T-RFLP profiles showed that for system A, the relative abundance of T-RF of 115, 118, 123, 471, 482 and 835 bp changed significantly. For system B, the T-RF of 123, 471, 750 and 835 bp changed significantly. The moving window analysis showed the average community change rates (every 15 days) of system A and system B were 21.5% +/- 5.7% and 22.6% +/- 5.0% respectively. Based on the Lorenz curves, it was observed for system A that 20% of the species corresponded with 43% to 50% of the cumulative relative abundance. For system B, 20% of the species corresponded with 47% to 52% of the cumulative relative abundance. In conclusion, the results of this study clearly revealed that, in the full-scale wastewater treatment systems, the functional stability did not correlate to the stable bacterial communities.

[Immobilization of lignin peroxidase on spherical mesoporous material].

The spherical mesoporous particles with two-dimensional (2D) hexagonal mesopores in diameter up to 11.6 nm was fabricated in acetic acid/sodium buffer solution (pH = 3.5) by using tetramethoxysilane (TMOS) as silica source, Pluronic P123 as template and 1,3,5-triisopropylbenzene (TIPB) as swelling agent. Then the mesoporous particles were employed as carriers for the immobilization of lignin peroxidase (LiP). The effect of immobilization time, the amount of added enzyme on the immobilized enzyme amount and activities were investigated. The characteristic and stability of immobilized LiP were also studied. The results showed that, as the mass ratio of enzyme (E) and mesoporous material (MS) was 76.8 mg/g,immobilizing time was 12 h, the largest immobilized enzyme amount (8.87 mg/g) and highest apparent activity (41.45 U/mg) of immobilization LiP were achieved. Comparing with free LiP, the optimum pH and temperature of the immobilized LiP were almost the same, while whose pH stability and thermal stability were significantly improved. No obvious activity loss was observed for the immobilized LiP after 7 weeks storage at 4 degrees C. After 6 times of usage, almost 30% of the initial activity could still remain.

[Community structures of ammonia-oxidizing bacteria in activated sludge of wastewater treatment plants].

In order to investigate the community structure of ammonia oxidizing bacteria (AOB) in wastewater treatment plants and the influence of treatment process and system scale on AOB community. We investigated AOB communities in activated sludges collected from 9 wastewater treatment systems, using specific PCR followed by terminal restriction fragment length polymorphism (T-RFLP), cloning, and sequencing of amoA genes. The T-RFLP fingerprints analysis shows that there are some differences among the AOB community structures from different wastewater treatment systems, and the dominant terminal restriction fragments (T-RFs) are 354, 491 and 291 bp. The T-RFLP profiles and cluster analysis may also indicate the AOB structures are slightly affected by the treatment process, while they are influenced by the system scale to some extent. Phylogenetic analysis of cloned amoA gene shows clearly that all the dominant AOB in the systems were Nitrosomonas spp., not Nitrosospira spp.. The reason may be explained as that Nitrosomonas spp. have higher micromax, than Nitrosomonas spp.. This growth advantage may favor the Nitrosomonas spp. rather than Nitrosomonas spp. being prevail in activated sludge.

[Degradation of acid blue 45 in a white-rot fungi reactor operated under non-sterile conditions].

To operate the white-rot fungal reactor under non-sterile conditions has important significance for its practical applications. Ozone (0.0144 mg/min) was selected as the bactericide to control the bacterial contamination in a white-rot fungal reactor. The control effect on bacteria, enzyme production, pH variation and decolorization of a recalcitrant dye (acid blue 45) in the reactor were studied during continuously operating the reactor. It was found that, the contamination bacteria were controlled at under a number of 1 x 10(5)-1 x 10(6) CFU/mL and the inactivation rate of contaminating bacteria in white-rot fungal reactor was close to 99%. The Phanerochaete chrysosporium continuously produced MnP with the maximum residual activity of 50 U/L and degraded the acid blue 45 with a rate of 40%-80%. At around pH 6, the system still had the capability to continuously produce MnP and degrade the acid blue 45. To keep the growth of white-rot fungi and production of MnP for a long-term operation is a next step point to be solved.

[Effect of ozone on membrane fouling in water and wastewater treatment: a research review].

As a high efficient water and wastewater treatment technology, membrane filtration has been mainly used in wastewater treatment as membrane bioreactor, in reclaiming secondary effluent,treating surface water and potable water, and etc. Membrane fouling is a main obstacle to the wide application of membrane technology. Ozone has strong oxidizing power and has been utilized widely in water and wastewater treatment. In recent years, researches on combined process of ozone-membrane filtration are increasing. This paper does reviews and analysis of these researches. It is noticed that there has been a few of researches on the ozone treatment plus MBR process. Pre-ozonation of feed to MBR and slight ozonation of the mixed liquid in MBR may be used to relieve membrane fouling.Combined processes of ozone-membrane filtration can be divided into three classes in terms of the function of ozone and the system configuration: (1) cleaning the fouled membrane with ozone; (2) separate ozone-membrane filtration process; (3) integrated ozone-membrane filtration process. Although most reports supported that ozonation can control membrane fouling development,there were contrary results. At present, researches on the mechanisms of ozone's effect on membrane fouling control concentrated on the change of organic composition of the filtration influent under ozonation, however, particulate substances, microbial and inorganic substances may also be affected and then play roles in membrane fouling, depending on source water quality and process configuration. Moreover, there have not been common parameters to evaluate the ozone diffusion equipment and efficiency. The authors suggest that further researches should emphasize on integrated ozone-membrane process, and more attention should be paid to the cost-effectiveness of the combined process.

[Evaluation of estrogenicity of sewage samples from Beijing, China].

The recombinant yeast estrogen screen (YES) was used to evaluate the estrogenic activities of sewage samples from every step of three sewage treatment plants (STPs) in Beijing, China. Meanwhile, eight target endocrine-disrupting compounds (EDCs) in corresponding sewage samples were simultaneously analyzed by GC/MS system to further elucidate the variations of their estrogenic activities. The results indicated that estrogenic activities could be considerably reduced by STPs with the rate of 82.2%-97.0%. However,they were still detected in the effluent samples with the values of EEQs (expressed as E2 equivalents) 2.6-16.0 ng/L. In addition,STPs could not completely remove the target EDCs according to GC/MS analysis. The average concentration of 17alpha-E2 in the effluent was the lowest with the value of 13.5 ng/L, and the concentration of BPA (106.4 ng/L) was the largest. The estrogenic activities of effluent samples were mainly composed of steroid estrogens. Consequently, potential environmental risk can be caused by the discharge of effluents from STPs. How to effectively remove steroid estrogens during sewage treatment is, therefore, of a major concern.

[Kinetics model for batch culture of white rot fungus].

In order to understand ligninolytic enzymes production process during culture of white rot fungus, accordingly to direct the design of fermentation process, a kinetics model was built for the batch culture of Phanerochaete chrysosporium. The parameters in the model were calibrated based on the experimental data from free and immobilized culture separately. The difference between each variable's values calculated based on kinetics model and experimental data is within 15%. Comparing parameters for the free and the immobilized culture, it is found that maximum biomass concentrations are both 1.78 g/L; growth rate ratio of immobilized culture (0.6683 d(-1)) is larger than that of free culture (0.5144 d(-1)); very little glucose is consumed for biomass growth in free culture while in immobilized culture much glucose is used and ammonium nitrogen is consumed at a greater rate. Ligninolytic enzymes production process is non-growth related; fungal pellets can produce MnP (231 U/L) in free culture with a production rate of 115.8 U x (g x d)(-1) before peak and 26.1 U x (g x d)(-1) after peak, thus fed-batch is a possible mode to improve MnP production and fermentation efficiency. MnP (410 U/L) and LiP (721 U/L) can be produced in immobilized culture, but MnP and LiP production rate decrease from 80.1 U x (g x d)(-1) and 248.9 U x (g x d)(-1) to 6.04 U x (g x d)(-1) and 0 U x (g x d)(-1), respectively, indicating a proper feed moment is before the enzymes peak during fed-batch culture.

[White rot fungi biofilter treating waste gas containing chlorobenzene].

A novel gas-solid phase bioreactor, using bamboo as support material, inoculated with white rot fungi Phanerochaete chrysosporium was established to treat waste gas containing chlorobenzene. The performance of P. chrysosporium bioreactor was examined under different conditions. Results showed that the maximum removal efficiency of nearly 80% (average removal efficiency of 50%) could be reached under the condition of chlorobenzene inlet concentration of 200-1500 mg/m3 and the empty bed retention time (EBRT) of 122 s. While the maximum chlorobenzene removal rate of 94 g/(m3 x h) [average removal efficiency o f 60 g/(m3 x h)] had been achieved within chlorobenzene inlet concentration of 500- 1500 mg/m3 and at the flow rate of 0.5 m3/h. Furthermore, the removal rates of the bioreactor at different flow rates were also examined, suggesting that the response of removal rate to the change of inlet loading was dependent on the flow rate. Lower flow rate could promote the extent of removal rate enhancement compared to the higher flow rate. Moreover, the profile of chlorobenzene concentration along the height of the biofilter showed a nonlinear decrease trend.

Biodegradation of gaseous chlorobenzene by white-rot fungus Phanerochaete chrysosporium.

OBJECTIVE: To evaluate the effect of white rot fungus Phanerochaete chrysosporium on removal of gaseous chlorobenzene. METHODS: Fungal mycelium mixed with a liquid medium was placed into airtight bottles. A certain amount of chlorobenzene was injected into the headspace of the bottles under different conditions. At a certain interval, the concentrations in the headspace were analyzed to evaluate the degradation of chlorobenzene by P. chrysosporium. RESULTS: The degradation effects of P. chrysosporium on chlorobenzene under different conditions were investigated. The difference in the optimum temperature for the growth of the fungi and chlorobenzene degradation was observed. The data indicated that a lower temperature (28 degrees C) would promote the degradation of chlorobenzene than the optimum temperature for the growth of the fungi (37 degrees C). A low nitrogen source concentration (30 mg N/L) had a better effect on degrading chlorobenzene than a high nitrogen source concentration (higher than 100 mg N/L). A high initial concentration (over 1100 mg/m3) of chlorobenzene showed an inhibiting effect on degradation by P. chrysosporium. A maximum removal efficiency of 95% was achieved at the initial concentration of 550 mg/m3. CONCLUSION: P. chrysosporium has a rather good ability to remove gaseous chlorobenzene. A low nitrogen source concentration and a low temperature promote the removal of chlorobenzene by P. chrysosporium. However, a high initial chlorobenzene concentration can inhibit chlorobenzene degradation.

[Effect of trace element on the growth of white rot fungus and suppressing yeast in liquid medium].

Growth of white rot fungus in liquid medium with different trace element and effect of these medium on suppressing the growth of yeast under non-sterile were investigated in agitated Erlenmeyer flasks. Results showed that the size and amount of mycelium pellets in liquid medium with trace element and Fe 3.5 mg/L was more than that with trace element but no iron and without any trace element under sterile condition; moreover, the size and amount of mycelium pellets increased with the rise of the concentration of trace element and iron. In addition, if applying the mode of incubating under sterile condition and degrading under non-sterile condition, the decolorization of reactive brilliant red K-2BP in liquid medium without any trace element and with trace element but no iron was similar to that of degrading under sterile condition; but the decolorization in liquid medium with trace element and Fe 3.5 mg/L was less than the results under sterile condition. Through observing under microscope, it was found that the amount of yeast in liquid medium with Fe 3.5 mg/L was more than the medium without any trace element and with trace element but no iron. Therefore, trace element, especially iron, plays a very important role in suppressing yeast in liquid medium; furthermore, iron also is necessary to the growth of whit rot fungus. So that, to control the concentration of iron in liquid medium could reduce the invasion of yeast under non-sterile condition as long as it does influence the growth of white rot fungus.

Effect of a suspended carrier on membrane fouling in a submerged membrane bioreactor.

In an attempt at membrane fouling control, a kind of cylindrical plastic suspended carrier was added in a submerged membrane bioreactor (SMBR) and its effect was investigated in this study. According to the transmembrane pressure (TMP) profiles and the sludge characteristics in comparative runs with and without suspended carriers, it was found that the suspended carriers added in SMBR had two effects on membrane fouling: one was the positive effect of mechanically scouring the membrane surface and the other was the negative effect of breaking up sludge flocs. Sludge particle size distribution change was mainly responsible. It was suggested to apply the suspended carrier at higher MLSS concentration and lower carrier dose based on the consideration for retarding sludge breakage caused by the carrier. The experiment was conducted under higher MLSS (8 gL(-1)) and lower carrier dose (carrier volume/total volume = 10/). The TMP increase was effectively retarded by added suspended carriers compared to the system without addition of the carriers. The effect of suspended carriers on membrane fouling at high MLSS concentration was verified.

[Production of ligninolytic enzymes in bioreactor].

Production of ligninolytic enzymes under nitrogen limited conditions(C/N = 56/2.2) was studied in a 5-L stirred tank bioreactor with a working volume of 2 L for obtaining higher production of ligninolytic enzymes by white rot fungus Phanerochaete chrysosporium BKM-F-1767 and its control strategy. Results show that the manganese peroxidase (MnP) and laccase (Lac) reached peak at the sixth day and the seventh day, respectively, and the variation of them with time in a batch cultivation are similar to the results by agitated Erlenmeyer flasks; however higher enzyme activity was not achieved by applying a fed-batch strategy, in which nitrogen limited medium was fed to the reactor. In addition, variation of pH during cultivation was related to the growth of P. chrysosporium and enzymes production during both batch and fed-batch cultivation. The pH value of liquid medium began to decline when the enzyme activity occurred in the system, and the decline became more and more slow along with the decrease of enzyme activity at the end of fermentation. So, pH would be as a control parameter to find out the growth of P. chrysosporium and enzymes production during incubating P. chrysosporium. However, fed-batch strategy still need further study.