First report of toxic Cylindrospermopsis raciborskii and Raphidiopsis mediterranea (Cyanoprokaryota) in Egyptian fresh waters

Cylindrospermopsis raciborskii, a potentially toxic and highly adaptable freshwater cyanobacterium, was believed to have been misidentified in the Nile at the end of the 19th century. This study reports the presence of Cylindrospermopsis raciborskii and Raphidiopsis mediterranea for the first time in Egyptian fresh waters since that time. Cylindrospermopsis raciborskii appeared in the El-Dowyrat fish pond during May 2002, when bottom waters reached, as a result of climatic change, sufficiently high temperatures to allow the germination of its akinetes in the sediments. Both C. raciborskii and R. mediterranea showed seasonal variations, with highest densities recorded in August of each year. The count of the two species correlated positively with pH, temperature and conductance, and negatively with nutrients, during the study period. The densities of C. raciborskii and R. mediterranea varied significantly along the depth profile of this pond, with peaks obtained at 1 and 0.5m, respectively. Isolates of C. raciborskii and R. mediterranea from this pond exhibited toxicity to Artemia salina, Daphnia magna and mice. Cylindrospermopsis raciborskii extracts had hepatotoxic effects on mice, but R. mediterranea extracts showed neurotoxic effects on mice. The identification of toxic C. raciborskii and R. mediterranea in this pond should be considered during the monitoring of cyanobacteria in drinking and recreational water sources in Egypt.     

Temporal and spatial distribution of planktic cyanobacteria in Lake Kastoria, Greece, a shallow, urban lake

The temporal and spatial distribution of planktic cyanobacteria and some environmental parameters were studied in the shallow, urban Lake Kastoria, Greece from June 1996 to June 1997. Water temperature varied from 6–27 °C, pH from 7.5–8.9 and dissolved O₂ concentration from 0.7–12 mg m^-3 10^-3. The mean annual Chl a concentration was 83 mg Chl a m^-3 indicative of the eutrophic-hypertrophic state of the lake. Cyanobacterial biomass ranged from 11–238 g FW m^-3, constituting about 90% of the total phytoplankton biomass throughout the year. Cyanobacterial biomass was non-uniformly distributed both vertically and horizontally from August to November 1996 and resulted mainly from the distribution of Microcystis. Seven cyanobacterial taxa were reported for the first time in Lake Kastoria. Six taxa were dominant: Microcystis aeruginosa, M. flos-aquae, M. novacekii. Limnothrix redekei, Anabaena sp. and Cylindrospermopsis raciborskii. The dominant cyanobacterial taxa can be grouped on the basis of their distribution patterns (1) Microcystis species: maximum biomass occurring at pH > 8, temperature 12–17 °C, depth < 0.2 m; (2) Anabaena sp. and Cylindrospermopsis raciborskii : maximum biomass at temperatures 23–26 °C; (3) Limnothrix redekei : maximum biomass at temperatures 6–27 °C. Usually, non-uniform, vertical distributions of cyanobacterial biomass were associated with the formation of temperature, pH and O₂ gradients. L. redekei was considered to be a key lake organism since it contributed up to 59% of the cyanobacterial biomass. Interestingly, three of the dominant cyanobacteria Microcystis aeruginosa, Anabaena sp. and Cylindrospermopsis raciborskii belong to genera that include toxin-producing species.     

Facultative diazotrophy increases Cylindrospermopsis raciborskii competitiveness under fluctuating nitrogen availability

Relative fitness of three bloom-forming and potentially toxic cyanobacteria from the subtropical St. John's River, Florida was investigated under a range of nutrient conditions, during a bloom dominated by Cylindrospermopsis raciborskii. Nitrogen (N) was the primary nutrient limiting phytoplankton primary productivity and biomass. Phytoplankton biomass was also enhanced by phosphorus (P) added either alone or jointly with N, suggesting different components of the phytoplankton experienced distinct nutrient limitations. Based on quantitative PCR, the diazotrophic cyanobacteria Anabaena sp. and C.?raciborskii were responsible for the primary production response to P additions, while the nondiazotrophic Microcystis aeruginosa appeared to benefit from N released from the diazotrophs. Cylindrospermopsis raciborskii maintained high net growth rates under diazotrophic and nondiazotrophic conditions, while Anabaena sp. growth was significantly reduced under DIN enrichment. C.?raciborskii appears to be a generalist with regard to N source, a lifestyle traditionally not considered a viable ecological strategy among diazotrophs. Using facultative diazotrophy, C.?raciborskii gains a growth advantage under fluctuating DIN conditions, such as systems that are under the influence of anthropogenic N loading events. The described niche differentiation may be a key factor explaining the recent global expansion of C.?raciborskii.     

Separation and identification of soybean leaf proteins by two-dimensional gel electrophoresis and mass spectrometry

To establish a proteomic reference map for soybean leaves, we separated and identified leaf proteins using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and mass spectrometry (MS). Tryptic digests of 260 spots were subjected to peptide mass fingerprinting (PMF) by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) MS. Fifty-three of these protein spots were identified by searching NCBInr and SwissProt databases using the Mascot search engine. Sixty-seven spots that were not identified by MALDI-TOF-MS analysis were analyzed with liquid chromatography tandem mass spectrometry (LC-MS/MS), and 66 of these spots were identified by searching against the NCBInr, SwissProt and expressed sequence tag (EST) databases. We have identified a total of 71 unique proteins. The majority of the identified leaf proteins are involved in energy metabolism. The results indicate that 2D-PAGE, combined with MALDI-TOF-MS and LC-MS/MS, is a sensitive and powerful technique for separation and identification of soybean leaf proteins. A summary of the identified proteins and their putative functions is discussed.     

Seasonal dynamics and toxicity of Cylindrospermopsis raciborskii in Lake Guiers (Senegal, West Africa)

Cylindrospermopsis raciborskii is a toxic bloom-forming cyanobacterium that occurs at tropical and temperate latitudes. Despite several reports from Africa, no data were previously available about its dynamics or toxic potential there. We therefore carried out a 1-year survey of the dynamics of C. raciborskii in the main water reservoir in Senegal, Lake Guiers. Cylindrospermopsis raciborskii never formed a bloom in this lake during the period studied, but was dominant during the dry season. The only observed bloom-forming species was a diatom, Fragilaria sp., which displayed a seasonal pattern contrary to that exhibited by C. raciborskii. Principal component analysis applied to environmental and phytoplankton data showed that high C. raciborskii biomasses were mainly related to high temperature and water column stability. Tests for C. raciborskii species-related toxicity and/or toxin synthesis were performed on 21 isolated clones. All the strains isolated tested negative in mouse toxicity bioassays, toxin analysis (MS/MS) and tests for known cylindrospermopsin genes (ps, pks). The limited number of isolates studied, and the occurrence of toxic and nontoxic clones in natural cyanobacterial populations, mean that we cannot conclude that there is no C. raciborskii-associated health risk in this drinking water reservoir.     

Carbon acquisition characteristics of six microalgal species isolated from a subtropical reservoir: potential implications for species succession

CO ₂ levels in freshwater systems can fluctuate widely, potentially influencing photosynthetic rates and growth of phytoplankton. Given the right conditions, this can lead to bloom formation and affect water quality. This study investigated the acquisition of dissolved inorganic carbon (DIC ) by six species of microalgae, a cyanobacterium Cylindrospermopsis raciborskii , the diatoms Cyclotella sp., Nitzschia sp., and the green algae Stichococcus sp., Staurastrum sp., and Monoraphidium sp., all isolated from a subtropical reservoir in Australia. Carbon acquisition characteristics, specifically the affinity for DIC , internal pH , and internal DIC concentrations were measured. Affinities for CO ₂ () ranged between 0.7 and 6 μM CO ₂. This was considerably lower than air‐equilibrated surface water CO ₂ concentrations, and below reported affinities for CO ₂ of RuBisCO suggesting operation of active carbon dioxide concentrating mechanisms (CCM s) in all species. Internal pH was lowest for Cyclotella sp. at 7.19, and highest for Staurastrum sp., at 7.71. At 180 μM external DIC , ratios of internal:external CO ₂ ranged from 2.5 for Nitzschia sp. to 14 in C. raciborskii . Internal HCO ₃^? concentration showed a linear relationship with surface area to biovolume ratio (SA :Vol). We hypothesized that species with a higher SA :Vol suffer more from diffusive escape of CO ₂, thus storage of DIC as bicarbonate is favored in these strains. For C. raciborskii , under stratified summer conditions, its strong CCM , and resilient photosynthetic characteristics may contribute to its bloom forming capacity.     

Evaluation of three different protocols of protein extraction for Arabidopsis thaliana leaf proteome analysis by two-dimensional electrophoresis

This work was performed to compare three precipitation protocols of protein extraction for 2-DE proteomic analysis using Arabidopsis leaf tissue: TCA-acetone, phenol, and TCA-acetone-phenol. There were no statistically significant differences in protein yield between the three methods. Samples were subjected to 2-DE in the 5 to 8 pH and 14-80 kDa ranges. The TCA-acetone-phenol protocol provided the best results in terms of spot focusing, resolved spots, spot intensity, unique spots detected, and reproducibility. In all, 93 qualitative or quantitative statistically significant differential spots were found between the three protocols. The 2-DE map of TCA-acetone-phenol extracts presented more resolved spots above 40 kDa, with no pI-dependent differences observed between the three protocols. 54 spots were selected for trypsin digestion, and the peptides were analyzed by MALDI-TOF-TOF MS. After database search using peptide mass fingerprinting, and MS/MS combined search, 30 proteins were identified, the proteins from chloroplastic photosynthetic and carbohydrate metabolism being those most highly represented. From these data, we were able to conclude that each extraction protocol had its main features. Considering this, the workflow of any standard comparative proteomic experiment should include the optimization and adaptation of the protein extraction protocol to the plant tissue and to the particular objective pursued.     

N-苯基-2-萘胺对拟柱胞藻生长、抗氧化酶及光合系统Ⅱ的影响

为探究化感物质对入侵水华蓝藻-拟柱胞藻(Cylindrospermopsis raciborskii)的影响, 研究对不同浓度(0、0.05、0.10、0.50、1.00和2.00 mg/L)N-苯基-2-萘胺处理下拟柱胞藻的生长、抗氧化酶活性及叶绿素荧光诱导动力学进行了测定。结果表明, 藻叶绿素a含量随N-苯基-2-萘胺浓度升高而显著降低, 浓度大于0.50 mg/L时, 拟柱胞藻叶绿素a含量显著低于对照。72h, N-苯基-2-萘胺对拟柱胞藻的EC50为1.02 mg/L。叶绿素荧光诱导动力学参数表明, 浓度小于0.50 mg/L时, 拟柱胞藻的最大光化学效率(P0)、单位面积上有活性的反应中心(RC/CS0)、以吸收光能为基础的性能指数(PIABS)及以单位面积为基础的性能指数(PICS)均显著高于对照, 而反映2ms时有活性的反应中心的开放程度的0则显著低于对照组。当N-苯基-2-萘胺浓度大于0.50 mg/L时, 以上参数均呈现相反趋势。这表明低浓度N-苯基-2-萘胺能提高拟柱胞藻的光合效率, 而在高浓度时, 拟柱胞藻PSⅡ有活性的反应中心减少, 电子传递受到阻碍, 光合效率下降。与对照相比, 在浓度高于0.50 mg/L条件下, 拟柱胞藻的过氧化氢酶(CAT)、过氧化物酶(SOD)及谷胱甘肽过氧化物酶(GPX)的活性显著增加, 表明拟柱胞藻能产生大量的抗氧化酶来减少N-苯基-2-萘胺的损伤。这些结果表明N-苯基-2-萘胺可能通过抑制PSⅡ的电子传递和减少其有活性的反应中心来影响拟柱胞藻的生长, 也暗示了N-苯基-2-萘胺能作为潜在的抑藻物质来控制拟柱胞藻水华。       

Induction of polyunsaturated fatty-acid synthesis enhances tolerance of a cyanobacterium, Cylindrospermopsis raciborskii, to low-temperature photoinhibition

Acyl-lipid desaturation introduces double bonds (unsaturated bonds) at specifically defined positions of fatty acids that are esterified to the glycerol backbone of membrane glycerolipids. Desaturation pattern of the glycerolipids of Cylindrospermopsis raciborskii (C. raciborskii), a filamentous cyanobacterial strain, was determined in cells grown at 35 degrees C and 25 degrees C. The lowering of the growth temperature from 35 degrees C to 25 degrees C resulted in a considerable accumulation of polyunsaturated octadecanoic fatty acids in all lipid classes. Lipid unsaturation of C. raciborskii was also compared to Synechocystis PCC6803. In C. raciborskii cells, a shift in growth temperature induced a much more pronounced alteration in the desaturation pattern of all lipid classes than in Synechocystis PCC6803. The tolerance to low-temperature photoinhibition of the C. raciborskii cells grown at 25 degrees C and 35 degrees C was also compared to the tolerance of Synechocystis cells grown at the same temperatures. Lower growth temperature increased the tolerance of C. raciborskii cells but not that of Synechocystis cells. These results strengthen the importance of polyunsaturated glycerolipids in the tolerance to environmental stresses and may give a physiological explanation for the determinative role of C. raciborskii strain in algal blooming in the Lake Balaton (Hungary).     

Influence of Biotic and Abiotic Factors on the Allelopathic Activity of the Cyanobacterium Cylindrospermopsis raciborskii Strain LEGE 99043

Allelopathy is considered to be one of the factors underlying the global expansion of the toxic cyanobacterium Cylindrospermopsis raciborskii. Although the production and release of allelopathic compounds by cyanobacteria is acknowledged to be influenced by environmental parameters, the response of C. raciborskii remains generally unrecognized. Here, the growth and allelopathic potential of C. raciborskii strain LEGE 99043 towards the ubiquitous microalga Ankistrodesmus falcatus were analyzed under different biotic and abiotic conditions. Filtrates from C. raciborskii cultures growing at different cell densities displayed broad inhibitory activity. Moreover, higher temperature, higher light intensity as well phosphate limitation further enhanced this activity. The distinct and comprehensive patterns of inhibition verified during the growth phase, and under the tested parameters, suggest the action of several, still unidentified allelopathic compounds. It is expectable that the observed increase in allelopathic activity can result in distinct ecological advantages to C. raciborskii.     

拟柱孢藻N8全基因组测序及磷吸收转运通路的比较分析

为从分子水平解析热带特征性蓝藻-拉氏拟柱孢藻(Cylindrospermopsis raciborskii,简称拟柱孢藻)环境适应机制,研究以广东省镇海水库中分离的拟柱孢藻藻株N8为材料,采用PacBio单分子实时测序技术(Single Molecule Real-Time, SMRT)进行测序,并初步进行全基因组特征的比较分析。结果显示拟柱孢藻N8全基因组大小为3.857 Mb, GC含量为40.13%,预测编码基因个数为3598个,在COG、KEGG和GO数据库中注释到的基因数分别为2429、1664和2244个。N8藻株与美国国立生物技术信息中心(NCBI)上公布的27株拟柱孢藻基因组大小和GC含量基本一致,但N8的编码基因数最多。基于拟柱孢藻全基因组单拷贝基因的系统进化树表明N8藻株与韩国藻株GIHE 2018的亲缘关系最近。N8藻株的基因组中未发现拟柱孢藻毒素(CYN)和石房蛤毒素(STX)合成酶基因簇,表明该藻株不产蓝藻毒素CYN和STX。对N8藻株和其他7藻株的磷吸收转运通路比较分析表明,这些藻株均拥有较为完整的磷吸收转运基因(双组分调节系统、低亲和力无机磷转运基因、高亲...     

The Smallest Known Genomes of Multicellular and Toxic Cyanobacteria: Comparison,Minimal Gene Sets for Linked Traits and the Evolutionary Implications

Cyanobacterial morphology is diverse, ranging from unicellular spheres or rods to multicellular structures such as colonies and filaments. Multicellular species represent an evolutionary strategy to differentiate and compartmentalize certain metabolic functions for reproduction and nitrogen (N₂) fixation into specialized cell types (e.g. akinetes, heterocysts and diazocytes). Only a few filamentous, differentiated cyanobacterial species, with genome sizes over 5 Mb, have been sequenced. We sequenced the genomes of two strains of closely related filamentous cyanobacterial species to yield further insights into the molecular basis of the traits of N₂ fixation, filament formation and cell differentiation. Cylindrospermopsis raciborskii CS-505 is a cylindrospermopsin-producing strain from Australia, whereas Raphidiopsis brookii D9 from Brazil synthesizes neurotoxins associated with paralytic shellfish poisoning (PSP). Despite their different morphology, toxin composition and disjunct geographical distribution, these strains form a monophyletic group. With genome sizes of approximately 3.9 (CS-505) and 3.2 (D9) Mb, these are the smallest genomes described for free-living filamentous cyanobacteria. We observed remarkable gene order conservation (synteny) between these genomes despite the difference in repetitive element content, which accounts for most of the genome size difference between them. We show here that the strains share a specific set of 2539 genes with >90% average nucleotide identity. The fact that the CS-505 and D9 genomes are small and streamlined compared to those of other filamentous cyanobacterial species and the lack of the ability for heterocyst formation in strain D9 allowed us to define a core set of genes responsible for each trait in filamentous species. We presume that in strain D9 the ability to form proper heterocysts was secondarily lost together with N₂ fixation capacity. Further comparisons to all available cyanobacterial genomes covering almost the entire evolutionary branch revealed a common minimal gene set for each of these cyanobacterial traits.     

Production and purification of agarase from a marine agarolytic bacterium Agarivorans sp. HZ105

Aims:  Isolation and characterization of an agarase-producing bacterium Agarivorans sp. HZ105.
Methods and Results:  An agarase-producing bacterium strain HZ105 had been isolated from marine sediment sample. Based on phylogenetic analysis of the 16S rRNA gene sequence and phenotypic analysis, as well as biochemical analyses, this strain was named Agarivorans sp. HZ105. Effect of pH, NaCl on the growth and agarase production of strain HZ105 was studied. Strain HZ105 produced three extracellular agarases which were purified to homogeneity from bands in the PAGE gel. Two agarases of these three had a molecular mass of 54, 58 kDa, respectively. And the MS and MS/MS spectra were used to identify the agarases.
Conclusions:  The MS spectra result showed that the agarases of strain HZ105 should be beta-agarase and belong to the family 50 of glycosyl hydrolases. The agarases could keep stable activity at room temperature.
Significance and Impact of the Study:  The strain HZ105 was useful to produce stable agarases. The solution produced by agar's degradation in the agar plates was first reported to be used for purification of agarase. Agarases were purified to homogeneity directly from the PAGE gel without stained by Coomassie brilliant blue.     

镇海水库拟柱孢藻的分离鉴定和氮磷对其生长的影响

以分离自广东省镇海水库的拟柱孢藻N8为对象, 探究其在不同磷浓度及氮磷浓度组合下的生长情况。结果表明, 拟柱孢藻N8对磷的适应范围很宽, 在0.025.12 mg/L磷浓度下均能生长, 最适生长磷浓度范围为0.165.12 mg/L, 磷浓度的升高能显著延长拟柱孢藻的对数生长期和提高生物量。动力学分析表明, 拟柱孢藻N8有较低的KSP值, 对磷元素的亲和性较高, 在磷营养贫乏的环境下更容易形成优势。在氮磷组合实验中, 低氮(0.5 mg/L)显著抑制拟柱孢藻的生长, 且这种生长抑制不受磷浓度的影响; 而在低磷(0.04 mg/L)条件下, 水体中氮浓度的增加会显著促进拟柱孢藻的生长, 拟柱孢藻在高氮中磷和高氮高磷下的生长显著优于其他氮磷组合条件。研究表明, 广东省水库拟柱孢藻的生长受磷的限制较弱, 氮是其生长的决定因子。       

Salt-responsive outer membrane proteins of Vibrio anguillarum serotype O1 as revealed by comparative proteome analysis

Aims:  Vibrio anguillarum is a universal marine pathogen causing vibriosis. Vibrio anguillarum encounters different osmolarity conditions between seawater and hosts, and its outer membrane proteins (OMPs) play a crucial role in the adaptation to changes of the surroundings. In the present study, proteomic approaches were applied to investigate the salt-responsive OMPs of V. anguillarum .
Methods and Results:  Lower salinity (0·85% NaCl) is more suitable for growth, survival and swimming motility of the bacterium. Comparative two-dimensional electrophoresis (2-DE) analysis reveals six differentially expressed protein spots among three different salinities, which were successfully identified as OmpU, maltoporin, flagellin B, Omp26La, Omp26La and OmpW respectively.
Conclusions:  OmpW and OmpU were highly expressed at 3·5% salinity, suggesting their role in the efficient efflux of NaCl. Maltoporin was downregulated in higher salinity, indicating that higher osmolarity inhibits carbohydrate transport and bacterial growth. Omp26La, the homologue of OmpV, functions as a salt-responsive protein in lower salinity.
Significance and Impact of the Study:  To the best of our knowledge, this is the first report describing salt stress-responsive proteins of V. anguillarum using proteomic approaches. Our results provide a useful strategy for delineating the osmoregulatory mechanism of the marine pathogens.     

An investigation of plasma collection, stabilization, and storage procedures for proteomic analysis of clinical samples

In order to evaluate the critical components of the process necessary to preserve clinical plasma samples collected at research sites for proteomic analysis, various collection and preservation protocols with controlled experimentation were evaluated. The presence of a protease inhibitor cocktail (PIC) included in the blood draw tube would stabilize the plasma proteins was hypothesized. To test this hypothesis, four plasma samples from each of 14 volunteers were collected. Samples were treated following a standard protocol that included PIC or were subjected to various processing treatments that included time, temperature, different anticoagulants, and the absence of PIC. Large format two dimensional-polyacrylamide gel electrophoresis (2D-PAGE) proteomic analysis and enzyme immunoassay (EIA) were used to detect differences between the treatment groups. A novel 2D-PAGE quality scoring method was developed to determine global differences in the treatment groups, wherein a rating scale questionnaire was used to quantify the quality of each 2D-PAGE gel. The data generated from EIAs, classical 2D-PAGE image analysis and 2D-PAGE quality scoring, each generated similar results. Inclusion of protease inhibitor cocktail in the sample tubes, provided stable and reliable human plasma samples that yielded reproducible results by proteomic analysis. When PIC was included, samples retained stability under less stringent processing, such that refrigeration for several hours before processing or one freeze-thaw cycle had little detrimental effect. We demonstrated that samples without PIC, from either heparin or ethylenediaminetetraacetic acid (EDTA) plasma tubes, gave results that varied significantly from the control samples. Also, even with PIC present in blood tubes, we found it was important to quickly decant the separated plasma from the cellular components found in the blood tubes following centrifugation, as prolonged exposure again yielded different results from the standard procedure.     

What drives the distribution of the bloom-forming cyanobacteria Planktothrix agardhii and Cylindrospermopsis raciborskii?

The cyanobacteria Planktothrix agardhii and Cylindrospermopsis raciborskii are bloom-forming species common in eutrophic freshwaters. These filamentous species share certain physiological traits which imply that they might flourish under similar environmental conditions. We compared the distribution of the two species in a large database (940 samples) covering different climatic regions and the Northern and Southern hemispheres, and carried out laboratory experiments to compare their morphological and physiological responses. The environmental ranges of the two species overlapped with respect to temperature, light and total phosphorus (TP); however, they responded differently to environmental gradients; C.?raciborskii biovolume changed gradually while P. agardhii shifted sharply from being highly dominated to a rare component of the phytoplankton. As expected, P.?agardhii dominates the phytoplankton with high TP and low light availability conditions. Contrary to predictions, C.?raciborskii succeeded in all climates and at temperatures as low as 11?°C. Cylindrospermopsis raciborskii had higher phenotypic plasticity than P.?agardhii in terms of pigments, individual size and growth rates. We conclude that the phenotypic plasticity of C.?raciborskii could explain its ongoing expansion to temperate latitudes and suggest its future predominance under predicted climate-change scenarios.     

Characterization of a novel thermophilic, cellulose-degrading bacterium Paenibacillus sp. strain B39

Aims:  The aims of this study were to identify and characterize the novel thermophilic, cellulose-degrading bacterium Paenibacillus sp. strain B39.
Methods and Results:  Strain B39 was closely related to Paenibacillus cookii in 16S rRNA gene sequence. Nonetheless, this isolate can be identified as a novel Paenibacillus sp. with respect to its physiological characteristics, biochemical reactions, and profiles of fatty acid compositions. A cellulase with both CMCase and avicelase activities was secreted from strain B39 and purified by ion-exchange chromatography. By sodium dodecyl sulfate–polyacrylamide gel electrophoresis analysis, the molecular weight of B39 cellulase was determined as 148 kDa, which was much higher than other cellulases currently reported from Paenibacillus species. The enzyme showed a maximum CMCase activity at 60°C and pH 6·5. Addition of 1 mmol l⁻¹ of Ca²⁺ markedly enhanced both CMCase and avicelase activities of the enzyme.
Conclusions:  We have identified and characterized a novel thermophilic Paenibacillus sp. strain B39 which produced a high-molecular weight cellulase with both CMCase and avicelase activities.
Significance and Impact of the Study:  Based on the ability to hydrolyse CMC and avicel, the cellulase produced by Paenibacillus sp. strain B39 would have potential applications in cellulose biodegradation.