Exploring Bacillus amyloliquefaciens strain OM81 for the production of polyhydroxyalkanoate (PHA) bioplastic using olive mill wastewater.

In this study, bacterial strains isolated from olive oil mill wastewater assigned to Bacillus (n = 4) and Klebsiella (n = 1) genera, were evaluated for their ability to accumulate intracellular PHA granules using Sudan Black staining. A maximum PHA production of 0.14 g/L (i.e., 30.2% wt./wt. in dry biomass) was observed in Bacillus amyloliquefaciens strain OM81 after 72 h of incubation in the presence of 2% glucose (synthetic medium). To reduce bioplastic production costs and recover a polluting product, olive mill wastewater was tested as a carbon source. In this context, the maximum growth (1.45 g/L) was observed in the presence of 50% olive mill wastewater. After extracting the biopolymers with chloroform, quantitative and qualitative analyses were conducted using Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA). FTIR showed an absorption band at 1730 cm-1 assigned to the elongation of the PHB carbonyl groups. This approach offers a dual benefit of reducing pollution and bioplastic production costs. The Bacillus amyloliquefaciens strain OM81 showed promising results for PHAs production, making it a potential candidate for further investigation. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03808-4.

Physicochemical characterization of chitin extracted by different treatment sequences from an edible insect.

Chitin present in the shell of edible insects is a potential source of chitin, lipids, and proteins, and it exerts various biological activities. Thus far, only a few studies have focused on the use of chitin as a source of high-protein-diet oligosaccharides. The use of insect chitin for the production of high-protein-diet oligosaccharides can lessen the reliance on diet crops. Moreover, although chitin composition in Tenebrio molitor larva, pupa, and adult has been extensively investigated, chitin extraction from T. molitor larval whole body and exuvium has received poor attention. The present study compared the effectiveness of two techniques for extracting high-protein-diet chitin oligosaccharide from an edible insect (T. molitor). Two different extraction sequences of chitin from the larval stage (molitor stage larvae) and adult stage (molitor stage adult) of edible T. molitor were investigated. Two processing steps were employed: (a) deproteinization (DEP) and (b) demineralization (DEM) treatments. Differences in the order, conditions, and period of their application resulted in two different chitin extraction procedures. The viscosity, degree of polymerization, and crystallinity index of the chitin extracted using the two procedures were measured, and its chemical components (chitin, ash, protein, fat, and moisture contents) were determined. T. molitor adults and larvae treated sequentially with DEM-DEP demonstrated the greatest yield of approximately 14.62 % ± 0.15 and 6.096 % ± 0.10 %, respectively. By contrast, when treated sequentially with DEP-DEM, the recorded yields were 10.96 % ± 0.18 and 5.31 % ± 0.38, respectively. Differences in the degree of deacetylation between both methods were observed. Additionally, Fourier transform infrared spectroscopy and X-ray diffractometry of the extracted chitin along with a commercial sample revealed consistent chain conformation, mean hydrogen bonding, and crystallinity index. In this way, residues produced by farmed edible insects can be recovered and used as a novel source of chitin.

Efficient Wastewater Treatment and Removal of Bisphenol A and Diclofenac in Mesocosm Flow Constructed Wetlands Using Granulated Cork as Emerged Substrate.

Constructed wetlands (CWs) are considered as low-cost and energy-efficient wastewater treatment systems. Media selection is one of the essential technical keys for their implementation. The purpose of this work was essentially to evaluate the removal efficiency of organic pollution and nitrogen from municipal wastewater (MWW) using different selected media (gravel/gravel amended with granulated cork) in mesocosm horizontal flow constructed wetlands (HFCWs). The results showed that the highest chemical oxygen demand (COD) and ammonium nitrogen removal of 80.53% and 42%, respectively, were recorded in the units filled with gravel amended with cork. The influence of macrophytes (Phragmites australis and Typha angustifolia) was studied and both species showed steeper efficiencies. The system was operated under different hydraulic retention times (HRTs) i.e., 6 h, 24 h, 30 h, and 48 h. The obtained results revealed that the COD removal efficiency was significantly enhanced by up to 38% counter to the ammonium rates when HRT was increased from 6 h to 48 h. Moreover, the removal efficiency of two endocrine-disrupting compounds (EDCs) namely, bisphenol A (BPA) and diclofenac (DCF) was investigated in two selected HFCWs, at 48 h HRT. The achieved results proved the high capacity of cork for BPA and DCF removal with the removal rates of 90.95% and 89.66%, respectively. The results confirmed the role of these engineered systems, especially for EDC removal, which should be further explored.

Antiviral activity of Inonotusin A an active compound isolated from Boletus bellinii and Boletus subtomentosus.

Mushrooms produce various classes of secondary metabolites that could be used as antivirals in the future. The aim of this study was to determine the antiviral activity of methanolic extracts obtained from two edible mushrooms, Boletus bellinii (B. bellinii) and Boletus subtomentosus (B. subtomentosus), collected from the north forests of Tunisia, against Herpes Simplex Virus type 2 and Coxsackie Virus B type 3. In vitro micro-inhibition assays and cytotoxicity screening were performed on Vero cells. The tested Boletus methanolic extracts were found to be non-cytotoxic at high doses (50% cytotoxic concentration - CC50 > 1 mg/mL) and exhibited relevant viral inhibition with 50% inhibitory concentration, i.e., IC50 of 3.60 ± 0.66 µg/mL and 35.70 ± 7.42 µg/mL for B. bellinii, and 5.67 ± 1.02 µg/mL and 56.88 ± 9.56 µg/mL for B. subtomentosus, against HSV-2 and CVB-3, respectively. Interestingly, Boletus methanolic extracts showed high selectivity index (SI) values against both viruses, with the highest values against HSV-2 (SI > 800). Both viral strains were inhibited when treated with extracts during the early stages of virus replication. Inonotusin A was isolated and identified as the compound responsible for these activities. The latter is a novel antiviral agent that may have clinical utility or serve as a lead compound for further development. This study is the first attempt to investigate the antiviral activity of inonotusin A, isolated from the genus Boletus. The information from the present work should be a valuable reference for future studies on the antiviral activity of inonotusin A.

Optimization of thermostable proteases production under agro-wastes solid-state fermentation by a new thermophilic Mycothermus thermophilus isolated from a hydrothermal spring Hammam Debagh, Algeria.

The present work investigates for the first time the presence and isolation of the thermophilic fungi from hydrothermal spring situated at the locality of Guelma, in the Northeast of Algeria. The production of the thermostable proteases and the optimization of culture conditions under agro-wastes solid-state fermentation to achieve optimal production capacity were explored. A statistical experimental approach consisting of two designs was used to determine the optimum culture conditions and to attain the greatest enzyme production. Besides, different agricultural wastes were initially evaluated as a substrate, whereby wheat bran was selected for enzyme production by the isolate under solid-state conditions. The isolate thermophilic fungi were identified as Mycothermus thermophilus by sequencing the ITS region of the rDNA (NCBI Accession No: MK770356.1). Among the various screened variables: the temperature, the inoculum size, and the moisture were proved to have the most significant effects on protease activity. Employing two-level fractional Plackett-Burman and a Box-Behnken designs statistical approach helped in identifying optimum values of screened factors and their interactions. The analysis showed up 6.17-fold improvement in the production of proteases (~1187.03 U/mL) was achieved under the optimal conditions of moisture content 47%, inoculum 5 × 105 spores/g, and temperature at 42 °C. These significant findings highlight the importance of the statistical design in isolation of Mycothermus thermophilus species from a specific location as well as identifying the optimal culture conditions for maximum yield.

Innovative Research Approaches to Cope with Water Security in Africa.

To achieve a water-secure world, water management should be approached from a multidimensional and integrative perspective, addressing the water-related issues of health, household supply, economics, the environment, and resilience to water-related and climate change hazards. Although water security has significantly improved since 2000 in Africa, there are still vast inequalities in access to water suitable in terms of quantity and quality, especially in rural areas. To achieve water-related sustainable development of African economies, a broad scope of innovative technological and management solutions is required, involving governments, research institutions, private sector parties, and civil society. This special series, composed of 8 papers, illustrates a selection of the most relevant results achieved by the 7 research projects selected and financed by the European Union under 2 dedicated Horizon 2020 calls in 2015: Water-5b-2015 "A coordination platform" and Water-5c-2015 "Development of water supply and sanitation technology, systems and tools, and/or methodologies." The innovations presented in this special series include both technological advancements and w'ater management approaches, given that the development of water-related technologies in developing countries needs to be integrated into water management strategies and economic instruments. This special series aims to help policy makers take informed decisions on how to implement innovative approaches to increase water security in African countries. Integr Environ Assess Manag 2020;16:853-855. © 2020 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Decision Support Tool for Water Reclamation Beyond Technical Considerations-Egyptian, Moroccan, and Tunisian Case Studies.

While the Middle East and North African (MENA) region is facing challenges to sustain water security, water reclamation has received increasing consideration as a favorable mitigating solution. Despite the availability of adequate technologies, economic, political, legal, social, and environmental constraints often hamper stakeholders, and especially decision makers, from exploiting the existing potential into solution implementation. In the present paper, a comprehensive assessment for water reclamation and reuse was developed. This assessment consisted of 4 objectives: 1) apply a decision-support tool (DST) for water reclamation potential for municipal wastewater, 2) apply a DST for simulating and estimating the lifecycle costs of project-related technologies for water reclamation (municipal and industrial wastewater, as well as drainage canal water), 3) assess the national-level conditions for water reuse with a multicriteria decision analysis (MCA), and 4) establish exemplary strategies, barriers, and measures for water reuse. The present MCA considered 6 thematic subjects: policy and institution, economy, society, water management, legislation, and environment. The assessment was applied to food and nonfood crop irrigation in Egyptian, Moroccan, and Tunisian case studies. For all defined case studies, adapted treatment trains that could treat wastewater to the desired quality at reasonable costs were identified and are presented in the present paper. Results showed that technological options are available for water reuse, but the concept is not widely implemented in Egypt, Morocco, and Tunisia. The present paper identifies key barriers and drivers for the implementation of water reclamation for irrigation. In particular, the considered countries showed different characteristics regarding efficient water management, water pricing, subsidies and wastewater tariffs, implementation of monitoring and reporting systems, or legal aspects regarding the use of reclaimed water for food crop irrigation. Further exploration of case studies on high potential water reuse and financially affordable wastewater reclamation, particularly case studies that explore the impacts of policies and practices across countries, would be useful for helping the MENA region improve their water security situation. Integr Environ Assess Manag 2020;16:885-897. © 2020 SETAC.

Treatment of olive mill wastewater through employing sequencing batch reactor: performance and microbial diversity assessment.

This work describes the performance of a sequencing batch reactor (SBR) and the involvement of a novel reconstituted bacterial consortium in olive mill wastewater (OMW) treatment. The organic loading rate applied to the SBR was serially increased in terms of initial COD from 10 to 75 g L-1 to allow gradual acclimatization of activated sludge to high concentrations of toxic compounds in OMW. After the acclimatization period, up to 60% of the total COD content were effectively biodegraded from OMW at 75 g L-1 COD within 30 day hydraulic retention time. The diversity and community composition of cultivable bacteria participating in the aerobic process of treating OMW were further assessed. A total of 91 bacterial strains were isolated from the reactor and analyzed by amplification of the 16S-23S rRNA internal transcribed spacer (ITS) region and by 16S rRNA gene sequencing. The most abundant phylum was Firmicutes (57.1%) followed by Proteobacteria (35.2%) and Actinobacteria (7.7%). The use of the Biolog® Phenotype Microarray system to evaluate the ability of isolated strains to utilize OMW phenolic compounds is reported in this work for the first time. Interestingly, results showed that all species tested were able to utilize phenolics as sole carbon and energy sources. The removals of COD and phenolics from undiluted OMW by the reconstituted bacterial consortium were almost similar to those obtained by the acclimatized activated sludge, which suggest that cultivable bacteria play the major role in OMW biodegradation. Phytotoxicity assays using tomato seeds showed a significant improvement of seed germination values for treated OMW. Our overall results suggest that the novel developed bacterial consortium could be considered as a good prospect for phenolics-rich wastewaters bioremediation applications.

Integrated technological and management solutions for wastewater treatment and efficient agricultural reuse in Egypt, Morocco, and Tunisia.

Mediterranean-African countries (MACs) face a major water crisis. The annual renewable water resources are close to the 500 m3 /capita threshold of absolute water scarcity, and water withdrawals exceed total renewable water resources by 30%. Such a low water availability curbs economic development in agriculture, which accounts for 86% of freshwater consumption. The analysis of the current situation of wastewater treatment, irrigation, and water management in MACs and of the research projects targeted to these countries indicates the need for 1) an enhanced capacity to analyze water stress, 2) the development of water management strategies capable of including wastewater reuse, and 3) development of locally adapted water treatment and irrigation technologies. This analysis shaped the MADFORWATER project (www.madforwater.eu), whose goal is to develop a set of integrated technological and management solutions to enhance wastewater treatment, wastewater reuse for irrigation, and water efficiency in agriculture in Egypt, Morocco, and Tunisia. MADFORWATER develops and adapts technologies for the production of irrigation-quality water from drainage canals and municipal, agro-industrial, and industrial wastewaters and technologies for water efficiency and reuse in agriculture, initially validated at laboratory scale, to 3 hydrological basins in the selected MACs. Selected technologies will be further adapted and validated in 4 demonstration plants of integrated wastewater treatment and reuse. Integrated strategies for wastewater treatment and reuse targeted to the selected basins are developed, and guidelines for the development of integrated water management strategies in other basins of the 3 target MACs will be produced. The social and technical suitability of the developed technologies and nontechnological tools in relation to the local context is evaluated with the participation of MAC stakeholders and partners. Guidelines on economic instruments and policies for the effective implementation of the proposed water management solutions in the target MACs will be developed. Integr Environ Assess Manag 2018;14:447-462. © 2018 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals, Inc. on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Isolation and Characterization of Thermophilic Bacteria from Jordanian Hot Springs: Bacillus licheniformis and Thermomonas hydrothermalis Isolates as Potential Producers of Thermostable Enzymes.

The aim of this study was the isolation and characterization of thermophilic bacteria from hot springs in Jordan. Ten isolates were characterized by morphological, microscopic, biochemical, molecular, and physiological characteristics. Sequencing of the 16S rDNA of the isolates followed by BLAST search revealed that nine strains could be identified as Bacillus licheniformis and one isolate as Thermomonas hydrothermalis. This is the first report on the isolation of Thermomonas species from Jordanian hot springs. The isolates showed an ability to produce some thermostable enzymes such as amylase, protease, cellulose, gelatins, and lecithin. Moreover, the UPGMA dendrogram of the enzymatic characteristics of the ten isolates was constructed; results indicated a high phenotypic diversity, which encourages future studies to explore further industrial and environmental applications.

Newly isolated yeasts from Tunisian microhabitats: Lipid accumulation and fatty acid composition.

Newly isolated yeasts from different Tunisian microhabitats, such as soil, milk, olive brine, vinegar, and from olive mill wastewater-contaminated biotopes were extensively studied for their biochemical arsenal and morphological features, i.e. cell, ascospore, and lipid body morphology. All strains were classified into the Ascomycota phylum. However, they showed great functional diversity, including different morphological and biochemical features, lipid production ability, and fatty acid profiles. Accordingly, the strains were placed in three different groups: Group I, which includes Candida species; Group II (Pichia and related); and Group III (Kluyveromyces marxianus strain CC1). Group I and II were characterized by a high percentage of oleic acid (41.6-65.3% of total lipids) while in Group III, linoleic acid was the major fatty acid (37.2%). Members of Group I and II were further grouped into subgroups according to their fatty acid composition. Among the newly isolated strains, Pichia etchellsii BM1 was able to accumulate around 25% wt/wt lipid per dry cell mass and thus characterized as oleaginous. Some other strains, such as Candida metapsilosis strain EL2, C. parapsilosis strain LV2, C. pararugosa strain BM24, and K. marxianus strain CC1, which are able to produce extracellular lipases, may be of interest for specific environmental applications and/or for the production of novel lipases.

Biosynthesis of single-cell biomass from olive mill wastewater by newly isolated yeasts.

The aim of this study was to assess the potential of newly isolated yeast strains Schwanniomyces etchellsii M2 and Candida pararugosa BM24 to produce yeast biomass on olive mill wastewater (OMW). Maximum biomass yield was obtained at 75% (v/v) OMW, after 96 h of incubation at 30 °C and 5% (v/v) inoculum size. The optimal carbon/nitrogen (C/N) ratio was in the range of 8:1 to 10:1, and ammonium chloride was selected as the most suitable nitrogen source. Under these conditions, a maximum biomass production of 15.11 and 21.68 g L(-1) was achieved for Schwanniomyces etchellsii M2 and Candida pararugosa BM24, respectively. Proteins were the major constituents of yeast cells (35.9-39.4% dry weight), lipids were 2.8-5% dry weight, and ash ranged from 4.8 to 9.5 % dry weight. Besides biomass production, yeast strains were also able to reduce toxicity and polluting parameter levels of the spent OMW-based medium. The practical results presented show that pH rose from initial value of 5.5 to 7.24-7.45 after fermentation. Approximately 23.1-41.4% of the chemical oxygen demand (COD) and 15.4-19.2% of the phenolic compounds were removed. The removal of phenolic compounds was associated with their biodegradation and their partial adsorption on yeast cells.

Pseudomonas extremorientalis BU118: a new salt-tolerant laccase-secreting bacterium with biotechnological potential in textile azo dye decolourization.

The present investigation focused on screening of a new potent strain for laccase production and optimizing the process parameters to achieve the maximum enzymatic decolourization of textile azo dye Congo red. Seven hydrocarbonoclastic bacterial strains were selected as positive in laccase production in solid medium using 2,6 dimethoxyphenol as an enzyme activity indicator. The best enzyme producer Pseudomonas extremorientalis BU118 showed a maximum laccase activity of about 7000 U/L of wheat bran under solid-state conditions. The influence of different concentrations of dye, enzyme, salt and various incubation times on Congo red decolourization was studied using response surface methodology to find the optimum conditions required for maximum decolourization by P. extremorientalis laccase. The enzyme exhibited a remarkable colour removal capability over a wide range of dye and salt concentrations. The above results show the potential use of this bacterial laccase in the biological treatment of the textile effluent.

Pentachlorophenol degradation by Janibacter sp., a new actinobacterium isolated from saline sediment of arid land.

Many pentachlorophenol- (PCP-) contaminated environments are characterized by low or elevated temperatures, acidic or alkaline pH, and high salt concentrations. PCP-degrading microorganisms, adapted to grow and prosper in these environments, play an important role in the biological treatment of polluted extreme habitats. A PCP-degrading bacterium was isolated and characterized from arid and saline soil in southern Tunisia and was enriched in mineral salts medium supplemented with PCP as source of carbon and energy. Based on 16S rRNA coding gene sequence analysis, the strain FAS23 was identified as Janibacter sp. As revealed by high performance liquid chromatography (HPLC) analysis, FAS23 strain was found to be efficient for PCP removal in the presence of 1% of glucose. The conditions of growth and PCP removal by FAS23 strain were found to be optimal in neutral pH and at a temperature of 30 °C. Moreover, this strain was found to be halotolerant at a range of 1-10% of NaCl and able to degrade PCP at a concentration up to 300 mg/L, while the addition of nonionic surfactant (Tween 80) enhanced the PCP removal capacity.

Diversity and enzymatic profiling of halotolerant micromycetes from Sebkha El Melah, a Saharan salt flat in southern Tunisia.

Twenty-one moderately halotolerant fungi have been isolated from sample ashes collected from Sebkha El Melah, a Saharan salt flat located in southern Tunisia. Based on morphology and sequence inference from the internal transcribed spacer regions, 28S rRNA gene and other specific genes such as ß-tubulin, actin, calmodulin, and glyceraldehyde-3-phosphate dehydrogenase, the isolates were found to be distributed over 15 taxa belonging to 6 genera of Ascomycetes: Cladosporium (n = 3), Alternaria (n = 4), Aspergillus (n = 3), Penicillium (n = 5), Ulocladium (n = 2), and Engyodontium (n = 2). Their tolerance to different concentrations of salt in solid and liquid media was examined. Excepting Cladosporium cladosporioides JA18, all isolates were considered as alkali-halotolerant since they were able to grow in media containing 10% of salt with an initial pH 10. All isolates were resistant to oxidative stresses and low temperature whereas 5 strains belonging to Alternaria, Ulocladium, and Aspergillus genera were able to grow at 45°C. The screening of fungal strains for sets of enzyme production, namely, cellulase (CMCase), amylase, protease, lipase, and laccase, in presence of 10% NaCl, showed a variety of extracellular hydrolytic and oxidative profiles. Protease was the most abundant enzyme produced whereas laccase producers were members of the genus Cladosporium.

Hydrocarbonoclastic bacteria isolated from petroleum contaminated sites in Tunisia: isolation, identification and characterization of the biotechnological potential.

Petroleum hydrocarbons are important energy resources used by industry and in our daily life, whose production contributes highly to environmental pollution. To control such risk, bioremediation constitutes an environmentally friendly alternative technology that has been established and applied. It constitutes the primary mechanism for the elimination of hydrocarbons from contaminated sites by natural existing populations of microorganisms. In this work, a collection of 125 strains, adapted to grow on minimal medium supplemented with crude oil, was obtained from contaminated sediments and seawater from a refinery harbor of the Bizerte coast in the North of Tunisia. The diversity of the bacterial collection was analyzed by amplification of the internal transcribed spacers between the 16S and the 23S rRNA genes (ITS-PCR) and by 16S rRNA sequencing. A total of 36 distinct ITS haplotypes were detected on agarose matrix. Partial 16S rRNA gene sequencing performed on 50 isolates showed high level of identity with known sequences. Strains were affiliated to Ochrabactrum, Sphingobium, Acinetobacter, Gordonia, Microbacterium, Brevundimonas, Novosphingobium, Stenotrophomonas, Luteibacter, Rhodococcus, Agrobacterium, Achromobacter, Bacilllus, Kocuria and Pseudomonas genera. Acinetobacter and Stenotrophomons were found to be the most abundant species characterized by a marked microdiversity as shown through ITS typing. Culture-independent approach (DGGE) showed high diversity in the microbial community in all the studied samples with a clear correlation with the hydrocarbon pollution rate. Sequencing of the DGGE bands revealed a high proportion of Proteobacteria represented by the Alpha and Gamma subclasses. The predominant bacterial detected by both dependent and independent approaches were the Proteobacteria. The biotechnological potential of the isolates revealed a significant production of biosurfactants with important emulsification activities useful in bioremediation. The highest emulsification activity was detected in Pseudomonas geniculata with 52.77% of emulsification. Our overall results suggest that the obtained bacterial isolates may constitute potential candidates for bioremediation and can be useful for biotechnological applications.

Improving the nutritive value of Olive Cake by solid state cultivation of the medicinal mushroom Fomes fomentarius.

Olive Cake (OC) generated by the olive oil industries, well implanted in Tunisia, represents a major disposal and potentially severe pollution problem. This work presents the study of bioconversion of OC in solid state fermentation with the medicinal mushroom, Fomes fomentarius so as to upgrade its nutritional values and digestibility for its use as ruminants feed. The fungus was cultured on OC for 7-30 d, and subsequently the chemical composition, lignocellulolytic enzyme activities and in vitro digestibility of the resultant substrate were determined. The results obtained showed an increase in the crude protein ranging from 6% to 22% for the control and for treated OC, respectively. Significant (P<0.05) decreases in the values of neutral detergent fiber (hemicelluloses, cellulose and lignin), acid detergent fiber (lignin and cellulose) and acid detergent lignin were detected (23%, 13% and 10%, respectively). The estimated in vitro digestibility improved from 9% (control) to 25% (treated OC). The present findings revealed F. fomentarius to be an efficient organism for lignocellulolytic enzymes production and simultaneous enhancement in crude protein and in vitro digestibility of OC.

Comparative study of olive oil mill wastewater treatment using free and immobilized Coriolopsis polyzona and Pycnoporus coccineus.

The efficiency of the two white-rot fungi Pycnoporus coccineus and Coriolopsis polyzona in the Olive Oil Mill Wastewater (OOMW) treatment was investigated. Both fungi were active in the decolourisation and COD removal of OOMW at 50 g/L COD, but only the first fungus remains effective on the crude effluent (COD=100 g/L). Moreover P. coccineus was less affected by oxygen supplementation and exhibited a high tolerance to agitation in comparison to C. polyzona. However, it required a nitrogen supplementation to obtain faster and higher COD removal. To overcome the negative effect of agitation on fungi growth and efficiency, immobilisation of C. polyzona and P. coccineus in polyurethane foam was applied. The immobilized system showed better COD decreases during three consecutive batches without remarkable loss of performances. The results obtained in this study suggested that immobilized C. polyzona and especially immobilized P. coccineus might be applicable to a large scale for the removal colour and COD of OOMW.

Purification and characterization of a highly thermostable esterase from the actinobacterium Geodermatophilus obscurus strain G20.

Intracellular thermostable esterase produced by the extremophilic Geodermatophilus obscurus G20 was purified to homogeneity by a heat treatment, followed by an anion-exchange chromatography, and then characterized. The molecular weight of the purified enzyme determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was shown to be approximatively 55 kDa. The enzyme showed an optimal activity between pH 8.0 and 9.0 and was stable in the pH range 7.0-10.0. Moreover, it is highly thermostable, with a residual activity greater than 90% after incubation at 80 °C for more than 10 h. The enzyme showed preference for esters of p -nitrophenol with short chain fatty acid. When the p -nitrophenyl acetate (C2) was used as substrate, the Michaelis-Menten constant (K(m) ) and maximum velocity for the reaction (V(max) ) of esterase were 400 µM and 2500 U/mg protein, respectively. The effect of phenylmethanesulphonyl fluoride (PMSF), a serine-specific inhibitor, on the enzyme activity suggested that the thermostable esterase belong to the serine hydrolase group. Because of its high thermostability, activity at alkaline pH, tolerance to methanol and various metal ions and specificity for short chain fatty acids, this enzyme showed high potential for use in biocatalysis. (© 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim).

Isolation and characterization of a new dimeric esterase from Lactobacillus sakei 23K.

The M(w) of a Lactobacillus sakei intracellular esterase, determined by gel filtration, was compared to those obtained from SDS-PAGE or MALDI-TOF, pointing to a dimeric structure. Its N-terminal sequence and peptide mass fingerprint suggest that it is the putative LSA044 protein from L. sakei 23K genome.