Manel Hamza

Enzymatic hydrolysis of olive wastewater for hydroxytyrosol enrichment

Aspergillus niger broth culture on wheat bran was assessed for olive wastewater (OW) hydrolysis in order to release hydroxytyrosol (HT). The enzyme profiles of this culture broth gave essentially (IU/L): 3000 β-glucosidase and 100 esterase. Hydrolysis activity of A. niger enzyme preparation was evaluated by using three substrates: raw OW, phenolic fraction extracted from OW by ethyl acetate and its corresponding exhausted fraction. Large amounts of free simple phenolics were released from exhausted fraction and raw OW after enzymatic treatment. HPLC analyses show that HT was the main phenolic compound. One step of ethyl acetate extraction of hydrolysed OW allowed the recovery of 0.8 g of HT per litre of OW. The antioxidant activity of extracts from OW and exhausted fraction, measured by DPPH method, was drastically enhanced after hydrolysis treatment. This study demonstrates that hydrolysed OW is a potential source of bioactive phenolic compounds with promising applications in food and pharmaceutical industries.

Evaluation of ultrasonic, acid, thermo-alkaline and enzymatic pre-treatments on anaerobic digestion of Ulva rigida for biogas production

Pre-treatment of macroalgae has received considerable research globally due to its influence on the technical, economic and environmental sustainability of algae biogas production. Some of the most promising pre-treatment methods require the application of chemicals, enzymatic, and mechanical. This study focused on these pre-treatments of Ulva rigida for biogas production. The evaluation of different pre-treatment in terms of reducing sugar yields demonstrates that 3.62, 2.88, 2.53 and 7.3g/L of reducing sugar was obtained in acid catalysis, thermoalkaline, ultrasonication and enzymatic pre-treatment, respectively. However in crude macroalgae only 0.6g/L of reducing sugar was given. After anaerobic digestion, the enzymatic hydrolysis was demonstrated the best biogas yield than other pre-treatment which reached 626.5mL/gCODint with 62.65% of biodegradability. The best demonstrated method which uses crude broth of Aspergillus niger showed an effective and environmentally friendly strategy for enhancing the biogas production yields after the anaerobic digestion.

Changes in the content of bioactive polyphenolic compounds of olive mill wastewater by the action of exogenous enzymes.

The aim behind the present research is to develop an enzymatic treatment for olive mill wastewater (OMW) to release high amounts of simple phenolics having high antioxidant value. OMW was hydrolyzed by a mixed enzyme preparation rich in β-glucosidase produced by Aspergillus niger . This research shows that A. niger β-glucosidase played a major role in the release of simple phenolic compounds from OMW. These compounds were recovered by ethyl acetate extraction and identified by HPLC and LC-MS. The main identified phenolic compound is hydroxytyrosol. The results of enzymatic hydrolysis of OMW under optimum conditions indicated a maximum hydroxytyrosol concentration of 2.9 g L(-1) compared to 0.015 g L(-1) contained in the control (test without added enzyme). The above results prove that OMW is a potential substrate for producing hydroxytyrosol through enzymatic hydrolysis of its glycosides.

Automated Significant Load Testing for WS-BPEL Compositions

Web service compositions must provide services to hundreds even thousands of users concurrently. These applications must be load tested to ensure that they can function properly under high load. We propose in this paper a solution for load testing of WS-BPEL compositions. It is based on Timed Automata as model for specifying requirements to test under various load conditions, a distributed testing architecture, an algorithm for online load test generation and execution, and finally an automated log analysis technique. We also illustrate our approach by means of a case study.

Production and characterization of enzymatic cocktail produced by Aspergillus niger using green macroalgae as nitrogen source and its application in the pre-treatment for biogas production from Ulva rigida

Marine macroalgae are gaining more and more importance as a renewable feedstock for durable bioenergy production, but polysaccharides of this macroalgae are structurally complex in its chemical composition. The use of enzymatic hydrolysis may provide new pathways in the conversion of complex polysaccharides to fermentable sugars. In this study, an enzymatic cocktail with high specificity was first isolated from Aspergillus niger using the green macroalgae Ulva rigida as nitrogen source. The cocktail is rich on β-glucosidase, pectinase and carboxy-methyl-cellulase (CMCase). The highest activity was obtained with β-glucosidase (109IUmL(-1)) and pectinase (76IUmL(-1)), while CMCase present the lowest activity 4.6IUmL(-1). The U. rigida pre-treatment with this enzymatic cocktail showed high rate of reduced sugar release, and could bring promising prospects for enzymatic pre-treatment of the biogas production from U. rigida biomass which reached 1175mLgCODint(-1).

WSCLT: A Tool for WS-BPEL Compositions Load Testing

This paper addresses the load testing of WS-BPEL compositions. For that, we developed WSCLT tool, which takes as input a specification of the composition under test, expressed as a Timed Automaton, and considers various parameters such as the number of requests to handle simultaneously. Our WSCLT tool injects this load in the application and monitors the sequence of requests, invocations and responses between the components. This log is then analyzed by the tool to separate the actions corresponding to each instance and to check that they follow legitimate paths. A global report is then issued regarding all concurrent instances. We illustrate how to use our prototype tool by means of a case study.

Extracellular hydrolytic enzymes produced by halophilic bacteria and archaea isolated from hypersaline lake

The screening of bacteria and archaea from Chott El Jerid, a hypersaline lake in the south of Tunisia, led to the isolation of 68 extremely halophilic prokaryotes growing in media with 15–25% of salt. Assessment of 68 partial 16S rRNA analyzed by amplified rDNA restriction analysis (ARDRA) revealed 15 different bacterial and archaeal taxonomic groups. Based on ARDRA results, phenotypic and hydrolytic activity tests, 20 archaeal and 6 bacterial isolates were selected for sequencing. The halophilic isolates were identified as members of the genera: Salicola, Bacillus, Halorubrum, Natrinema and Haloterrigena. Most of these isolates are able to produce hydrolytic enzymes such as amylase, protease, lipase, cellulase, xylanase, pectinase and some of them showed combined activities. Natrinema genus is an excellent candidate for lipase production. These results indicated that the extremely halophilic archaea and bacteria from Chott El Jerid are a potential source of hydrolytic enzymes and may possess commercial value.

The Possibility of Recovering of Hydroxytyrosol from Olive Milling Wastewater by Enzymatic Bioconversion

This chapter discusses an innovative approach to obtain liquid fractions from olive mill wastewater (OMW) rich in hydroxytyrosol. The method is based on bioconversion combined with membrane separation techniques. An enzymatic bioconversion of three types of OMW was tested. TheS total volumes of OMW are 15 and 40 L. The reaction was monitored in mechanically stirred systems for 2 h at 50°C. Maximum hydroxytyr‐ osol concentrations of about 1.53, 0.83 and 0.46 g/L in the presence of 5 IU Aspergillus niger β‐glucosidase per milliliter from North OMW and South OMW were procured by two different olive millings, which are milling super press (MSP) and milling continuous chain (MCC), respectively. Enzymatic pretreatment was followed by two tangential flow membrane separation stages, microfiltration (MF) and ultrafiltration (UF). The ultrafiltration permeate was concentrated by evaporation at 45°C for 2 h. The latter exhibited a chemical oxygen demand (COD) level of 48.44 g/L. The UF permeate dehydration increased the hydroxytyrosol concentration to 7.2 g/L. A new natural product that contains some minerals beneficial to health and devoid of heavy metals or chemicals was obtained by this innovative work which describes an environmentally friendly process at pilot‐scale.