A novel multifunctional cellulolytic enzyme screened from metagenomic resources representing ruminal bacteria

Metagenomic resources representing ruminal bacteria were screened for novel exocellulases using a robotic, high-throughput screening system, the novel CelEx-BR12 gene was identified and the predicted CelEx-BR12 protein was characterized. The CelEx-BR12 gene had an open reading frame (ORF) of 1140 base pairs that encoded a 380-amino-acid-protein with a predicted molecular mass of 41.8 kDa. The amino acid sequence was 83% identical to that of a family 5 glycosyl hydrolase from Prevotella ruminicola 23. Codon-optimized CelEx-BR12 was overexpressed in Escherichia coli and purified using Ni–NTA affinity chromatography. The Michaelis–Menten constant (K_m value) and maximal reaction velocity (V_max values) for exocellulase activity were 12.92 μM and 1.55 × 10⁻⁴ μmol min⁻¹, respectively, and the enzyme was optimally active at pH 5.0 and 37 °C. Multifunctional activities were observed against fluorogenic and natural glycosides, such as 4-methylumbelliferyl-β-d-cellobioside (0.3 U mg⁻¹), CMC (105.9 U mg⁻¹), birch wood xylan (132.3 U mg⁻¹), oat spelt xylan (67.9 U mg⁻¹), and 2-hydroxyethyl-cellulose (26.3 U mg⁻¹). Based on these findings, we believe that CelEx-BR12 is an efficient multifunctional enzyme as endocellulase/exocellulase/xylanase activities that may prove useful for biotechnological applications.     

Assessment of olive-mill wastewater as a growth medium for lipase production by Candida cylindracea in bench-top reactor

Olive-mill wastewater (OMW) was investigated for its suitability to serve as a medium for lipase production by Candida cylindracea NRRL Y-17506. The OMW that best supported enzyme production was characterized by low COD and low total sugars content. In shake flask batch cultures, OMW supplementation with 2.4 g l⁻¹ NH₄Cl and 3 g l⁻¹ olive oil led to an enzyme activity of about 10 U ml⁻¹. The addition of glucose or malt extract and supplements containing organic N (e.g., peptone, yeast extract) either depressed or did not affect the enzyme production. Further experiments were then performed in a 3-l stirred tank reactor to assess the impact of medium pH and stirring speed on the yeast enzyme activity. The lipase activity was low (1.8 U ml⁻¹) when the pH was held constant at 6.5, significantly increased (18.7 U ml⁻¹) with uncontrolled pH and was maximum (20.4 U ml⁻¹) when the pH was let free to vary below 6.5. A stirring regime, that varied depending on the dissolved oxygen concentration in the medium, both prevented the occurrence of anoxic conditions during the exponential growth phase and enabled good lipase production (i.e., 21.6 U ml⁻¹) and mean volumetric productivity (i.e., 123.5 U l⁻¹ h⁻¹).     

Synergistic production of L-arabinose from arabinan by the combined use of thermostable endo- and exo-arabinanases from Caldicellulosiruptor saccharolyticus

The optimum conditions for the production of l-arabinose from debranched arabinan were determined to be pH 6.5, 75 °C, 20 g l⁻¹ debranched arabinan, 42 U ml⁻¹ endo-1,5-α-l-arabinanase, and 14 U ml⁻¹ α-l-arabinofuranosidase from Caldicellulosiruptor saccharolyticus and the conditions for sugar beet arabinan were pH 6.0, 75 °C, 20 g l⁻¹ sugar beet arabinan, 3 U ml⁻¹ endo-1,5-α-l-arabinanase, and 24 U ml⁻¹ α-l-arabinofuranosidase. Under the optimum conditions, 16 g l⁻¹l-arabinose was obtained from 20 g l⁻¹ debranched arabinan or sugar beet arabinan after 120 min, with a hydrolysis yield of 80% and a productivity of 8 g l⁻¹ h⁻¹. This is the first reported trial for the production of l-arabinose from the hemicellulose arabinan by the combined use of endo- and exo-arabinanases.     

Hydrolysis of the soluble fluorescent molecule carboxyumbelliferyl-beta-D-glucuronide by E. coli beta-glucuronidase as applied in a rugged, in situ optical sensor

Techniques utilizing β-glucuronidase (GUS) activity as an indicator of Escherichia coli (E. coli) presence use labeled glucuronides to produce optical signals. Carboxyumbelliferyl-β-d-glucuronide (CUGlcU) is a fluorescent labeled glucuronide that is soluble and highly fluorescent at natural water pHs and temperatures and, therefore, may be an ideal reagent for use in an in situ optical sensor. This paper reports for the first time the Michaelis-Menten kinetic parameters for the binding of E. coli GUS with CUGlcU as K_m = 910 μM, V_max = 41.0 μM min⁻¹, V_max/K_m 45.0 μmol L⁻¹ min⁻¹, the optimal pH as 6.5 ± 1.0, optimal temperature as 38 °C, and the Gibb's free energy of activation as 61.40 kJ mol⁻¹. Additionally, it was found CUGlcU hydrolysis is not significantly affected by heavy solvents suggesting proton transfer and solvent addition that occur during hydrolysis are not limiting steps. Comparison studies were made with the more common fluorescent molecule methylumbelliferyl-β-d-glucuronide (MUGlcU). Experiments showed GUS preferentially binds to MUGlcU in comparison to CUGlcU. CUGlcU was also demonstrated in a prototype optical sensor for the detection of E. coli. Initial bench testing of the sensor produced detection of low concentrations of E. coli (1.00 × 10³ CFU/100 mL) in 230 ± 15.1 min and high concentrations (1.05 × 10⁵ CFU/100 mL) in 8.00 ± 1.01 min.     

Characterization of alkaline thermostable keratinolytic protease from thermoalkalophilic Bacillus halodurans JB 99 exhibiting dehairing activity

A thermostable alkaline protease produced from Bacillus sp. JB 99 exhibited significant keratinolytic and dehairing activity. The enzyme was purified by ammonium sulphate precipitation followed by CM-cellulose and Sephadex G-100 chromatography and resulted in 13.6 fold purification with 23.8% of recovery. The specific activity of purified enzyme was 2989.6 U mg^−l. Purified protease had a molecular weight of 29 kDa and appeared as a single band. Gelatin zymogram analysis also revealed a clear hydrolytic zone, which corresponded to the band obtained with SDS-PAGE. The optimum pH and temperature for the keratinolytic activity was pH 11.0 and 70 °C respectively and half life of protease was 70 °C for 4 h. N-terminal amino acid sequence of purified enzyme exhibited extensive homology with other thermostable alkaline proteases and inhibition by PMSF indicated serine type of protease. The K_m and V_max of protease for keratin substrate were 3.8 ± 0.5 mg ml⁻¹ and 15.1 ± 1.6 ??m min⁻¹ mg⁻¹ and casein were 3.3 ± 0.4 mg ml^−l and 15.6 ± 0.9 ??m min⁻¹ mg⁻¹ respectively. The enzyme efficiently dehaired buffalo and goat hide without damaging the collagen layer, which makes it a potential candidate for application in leather industry to avoid pollution problem associated with the use of chemicals in the industry. The enzyme also degraded chicken feathers in presence of reducing agent which can help poultry industry in management of keratin-rich waste and obtaining value added products.     

Urea uptake by the scleractinian coral Stylophora pistillata

Urea can be one of the major sources of nitrogen for phytoplankton, but little is known about its importance for corals. Experiments were therefore designed to assess the uptake rates of urea by the scleractinian coral Stylophora pistillata; ¹⁵N-urea was used to follow the incorporation of nitrogen into the zooxanthellae and animal tissue. The uptake kinetics of urea in the tissue of S. pistillata showed that there is a concentration-dependent uptake of urea. The transport of urea was composed of a linear component (diffusion) at concentrations higher than 6 μmol N-urea l^− 1 and an active carrier-mediated component, at lower concentrations. The value of the carrier affinity (K_m = 1.05 μmol urea l^− 1) indicates a good adaptation of the corals to low levels of urea in seawater. At the in situ concentration of ca. 0.2 μmol N-urea l^− 1, the uptake rate was equal to 0.1 nmol N h^− 1 cm^− 2. Urea uptake was at least four times higher in the animal than in the algal fraction, and five times higher when corals were incubated in the light than in the dark. These results could be explained by the involvement of urea in the calcification process, which is also enhanced by light. Comparison of urea uptake rates with nitrate or ammonium uptake rates for the same S. pistillata species, at in situ concentrations, showed that urea is preferred to nitrate and may therefore be an important source of nitrogen for scleractinian corals.     

Denitrifying sulfide removal and carbon methanogenesis in a mesophilic, methanogenic culture

The inhibitory effects of 90-189 mg l⁻¹ of sulfide and 25-75 mg-N l⁻¹ of nitrate on methanogenesis were investigated in a mixed methanogenic culture using butyrate as carbon source. In the initial phase of 90 mg l⁻¹ S²⁻ test, autotrophic denitrification of nitrate occurred with sulfide as the electron donor. Then the sulfate-reducing strains converted the produced sulfur back to sulfide via heterotrophic oxidation pathway. Methanogenesis was not markedly inhibited when 90 mg l⁻¹ of sulfide was dosed alone. When 25-75 mg-N l⁻¹ of nitrate was presented, initiation of methanogenesis was seriously delayed. Nitrogen oxides (NO_x), the intermediates for nitrate reduction via denitrification pathway, inhibited methanogenesis. The 90 mg l⁻¹ of sulfide favored heterotrophic dissimilatory nitrate reduction to ammonia (DNRA) pathway for nitrate reduction. Possible ways of maximizing methane production from an organic carbon-rich wastewater with high levels of sulfide and nitrate were discussed.     

Anaerobic degradation of tetrachlorobisphenol-A in river sediment

This study investigated the anaerobic degradation of tetrachlorobisphenol-A (TCBPA) in sediment samples collected at three sites along the Erren River in southern Taiwan. TCBPA anaerobic degradation half-lives (t_1/2) in the sediment were 12.6, 16.9 and 21.7 d at concentrations of 50, 100, and 250 ??g g⁻¹, respectively. TCBPA (50 ??g g⁻¹) anaerobic degradation half-lives (t_1/2) in the sediment were 10.1, 11.8, 11.0, 11.6, 10.8, 9.1, 8.5, 18.2, 19.3, and 16.1 d by the addition of yeast extract (5 mg l⁻¹), cellulose (0.96 mg l⁻¹), sodium chloride (1%), brij 30 (130 mg l⁻¹), brij 35 (43 mg l⁻¹), rhamnolipid (55 ??M), surfactin (91 ??M), phthalic esters (2 mg l⁻¹), nonylphenol (2 mg l⁻¹), and heavy metals (2 mg l⁻¹), respectively. The degradation rate of TCBPA was enhanced by the addition of yeast extract, cellulose, sodium chloride, brij 30, brij 35, rhamnolipid, or surfactin. However, it was inhibited by the addition of phthalic esters, nonylphenol, or heavy metals. Also noted was the presence of dichlorobisphenol-A and bisphenol-A, two intermediate products resulting from the anaerobic degradation of TCBPA accumulated in the sediments.     

The use of Fourier transform infrared spectroscopy to assay for urease from Pseudomonas aeruginosa and Canavalia ensiformis

A novel assay method was investigated for urease (EC 3.5.1.5) from Pseudomonas aeruginosa and Canavalia ensiformis by Fourier transform infrared spectroscopy. This enzyme catalyzed the hydrolysis of urea in phosphate buffer in deuterium oxide (²H₂O). The intensities of the bicarbonate bands maxima at 1625 and 1365 cm⁻¹ and of the amide I band at 1605 cm⁻¹ were measured as a function of time to study the kinetics of urea hydrolysis. The extinction coefficients ε of urea and bicarbonate were determined to be 0.72, 0.48, and 0.56 mM⁻¹ cm⁻¹ at 1625, 1605, and 1365 cm⁻¹, respectively. The initial velocity is proportional to the enzyme concentration by using the ureases from both C.ensiformis and P. aeruginosa. The kinetic constants (V_max, K_m, and K_cat) determined by Lineweaver-Burk plot were 532.2  U mg⁻¹ protein, 6.4 mM, and 806.36 s⁻¹, respectively. These data are in agreement with the results obtained by a spectrophotometric method using a linked assay based on glutamate dehydrogenase in aqueous media. Therefore, this spectroscopic method is highly suited to assay for urease activity and its kinetic parameters by using either cell-free extracts or purified enzyme preparations with an additional advantage of performing a real-time measurement of urease activity.     

Lactic acid production by Lactobacillus sp. RKY2 in a cell-recycle continuous fermentation using lignocellulosic hydrolyzates as inexpensive raw materials

Continuous lactic acid fermentations were conducted using lignocellulosic hydrolyzates and corn steep liquor as inexpensive raw materials. Lactic acid concentrations decreased with increases in the dilution rate, whereas the residual substrate concentrations increased. However, lactic acid yields were maintained at more than 0.90 g g⁻¹ over all cases experimented. The cell-recycle cultivation system exerted positive effects on fermentation efficiency, including volumetric productivity, which is attributable to the retention of cells in the bioreactor. The cell-recycle continuous fermentation of lignocellulosic hydrolyzates yielded a lactic acid productivity of 6.7 g l⁻¹ h⁻¹ for a dilution rate of 0.16 h⁻¹ using 30 g l⁻¹ of corn steep liquor and 1.5 g l⁻¹ of yeast extract as nutrients. The productivity (6.7 g l⁻¹ h⁻¹) acquired by the cell-recycle continuous fermentation of lignocellulosic hydrolyzates was 1.6 times higher than the lactic acid productivity yielded in the continuous fermentation without cell-recycle system.     

Effect of substituent dtp to optical properties of heterobimetallic M/Ag/S nest-shaped clusters (M = Mo, W)

Clusters [MoS₄Ag₃(PPh₃)₃{S₂P(OPrⁱ)₂}] (1), [WS₄Ag₃(PPh₃)₃{S₂P(OPrⁱ)₂}] (2) and [WOS₃Ag₃(PPh₃)₃{S₂P(OPrⁱ)₂}] (3) were synthesized by the reaction of (NH₄)₂MoS₄/(NH₄)₂WS₄, (NH₄)₂WOS₃ with Ag[S₂P(OPrⁱ)₂]. Their structures have been characterized by X-ray diffraction. The clusters consist of a distorted tetrahedral MS₄ (or MOS₃) (M = Mo, W) with three Ag atoms and three sulfur atom bridges (Fig. 1), and resemble roughly that of cubane-like clusters. The nonlinear optical (NLO) properties were studied with an 8 ns pulsed laser at 532 nm. Its optical response to the incident light exhibits good optical absorptive and refractive effects, with α₂ = 1.56 × 10⁻¹⁰ m W⁻¹, n₂ = 3.87 × 10⁻¹⁷ m² W⁻¹ for cluster 1; α₂ = 1.33 × 10⁻¹⁰ m W⁻¹, n₂ = 6.52 × 10⁻¹⁷ m² W⁻¹for cluster 2; and α₂ = 2.54 × 10⁻¹⁰ m W⁻¹, n₂ = 4.07 × 10⁻¹⁷ m² W⁻¹ for cluster 3 for a 1.56 × 10⁻⁴ mol dm⁻³ CH₂Cl₂ solution.     

A batch decolorization and kinetic study of Reactive Black 5 by a bacterial strain Enterobacter sp. GY-1

An Enterobacter strain (GY-1) with high activity of decolorization of Reactive Black 5 (RB 5) was isolated from textile wastewater treating sludge. The kinetic characteristics of dye decolorization by the strain GY-1 were determined quantitatively using the diazo dye, RB 5. Effects of different operation parameters (inoculum size, pH, temperature and salinity) and various electron donors on decolorization of the azo dye by GY-1 were systematically investigated to reveal the primary factors that determine the performance of the azo dye decolorization. The decolorization of RB 5 was attributed to extracellular enzymes. A kinetic model was established giving the dependence of decolorization rate on cell mass concentration (first order). Decolorization rate increased with increasing temperature from 20 to 35 °C, which can be predicted by Arrhenius equation with the activation energy (E_a) of 8.50 kcal mol⁻¹ and the frequency factor (A₀) of 6.28 × 10⁷ mg l g MLSS⁻¹ h⁻¹. Michaelis-Menten kinetics and Eadie-Hofstee plot were used to determine V_max, 1.05 mg l⁻¹ h⁻¹ and K_m, 24.06 mg l⁻¹.     

Silver nanoclusters stabilized with denatured fish sperm DNA and the application on trace mercury ions detection

In this study, fluorescent silver nanoclusters (Ag NCs) were synthesized using denatured fish sperm DNA as the template. In contrast to other methods, this method did not use artificial DNA as the template. After their reaction with denatured fish sperm DNA, Ag⁺ ions were reduced by NaBH₄ to form Ag NCs. The Ag NCs showed a strong fluorescence emission at 650 nm when excited at 585 nm. The fluorescence intensity increased fourfold at pH 3.78, controlled with Britton–Robinson buffer solution. The fluorescence of the Ag NCs was quenched in the presence of trace mercury ions (Hg²⁺) in a weakly acidic medium and nitrogen atmosphere. The extent of the fluorescence quenching of Ag NCs strongly depends on the Hg²⁺ ion concentration over a linear range from 2.0 nmol L^?1 to 3.0 μmol L^?1. The detection limit (3σ/k) for Hg²⁺ was 0.7 nmol L^?1. Thus, a sensitive and rapid method was developed for the detection of Hg²⁺ ions.     

Inhibition studies with anions and small molecules of two novel β-carbonic anhydrases from the bacterial pathogen Salmonella enterica serovar Typhimurium

Two new β-carbonic anhydrases (CAs, EC 4.2.1.1) from the bacterial pathogen Salmonella enterica serovar Typhimurium, stCA 1 and stCA 2, were characterized kinetically. The two enzymes possess appreciable activity as catalysts for the hydration of CO₂ to bicarbonate, with k_cat of 0.79 × 10⁶ s⁻¹ and 1.0 × 10⁶ s⁻¹, and k_cat/K_m of 5.2 × 10⁷ M⁻¹ s⁻¹ and of 8.3 × 10⁷ M⁻¹ s⁻¹, respectively. A large number of simple/complex inorganic anions as well as other small molecules (sulfamide, sulfamic acid, phenylboronic acid, phenylarsonic acid, dialkyldithiocarbamates) showed interesting inhibitory properties towards the two new enzymes, with several low micromolar inhibitors discovered. As many strains of S. enterica show extensive resistance to classical antibiotics, inhibition of the β-CAs investigated here may be useful for developing lead compounds for novel types of antibacterials.     

Skeletal muscle glycogen phosphorylase is irreversibly inhibited by mercury: Molecular,cellular and kinetic aspects

Muscle glycogen phosphorylase (GP) plays an important role in muscle functions. Mercury has toxic effects in skeletal muscle leading to muscle weakness or cramps. However, the mechanisms underlying these toxic effects are poorly understood. We report that GP is irreversibly inhibited by inorganic (Hg²⁺) and organic (CH₃Hg⁺) mercury (IC₅₀ = 380 nM and k_inact = 600 M⁻¹ s⁻¹ for Hg²⁺ and IC₅₀ = 43 μM and k_inact = 13 M⁻¹ s⁻¹ for CH₃Hg⁺) through reaction of these compounds with cysteine residues of the enzyme. Our data suggest that the irreversible inhibition of GP could represent one of the mechanisms that contribute to mercury-dependent muscle toxicity.     

Ethanol production from sweet sorghum juice using very high gravity technology: Effects of carbon and nitrogen supplementations

Ethanol production from sweet sorghum juice by Saccharomyces cerevisiae NP01 was investigated under very high gravity (VHG) fermentation and various carbon adjuncts and nitrogen sources. When sucrose was used as an adjunct, the sweet sorghum juice containing total sugar of 280 g l⁻¹, 3 g yeast extract l⁻¹ and 5 g peptone l⁻¹ gave the maximum ethanol production efficiency with concentration, productivity and yield of 120.68 ± 0.54 g l⁻¹, 2.01 ± 0.01 g l⁻¹ h⁻¹ and 0.51 ± 0.00 g g⁻¹, respectively. When sugarcane molasses was used as an adjunct, the juice under the same conditions gave the maximum ethanol concentration, productivity and yield with the values of 109.34 ± 0.78 g l⁻¹, 1.52 ± 0.01 g l⁻¹ h⁻¹ and 0.45 ± 0.01 g g⁻¹, respectively. In addition, ammonium sulphate was not suitable for use as a nitrogen supplement in the sweet sorghum juice for ethanol production since it caused the reduction in ethanol concentration and yield for approximately 14% when compared to those of the unsupplemented juices.     

Heterobimetallic M/Zn (M = Cu, Ni) complexes with open-chain N- and N,O-ligands: Template synthesis from metal powders and supramolecular crystal structures

A series of new heterometallic Cu^IIZn^II and Ni^IIZn^II complexes with N- and N,O open-chain multidentate ligands (L¹ = 4,6,6-trimethyl-1,9-diamino-3,7-diazanon-3-ene; L² = 3,7-bis(2-aminoethyl)-1,3,5,7-tetraazabicyclo[3.3.1]nonane; L³ = 1,15-dihydroxy-7,9,9-trimethyl-3,6,10,13-tetraazapentadec-6-ene and L⁴ = 1-hydroxy-9-oxy-4,6,6-trimethyl-3,7-diazanon-3-ene) have been prepared through the “direct template synthesis” approach, which is a combination of classical template reactions of amines with acetone/formaldehyde and the “direct synthesis” method based on using elemental metals as starting materials. There is a significant decrease in the reaction time when the “direct synthesis” method is used compared to the conventional template condensation methods. X-ray crystallographic analyses of the complexes with the general formula M(L)ZnX₄ and [CuL⁴ZnCl₃]₂ (M = Cu²⁺, Ni²⁺; L = L¹-L³; X = Cl⁻, NCS⁻) reveal the presence of long intermolecular distance interactions, such as semi-coordination, S?S and H-bonding, in their crystal organization.     

Two-stage anaerobic digestion of biodegradable municipal solid waste using a rotating drum mesh filter bioreactor and anaerobic filter

A rotating drum mesh filter bioreactor (RDMFBR) with a 100 μm mesh coupled to an anaerobic filter was used for the anaerobic digestion of biodegradable municipal solid waste (BMW). Duplicate systems were operated for 72 days at an organic loading rate (OLR) of 7.5 gVS l⁻¹ d⁻¹. Early in the experiment most of the methane was produced in the 2nd stage. This situation gradually reversed as methanogenesis became established in the 1st stage digester, which eventually produced 86–87% of the total system methane. The total methane production was 0.2 l g⁻¹ VS_added with 60–62% volatile solids destruction. No fouling was experienced during the experiment at a transmembrane flux rate of 3.5 l m⁻² h⁻¹. The system proved to be robust and stably adjusted to a shock loading increase to 15 gVS l⁻¹ d⁻¹, although this reduced the overall methane production to 0.15 l g⁻¹ VS_added.     

Production of keratinolytic proteases through bioconversion of feather meal by the Amazonian bacterium Bacillus sp. P45

Extracellular keratinase production by the feather-degrading Amazonian isolate Bacillus sp. P45 was evaluated with various growth substrates. Higher enzyme production occurred with feather meal (FM) in comparison to casein, gelatin, and cheese whey, suggesting the specificity of this strain for the utilization of keratinous substrates. Supplementation of FM medium with carbohydrates reduced enzyme production, probably due to catabolite repression. Increased keratinase yield was achieved when NH₄Cl was added to FM medium. The effects of FM and NH₄Cl concentrations on enzyme production were investigated using a 2² central composite design. Feather meal was the most significant parameter, while NH₄Cl concentrations resulted in slight differences in enzyme yield. In the range studied, optimal concentrations of FM and NH₄Cl were 43-50 g l⁻¹ and 1.8-8.6 g l⁻¹, respectively, resulting in an effective low-cost medium for the production of keratinolytic protease. Crude keratinase showed maximum activity at 50 °C and pH 7.0, and was strongly inhibited by EDTA, indicating the importance of metal ions for activity/stability. The crude keratinase from mesophilic Bacillus sp. P45 could potentially be used in the bioconversion of recalcitrant keratinous wastes through an environmentally friendly and energy-saving process, producing protein hydrolysates with commercial value for utilization as animal feed and fertilizers.     

Electrochemical monitoring of chlorhexidine digluconate effect on polyelectrolyte immobilized bacteria and kinetic cell adhesion

The electrochemical impedance spectroscopy (EIS) technique has been used as a sensitive method to explore the effect of antibacterial molecules on immobilized bacteria and biofilm formation. In this work, we describe the electrochemical spectroscopy as a powerful method to monitor the effect of Chlorhexidine Digluconate (CHX-Dg) on polyelectrolyte immobilized Escherichia coli K12 MG1655 and the kinetics of cell adhesion on gold electrodes. The experimental impedance data were modelised with a Zview program to find the best equivalent electrical circuit and analyse its parameter's properties. Polyelectrolyte multilayer formation on the electrode surface and bacteria immobilization greatly increased the electron-transfer resistance (R_et) and reduced the constant phase element (CPE_dl). The effect of CHX-Dg was studied in a 0.5 × 10⁻⁴ mmol l⁻¹ to 0.5 mmol l⁻¹ range. The relation between the evolution of R_et and CHX-Dg concentration was found to be negatively correlated. When CHX-Dg was added, the electrochemical monitoring of the bacterial kinetic adhesion showed that the electrode's capacity (C_P) variation remained stable, demonstrating that the addition of CHX-Dg in the broth inhibited bacterial adhesion.