Hexavalent Chromium Reduction by Using Ca-Alginate Immobilized Bacillus sp.

in an Internal Loop Airlift Reactor (ILALR) Document By: Bharadwaj Visit my website www.Engineeringpapers.blogspot.com More Papers and Presentations available on above site
Abstract Extensive use of chromium in industries such as leather tanning, metallurgical, electroplating etc., resulted in industrial wastes containing hexavalent chromium (Cr(VI)). Toxic Cr(VI) ions cause physical discomfort and sometimes life-threatening illness including irreversible damage to vital body system. Compared to Cr(VI), Cr(III) is non-toxic and, due to its lower environmental mobility, exhibits limited environmental impact. For this reason, the reduction of Cr(VI) to Cr(III) remains as a primary method for the treatment of Cr(VI) containing wastes. Microbial techniques developed to treat chromium contaminated wastewater were found to be economic. The present study presents microbial reduction of (Cr(VI)) using caalginate immobilized Bacillus sp. in a lab scale internal loop airlift reactor. Cr(VI) reduction was observed at various air flowrates such as 4, 8, and 12 l/min. Complete reduction was obtained after 222 h at pH 7.0, ambient temperature, and 12 l/min air flowrate with 200 g beads loading for 200 mg/l of initial concentration of Cr(VI). The increase in gas flowrate increased the mass transfer rate and in turn increase in reduction rate was observed. Key words: chromium reduction, immobilization, gas flow, air lift reactor. 1.0 Introduction Modern industrialization makes the environment highly polluted and especially heavy metals contamination is considered as a major environmental problem. The industries like electroplating, leather tanning, textile dyeing, and metal processing industries released wastewaters containing toxic hexavalent chromium (Cr(VI)) have permanent toxic effects to aquatic system and human beings. Chromium exists in several oxidation states viz., from Cr(II) to Cr(VI) and among the various forms, Cr(VI) is most toxic and carcinogenic due to its high solubility in water. Moreover, Cr(VI) has rapid permeability through biological membranes and subsequent interaction with intracellular proteins and nucleic acids [1]. The conventional methods like chemical reduction, reverse osmosis, ion exchange, and adsorption

on activated carbon are available to remove Cr(VI) from wastewaters. However, these methods have many serious disadvantages such as high cost and generation of toxic sludge [2-3]. Biological reduction is an alternate method to over come the aforementioned problems. Bacterial reduction of Cr(VI) to Cr(III) using sewage sludge was first observed by Korenkov [4]. Many bacterial species have been reported to reduce Cr(VI) and most of which have been isolated from indigenous microbial population[5,6]. The Cr(VI) reducing bacterial species reported in the literature include Pseudomonas sp. [7], Microbacterium [8], Desulfo vibrio [9], Enterobacter sp. [10], Escherichia coli [11]. From the last decade, Cr(VI) reduction using immobilized bacteria in various bioreactors was developed and such reactors offer many advantages. The reactors reported for the use reduction of Cr(VI) include membrane [12], continuous-flow [13], fixed film [14], and continuous stirred tank [15]. Recently, the airlift loop reactor (ALR) has been extensively used for fermentations and wastewater treatment processes, because of its advantages such as simple construction and operation, low investment and operating costs, very fine gas dispersion, definitely directed circulation flow, high mixing and mass-transfer performance, and relatively low power requirement [16]. In the present scenario, airlift bioreactors have become popular and employed for the biodegradation of various pollutants such as p-xylene and naphthalene [17], phenol [18] and quinoline [19]. In the present study, the reduction of hexavalent chromium (Cr(VI)) using ca-alginate immobilized Bacillus sp. in a lab scale internal loop airlift reactor was performed. The effect of air flowrate, initial concentration of Cr(VI). The increase in gas flowrate increased the mass transfer rate and in turn increase in reduction rate was observed. 2.0 Materials and Methods 2.1 Microorganism and culture conditions Microorganism used in this study was isolated from tannery effluent contaminated environment. The maximum chromium resistance isolate was inoculated with fresh nutrient broth and purified by streak plate technique. The potential isolate was identified by colony morphology, cell morphology and biochemical tests and isolated bacterium, identified as Bacillus sp., reduced Cr(VI) effectively. 2.2 Immobilized cells preparation Alginate was dissolved in boiling water and autoclaved at 121C for 15 min. Bacillus sp. cells harvested during the mid-logarithmic growth phase were centrifuged (6000 rpm, 10

min) and resuspended in 15 ml of saline. Then the cells obtained were added into 100 ml of sterilized alginate solution. This alginate/cell mixture (with stirring) was extruded drop by drop into sterile 3% CaCl2 solution through nozzle. Gel beads were hardened by resuspending into a fresh 0.01M CaCl2 solution for 24 h. 2.4 Cr(VI) reduction using immobilized cells in an ILALR An internal loop airlift reactor fabricated using glass with a working volume of 2.5 L was used in this study and schematic diagram of the reactor is shown in Fig.1. The reactor consists of two concentric tubes, where the inner tube is removable draft tube (35 cm x 3.5 cm) and the external tube has dimensions of 45 cm x 10 cm. The air was sparged through a sparger into the reactor at the bottom. The sparger was placed inside the draft tube.

Fig. 1 Schematic diagram of the ILALR for Cr(VI) reduction using immobilized Bacillus sp. 2.3 Analytical Methods Chromium reduction activity was estimated using hexavalent chromium specific colorimetric reagent 1,5-diphenylcarbozide (DPC), 0.25% (w/v), prepared with acetone to minimize deterioration [8]. The reaction mixture was set up in 10 ml volumetric flasks as follows: 200 l of sample was made up to 1ml using distilled water, which was followed by the addition of 330 l of 6M H2SO4 and 400 l of DPC. Spectrophotometric measurements were made immediately at 540 nm. 3.0 Results and Discussion

The colony morphology, cell morphology and biochemical test results of the isolated species are presented in Table 1 and from the results isolated species was identified as Bacillus sp. The size of the beads was determined using water displacement method and found as 0.22 cm. The viability of the cells within the bead was estimated using agar spread plate colony counting method and 2.6 x 109 cfu/ml was observed. Table 1 Cell morphology, biochemical test results for the isolate Analysis Gram staining Endospore staining Motility Morphology Anaerobic growth Catalase test Indole test Lactose fermentation test Urease Lactose Amylase test Nitrate redcution test Voges-Proskquer Citrate test Growth in 10% NaOH MR/VP test +: Positive; - : Negative Results + + + rod + + + + + + + +/-

3.1 Effect of incubation time and initial concentration on Cr(VI) reduction The effect of incubation time on Cr(VI) reduction was studied with various initial concentration of Cr(VI) ranging from 10-200 mg/l as shown in Fig.2. The Cr(VI) reduction rate was faster and complete reduction was obtained within 175 h for the concentration ranging from 10-100 mg/l while longer incubation time was required for 200mg/l. The results showed that the time required for the complete reduction of Cr(VI) increased with an increase in incubation time. Similar results were reported by Philip et al., [20] and Pattanapipitpaisal et al., [13].

100

80 % of Cr(VI) reduction

60
10 m g/l 25 m g/l 50 m g/l 75 m g/l 100 m g/ l 200 m g/ l

40

20

0 0 25 50 75 1 0 0 1 2 5 1 5 0 17 5 20 0 2 2 5 T im e (h )

Fig. 2 Effect of incubation time on Cr(VI) reduction using immobilized Bacillus sp. in an ILALR. [conditions: pH-7, Air flow rate-12 l/min, Bead size-0.22 cm and T-ambient] 3.3 Effect of air flow rate on Cr(VI) reduction The effect of air flow rate on Cr(VI) reduction rate is shown in Fig. 3. Significant variation in Cr(VI) reduction rate with air flow rate was observed and this may be due to the variations in mass transfer rates at lower and higher air flow rates. At high velocity, the external mass transfer resistance become negligible [22]. Cr(VI) reduction rate was increased with an increase in air flow rate and the % Cr(VI) reduction observed for the air flow rates 4, 8 and 12 l/min were 94, 95.85, and 100% respectively. Benyahia and Polomarkaki [23] reported that the air flow rate increased the nitrification rate.

0.014

0.012
-1

) Reduction rate (h 0.01 0.008 0.006 0.004 0 4 8 Flow rate (l/min) 12 16

Fig. 4 Effect of Air flow rate on Cr(VI) reduction using immobilized Bacillus sp. in an ILALR. [conditions: pH-7, C0-200mg/l, Bead size-0.22 cm and T-ambient]

4.0 Conclusions The present study demonstrated that the Cr(VI) reduction using alginate immobilized Bacillus sp. in an ILALR has been influenced by operating parameters such as initial concentration, air flow rate and incubation time. The reduction rate was decreased with an increase in initial concentration of Cr(VI). As the incubation time increases, the reduction also increases. The reduction rate was increased with an increase in air flow rate. 5.0 References [1] S.P. Kamaludeen, Megharaj, M, Juhasz, A.L, Sethunathan N, Naidu, R. (2003). Chromium microorganism interactions in soil: remediation implications. Rev Environ Contaim. Toxicol. 178, 93164. [2] M.Y. Arica, G. Bayramoglu, (2005). Cr(VI) biosorption from aqueous solutions using free and immobilized biomass of Lentinus sajor-caju: Preparation and kinetic characterization, Colloids and Surfaces A: Physicochem. Esngg. Aspe. 253, 203211. [3] Y. Sahin, A. Ozturk, (2005). Biosorption of chromium (VI) ions from aqueous solution by the bacterium Bacillus thuringiensis, Pro. Biochem. 40, 18951901.

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Document By: Bharadwaj Visit my website www.Engineeringpapers.blogspot.com More Papers and Presentations available on above site

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