Process Biochemistry 40 (2005) 30503054 www.elsevier.

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The inuence of poly-vinyl-alcohol (PVA) characteristics on the physical stability of encapsulated immobilization media for advanced wastewater treatment
In-Soung Chang a,*, Chi-Il Kim a, Byeong-Uk Nam b
b a Department of Environmental Engineering, Hoseo University, Asan 336-795, South Korea Department of Applied Chemical Engineering, Korea University of Technology and Education, Chonan 330-708, South Korea

Received 21 June 2004; received in revised form 20 August 2004; accepted 21 February 2005

Abstract Immobilization of biomass by poly-vinyl-alcohol (PVA) encapsulation is an effective alternative for advanced wastewater treatment. The application of PVA immobilization has been gaining importance in the last few decades, but little information is available about the effect of PVA characteristics on physical properties as an encapsulation media. This study investigated the effect of PVA characteristics (such as saponication ratio and molecular weight) on the solubility of the immobilized PVA gels. In addition, the effect of ller-addition on the solubility of PVA was investigated too. As the saponication ratio (SR) of PVA increased, the solubility decreased. The solubility of the PVA gels decreased as the molecular weight (MW) of PVA increased for the fully saponicated PVAs (F series), whereas the solubility of the partially saponicated PVAs (P series) decreased as the molecular weight decreased. These phenomena could be explained by inter- and intramolecular hydrogen bonding between PVA molecules. The addition of powdered activated carbon (PAC) to PVA lessened the solubility signicantly. For example, the solubility of P24 was reduced to 62%. In the case of immobilization with biomass (both of activated sludge and nitriers), the overall solubility tendency of each PVA was not changed (P24 > P17 > F05 > F17). # 2005 Elsevier Ltd. All rights reserved.
Keywords: Encapsulation; Immobilization; Nitrier; Poly-vinyl-alcohol; Solubility; Wastewater

1. Introduction Recently, interest in nitrication and/or denitrication has increased due to stringent legislative pressure. The need for nitrogen removal in wastewater treatment arises from water quality concerns over the eutrophication of the receiving water body. Nitrogen removal is also required for groundwater recharge or other reuse applications such as agricultural and recreational irrigation. Nitrication is usually accomplished by various kinds of biological nutrients removal (BNR) processes in advanced wastewater treatment plants, where suspended biomass metabolizes the organic carbon and nutrients. The commercial BNR processes have been reported to be an
* Corresponding author. Tel.: +82 41 540 5744; fax: +82 41 540 5748. E-mail address: cis@ofce.hoseo.ac.kr (I.-S. Chang). 1359-5113/$ see front matter # 2005 Elsevier Ltd. All rights reserved. doi:10.1016/j.procbio.2005.02.009

economical form of treatment. However, the principle limitations of these kinds of processes lie in the slowgrowing nitrifying bacteria that require a long sludge retention time. Immobilization techniques provide an alternative solution. They maintain high cell concentrations and their application for wastewater treatment has been gaining importance in the last few decades [1]. Enhanced nitrication can be expected from the immobilization techniques due to the high cell concentration in the media. The immobilization system also allows for an easier solidliquid separation in a settling tank. Furthermore, the sensitivity to temperature of the nitrication process is less in the case of the immobilized bacteria [2], which is good for many BNR plants suffering from cold temperature in the winter. There are two types of immobilization technique: attachment and encapsulation. Immobilization by attach-

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ment can be obtained by spontaneous biomass adhesion onto porous support media such as polyurethane foam and inorganic matrix. Encapsulation is accomplished using porous polymeric materials such as alginate, agar, polyacrylamide, carrageenan, cellulose acetate and poly-vinylalcohol (PVA) to conne the migration of microorganisms [35]. The porous structure of polymers allows substrates and oxygen to diffuse into the internal pores where nitrication is carried out by the entrapped cells. Attachment has been the preferred method of immobilization for wastewater treatment. However, much higher cell concentrations can be obtained by encapsulation than by attachment, although the attachment method requires no chemical additions. It has been reported that nitrication rates were generally higher for PVA encapsulation systems than for the attachment systems [6]. The most widely used material for encapsulation is PVA because the freezingthawing method is a simple technique, which does not involve chemical initiation. Many studies have been carried out with PVA encapsulation for nitrication [710]. To treat wastewater using the encapsulation technique, PVA as support material needs to satisfy physical stability requirements. For example, solubility, biodegradability, diffusivity and mechanical stability are priority concerns. In particular, the solubility of PVA to water should be lessened because PVA is easily dissolved in water. PVA has a hydrophilic nature due to the hydroxyl functional groups in its molecular structure. The effect of PVA characteristics on solubility should be considered. For example, the saponication ratio (SR) and the molecular weight (MW) of PVA can inuence the solubility of the encapsulated media. However, there is little information available on the effect of PVA characteristics on physical stabilities as an encapsulation media. This study focused on investigating the effect of PVA characteristics on the solubility of the encapsulated media.

polymerisation is 1700, thus the MW of P17 and F17 are both 74,800 Da (=1700 44 Da, 44 is the MW of the repeating unit of monomer). The MW of the P24 is calculated as 105,600 Da in the same way. A 25% (w/w) aqueous PVA solution was prepared and then, the solution was frozen to 5 8C for 2 h. The frozen PVA was cut into small cubes (approximately 1 cm3). To obtain PVA gels, the cubes were immersed in boric acid and subjected to agitation for 24 h. This gave PVA crosslinked to boron. Finally, the PVA gels were washed with distilled water. At the initial stage of PVA preparation, 0.4% (w/w) powdered activated carbon (PAC) was mixed with the PVA solution to verify whether ller-addition to PVA is benecial to the solubility of the PVA gels or not. The median size of the PAC is 22.5 mm. Activated sludge and nitrifying bacteria suspension cultivated in the laboratory were used for the encapsulation with the PVA solution. Both suspensions had been acclimated to a synthetic wastewater for 2 months to achieve a steady-state condition prior to the PVA encapsulation. Using a ll and draw technique, aeration and mixing were provided through a porous stone diffuser delivering compressed air. The average size of the activated sludge and nitrifying bacteria was 103 and 57.8 mm, respectively. The biomass concentration of the encapsulated PVA was adjusted to 0.5% (w/w). The solubility of the PVA gels in water was tested as follows: the immobilized PVA gels were immersed in 1 l of water and agitated with a velocity gradient (G) of 155 s1. The water was sampled periodically. Total organic carbon (TOC) of the sampled water was measured to monitor how much of the PVA was dissolved. Therefore, the solubility of PVA gels was expressed by the TOC concentration. TOC was measured with a TOC analyser (Phoenix8000, Tekmar Dohrmann, USA). An infrared spectra of PVA was obtained by FTIR (Magna-IR550, NICOLET, USA).

2. Material and methods Five PVAs with different SR and MW were used in this study. PVAs were provided by Tong-Yang Chemicals Inc. (South Korea). Table 1 shows the characteristics of each PVA. P stands for a partially saponicated PVA (SR is 87 mol%) whereas F stands for a fully saponicated PVA (SR is 98 mol%). The number 17 means that the degree of
Table 1 Characteristics of PVA used in this study PVA Characteristics Saponication ratio (mol%) P05 P17 P24 F05 F17 87.1 87.1 87.3 98.6 98.5 Molecular weight (Da) 22,000 74,800 105,600 22,000 74,800

3. Results and discussion 3.1. Effect of SR and MW of PVA on the solubility Fig. 1 shows the effect of the saponication ratio and the molecular weight of PVA on the solubility of the gels. Regardless of each PVA, TOC concentration increased gradually along with time and nally reached a plateau. The lower the SR the PVA, the higher the TOC, i.e., the solubility of the P series (P17 and P24) was much higher than that of the F series (F05 and F17). The PVA with the lowest SR and MW, P05, was dissolved in water entirely at the preparation stage, so data are not shown in this gure. The effect of SR on the solubility could be attributed to hydrogen bonding (Fig. 2). In the molecular structure, PVA with the lower SR (P series) has side branches consisting of 87% of the hydroxyl group (OH) and 13% of the acetyl

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Fig. 1. TOC variation of the sampled water from different kinds of PVA gels. Fig. 3. FTIR spectra of partially saponicated PVA (P17) and fully saponicated PVA (F17).

(OCOCH3) group. But the side chains of the PVA with greater SR (F series) consist almost entirely of the hydroxyl group (98.5%) and only 1.5% of the side chains belong to the acetyl group. Therefore, hydrogen bonding between intermolecular hydroxyl groups for the F series could be much stronger than that of the P series. Two different types of hydrogen bonding compete with each other in the water solution. The rst is the intermolecular hydrogen bonding in PVA molecules and the other is hydrogen bonding between water and hydroxyl groups in PVA molecules. If the entire strength of hydrogen bonding between water and PVA exceeds the strength of hydrogen bonding between the hydroxyl groups in PVA (inter-molecular hydrogen bonding), PVA could be dissolved in water. Consequently, strong hydrogen bonding between inter-molecules makes the F-series PVA difcult to dissolve in water. In contrast, the P series, having relatively weak hydrogen bonding between inter-molecules, are more easily dissolved in water than the F series.

FTIR analysis (Fig. 3) conrmed the difference in the molecular structure between P and F series. The FTIR spectrum for P series shows the C O carbonyl group at near 1750 cm1 whereas that of the F series shows a very weak signal of C O, indicating that the F series has a high potential for an inter-molecular hydrogen bonding. On the other hand, the effect of MW on solubility is not easy to understand. As the MW of the F series increased, the solubility decreased. However, the solubility of the P series decreased as the MW decreased (Fig. 1). Generally speaking, linear polymer molecules could be entangled as the degree of polymerisation increased. Therefore, the probability of intra-molecular hydrogen bonding between hydroxyl groups in side branches could be increased as the MW increased. Therefore, these kinds of intra-molecular interactions make the PVA difcult to

Fig. 2. Inter-molecular hydrogen bonding for the F and P series PVAs. Inter-molecular hydrogen bonding between (a) fully saponicated PVA molecules (F series) and (b) partially saponicated PVA molecules (P series); dashed lines (- - -) indicate the hydrogen bonding between PVA molecules.

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dissolve in water. This explanation could be feasible for the solubility behaviour of the F series; the solubility of F05 (MW = 22,000) is higher than that of F17 (MW = 74,800). But it cannot explain the data of the P series, i.e., the solubility of P24 (MW = 105,600) is higher than that of the P17 (MW = 74,800). This unusual and unexpected solubility data for the P series might be explained as follows: hydroxyl groups in the P series may not be sufcient to develop intra-molecular hydrogen bonding because the P series has a relatively smaller number of hydroxyl groups than the F series. Therefore, the intra-molecular hydrogen bonding for the P series might be a less important factor for determining solubility. P24 has a longer chain than P17 because of its MW. Although both have the same proportion of hydroxyl groups (same SR), the total number of hydroxyl groups of P24 of each molecule is greater than that of P17. If inter-molecular hydrogen bonding exists equally for both P17 and P24, the probability of hydrogen bonding between P24 and water would be greater than between P17 and water. This hypothesis could explain why P24 exhibits greater solubility than P17. Further studies, beyond the scope of this study, are needed to entirely explain this phenomenon. 3.2. Effect of carbon addition on the solubility To improve the mechanical properties of synthetic polymers, inorganic llers are added during the preparation of polymers. Powdered activated carbon is selected as a ller material in this study because it can provide many hidingsites and habitats for biomass. Fig. 4 shows the effect of PAC addition on the solubility of PVA gels. Compared to the data of PAC-free PVA (Fig. 1), all of the PVAs with PAC exhibit a low TOC concentration. That is, an addition of PAC to PVA gels leads to substantial reduction of the PVA solubility. For example, TOC concentration of P24 without PAC was 6310 mg/l (Fig. 1) after 30 h of agitation, whereas the TOC of the PAC-added P24 was 2410 mg/l under the same conditions. About 62% of the solubility was reduced due to the added PAC. Even though PAC was added, the solubility tendency with respect

to the SR and MW was unchanged, i.e., P24 had the greatest solubility and F17 showed the least solubility. The reduced solubility is due to the steric-hindrance of carbon particles. Carbon particles could effectively block the hydrogen bonding between water and hydroxyl groups in PVA molecules. In addition, carbon particles could be intercalated into the PVA molecules. This situation resulted in the reduced solubility of the PAC-added PVA. The structure of PVA gels should be sufciently porous to provide sufcient channels for gas- and nutrients-transfer, which enables the immobilized biomass to metabolite substrates. However, PAC inside the PVA gels could be a barrier for the transportation of oxygen and substrates. Therefore, increased dosage of PAC for the purpose of enhancing the physical property of PVA gels could lead to mass-transfer limitation. Further studies on the optimum PAC dosage to PVA are needed. 3.3. Immobilization of biomass The solubility of the PVA gels encapsulating activated sludge is plotted along with time in Fig. 5. The PVA gels were prepared with the mixed liquor of activated sludge suspension and PAC. For the F series, there was no signicant difference in solubility between the encapsulating PVAs and the biomass-free PVAs. But for the P series, the solubility of the immobilized PVAs is a little higher than those of the biomass-free PVAs. And the solubility tendency based on the SR and MW was unchanged (P24 had the greatest solubility and F17 showed the least solubility). Solubility variation of the PVA gels immobilized with nitrifying bacteria is shown in Fig. 6. The nitriers-PVA gels were prepared with the same concentration (0.5%, w/w) and the same preparation method that was used for the immobilization of the activated sludge. The solubility for the P series was a little lower than those of the activated sludge-PVA gels, but the solubility of each F-PVA was greater than those of the activated sludge-PVA gels. The physiological difference between activated sludge and nitriers such as size, zeta potential and hydrophobicity/ hydrophilicity could make them have different solubility. In

Fig. 4. TOC variation of the sampled water from different kinds of PVA gels with activated carbon addition.

Fig. 5. TOC variation of the sampled water from different kinds of PVA gels encapsulating activated sludge and PAC.

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- In the case of encapsulation of activated sludge and nitriers with PVA, the effect of the saponication ratio and the molecular weight on the solubility was the same as the results obtained without biomass encapsulation. Consequently, the PVA solubility obeys the following order: P24 > P17 > F05 > F17.

References
[1] Tyagi RD, Vembu K, editors. Wastewater treatment by immobilized cells. CRC Press; 1990. [2] Wijffels RH, Englund G, Hunik JH, Leenen EJTM, Bakketun A, Gunter A, et al. Effects of diffusion limitation on immobilized nitrifying microorganisms at low temperatures. Biotechnol Bioeng 1995;45(1):19. [3] Yang PY, Nitisorvut S, Wu JS. Nitrate removal using a mixed-culture entrapped microbial cell immobilization process under high salt conditions. Water Res 1995;29(6):152532. [4] Kim SK, Kong I, Lee BH, Kang LK, Lee MG, Suh KH. Removal of ammonium-N from a recirculation aquacultural system using an immobilized nitrier. Aquacult Eng 2000;21:13950. [5] Leenen EJTM, Santos VAP, Grolle KCF, Tramper J, Wijffels R. Characteristics of and selection criteria for support materials for cell immobilization in wastewater treatment. Water Res 1996;30(12): 298596. [6] Rostron WM, Stuckey DC, Young AA. Nitrication of high strength ammonia wastewater: comparative study of immobilization media. Water Res 2001;35(5):116978. [7] Asano H, Myoga H, Asano M, Toyao M. Nitrication treatability of whole miroorganisms immobilized by the PVA-freezing method. Water Sci Technol 1992;26(911):103746. [8] Chen KC, Lee SC, Chin SC, Houng JY. Simultaneous carbon nitrogen removal in wastewater using phosphorylated PVAimmobilized microorganisms. Enzyme Microb Technol 1998;23: 31120. [9] Seo JK, Jung IH, Kim MR, Kim BJ, Nam SW, Kim SK. Nitrication performance of nitriers immobilized in PVA for a marine recirculating aquarium system. Aquacult Eng 2001;24: 18194. [10] Vogelsang C, Golembiewski K, Ostgaard K. Effect of preservation of gel entrapped nitrifying sludge. Water Res 1999;33(1):1648.

Fig. 6. TOC variation of the sampled water from different kinds of PVA gels encapsulating nitriers and PAC.

spite of the encapsulation of biomass (both of activated sludge and nitriers), F17 PVA exhibited the lowest solubility and the overall solubility tendency of each PVA was not changed, P24 > P17 > F05 > F17.

4. Conclusion This study investigated the effect of PVA characteristics on the physical properties as an encapsulation media used for advanced wastewater treatment and the following conclusions were drawn: - As the saponication ratio increased, the solubility of the PVA gels to water decreased. - The molecular weight of PVA affected the solubility for both partially and fully saponicated PVAs. - These phenomena could be explained by inter- and intramolecular hydrogen bonding. - Addition of PAC gave PVA a benecial inuence on the physical property, i.e., the solubility of the PVA gels decreased.