Kanchana NONTHASEN, Weera PIYATHEERAWONG, Pornthap THANONKEO

Efficient entrapment of Kluyveromyces marxianus DBKKUY-103 in polyvinyl alcohol hydrogel for ethanol production from sweet sorghum juice

The aim of this investigation was to optimize immobilization conditions to entrap Kluyveromyces marxianus DBKKUY-103 during ethanol fermentation from sweet sorghum juice using a response surface methodology. The effects and interactions of variables involved in cell entrapment using polyvinyl alcohol and sodium alginate were studied through a fractional factorial design. The results suggested that the primary variables involved in cell entrapment that significantly affected ethanol fermentation were sodium sulfate concentration, polyvinyl alcohol, and sodium alginate contents (P < 0.01). Subsequently, these variables were optimized for the preparation of cell entrapment based on response surface methodology and using central composite design. The results indicate that cell entrapment can achieve an ethanol concentration of 81.56 g L-1 under the following optimal conditions: bead gel diameter of 4.0 mm, calcium chloride concentration of 0.3 M, polyvinyl alcohol content of 10.09% (w v-1), sodium alginate content of 3.39% (w v-1), and sodium sulfate concentration of 0.44 M. Furthermore, statistical analysis of the model reveals that it adequately fit the experimental data (P < 0.01, R2 = 0.9879). Therefore, this model can be used to efficiently prepare entrapped cells for ethanol production.

Anahtar Kelimeler:

Cell entrapment, Kluyveromyces marxianus, ethanol fermentation, sweet sorghum juice, polyvinyl alcohol, sodium alginate, interpenetrating polymer networks, response surface methodology, central composite design

Efficient entrapment of Kluyveromyces marxianus DBKKUY-103 in polyvinyl alcohol hydrogel for ethanol production from sweet sorghum juice

Keywords:

Cell entrapment, Kluyveromyces marxianus, ethanol fermentation, sweet sorghum juice, polyvinyl alcohol, sodium alginate, interpenetrating polymer networks, response surface methodology, central composite design,

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80 g L–1
Phisalaphong et al., 2007 S. cerevisiae DTN
Alginate-maize stem disks Sugar beet juice
pH 5, 30 °C for 48 h 69.18 g L–1
Razmovski and Vučurović, 2013
K. marxianus DMKU 3-1042 Alginate-loofa matrix Cane juice
pH 0, 37 °C for 72 h 70.0 g L–1
Eiadpum et al., 2012
K. marxianus DBKKUY-103
PVA – alginate hydrogel
Sweet sorghum juice pH 5.0, 40 °C for 84 h 81.56 g L–1 This work References
Abdel-Banat BM, Hoshida H, Ano A, Nonklang S, Akada R (2010). High-temperature fermentation: how can processes for ethanol production at high temperatures become superior to the traditional process using mesophilic yeast? Appl Microbiol Biot 85: 861–867.
Awad GE, Amer H, El-Gammal EW, Helmy WA, Esawy MA, Elnashar MM (2013). Production optimization of invertase by Lactobacillus brevis Mm-6 and its immobilization on alginate beads. Carbohyd Polym 93: 740–746.
Bai FW, Anderson WA, Young MM (2008). Ethanol fermentation technologies from sugar and starch feedstocks. Biotechnol Adv 26: 89–105.
Behera S, Kar S, Mohanty RC, Ray RC (2010a). Ethanol fermentation of mahula (Madhuca latifolia) flowers using free and immobilized bacteria Zymomonas mobilis MTCC 92. Biologia 65: 416–421.
Behera S, Mohanty RC, Ray RC (2010b). Comparative study of bio- ethanol production from mahula (Madhuca latifolia L.) flowers by Saccharomyces cerevisiae cells immobilized in agar and Ca- alginate matrices. Appl Energ 87: 96–100.
Bisht D, Yadav SK, Darmwal NS (2013). Optimization of immobilization conditions by conventional and statistical strategies for alkaline lipase production by Pseudomonas aeruginosa mutant cells: scale-up at bench-scale bioreactor level. Turk J Biol 37: 392–404.
Demir H, Göğüş N, Tarı C, Heerd D, Lahore MF (2012). Optimization of the process parameters for the utilization of orange peel to produce polygalacturonase by solid-state fermentation from an Aspergillus sojae mutant strain. Turk J Biol 36: 394–404.
Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956). Colorimetric method for determination of sugars and related substances. Anal Chem 28: 350–356.
Eiadpum A, Limtong S, Phisalaphong M (2012). High-temperature ethanol fermentation by immobilized coculture of Kluyveromyces marxianus and Saccharomyces cerevisiae. J Biosci Bioeng 114: 325–329.
Fonseca GG, Heinzle E, Wittmann C, Gombert AK (2008). The yeast Kluyveromyces marxianus and its biotechnological potential. Appl Microbiol Biot 79: 339–354.
Hashimoto S, Furukawa K (1987). Immobilization of activated sludge by PVA-boric acid method. Biotechnol Bioeng 30: 52–59.
Idris A, Suzana W (2006). Effect of sodium alginate concentration, bead diameter, initial pH and temperature on lactic acid production from pineapple waste using immobilized Lactobacillus delbrueckii. Process Biochem 41: 1117–1123.
Kourkoutas Y, Bekatorou A, Banat IM, Marchant R, Koutinas AA (2004). Immobilization technologies and support materials suitable in alcohol beverages production: a review. Food Microbiol 21: 377–397.
Kumar SS, Kumar MS, Siddavattam D, Karegoudar TB (2012). Generation of continuous packed bed reactor with PVA- alginate blend immobilized Ochrobactrum sp. DGVK1 cells for effective removal of N,N-dimethylformamide from industrial effluents. J Hazard Mater 199–200: 58–63.
Limtong S, Sringiew C, Yongmanitchai W (2007). Production of fuel ethanol at high temperature from sugar cane juice by a newly isolated Kluyveromyces marxianus. Bioresource Technol 98: 3367–3374.
Makas YG, Kalkan NA, Aksoy S, Altinok H, Hasirci N (2010). Immobilization of laccase in kappa-carrageenan based semi- interpenetrating polymer networks. J Biotechnol 148: 216–220.
Montgomery DC (2005). Design and Analysis of Experiments. 6th ed. New York, NY, USA: John Wiley.
Mussatto SI, Dragone G, Guimarães PM, Silva JP, Carneiro LM, Roberto IC, Vicente A, Domingues L, Teixeira JA (2010). Technological trends, global market, and challenges of bio- ethanol production. Biotechnol Adv 28: 817–830.
Myers RH, Montgomery DC, Anderson-Cook CM (2009). Response Surface Methodology: Process and Product Optimization Using Designed Experiments. 3rd ed. New York, NY, USA: John Wiley.
Nikolić S, Mojović L, Pejin D, Rakin M (2010). Production of bioethanol from corn meal hydrolyzates by free and immobilized cells of Saccharomyces cerevisiae var. ellipsoideus. Biomass Bioenerg 34: 1449–1456.
Öngen G, Sargın S, Üstün Ö, Kutlu C, Yücel M (2012). Dipeptidyl peptidase IV production by solid state fermentation using alternative fungal sources. Turk J Biol 36: 665–671.
Phisalaphong M, Budiraharjo R, Bangrak P, Mongkolkajit J, Limtong S (2007). Alginate-loofa as carrier matrix for ethanol production. J Biosci Bioeng 104: 214–217.
Pulat M, Akalın GO, Karahan ND (2014). Lipase release through semi-interpenetrating polymer network hydrogels based on chitosan, acrylamide, and citraconic acid. Artif Cells Nanomed Biotechnol 42: 121–127.
Pulat M, Kahraman AS, Tan N, Gümüşderelioğlu M (2013). Sequential antibiotic and growth factor releasing chitosan- PAAm semi-IPN hydrogel as a novel wound dressing. J Biomat Sci-Polym E 24: 807–819.
Quintana MG, Dalton H (1998). Production of toluene cis-diol by immobilized Pseudomonas putida UV4 cells in barium alginate beads. Enzyme Microb Tech 22: 713–720.
Ratnavathi CV, Suresh K, Kumar BS, Pallavi M, Komala VV, Seetharama N (2010). Study on genotypic variation for ethanol production from sweet sorghum juice. Biomass Bioenerg 34: 947–952.
Razmovski R, Vučurović V (2011). Ethanol production from sugar beet molasses by S. cerevisiae entrapped in an alginate-maize stem ground tissue matrix. Enzyme Microb Tech 48: 378–385.
Razmovski R, Vučurović V (2013). Alcoholic fermentation by yeast immobilized on maize stem disks filled with Ca-alginate. Rom Biotech Lett 18: 8873–8882.
Sheng ZL, Zhong WW, Long WJ (2007). Immobilization of activated sludge using improved polyvinyl alcohol (PVA) gel. J Environ Sci 19: 1293–1297.
Sperling LH, Hu R (2003). Interpenetrating polymer networks. In: Utracki LA, editor. Polymer Blends Handbook. Dordrecht, the Netherlands: Kluwer Academic Publishers, pp. 417–447.
Takei T, Ikeda K, Ijima H, Kawakami K (2011). Fabrication of poly (vinyl alcohol) hydrogel beads crosslinked using sodium sulfate for microorganism immobilization. Process Biochem 46: 566–571.
Wang P, Chen Z, Li J, Wang L, Gong G, Zhao G, Liu H, Zheng Z (2013). Immobilization of Rhizopus oryzae in a modified polyvinyl alcohol gel for L(+)-lactic acid production. Ann Microbiol 63: 957–964.
Wirawan F, Cheng CL, Kao WC, Lee DJ, Chang JH (2012). Cellulosic ethanol production performance with SSF and SHF processes using immobilized Zymomonas mobilis. Appl Energ 100: 19– 26.
Wu KY, Wisecarver KD (1992). Cell immobilization using PVA cross linked with boric acid. Biotechnol Bioeng 39: 447–449.
Wu Z, Liu Y, Liu H, Xia Y, Shen W, Hong Q, Li S, Yao H (2012). Characterization of the nitrobenzene-degrading strain Pseudomonas sp. a3 and use of its immobilized cells in the treatment of mixed aromatics wastewater. World J Microb Biot 28: 2679–2687.
Yu J, Zhang X, Tan T (2007). A novel immobilization method of Saccharomyces cerevisiae to sorghum bagasse for ethanol production. J Biotechnol 129: 415–420.
Zain NAM, Suhaimi MS, Idris A (2011). Development and modification of PVA–alginate as a suitable immobilization matrix. Process Biochem 46: 2122–2129.
Zhang LS, Wu WZ, Wang JL (2007). Immobilization of activated sludge using improved polyvinyl alcohol (PVA) gel. Environ Sci 19: 1293–1297.