Optimization of process parameters for xylanase production ... · Optimization of process parameters for xylanase production by Bacillus sp. in submerged fermentation Muhammad Irfan
This document is posted to help you gain knowledge. Please leave a comment to let me know what you think about it! Share it to your friends and learn new things together.
Transcript
w.sciencedirect.com
J o u rn a l o f R a d i a t i o n R e s e a r c h and A p p l i e d S c i e n c e s 9 ( 2 0 1 6 ) 1 3 9e1 4 7
HOSTED BY Available online at ww
ScienceDirectJournal of Radiation Research and Applied
Sciencesjournal homepage: http : / /www.elsevier .com/locate/ j r ras
Optimization of process parameters for xylanaseproduction by Bacillus sp. in submergedfermentation
Muhammad Irfan a,b,*, Umar Asghar b, Muhammad Nadeem b,Rubina Nelofer b, Quratulain Syed b
a Department of Zoology, University of the Punjab, New Campus Lahore, 54590, Pakistanb Food & Biotechnology Research Center (FBRC), Pakistan Council of Scientific and Industrial Research (PCSIR),
Fig. 4 e Effect of substrate concentration on xylanase production by Bacillus sp. in submerged fermentation (initial pH 8, 48 h
period). Values presented were the means of triplicates and different letters differ significantly at p < 0.05.
Fig. 5 e Effect of inoculum size on xylanase production by Bacillus sp. in submerged fermentation (initial pH 8, 48 h period).
Values presented were the means of triplicates and different letters differ significantly at p < 0.05.
J o u rn a l o f R a d i a t i o n R e s e a r c h and A p p l i e d S c i e n c e s 9 ( 2 0 1 6 ) 1 3 9e1 4 7 143
3.6. Effect of additional carbon sources on xylanaseproduction
Xylanase production was further enhanced by supplementing
the fermentation medium with suitable additional carbon
source. Table 1 depicts that sucrose and xylose at concentra-
tion of 0.5% were the best xylanase inducer by B. subtilis BS04
and B. megaterium BM07 in submerged fermentation respec-
tively. Any other carbon sources like fructose, glucose, CMC
Table 1 e Effect of supplementation of additional carbonsources on xylanase production by Bacillus sp. insubmerged fermentation (initial pH 8, 48 h period).
Sr. No. Carbon source Xylanase activity (IU/ml)
B. subtilis B. megaterium
1 Control 19.96 ± 1.55a 16.63 ± 1.18a
2 Xylose 48.30 ± 0.61e 43.06 ± 1.10e
3 Fructose 38.80 ± 1.25d 26.60 ± 1.21b
4 Sucrose 54.80 ± 0.52f 39.27 ± 0.94d
5 Glucose 36.93 ± 1.10d 34.90 ± 1.15c
6 CMC 26.23 ± 1.08c 18.16 ± 1.04a
7 Arabinose 23.20 ± 0.72b 25.67 ± 1.53b
Values presentedwere themeans of triplicates and different letters
differ significantly at p < 0.05.
(carboxymethyl cellulose) and arabinose gave less enzyme
production. Azeri, Tamer, and Oskay (2010) isolated different
strains of Bacillus sp. and all exhibit maximum xylanase pro-
duction by using birchwood xylan as a carbon source. Saleem
et al. (2002) reported that supplementation of sucrose to the
fermentation medium significantly enhance the xylanase
production by B. subtilis. In 96 h of fermentation, wheat bran
was best carbon source for xylanase production by Strepto-
myces sp. (Sharma & Bajaj, 2005).
3.7. Supplementation of nitrogen sources
Among all the tested inorganic and organic nitrogen sources,
tryptone and (NH4)2SO4 are best for B. subtilis BS04 while KNO3
and malt extract for B. megaterium BM07 for xylanase syn-
thesis in submerged fermentation as shown in Table 2.
Therefore, these nitrogen sources were selected as optimum
for respective bacterial strains. Earlier studies reported that
organic nitrogen sources have been found to stimulate xyla-
nase production in Bacillus species (Battan et al., 2007). Yeast
extract þ tryptone and yeast extract þ NH4NO3 have stimu-
latory effect on xylanase production from B. mojavensis AG137
in submerge fermentation (Sepahy et al., 2011). Sharma and
Bajaj (2005) isolated different species of Streptomyces sp. and
reported that best xylanase production was observed with
J o u r n a l o f R a d i a t i o n R e s e a r c h and A p p l i e d S c i e n c e s 9 ( 2 0 1 6 ) 1 3 9e1 4 7146
study indicated that nutrients and cultural properties played a
pivotal role in enzyme production. The optimization of all the
process parameters are being considered as pre-requisites to
make the process of enzyme production cost effective at large
scale.
r e f e r e n c e s
Annamalai, N., Thavasi, R., Jayalakshmi, S., &Balasubramanian, T. (2009). Thermostable and alkalinetolerant xylanase production by Bacillus subtilis isolated frommarine environment. Indian Journal of Biotechnology, 8, 291e297.
Azeri, C., Tamer, A. U., & Oskay, M. (2010). Thermoactive cellulase-free xylanase production from alkaliphilic Bacillus strains usingvarious agro-residues and their potential in biobleaching ofkraft pulp. African Journal of Biotechnology, 9(1), 63e72.
Battan, B., Sharma, J., Dhiman, S. S., & Kuhad, R. C. (2007).Enhanced production of cellulase-free thermostable xylanaseby Bacillus pumilus ASH and its potential application in paperindustry. Enzyme and Microbial Technology, 41(6e7), 733e739.
Bernier, R., Desrochers, M., Jurasek, L., & Paice, M. G. (1983).Isolation and characterization of a xylanase from Bacillussubtilis. Applied and Environmental Microbiology, 46(2), 511e514.
Bilgrami, K. S., & Pandy, A. K. (1992). In E. S. K. Jain (Ed.), Industryand fermentation in introduction to biotechnology (pp. 149e165).
Chakrit, T., Khin, L. K., & Khanok, R. (2006). Purification ofxylanase from alkaliphilic Bacillus sp. K-8 by using corn huskcolumn. Process Biochemistry, 41(12), 2441e2445.
Coughlan,M. P., &Hazlewood, G. P. (1993). b-1,4-D-Xylan degradingenzyme systems: biochemistry, molecular biology andapplications. Biotechnology andApplied Biochemistry, 17, 259e289.
Couri, S., Terzi, S. D. C., Pinto, G. A. S., Freitas, S. P., &Costa, A. C. A. D. (2000). Hydrolytic enzyme production insolid-state fermentation by Aspergillus niger 3T5B8. ProcessBiochemistry, 36(3), 255e261.
de Souza, D. F., de Souza, C. G. M., & Peralta, R. M. (2001). Effect ofeasily metabolizable sugars in the production of xylanase byAspergillus tamarii in solid-state fermentation. ProcessBiochemistry, 36(8e9), 835e838.
Elegir, G., Szakacs, M., & Jeffries, T. W. (1994). Purification,characterization and substrate specificities of multiplexylanases from Streptomyces sp. strain B-12-2. Applied andEnvironmental Microbiology, 60(7), 2609e2615.
Gomes, D. J., Gomes, J., & Stiener, W. (1994). Production of highlythermostable xylanase by a wild strain of thermophilic fungusThermoascus aurantiacus and partial characterization of theenzyme. Journal of Biotechnology, 37, 11e22.
Gouri, M. I., & Malana, M. A. (2010). Production of carboxymethylcellulase from local isolate of Aspergillus species. PakistanJournal of Life and Social Sciences, 8(1), 1e6.
Gouda, M. K. (2000). Purification and partial characterization ofcellulose free xylanase produced in solid state and submergedfermentation by Aspergillus tamarii. Advances in Food Science, 22,31e37.
Gupta, U., & Kar, R. (2008). Optimization and scale up of cellulasefree endo xylanase production by solid state fermentation oncorn cob and by immobilized cells of a thermotolerantbacterial isolate. Jordan Journal of Biological Science, 1(3),129e134.
Gupta, U., & Kar, R. (2009). Xylanase production by a thermo-tolerant Bacillus species under solid-state and submergedfermentation. Brazilian Archives of Biology and Technology, 52(6),1363e1371.
Heck, J. X., Plinho, F., Marco, H., & Ayub, A. Z. (2002). Cellulase andxylanase production by isolated amazon Bacillus strains using
soybean industrial residue based solid-state cultivation.Brazilian Journal of Microbiology, 33, 213e218.
Hoq, M. M., Hempel, C., & Deckwer, W. D. (1994). Cellulase freexylanase by Thermomyces lanuginosus RT9: effects of aeration,agitation and medium components on production. Journal ofBiotechnology, 37, 49e58.
Immanuel, G., Dhanusha, R., Prema, P., & Palavesam, A. (2006).Effect of different growth parameters on endoglucanaseenzyme activity by bacteria isolated from coir retting effluentsof estuarine environment. International Journal of EnvironmentScience and Technology, 3(1), 25e34.
Irfan, M., Nadeem, M., Syed, Q., & Baig, S. (2012). Effect of mediumcomposition on xylanase production by Bacillus subtilis usingvarious agricultural wastes. American-Eurasian Journal ofAgriculture & Environmental Science, 12(5), 561e565.
Kalogeris, E., Christakopoulos, P., Kekos, D., & MacRis, B. J. (1998).Studies on the solid-state production of thermostableendoxylanases from Thermoascus aurantiacus, characterizationof two isozymes. Journal of Biotechnology, 60(3), 155e163.
Kamble, R. D., & Jadhav, A. R. (2012). Isolation, purification, andcharacterization of xylanase produced by a new species ofBacillus in solid state fermentation. International Journal ofMicrobiology, 2012, 8. http://dx.doi.org/10.1155/2012/683193(Article ID 683193).
Kapoor, M., Nair, L. M., & Kuhad, R. C. (2008). Cost-effectivexylanase production from free and immobilized Bacilluspumilus strain MK001 and its application in saccharification ofProsopis juliflora. Biochemical Engineering Journal, 38(1), 88e97.
Khandeparker, R., Verma, P., & Deobagkar, D. (2011). A novelhalotolerant xylanase from marine isolate Bacillus subtilischo40, gene cloning and sequencing. New Biotechnology, 28,814e821.
Kumar, S. S., Panday, D. D., & Naik, G. R. (2011). Purification andmolecular characterization of low molecular weight cellulase-free xylanase from thermoalkalophilic Bacillus spp. JB 99.World Journal of Science & Technology, 1, 09e16.
Li, Y., Lin, J., Meng, D., Lu, J., & Gu, G. (2006). Effect of pH,cultivation time and substrate concentration on theendoxylanase production by Aspergillus awamori ZH-26 undersubmerged fermentation using central composite rotarydesign. Food Technology and Biotechnology, 44(4), 473e477.
Monisha, R., Uma, M. V., & Murthy, V. K. (2009). Partialpurification and characterization of Bacillus pumilus xylanasefrom soil source. Kathmandu University Journal of ScienceEngineering and Technology, 5(II), 137e148.
Mrudula, S., & Shyam, N. (2012). Immobilization of Bacillusmegaterium MTCC 2444 by Ca-alginate entrapment method forenhanced alkaline protease production. Brazilian Archives ofBiology and Technology, 55(1), 135e144.
Murugan, S., Arnold, D., Pongiya, U. D., & Narayanan, P. M. (2011).Production of xylanase from Arthrobacter sp. MTCC 6915 usingsaw dust as substrate under solid state fermentation. EnzymeResearch, 2011, 7. http://dx.doi.org/10.4061/2011/696942 (ArticleID 696942).
Poorna, C. A., & Prema, P. (2006). Production and partialcharacterization of endoxylanase by Bacillus pumilus usingagro industrial residues. Biochemical Engineering Journal, 33(2),106e112.
Pratumteep, A., Sansernsuk, J., Nitisinprasert, S., &Apiraksakorn, J. (2010). Production, characterization andhydrolysation products of xylanase from Bacillus subtilisGN156. KKU Research Journal, 15, 343e350.
Rafi, S., Asghar, M., Yaqube, M., & Ghour, M. A. (1998). Productionof xylanase from corn stover by Arachniotus sp. Pakistan Journalof Biological Sciences, 1(4), 380e382.
Roy, N., & Rowshanul, H. M. (2009). Isolation and characterizationof xylanase producing strain of Bacillus cereus from soil. IranianJournal of Microbiology, 1, 49e53.
J o u rn a l o f R a d i a t i o n R e s e a r c h and A p p l i e d S c i e n c e s 9 ( 2 0 1 6 ) 1 3 9e1 4 7 147
Sa'-Pereira, P., Costa-Ferreira, M., & Aires-Barros, M. R. (2002).Enzymatic properties of a neutral endo-1,3(4)-b-xylanase Xyl IIfrom Bacillus subtilis. Journal of Biotechnology, 94, 265e275.
Sajitha, N., Vasanthabharathi, V., Lakshminarayanan, R., &Jayalakshmi, S. (2011). Amylase from an estuarine Bacillusmegaterium. Current Research Journal of Biological Sciences, 3(2),110e115.
Saleem, M., Akhtar, M. S., & Jamil, S. (2002). Production ofxylanase on natural substrates by Bacillus subtilis. InternationalJournal of Agricultural Biology, 4(2), 211e213.
Sanghi, A., Garg, N., Gupta, V. K., Mittal, A., & Kuhad, R. C. (2010).One-step purification and characterization of cellulase-freexylanase produced by alkalophilic Bacillus subtilis ASH.Brazilian Journal of Microbiology, 41, 467e476.
Sepahy, A. A., Ghazi, S., & Sepahy, M. A. (2011). Cost-effectiveproduction and optimization of alkaline xylanase byindigenous Bacillus mojavensis AG137 fermented onagricultural waste. Enzyme Research, 9. http://dx.doi.org/10.4061/2011/593624 (Article ID 593624).
Shabeb, M. S., Younis, A. M., Hezayen, F. F., & Nour-Eldien, M. A.(2010). Production of cellulase in low-cost medium by Bacillussubtilis KO strain. World Applied Science Journal, 8(1), 35e42.
Shaheen, M., Shah, A. A., Hameed, A., & Hasan, F. (2008).Influence of culture conditions on production and activity ofprotease from Bacillus subtilis bs1. Pakistan Journal of Botany,40(5), 2161e2169.
Sharma, B., & Bajaj, B. K. (2005). Production and partialcharacterization of alkali-tolerant xylanase from analkalophilic Streptomyces sp. CD3. Journal of Scientific andIndustrial Research, 64, 688e697.
Simphiwe, P., Ademola, B., Olaniran, O., & Pillay, B. (2011).Sawdust and digestive bran as cheap alternate substrates forxylanase production. African Journal of Microbiology Research,5(7), 742e752.
Srinivasan, M. C., & Meenakshi, V. R. (1999). Microbial xylanasefor paper industry. Current Science, 77(1), 137e142.
Veluz, G., Taksuo, K., Hiroshi, M., & Yusaku, F. (1999). ScreeningRhizopus sp. Journal of Faculty of Agriculture, 43, 419e423.