DİLŞAT NİGAR ÇOLAK, Kadriye İNAN BEKTAŞ, Müslüm TOKGÖZ, SABRİYE ÇANAKÇI, ALİ OSMAN BELDÜZ

Screening of Xylanase and Glucose Isomerase Producing Bacteria Isolated from Hot Springs in Turkey

The aim of this study was screening of xylanase and glucose isomerase producing thermophilic bacteria isolated from some hot springs located in Aegean Region of Turkey. Total sixty eight thermophilic isolates (Anoxybacillus, Brevibacillus, Geobacillus, Aneurinibacillus, Thermus, Paenibacillus and Proteobacter) were collected previously from these fields and identified based on 16S rDNA gene sequences. Isolates were screened by plate assay for determining the xylanase and glucose isomerase production abilities seperately in order to find new strains for industrial processes. After an incubation period of two days for xylanase and 5-6 days for glucose isomerase at 50-60 °C, positive isolates were determined. Enzyme producing isolates were confirmed by spectrophotometric measurements with crude enzyme extracts, birchwood xylan and glucose were used as substrates. Most of the isolates (fifty nine) were positive for xylan degradation while only sixteen of them were positive for glucose isomerase activity. Fourteen of the isolates showed both xylanase and glucose isomerase activity. None of the isolates belong to the genera Paenibacillus, Aneurinibacillus, and Proteobacter were glucose isomerase positive, although the glucose isomerase activity of Geobacillus isolates were notably high. Both xylanase and glucose isomerase activities were observed at 50-60 °C which is suitable for biotechnological applications.

PDF

___

P. Anbu, S. C. Gopinath, A. C. Cihan, and B. P. Chaulagain, “Microbial enzymes and their applications in industries and medicine,” Biomed. Res. Int., pp. 1–3, 2015.

M. E. Bruins, A. E. M. Janssen, and R. M. Boom, “Thermozymes and their applications,” Appl. Biochem. Biotechnol. vol 90, pp.155-186, 2001.

J. Eichler, “Biotechnological uses of archaeal enzymes,” Biotechnol. Adv. vol 19, pp. 261-278. 2001.

D. Covan, R. Daniel, and H. Morgan, “Thermophilic proteases: properties and applications,” Trends Biotechnol. vol 3, pp. 68-72, 1985.

D. A. Covan, “Thermophilic proteins: stability and function in aqueous and organic solvents,” Comp. Biochem. Physiol., Part A: Mol. Integr. Physiol. vol 118, pp.429-438, 1997.

A. Gupta, and S. K. Khare, “A protease stable in organic solvents from solvent tolerant strain Pseudomonas aeruginosa,” Bioresour. Technol. vol 97, pp. 1788-1793, 2006.

T. Collins, C. Gerday, and G. Feller, “Xylanases, xylanase families and extremophilic xylanases,” FEMS Microbiol. Reviews. vol 29, pp. 3–23, 2005.

N. Kulkarni, A. Shendye, and M. Rao, “Molecular and biotechnological aspects of xylanases,” FEMS Microbiology Reviews, vol. 23, no 4, pp. 411–456, 1999.

Q. K. Beg, M. Kapoor, L. Mahajan, and G. Hoondal, “Microbial xylanases and their industrial applications: a review,” Applied Microbiology and Biotechnology. vol 56, pp. 326–338, 2001.

A. Blanco, P. Diaz, J. Zueco, P. Parascandola, and F. I. J. Pastor, “A multidomain xylanase from a Bacillus sp. with a region homologous to thermostabilizing domains of thermophilic enzymes,” Microbiology, vol 145, pp. 2163– 2170. 1999.

C. Tachaapaikoon, K. L. Kyu, and K. Ratanakhanokchai, “Purification of xylanase from alkaliphilic Bacillus sp. K-8 by using corn husk column,” Process Biochemistry, vol 12, pp. 2441–2445, 2006.

S. B. Chidi, B. Godana, I. Ncube, Van E. J. Rensburg, A. Cronshaw, and E. K. “Abotsi Production, purification and characterization of celullase-free xylanase from Aspergillus terreus UL 4209,” African Journal of Biotechnology, vol 7, no 21, pp. 3939–3948, 2008.

S. Ahmed, S. Riaz, and A. Jamil, “Molecular cloning of fungal xylanases: an overview,” Applied microbiology and biotechnology, vol 84, no 1, pp. 19-35, 2009.

L. A. Van den Broek R. M. Lloyd, G. Beldman, J. C. Verdoes, B. V. McCleary, and A. G. Voragen, “Cloning and characterization of arabinoxylan arabinofuranohydrolase-d3 (Axhd3) from Bifi-dobacterium adolescetis DSM20083,” App. Microbiol. and Biotechnology. vol 67, pp. 641–647, 2005.

D. Verma, A. Anand, and T. Satyanarayana, “Thermostable and alkalistable endoxylanase of the extremely thermophilic bacterium Geobacillus thermodenitrificans TSAA1: cloning, expression, characteristics and its applicability in generating xylooligosaccharides and fermentable sugars,” App. Biochemistry and Biotechnology, vol 170, pp. 119-130, 2013.

K. Bouacem, et al. “Partial characterization of xylanase produced by Caldicoprobacter algeriensis, a new thermophilic anaerobic bacterium isolated from an Algerian hot spring,” Appl Biochem Biotechnol. vol 174, pp. 1969–1981, 2014.

N. Annamalai, R. Thavasi, S. Jayalakshmi, and T. Balasubramanian, “Thermostable and alkaline tolerant xylanase production by Bacillus subtilis isolated from marine environment,” Indian J Biotechnol. vol 8, pp. 291–297, 2009.

J. Bradner, M. Gillings, and K. Nevalainen, “Qualitative assessment of hydrolytic activities in Antarctic microfungi grown at different temperatures on solid media,” World J Microbiol Biotechnol. vol 15, pp. 131–132, 1999.

B. Sonnleitner, and A. Fiechter, “Advantages of using thermophiles in biotechnological processes: expectations and reality,” Trends in Biotechnology, no 1, pp. 74-80, 1983.

S. H. Bhosale, M. B. Rao, and V. V. Deshpande, “Molecular and industrial aspects of glucose isomerase,” Microbiol Rev. vol 60, pp. 280–300. 1996.

M. G. Wovcha, D. L. Steuerwald, and K. E. Brooks, “Amplification of D-xylose and Dglucose isomerase activities in Escherichia coli by gene cloning,” Appl. Environ. Microbiol. vol 45, pp. 1402–1404, 1983.

C. Bucke, In "Microbial Enzymes and Biotechnology" (Ed. W.M.Fogarty); Applied Science Pub., London; pp. 93-129, 1983.

Y. B. Tewari, and R. N. Goldberg, “Thermodynamics of the conversion of aqueous glucose to fructose,” Appl. Biochem. Biotechnol. vol 11, pp. 17–24, 1985.

F. H. Verhoff, G. Boguslawski, O. J. Lautero, S. T. Schlager, and Y. C. Jao, “Glucose isomerase,” In “H. Comprehensive biotechnology;” Balanch, W., Drew, S., Wang, D.I.C, Pergamon Press, Oxford, pp. 837–859, 1985.

K. Inan, “İzmir ve Aydın illerindeki bazı kaplıcalardan izole edilen termofilik bakteri izolatlarının moleküler taksonomisi ve D1021 izolatının glukoz izomerazının karakterizasyonu,” Doktora tezi. Karadeniz Teknik Üniversitesi, Fen Bilimleri Enstitüsü, Trabzon, 2011.

A. Gessesse, and B. A. Gashe, “Production of alkaline xylanase by an alkaliphilic Bacillus sp. Isolated from a soda lake,” Journal of applied Microbiology, vol 83, pp. 402-406, 1997.

G. L. Miller, “Use of dinitrosalicyclic acid reagent for determination of reducing sugars,” Anal Chem. vol 31, pp. 426-428, 1959.

C. Lee, L. Bhatnagar, B. C. Saha, Y. E. Lee, M. Takagi, T. Imanaka, M. Bagdasarian, and J. G. Zeikus, “Cloning and Expression of the Clostridium thermosulfurogenes Glucose Isomerase Gene in Escherichia coli and Bacillus subtilis,” Appl. Environ. Microbiol. vol 56, pp. 2638–2643, 1990.

Z. Dische, and E. A. Borenfreund, “New Spectrophotometric Method for the Detection and Determination of Ketosugars and Trioses,” J.Biol.Chem. vol 192, pp. 583- 587, 1951.

X. Huang, J. Lin, X. Ye, and G. Wang, “Molecular characterization of a thermophilic and salt-and alkaline-tolerant xylanase from Planococcus sp. SL4, a strain isolated from the sediment of a soda lake,” J Microbiol Biotechnol. vol 25, pp. 662–671, 2015.

S. Subramanian, and P. Prema, “Cellulasefree xylanases from Bacillus and other micro-organisms,” FEMS Microbiol Lett. vol 183, pp. 1–7, 2000.

S. Gupta, R. C. Kuhad, B. Bhushan, and G.S. Hoondal, “Improved xylanase production from a haloalkalophil-ic Staphylococcus sp. SG-13 using inexpensive agricultural residues,” World J Microbiol Biotechnol. vol 17, pp. 5–8, 2001.

D. Chapla, H. Patel, D. Madamwar, and A. Shah, “Assessment of a thermostable xylanase from Paenibacillus sp. ASCD2 for application in prebleaching of eucalyptus kraft pulp,” Waste Biomass Valor. vol 3, pp. 269–274, 2012.

L. Dahlberg, “Thermostable Xylanases from Rhodothermus marinus,” Ph.D. dissertation, Biotechnology, Lund University, 1996. [35] H. Chakdar, M. Kumar, K. Pandiyan, A. Singh, K. Nanjappan, P. L. Kashyap, and A. K. Srivastava, “Bacterial xylanases: Biology to biotechnology,” Biotech. vol 6, no 2, pp. 150, 2016.

D. Verma, and T. Satyanarayana, “Cloning, expression and applicability of thermoalkali- stablexylanaseof Geobacillus thermoleovorans in generating xylooligosaccharides from agro-residues,” Biores Technol. vol 107, pp. 333–338, 2012.

A. Bhalla, K. M. Bischoff, and R. K. Sani, “Highly thermostable xylanase production from a thermophilic Geobacillus sp. strain WsUcF1 utilizing lignocellulosic biomass,” Front Bioeng Biotechnol. vol 3, pp. 84, 2015.

M. Kacagan, S. Canakci, C. Sandalli, K. Inan, D. N. Colak, and A. O. Belduz, “Characterization of a xylanase from a thermophilic strain of Anoxybacillus pushchinoensis A8,” Biologia, Sec. Cellular and Molecular Biology, vol 63, no 5, pp. 599-606, 2008.

J. T. Ellis, and T. S. Magnuson, “Thermostable and alkalistable xylanases produced by the thermophilic bacterium Anoxybacillusflavithermus TWX YL3,” ISRN Microbiol. pp. 517-524, 2012.

R. O. Marshall, and E. R. Kooi, “Enzymatic conversion of D-glucose to Dfrucose,” Sci. vol 125, pp. 648–649, 1957.

S. A. Barker, and J. A. Shirley, “Glucose oxidase, Glucose dehydrogenase, Glucose isomerase, β-Galactosidase and Invertase,” In Microbial Enzymes and Bioconversion, A. H. Rose (eds.). San Francisco, London New York, Toronto and Sydney: Academic Press. pp. 173-226, 1980.

I. V. Ulezlo, A. V. Ananychev, and A. M. Bezborodov, “Glucose isomerase (xylose ketol isomerase),” Uspekhi Biologicheskoy Khimii (Moscow), vol 27, pp. 136-163, 1986.

C. A. Kent, and A. N. Emery, “The preparation of an immobilised glucose isomerase. I. The production and selected properties of the partially purified enzyme from Lactobacillus brevis,” Journal of Appl. Chem. And Biotechn. vol 23, no 9, pp. 689- 703, 1973.

M. Weber, M. J. Foglietti, and F. Percheron, “Purification of alpha-amylase by affinity chromatography on cross-linked starch,” Biochimie, vol 58, pp. 1299–1302, 1976.

J. Chauthaiwale, and M. Rao, “Production and purifi cation of extracellular D-xylose isomerase from an alkaliphilic, thermophilic Bacillus sp.” Appl. Environ. Microbiol. vol 60, pp. 4495–4499, 1994.

G. Boguslawski, and M. J. Rynski, “Novel strain of Bacillus licheniformis useful in production of glucose isomerase and method of screening Bacillus mutants for the ability to produce glucose isomerase in the absence of xylose,” U.S. patent 4,355,103. 1982.

C. A. Smith, M. Rangarajan, and B. S. Hartley, “D-Xylose (D-glucose) isomerase from Arthrobacter strain NRRL B3728,” Biochem. J. vol 277, pp. 255–261, 1991.

A. N. Moneke et al. “D-xylose isomerases from a newly isolated strain, Paenibacillus sp., and from Alcaligenes ruhlandii: isolation, characterization and immobilisation to solid supports,” Appl Micr B. vol 50, no 5, pp. 552-557, 1998.

H. Karaoglu, D. Yanmis, F. A. Sal, A. Celik, S. Canakci, and A. O. Belduz, “Biochemical characterization of a novel glucose isomerase from Anoxybacillus gonensis G2 T that displays a high level of activity and thermal stability,” J Mol Catal B: Enzym, vol 97, pp. 215–224, 2013.

L. Konak, Y. Kolcuoglu, E. Ozbek, A. Colak, and B. Ergenoglu, “Purification and characterization of an extremely stable glucose isomerase from Geobacillus thermodenitrificans Th2,” App. Biochem. and Microbiol. vol 50: pp. 25-29, 2014.

K. Dekker, H. Yamagata, K. Sakaguchi, and S. Udaka, “Xylose (glucose) isomerase gene from the thermophile Thermus thermophilus: cloning, sequencing, and comparison with other thermostable xylose isomerases,” J Bacteriol. vol 173, no 10, pp. 3078–3083, 1991.

K. Inan Bektas, A. Ozer, H. I. Guler, A. O Belduz, and S. Canakci, “Brevibacillus gelatini sp. nov., isolated from a hot spring,” Int. Journal of Systematic and Evolutionary Microbiology, vol 66, pp. 712-718, 2016.