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Journals / Osmaniye Korkut Ata Üniversitesi Fen Bilimleri Enstitüsü Dergisi / 2026 / Cilt: 9 Sayı: 4

Partial Characterisation of Acid-stable Cellulase Enzymes Produced by Bacillus Isolates

Bacillus İzolatları Tarafından Üretilen Asit-dirençli Selülaz Enzimlerinin Kısmi Karakterizasyonu

Cansu Nur Karaturna(ÇUKUROVA ÜNİVERSİTESİ, FEN BİLİMLERİ ENSTİTÜSÜ)0009-0005-2253-485X
B. Devrim Özcan(ÇUKUROVA ÜNİVERSİTESİ, ZİRAAT FAKÜLTESİ, ZOOTEKNİ BÖLÜMÜ)0000-0002-9198-656X
Elif Dikkaya(ÇUKUROVA ÜNİVERSİTESİ, ZİRAAT FAKÜLTESİ)
Pages
2369–2383
Publication Date
2026-09-09
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AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

Zymogram analysis determined the molecular weights of CNK1 and CNK2 cellulase enzymes to be approximately 21.1 and 231.9 kDa, respectively.

📌 3 Key Takeaways

🎯
Objective & Hypothesis:In this study, two acidophilic Bacillus sp. isolates exhibiting cellulase activity were obtained from soil.
📊
Key Findings & Data:CNK1 and CNK2, respectively.
💡
Conclusion & Impact:Zymogram analysis determined the molecular weights of CNK1 and CNK2 cellulase enzymes to be approximately 21.1 and 231.9 kDa, respectively.
🔬Design:Araştırma Makalesi
📍Scope:Comprehensive Sample
⚙️Variables:Bacillus sp. · İzolasyon · Asit-dirençli selülaz

Abstract

In this study, two acidophilic Bacillus sp. isolates exhibiting cellulase activity were obtained from soil. The isolates were designated as Bacillus sp. CNK1 and CNK2, respectively. CNK1 cellulase reached its maximum activity level at the 48th hour of the bacterium's growth period, while CNK2 cellulase reached its maximum activity at the 72nd hour. Both CNK1 and CNK2 cellulases exhibited optimal activity at 40 °C, with CNK1 cellulase showing optimal activity at pH 4.0 and CNK2 cellulase at pH 5.0. CNK1 cellulase was stimulated by CaCl2 and inhibited by MgCl2, CoCl2, EDTA, and SDS. Conversely, CNK2 cellulase was stimulated by CaCl2, CoCl2, MgCl2, and EDTA, but inhibited by SDS. Zymogram analysis determined the molecular weights of CNK1 and CNK2 cellulase enzymes to be approximately 21.1 and 231.9 kDa, respectively.

Özet

Bu çalışmada, topraktan selülaz aktivitesine sahip iki adet asidofil Bacillus sp. izolasyonu gerçekleştirilmiştir. İzolatlar sırasıyla Bacillus sp. CNK1 ve CNK2 olarak isimlendirilmişlerdir. CNK1 selülazı bakterinin üreme periyodunun 48. saatinde maksimum aktivite seviyesine ulaşırken, CNK2 selülazı 72. saatte ulaşmıştır. CNK1 ve CNK2 selülazlarının her ikisi de optimum aktivitelerini 40 °C’de gösterirken, CNK1 selülazı optimum aktivitesini pH 4.0’de, CNK2 selülazı ise 5.0’da göstermiştir. CNK1 selülazı CaCl2 tarafından stimüle edilirken, MgCl2, CoCl2, EDTA ve SDS tarafından inhibe edilmiştir. Buna karşılık, CNK2 selülazı CaCl2, CoCl2, MgCl2 ve EDTA tarafından stimüle edilirken, SDS tarafından inhibe edilmiştir. Zimogram analizi ile CNK1 ve CNK2 selülaz enzimlerinin moleküler ağırlıkları sırasıyla yaklaşık olarak 21.1 ve 231.9 kDa olarak belirlenmiştir.

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Cite This Article

Karaturna, C. N., Özcan, B. D., Dikkaya, E. (2026). Partial Characterisation of Acid-stable Cellulase Enzymes Produced by Bacillus Isolates. Osmaniye Korkut Ata Üniversitesi Fen Bilimleri Enstitüsü Dergisi, Cilt: 9 Sayı: 4, 2369–2383. https://doi.org/10.47495/okufbed.1494318

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References (61)

  1. Amanda W., Thompson VF., Goll DE. Interaction of calpastatin with calpain: a review. Biological Chemistry, 2004; 385: 465-472.
  2. Ariffin H., Abdullah N., Umi Kalsom MS., Shirai Y., Hassan MA. Production and characterization of cellulase by Bacillus pumilus EB3. International Journal of Engineering and Technology 2006; 3(1): 47-53.
  3. Arja MO. Cellulases in the textile industry. In Industrial Enzymes: Structure, function and applications, 2007; pp. 51-63. Dordrecht: Springer, Netherlands.
  4. Asha BM., Sakthivel N. Production, purification and characterization of a new cellulase from Bacillus subtilis that exhibit halophilic, alkalophilic and solvent-tolerant properties. Annals of Microbiology, 2014; 64(1): 1839-1848.
  5. Avanus K. Genetic variability of CAST gene in native sheep breeds of Turkey. Kafkas Üniversitesi Veteriner Fakültesi Dergisi, 2015; 21(6): 789-794.
  6. Averna M., Tullio RD., Passalacqua M., Salamini F., Ponteremoli S., Mellon E. Changes in intracellular calpstatin localisation are mediated by reversible phosphorylation. Biochemical Journal 2001; 354: 25-30.
  7. Azadian F., Badoei-Dalfard A., Namaki-Shoushtari A., Hassanshahian M. Purification and biochemical properties of a thermostable, haloalkaline cellulase from Bacillus licheniformis AMF-07 and its application for hydrolysis of different cellulosic substrates to bioethanol production. Molecular Biology Research Communications, 2016; 5(3): 143.
  8. Bader J., Albin A., Stahl U. Spore-forming bacteria and their utilisation as probiotics. Beneficial Microbes, 2012; 3(1): 67-75.
  9. Balasubramanian N., Toubarro D., Teixeira M., Simõs N. Purification and biochemical characterization of a novel thermo-stable carboxymethyl cellulase from Azorean isolate Bacillus mycoides S122C. Applied Biochemistry and Biotechnology 2012; 168: 2191-2204.
  10. Bhat M. Cellulases and related enzymes in biotechnology. Biotechnology Advances 2000; 18(5): 355-383.
  11. Bhat MK., Bhat S. Cellulose degrading enzymes and their potential industrial applications. Biotechnology Advances 1997; 15(3-4): 583-620.
  12. Brumm PJ., Hebeda RE., Teague WM. Purification and characterization of the commercialized, cloned Bacillus megaterium α‐amylase. Part I: Purification and Hydrolytic Properties. Starch‐Stärke, 1991; 43(8): 315-319.
  13. Burhan A., Nisa U., Gökhan C., Ömer C., Ashabil A., Osman G. Enzymatic properties of a novel thermostable, thermophilic, alkaline and chelator resistant amylase from an alkaliphilic Bacillus sp. isolate ANT-6. Process Biochemistry 2003; 38(10): 1397-1403.
  14. Caf Y. Valipour E., Arikan B. Study on cold-active and acidophilic cellulase (CMCase) from a novel psychrotrophic isolate Bacillus sp. K-11. International Journal of Current Microbiology and Applied Sciences 2014; 3(5): 16-25.
  15. Cherry JR., Fidantsef AL. Directed evolution of industrial enzymes: An update. Current Opinion in Biotechnology 2003; 14(4): 438–443.
  16. Duman Y., Karakuş YY., Sertel A., Polat F. Production, purification, and characterization of a thermo-alkali stable and metal-tolerant carboxymethylcellulase from newly isolated Bacillus methylotrophicus Y37. Turkish Journal of Chemistry 2016; 40(5): 802-815.
  17. Gessesse A., Gashe BA. Production of alkaline protease by an alkaliphilic bacteria isolated from an alkaline soda lake. Biotechnology Letters 1997; 19: 479-481.
  18. Goyal V., Mittal A., Bhuwal AK., Singh G., Yadav A., Aggarwal NK. Parametric optimization of cultural conditions for carboxymethyl cellulase production using pretreated rice straw by Bacillus sp. 313SI under stationary and shaking conditions. Biotechnology Research International, 2014; Article ID 651839, 7 pages, https://doi.org/10.1155/2014/651839.
  19. Kelly CT., Heffernan ME., Fogarty WM. A novel α-glucosidase produced by Bacillus amylolyticus. Biotechnology Letters 1980; 2: 351-356.
  20. Kırcı H. Selüloz türevleri ve kullanım yerleri. KSÜ Fen ve Mühendislik Bilimleri Dergisi 2001; 4: 119-130.
  21. Kim BK., Lee BH., Lee YJ., Jin IH., Chung CH., Lee JW. Purification and characterization of carboxymethylcellulase isolated from a marine bacterium, Bacillus subtilis subsp. subtilis A-53. Enzyme and Microbial Technology 2009; 44(6-7): 411-416.
  22. Kim CH., Kim DS. Extracellular cellulolytic enzymes of Bacillus circulans are present as two multipleprotein complexes. Applied Biochemistry and Biotechnology 1993; 42: 83-94.
  23. Kiran ÖE., Çömlekçioğlu U., Dostbil N. Bazı mikrobiyal enzimler ve endüstrideki kullanım alanları. Fen ve Mühendislik Dergisi 2006; 9: 12-19.
  24. Kocamaz S., Kocamaz D. Isolation and characterisatıon of acidic cellulase enzyme from native Bacillus sp. S-41 strain. Journal of Applied Biological Sciences 2019; 13(1): 01-05.
  25. Kozen FH., Netravali AN. Cellulosic fibers: A brief review. Sustainable Composites 2014; 143-180.
  26. Kuhad RC., Gupta R., Singh A. Microbial cellulases and their industrial applications. Enzyme Research 2011; 2011, 280696.
  27. Ladeira SA., Cruz E., Delatorre AB., Barbosa JB., Leal Martins ML. Cellulase production by thermophilic Bacillus sp: SMIA-2 and its detergent compatibility. Electronic Journal of Biotechnology 2015; 18(2): 110-115.
  28. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227(5259): 680-685.
  29. Lee KD., Kim J. Kim H. Isolation and characterization of Bacillus sp. KD1014 producing carboxymethyl-cellulase. Journal of Microbiology 1996; 34(4): 305-310.
  30. Lin L., Kan X., Yan H., Wang D. Characterization of extracellular cellulose-degrading enzymes from Bacillus thuringiensis strains. Electronic Journal of Biotechnology 2012; 15(3): 1-7.
  31. Listyaningrum NP., Sutrisno A., Wardani AK. Characterization of thermostable cellulase produced by Bacillus strains isolated from solid waste of carrageenan. IOP Conference Series: Earth and Environmental Science, International Conference on Biomass and Bioenergy, Yogyakarta, Indonesia, 2018.
  32. Lo HF., Lin LL., Chen HL., Hsu HH., Chang CT. Enzyme properties of a SDS-resistant Bacillus sp. TS-23 α-amylase produced by recombinant Escherichia coli. Process Biochemistry 2001; 36: 743-750.
  33. Lucas R., Robles A., García MT., Alvarez de Cienfuegos G., Gálvez A. Production, purification, and properties of an endoglucanase produced by the hyphomycete Chalara (Syn. Thielaviopsis) paradoxa CH32. Journal of Agricultural and Food Chemistry 2001; 49(1): 79-85.
  34. Lynd LR., Weimer PJ., van Zyl WH., Pretorius IS. Microbial cellulose utilization: Fundamentals and biotechnology. Microbiology and Molecular Biology Reviews 2002; 66(3): 506–577.
  35. Madika A., Ameh JB., Machido DA. Isolation and screening of Bacillus subtilis from soil for amylase production. UMYU Journal of Microbiology Research (UJMR) 2017; 2(2): 82-86.
  36. Marco ÉGD., Heck K., Martos ET., Van Der Sand ST. Purification and characterization of a thermostable alkaline cellulase produced by Bacillus licheniformis 380 isolated from compost. Anais da Academia Brasileira de Ciências 2017; 89(3 Suppl.): 2359-2370.
  37. Miller GL. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical Chemistry 1959; 31(3): 426-428.
  38. Miyagi T., Kaneichi K., Fukumura M., Karita S., Sakka K., Shimada K., Ohmiya K. Electro-transformation of Bacillus subtilis for expressing a Ruminococcus albus cellulase gene. Animal Science and Technology 1994; 65(5): 407-407.
  39. Özcan N., Cunningham C., Harris WJ. Cloning of a cellulase gene from the rumen anaerobe Fibrobacter succinogenes SD35 and partial characterization of the gene product. Letters in Applied Microbiology 1996; 22(1): 85-89.
  40. Rahman MS., Hasan MS., Nitai AS., Nam S., Karmakar AK., Ahsan MS., Shiddiky MJA, Ahmed MB. Recent developments of carboxymethyl cellulose. Polymers 2021; 13(8): 1345.
  41. Rawat R., Tewari L. Purification and characterization of an acidothermophilic cellulase enzyme produced by Bacillus subtilis strain LFS3. Extremophiles 2012; 16: 637-644.
  42. Ryu DD., Mandels M. Cellulases: biosynthesis and applications. Enzyme and Microbial Technology 1980; 2(2): 91-102.
  43. Sankaralingam S., Palpperumal S., Harinathan B., Kathiresan D., Shankar T., Sundarapandian S. Screening and characterization of cellulase by Bacillus megaterium isolated from marine sediments and its antimicrobial activity. Boletin Latinoamericano y del Caribe de Plantas Medicinales y Aromaticas 2018; 17(5): 646-653.
  44. Sethi A., Kovaleva ES., Slack JM., Brown S., Buchman GW. Scharf ME. A GHF7 cellulase from the protist symbiont community of Reticulitermes flavipes enables more efficient lignocellulose processing by host enzymes. Archives of Insect Biochemistry and Physiology 2013; 84(4): 175-193.
  45. Shahid ZH., Irfan M., Nadeem M., Syed Q., Qazi JI. Production, purification, and characterization of carboxymethyl cellulase from novel strain Bacillus megaterium. Environmental Progress & Sustainable Energy 2016; 35(6): 1741-1749.
  46. Shankar T., Isaiarasu L. Cellulase production by Bacillus pumilus EWBCM1 under varying cultural conditions. Middle-East Journal of Scientific Research 2011; 8(1): 40-45.
  47. Shankar T., Sankaralingam S., Balachandran C., Chinnathambi A., Nasif O., Alharbi SA., Sungwon P., Baskar K. Purification and characterization of carboxymethylcellulase from Bacillus pumilus EWBCM1 isolated from earthworm gut (Eudrilus eugeniae). Journal of King Saud University-Science 2021; 33(1): 101261.
  48. Shirai T., Ishida H., Noda JI., Yamane T., Ozaki K., Hakamada Y. Ito S. Crystal structure of alkaline cellulase K: Insight into the alkaline adaptation of an industrial enzyme. Journal of Molecular Biology 2001; 310(5): 1079-1087.
  49. Singh J., Batra N., Sobti RC. Purification and characterisation of alkaline cellulase produced by a novel isolate, Bacillus sphaericus JS1. Journal of Industrial Microbiology and Biotechnology 2004; 31(2): 51-56.
  50. Singhania, RR., Sukumaran RK., Patel AK., Larroche C., Pandey A. Advancement and comparative profiles in the production technologies using solid-state and submerged fermentation for microbial cellulases. Enzyme and Microbial Technology 2010; 46(7): 541-549.
  51. Sinoquet H., Thanisawanyangkura S., Mabrouk H., Kasemsap P. Characterization of the light environment in canopies using 3D digitising and image processing. Annals of Botany 1998; 82(2): 203-212.
  52. Sivakumar N., Al Zadjali A., Al Bahry S., Elshafie A., Eltayeb EA. Isolation and characterization of cellulolytic Bacillus licheniformis from compost. African Journal of Biotechnology 2016; 15(43): 2434-2446.
  53. Sukumaran RK., Singhania RR., Pandey A. Microbial cellulases-production, applications and challenges. Journal of Scientific & Industrial Research 2005; 64: 832-844.
  54. Sukumaran RK., Singhania RR., Mathew GM., Pandey A. Cellulase production using biomass feed stock and its application in lignocellulose saccharification for bioethanol production. Renewable Energy 2005; 34(2): 421–424.
  55. Takasaki Y. An amylase producing maltotetraose and maltopentaose from Bacillus circulans. Agricultural and Biological Chemistry 1983; 47(10): 2193-2199.
  56. Takasaki Y. An amylase producing maltotriose from Bacillus subtilis. Agricultural and Biological Chemistry 1985; 49(4): 1091-1097.
  57. Trivedi N., Gupta V., Kumar M., Kumari P., Reddy CRK., Jha B. An alkali-halotolerant cellulase from Bacillus flexus isolated from green seaweed Ulva lactuca. Carbohydrate Polymers 2011; 83(2): 891-897.
  58. Vijayaraghavan P., Vincent SGP. Purification and characterization of carboxymethyl cellulase from Bacillus sp. isolated from a paddy field. Polish Journal of Microbiology 2012; 61(1): 51-57.
  59. Wind RD., Buitelaar RM., Eggink G., Huizing HJ., Dijkhuizen L. Characterization of a new Bacillus stearothermophilus isolate: A highly thermostable α-amylase-producing strain. Applied Microbiology and Biotechnology 1994; 41: 155-162.
  60. Wiseman A. The application of enzymes in industry. In: Horwood, E. (Ed.), Handbook of Enzymes Biotechnology, Chichester-UK, 1987; 274-373.
  61. Wiseman A. Handbook of enzyme biotechnology. Ellis Horwood Limited. 1987.