Comparative Characterization of Crude and Recombinant Cellulase from Bacillus velezensis C37PLCA for Detergent Applications.

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Ceylan Yerebakan E., Nalçaoğlu Şenocak A., Beldüz A. O., Çanakçı S.

23rd Biotechnology Congress with International Participation, Ankara, Türkiye, 23 - 25 Ekim 2025, ss.97, (Özet Bildiri)

  • Yayın Türü: Bildiri / Özet Bildiri
  • Basıldığı Şehir: Ankara
  • Basıldığı Ülke: Türkiye
  • Sayfa Sayıları: ss.97
  • Karadeniz Teknik Üniversitesi Adresli: Evet

Özet

Comparative Characterization of Crude and Recombinant Cellulase from Bacillus velezensis C37PLCA for Detergent Applications

Esma CEYLAN¹, Aleyna NALÇAOGLU SENOCAK², Fatma Nur YILDIRIM1, Ali Osman BELDUZ¹, Sabriye CANAKCI¹

¹ Karadeniz Technical University, Faculty of Science, Department of Biology, Trabzon, TURKEY
² Karadeniz Technical University, Faculty of Science, Department of Molecular Biology and Genetics, Trabzon, TURKEY

esmaceylan@ktu.edu.tr

 

Cellulases are hydrolytic enzymes widely distributed among bacteria, fungi, and actinomycetes, catalyzing the hydrolysis of cellulose into soluble sugars. These enzymes are of considerable interest in the detergent industry due to their ability to enhance fabric care and remove cellulose-based residues. In detergents, cellulases contribute to stain removal, prevent fabric pilling, and improve overall wash performance, making them key components in modern laundry formulations. This study aimed to isolate, produce, and characterize a novel cellulase from Bacillus velezensis C37PLCA for potential detergent applications.

The crude enzyme was obtained by cultivating the bacterium in nutrient broth supplemented with 1% carboxymethyl cellulose (CMC) for three days, followed by recovery through centrifugation. The crude cellulase exhibited an optimum temperature of 60 °C and an optimum pH of 5.

For recombinant production, the cellulase gene was identified via genome analysis, amplified by PCR, cloned into pGEM-T Easy, subcloned into the extracellular expression vector pET20b, and expressed in Escherichia coli BL21. The recombinant enzyme was purified and confirmed by SDS-PAGE. It exhibited an optimum temperature of 60 °C and an optimum pH of 4. Thermal stability analysis revealed complete activity loss at 60-70 °C after 24 h, while more than 50% residual activity was retained at 25-50 °C. The enzyme maintained over 65% activity across pH 5-9 for up to 120 h.

Both crude and recombinant enzymes were evaluated for detergent compatibility. Comparative characterization highlighted functional similarities and differences between the two forms. Both enzymes were stable and active at 60 °C, and the recombinant form demonstrated broader pH tolerance. High thermal stability and pH tolerance are critical for detergent applications, as they ensure enzyme activity under harsh washing conditions. These findings underscore the potential of B. velezensis C37PLCA cellulase as a robust and effective candidate for incorporation into detergent formulations.

 

Keywords: Bacillus velezensis C37PLCA, cellulase, cloning, recombinant expression, detergent industry.

 

Acknowledgment: This work was supported by Scientific Research Projects Coordination Unit of Karadeniz Technical University. Project number: FBA-2023-10544