Cloning and Characterization of Cellulase and Xylanase Coding Genes from Anaerobic Fungus Neocallimastix sp GMLF1


Comlekcioglu U., Ozkose E., Tutus A., Akyol İ., Ekinci M. S.

INTERNATIONAL JOURNAL OF AGRICULTURE AND BIOLOGY, cilt.12, sa.5, ss.691-696, 2010 (SCI-Expanded) identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 12 Sayı: 5
  • Basım Tarihi: 2010
  • Dergi Adı: INTERNATIONAL JOURNAL OF AGRICULTURE AND BIOLOGY
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.691-696
  • Anahtar Kelimeler: Neocallimastix, Cellulase, Xylanase, Cereal, Kraft pulp, CATALYTIC DOMAINS, MOLECULAR-BIOLOGY, WHEAT-STRAW, PURIFICATION, ENZYME, CELLOBIOHYDROLASE, ENDOGLUCANASE, ENUMERATION, SEQUENCE, CELA
  • Ankara Üniversitesi Adresli: Hayır

Özet

An anaerobic fungus was isolated from the cattle feces and identified as Neocallimastix sp. by using morphological approaches. Cellulase, xylanase, beta-xylosidase and beta-glucosidase production were assayed from culture supernatants and maximal activities were found 6.99, 10.68, 2.72 and 3.24 U/mL, respectively. Cellulase and xylanase coding genes were isolated by using polymerase chain reaction and expressed in E. coli. Nucleotide sequencing showed that the cell A (1367 bp) and xyn I B (992 bp) had open reading frames encoding polypeptides of 393 and 259 amino acids, respectively. Cel 1A showed highest activity on lichenan and followed by carboxymethyl cellulose but Xyn1B was found to be only active on xylan. The optimal conditions were pH 6.0 for CellA and pH 6.5 for Xyn1B and 50 C for both enzymes. The enzymes were stabile at 40-50 degrees C but readily inactivation occurred at 60 degrees C. Cell A activity was enhanced more than 50% in the presence of 1 mM MnCl2, CoCl2 and dithiothreitol, however the same effect was only recorded for Xyn1B with MnCl2. Application studies showed that CellA was found to be active on cereal grains such as barley and oat. Bio-bleaching trials by using Xyn1B reduced the kappa numbers of wheat straw and eucalyptus kraft pulps. (c) 2010 Friends Science Publishers