Cloning, purification and characterization of an alkali-stable endoxylanase from thermophilic Geobacillus sp 71

ÇANAKÇI S., Cevher Z., Inan K., Tokgoz M., Bahar F., Kacagan M., ...Daha Fazla

WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY, cilt.28, sa.5, ss.1981-1988, 2012 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 28 Sayı: 5
  • Basım Tarihi: 2012
  • Doi Numarası: 10.1007/s11274-011-1000-3
  • Dergi Adı: WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.1981-1988
  • Anahtar Kelimeler: Endo acting, Endoxylanase, Geobacillus sp., Thermostable, HETEROLOGOUS EXPRESSION, THERMOSTABLE XYLANASE, MICROBIAL XYLANASES, MOLECULAR-CLONING, BACILLUS, MICROORGANISMS, BIOLOGY, SYSTEMS, GENE
  • Karadeniz Teknik Üniversitesi Adresli: Evet

Özet

The gene encoding a xylanase from Geobacillus sp. 71 was isolated, cloned, and sequenced. Purification of the Geobacillus sp 7.1 xylanase, XyzGeo71, following overexpression in E. coli produced an enzyme of 47 kDa with an optimum temperature of 75A degrees C. The optimum pH of the enzyme is 8.0, but it is active over a broad pH range. This protein showed the highest sequence identity (93%) with the xylanase from Geobacillus thermodenitrificans NG80-2. XyzGeo71 contains a catalytic domain that belongs to the glycoside hydrolase family 10 (GH10). XyzGeo71 exhibited good pH stability, remaining stable after treatment with buffers ranging from pH 7.0 to 11.0 for 6 h. Its activity was partially inhibited by Al3+ and Cu2+ but strongly inhibited by Hg2+. The enzyme follows Michaelis-Menten kinetics, with K-m and V-max values of 0.425 mg xylan/ml and 500 mu mol/min.mg, respectively. The enzyme was free from cellulase activity and degraded xylan in an endo fashion. The action of the enzyme on oat spelt xylan produced xylobiose and xylotetrose.