Expression of beta-(1,3-1,4)-glucanase gene of Orpinomyces sp GMLF18 in Escherichia coli EC1000 and Lactococcus lactis subsp cremoris MG1363
TURKISH JOURNAL OF BIOLOGY, cilt.35, sa.4, ss.405-414, 2011 (SCI-Expanded, Scopus, TRDizin)
- Yayın Türü: Makale / Tam Makale
- Cilt numarası: 35 Sayı: 4
- Basım Tarihi: 2011
- Doi Numarası: 10.3906/biy-0912-25
- Dergi Adı: TURKISH JOURNAL OF BIOLOGY
- Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, TR DİZİN (ULAKBİM)
- Sayfa Sayıları: ss.405-414
- Anahtar Kelimeler: Orpthomyces, beta-(1,3-1,4)-glucanase, licA, Lactococcus lactis, lichenan, LACTOBACILLUS-REUTERI, STREPTOCOCCUS-LACTIS, MOLECULAR-CLONING, BETA-GLUCANASE, CEREAL-GRAINS, RUMINAL FUNGI, SEQUENCE, XYLANASE, CELLULASES, DEGRADATION
- Ankara Üniversitesi Adresli: Hayır
Özet
A gene encoding beta-(1,3-1,4)-glucanase (licA) was amplified from Orpinomyces sp. GMLF18 and expressed in Escherichia colt. The DNA sequence of licA showed that the gene was 707 bp and encoded a protein with a molecular mass of 26 kDa that belongs to family glycosyl hydrolase 16. The main LicA activity was observed to be cell-associated for the licA containing transformant E. coli, and the enzyme expressed in E. coli showed the highest activity at pH 5.0-6.0 and at temperatures of 40-50 degrees C. The enzyme was found to be stable at 40 degrees C; however, 12% of LicA activity was lost at 50 degrees C in 20 min. The licA was then introduced into the facultative anaerobic bacterium Lactococcus lactis subsp. cremoris MG1363 by a stable recombinant plasmid, pIL253. Although the enzymatic activity was lower than that in E. coli, the gene encoding the fungal originated lichenase was successfully expressed in L. lactis.