Expression of β-(1,3-1,4)-glucanase gene of Orpinomyces sp GMLF18 in Escherichia coli EC1000 and Lactococcus lactis subsp cremoris MG1363

被引:5
作者
Comlekcioglu, Ugur [1 ]
Ozkose, Emin [1 ]
Akyol, Ismail [1 ]
Yazdic, Ferit Can [1 ]
Ekinci, Mehmet Sait [1 ]
机构
[1] Kahramanmaras Sutcu Imam Univ, Fac Agr, Dept Anim Sci, Biotechnol & Gene Engn Lab, Kahramanmaras, Turkey
关键词
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;
D O I
10.3906/biy-0912-25
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
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.
引用
收藏
页码:405 / 414
页数:10
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