Journal
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY
Volume 69, Issue 9, Pages 1711-1720Publisher
TAYLOR & FRANCIS LTD
DOI: 10.1271/bbb.69.1711
Keywords
animal cellulase; endo-beta-1,4-D-glucanase; termite; DNA shuffling; overexpression
Ask authors/readers for more resources
Among cellulase genes, those of animals are known for their difficulty in overexpression. We constructed a chimeric library by family shuffling of endo-beta-1,4-glucanase genes from four different termite species (Reticulitermes speratus, Nasutitermes takasagoensis, Coptotermes formosanus, and Coptotermes acinaciformis) sharing 78.5-96% homology in amino acid sequence. The constructed library was screened by Congo red plate assay combined with 96-well micro-enzyme assay, and clones showing enhanced CMCase activities were obtained. The mutated genes were overexpressed in Escherichia coli intracellularly as an active form. The endo-beta-1,4-glucanase (CMCase) activity in soluble fractions of E. coli harboring the mutant genes was 20-30 fold higher than that of wild-type genes. The mutant enzyme showed high activity against CMC and properties similar to those of the native enzymes.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available