InXy and SeXy, compact heterologous reporter proteins for mammalian cells.
AuthorsFluri DA, Kelm JM, Lesage G, Baba MD, Fussenegger M,
JournalBiotechnol Bioeng
PubMed ID17461419
'Mammalian reporter proteins are essential for gene-function analysis, drugscreening initiatives and as model product proteins for biopharmaceutical manufacturing. Bacillus subtilis can maintain its metabolism by secreting Xylanase A (XynA), which converts xylan into shorter xylose oligosaccharides. XynA is a family 11 xylanase monospecific for D-xylose containing substrates. Mammalian cells transgenic ... More
An ultrasensitive, continuous assay for xylanase using the fluorogenic substrate 6,8-difluoro-4-methylumbelliferyl beta-D-xylobioside.
AuthorsGe Y, Antoulinakis EG, Gee KR, Johnson I
JournalAnal Biochem
PubMed ID17241608
'We describe a fluorescence-based assay for the analysis of xylanase activity using a novel fluorogenic substrate, 6,8-difluoro-4-methylumbelliferyl beta-D-xylobioside (DiFMUX(2)). Generation of fluorescent 6,8-difluoro-4-methylumbelliferone (DiFMU) from the substrate corresponded directly to xylanase activity. High-performance liquid chromatography analysis of the digestion products showed that xylanase hydrolyzed DiFMUX(2) directly to DiFMU and xylobiose. ... More
Engineering highly thermostable xylanase variants using an enhanced combinatorial library method.
AuthorsHokanson CA, Cappuccilli G, Odineca T, Bozic M, Behnke CA, Mendez M, Coleman WJ, Crea R,
JournalProtein Eng Des Sel
PubMed ID21708791
A new directed evolution method was used to enhance the thermostability of the wild-type GH11 xylanase 2 (known as BD-11) from Hypocrea jecorina (Trichoderma reesei). Both Look-Through Mutagenesis (LTM™), which is a method for rapidly screening selected positions in the protein sequence for amino acids that introduce favorable properties, and ... More
Cellulose- and xylan-degrading thermophilic anaerobic bacteria from biocompost.
Nine thermophilic cellulolytic clostridial isolates and four other noncellulolytic bacterial isolates were isolated from self-heated biocompost via preliminary enrichment culture on microcrystalline cellulose. All cellulolytic isolates grew vigorously on cellulose, with the formation of either ethanol and acetate or acetate and formate as principal fermentation products as well as lactate ... More