| Products/Services Used | Details | Operation |
|---|---|---|
| Proteins, Expression, Isolation and Analysis | … The crude extract was applied to a 'High Affinity Ni-Charged Resin' column (GenScript, USA) and washed with 8× bed volumes of Washing buffer (20 mmol/L Tris-HCl, 20–100 mmol/L imidazole, 0.5 mol/L NaCl), and then eluted by using 10× bed volumes of Elution buffer (20 … | Get A Quote |
Microbial transglutaminase (MTG) gene (mtg) from H197 strain was cloned by PCR and mutated by deleting a specific 84 bp fragment using overlapping extension PCR. The mutant MTG and the wild MTG genes expressed by recombinant plasmid pET32a+- mutant mtg and pET32a+ -mtg， respectively， and were harvested by alternating freeze-thaw steps and purified by Ni column. The purified mutant MTG and the wild MTG exhibited 0.22 U/mg and 0.16 U/mg activity， respectively， and 0.69 U/mg and 0.54 U/mg activity， respectively， after activated by trypsin. The molecular weight of mutant MTG was estimated as 67 kDa by SDS-PAGE. Both MTGs showed optimum activity at pH 6-8 for hydroxamate formation from N-CBZ-Gln-Gly and... More
