Characterization of expression and stability of recombinant cystein-rich protein human MT1A from yeast.
Metallothionein (MT) is the protein that has been shown to bind heavy metals, scavenge free radicals, protect DNA from radiation damage, and alleviate disease symptoms. However, only very limited success has been achieved in expression and production of active recombinant metallothionein. In this study, human metallothionein 1A (hMT1A) was transformed into yeast Pichia pastoris for expression with secretion of the protein into the medium. The expression system was optimized to obtain the targeted protein in active form at 335 mg per litre culture. hMT1A showed the character of extreme instability in the experiment. High concentration, aeration and heavy metal ions are the main factors affecting hMT1A stability.
Jie
Li
L
National Key Laboratory of Protein Engineering, College of life sciences, Peking University, 100871, PR China.
Kaifeng
Shao
S
Dian
Yao
Y
Lin
An
A
Binggen
Ru
R
D002244:Carbon / Q000378:metabolism; D002712:Chlorides / Q000378:metabolism; D003545:Cysteine / Q000032:analysis / Q000378:metabolism*; D015870:Gene Expression; D006461:Hemolysis; D006639:Histidine / Q000378:metabolism; D006801:Humans; D008668:Metallothionein / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D017967:Zinc Compounds / Q000378:metabolism
D002712:Chlorides; D011994:Recombinant Proteins; D017967:Zinc Compounds; D006639:Histidine; D002244:Carbon; C016837:zinc chloride; D008668:Metallothionein; D003545:Cysteine