Immobilization of urease from pigeonpea (Cajanus cajan) on agar tablets and its application in urea assay.
The pigeonpea urease was immobilized on agar, a common gelling substance. The tablet strips were used as moulds to cast agar tablets of uniform shape and size. The time and temperature of solidification of agar was 6 min and 44 degrees C, respectively. The 5 % agar (w/v) and 0.019 mg protein/agar tablet yielded an optimum immobilization of 51.7%. The optimum pH was shifted through 0.2 U (from 7.3 to 7.5) towards basic side upon immobilization. The optimum temperature of soluble and immobilized urease was 30 degrees C and 60 degrees C, respectively, showing the improvement in thermal stability of urease. There was an increase in K m from 3.23 to 5.07 mM after immobilization. The half-lives of soluble and immobilized urease were 21 and 53 days, respectively, at pH 7.3 and 4 degrees C. The urea was estimated in different blood samples with the help of immobilized urease and the results were consistent with those from clinical pathology laboratory through an autoanalyzer (Zydus Co., Rome, Italy).
Mulagalapalli
Swati
S
School of Biotechnology, Faculty of Science, Banaras Hindu University, Varanasi, 221005, India.
Kumar
Sandeep
S
Kalathur
Ravi Charan Reddy
RC
Kayastha
Arvind M
AM
D000362:Agar / Q000737:chemistry*; D036561:Cajanus / Q000201:enzymology*; D002624:Chemistry, Clinical / Q000379:methods; D019963:Clinical Chemistry Tests / Q000379:methods; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D013696:Temperature; D014508:Urea / Q000032:analysis / Q000097:blood / Q000378:metabolism*; D014510:Urease / Q000737:chemistry / Q000378:metabolism*
D004800:Enzymes, Immobilized; D014508:Urea; D000362:Agar; D014510:Urease