Affinity covalent immobilization of glucoamylase onto ρ-benzoquinone-activated alginate beads: II. Enzyme immobilization and characterization.
A novel affinity covalent immobilization technique of glucoamylase enzyme onto ρ-benzoquinone-activated alginate beads was presented and compared with traditional entrapment one. Factors affecting the immobilization process such as enzyme concentration, alginate concentration, calcium chloride concentration, cross-linking time, and temperature were studied. No shift in the optimum temperature and pH of immobilized enzymes was observed. In addition, K (m) values of free and entrapped glucoamylase were found to be almost identical, while the covalently immobilized enzyme shows the lowest affinity for substrate. In accordance, V (m) value of covalently immobilized enzyme was found lowest among free and immobilized counter parts. On the other hand, the retained activity of covalently immobilized glucoamylase has been improved and was found higher than that of entrapped one. Finally, the industrial applicability of covalently immobilized glucoamylase has been investigated through monitoring both shelf and operational stability characters. The covalently immobilized enzyme kept its activity over 36 days of shelf storage and after 30 repeated use runs. Drying the catalytic beads greatly reduced its activity in the beginning but recovered its lost part during use. In general, the newly developed affinity covalent immobilization technique of glucoamylase onto ρ-benzoquinone-activated alginate carrier is simple yet effective and could be used for the immobilization of some other enzymes especially amylases.
Polymer Materials Research Department, Advanced Technologies and New Material Research Institute, Mubarak City for Scientific Research and Technology Applications, New Borg El-Arab City, Alexandria, Egypt. mohyeldinmohamed@yahoo.com
M S Mohy
MS
Eldin
E I
EI
Seuror
M A
MA
Nasr
H A
HA
Tieama
D000464:Alginates / Q000737:chemistry; D019843:Archaeal Proteins / Q000737:chemistry* / Q000378:metabolism; D016227:Benzoquinones / Q000737:chemistry; D055162:Biocatalysis; D002122:Calcium Chloride / Q000378:metabolism; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000378:metabolism; D017743:Equipment Reuse; D005782:Gels / Q000737:chemistry; D005087:Glucan 1,4-alpha-Glucosidase / Q000737:chemistry / Q000378:metabolism*; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008863:Microspheres; D013213:Starch / Q000378:metabolism; D013696:Temperature; D019603:Thermoplasmales / Q000737:chemistry; D017155:Wettability
D000464:Alginates; D019843:Archaeal Proteins; D016227:Benzoquinones; D004800:Enzymes, Immobilized; D005782:Gels; D006603:Hexuronic Acids; C004532:quinone; D020723:Glucuronic Acid; D013213:Starch; D005087:Glucan 1,4-alpha-Glucosidase; D002122:Calcium Chloride