Immobilization of Lipase to Chitosan Beads using a Natural Cross‐Linker

Abstract Genipin, a reagent of plant origin was used for the immobilization of lipase by cross‐linking to chitosan beads. The catalytic properties and operational and storage stabilities of the immobilized lipase were compared with the soluble lipase. Under optimum conditions, 198 µg protein was bound per g chitosan with a protein‐coupling yield of 35%. The hydrolytic activity was 10.8 U/g chitosan and the relative specific activity was 108%. The immobilized lipase showed better thermal and pH stabilities compared to the soluble form. The immobilized enzyme exhibited mass transfer limitations as reflected by a higher apparent Km value and a lower energy of activation. The immobilized enzyme retained about 74% of its initial activity after five hydrolytic cycles.

[1]  A. Kılınç,et al.  Immobilization of Pancreatic Lipase on Chitin and Chitosan , 2006, Preparative biochemistry & biotechnology.

[2]  Xiaoyan Yuan,et al.  Immobilization of cellulase in nanofibrous PVA membranes by electrospinning , 2005 .

[3]  J. Kumada,et al.  The continuous hydrolysis of geniposide to genipin using immobilized β-glucosidase on calcium alginate gel , 1987, Biotechnology Letters.

[4]  Ho-Shing Wu,et al.  Kinetics of tributyrin hydrolysis by lipase , 2004 .

[5]  K. Atia,et al.  Immobilization of β-amylase using polyacrylamide polymer derivatives , 2003 .

[6]  R. Fernández-Lafuente,et al.  Interfacial adsorption of lipases on very hydrophobic support (octadecyl-Sepabeads): Immobilization, hyperactivation and stabilization of the open form of lipases , 2002 .

[7]  A. Bigi,et al.  Stabilization of gelatin films by crosslinking with genipin. , 2002, Biomaterials.

[8]  R. Zhuo,et al.  Polymerization of Trimethylene Carbonate with High Molecular Weight Catalyzed by Immobilized Lipase on Silica Microparticles , 2002 .

[9]  H. Sung,et al.  In vivo evaluation of cellular and acellular bovine pericardia fixed with a naturally occurring crosslinking agent (genipin). , 2002, Biomaterials.

[10]  M. Reetz.  Lipases as practical biocatalysts. , 2002, Current opinion in chemical biology.

[11]  A. Tuncel,et al.  Immobilization of invertase onto crosslinked poly(p‐chloromethylstyrene) beads , 2002 .

[12]  Mahiran Basri,et al.  Immobilization of lipase on poly(N‐vinyl‐2‐pyrrolidone‐co‐styrene) hydrogel , 2001 .

[13]  Xiao Dong Chen,et al.  Immobilization of β-galactosidase on graphite surface by glutaraldehyde , 2001 .

[14]  H. Sung,et al.  Effects of heparin immobilization on the surface characteristics of a biological tissue fixed with a naturally occurring crosslinking agent (genipin): an in vitro study. , 2001, Biomaterials.

[15]  H. Sung,et al.  Synthesis and characterization of a novel chitosan‐based network prepared using naturally occurring crosslinker , 2000 .

[16]  Ann-Christine Albertsson,et al.  Novel pH-sensitive chitosan hydrogels: swelling behavior and states of water , 2000 .

[17]  Akkuş Çetinus S,et al.  Immobilization of catalase on chitosan film. , 2000, Enzyme and microbial technology.

[18]  S. Quirce,et al.  Glutaraldehyde‐induced asthma , 1999, Allergy.

[19]  H. Sung,et al.  Feasibility study of a natural crosslinking reagent for biological tissue fixation. , 1998, Journal of biomedical materials research.

[20]  B. C. Bhattacharyya,et al.  Immobilization of lipase using egg shell and alginate as carriers: optimization of reaction conditions , 1998 .

[21]  I. Maliszewska,et al.  Immobilization of lipase on various acrylic copolymers , 1997 .

[22]  N. Gandhi.  Applications of lipase , 1997 .

[23]  L. M. Simon,et al.  Immobilization and characterization of porcine pancreas lipase , 1997 .

[24]  S. Dharmsthiti,et al.  Lipase production by Aeromonas sobria LP004 in a medium containing whey and soybean meal , 1997 .

[25]  M. Basri,et al.  Enzymic synthesis of fatty esters by hydrophobic lipase derivatives immobilized on organic polymer beads , 1995 .

[26]  J. Serra,et al.  Immobilization of Candida rugosa lipase and some properties of the immobilized enzyme. , 1993, Enzyme and microbial technology.

[27]  S. Ponrathnam,et al.  Immobilization of penicillin G acylase on methacrylate polymers. , 1991, Journal of chemical technology and biotechnology.

[28]  P. Burge.  Occupational risks of glutaraldehyde. , 1989, BMJ.

[29]  Shigeyuki Nakamura,et al.  Genipin, a New Type of Protein Crosslinking Reagent from Gardenia Fruits , 1988 .

[30]  John F. Kennedy,et al.  Immobilised Enzymes and Cells , 1990 .

[31]  S. Gonçalo,et al.  Occupational contact dermatitis to glutaraldehyde , 1984, Contact dermatitis.

[32]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[33]  V. Surovtsev.  [Immobilized enzymes and cells]. , 1975, Uspekhi sovremennoi biologii.

[34]  E. Katchalski,et al.  A WATER-INSOLUBLE POLYANIONIC DERIVATIVE OF TRYPSIN. II. EFFECT OF THE POLYELECTROLYTE CARRIER ON THE KINETIC BEHAVIOR OF THE BOUND TRYPSIN. , 1964, Biochemistry.