title: Immobilization of Brassica oleracea chlorophyllase 1 (BoCLH1) and Candida rugosa lipase (CRL) in magnetic alginate beads: an enzymatic evaluation in the corresponding proteins.
abstract: Enzymes have a wide variety of applications in diverse biotechnological fields, and the immobilization of enzymes plays a key role in academic research or industrialization due to the stabilization and recyclability it confers. In this study, we immobilized the Brassica oleracea chlorophyllase 1 (BoCLH1) or Candida rugosa lipase (CRL) in magnetic iron oxide nanoparticles-loaded alginate composite beads. The catalytic activity and specific activity of the BoCLH1 and CRL entrapped in magnetic alginate composite beads were evaluated. Results show that the activity of immobilized BoCLH1 in magnetic alginate composite beads (3.36±0.469 U/g gel) was higher than that of immobilized BoCLH1 in alginate beads (2.96±0.264 U/g gel). In addition, the specific activity of BoCLH1 beads (10.90±1.521 U/mg protein) was higher than that immobilized BoCLH1 in alginate beads (8.52±0.758 U/mg protein). In contrast, the immobilized CRL in magnetic alginate composite beads exhibited a lower enzyme activity (11.81±0.618) than CRL immobilized in alginate beads (94.83±7.929), and the specific activity of immobilized CRL entrapped in magnetic alginate composite beads (1.99±0.104) was lower than immobilized lipase in alginate beads (15.01±1.255). A study of the degradation of magnetic alginate composite beads immersed in acidic solution (pH 3) shows that the magnetic alginate composite beads remain intact in acidic solution for at least 6 h, indicating the maintenance of the enzyme catalytic effect in low-pH environment. Finally, the enzyme immobilized magnetic alginate composite beads could be collected by an external magnet and reused for at least six cycles.
authors.affiliation: Department of Biological Science & Technology, I-Shou University, Kaohsiung 840, Taiwan.
authors.forename: Chih-Hui
authors.initials: CH
authors.lastname: Yang
authors.affiliation: Department of Biological Science & Technology, I-Shou University, Kaohsiung 840, Taiwan.
authors.forename: Chih-Chung
authors.initials: CC
authors.lastname: Yen
authors.affiliation: Department of Biological Science & Technology, I-Shou University, Kaohsiung 840, Taiwan.
authors.forename: Jen-Jyun
authors.initials: JJ
authors.lastname: Jheng
authors.affiliation: Department of Biomedical Engineering, I-Shou University, Kaohsiung 840, Taiwan.
authors.forename: Chih-Yu
authors.initials: CY
authors.lastname: Wang
authors.affiliation: Department of Biological Science & Technology, I-Shou University, Kaohsiung 840, Taiwan.
authors.forename: Sheau-Shyang
authors.initials: SS
authors.lastname: Chen
authors.affiliation: Department of Biological Science & Technology, I-Shou University, Kaohsiung 840, Taiwan.
authors.forename: Pei-Yu
authors.initials: PY
authors.lastname: Huang
authors.affiliation: The School of Chinese Medicine for Post-Baccalaureate, I-Shou University, No.8, Yida Road, Jiaosu Village Yanchao District, Kaohsiung 82445, Taiwan. huangks@isu.edu.tw.
authors.forename: Keng-Shiang
authors.initials: KS
authors.lastname: Huang
authors.affiliation: Department of Biological Science & Technology, I-Shou University, Kaohsiung 840, Taiwan. shawjf@isu.edu.tw.
authors.forename: Jei-Fu
authors.initials: JF
authors.lastname: Shaw
mesh_terms: D000464:Alginates / Q000737:chemistry; D001709:Biotechnology; D001937:Brassica / Q000201:enzymology; D002175:Candida / Q000201:enzymology; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry* / Q000378:metabolism; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry*; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D008049:Lipase / Q000737:chemistry* / Q000378:metabolism; D060328:Magnetic Phenomena; D013696:Temperature
chemical_list: D000464:Alginates; D004800:Enzymes, Immobilized; D006603:Hexuronic Acids; D020723:Glucuronic Acid; D002265:Carboxylic Ester Hydrolases; C019875:chlorophyllase; D008049:Lipase
references.citation: Int J Pharm. 2014 Mar 25;463(2):155-60
references.pmid: 23988565
references.citation: Biotechnol Bioeng. 1988 Apr 20;31(6):628-33
references.pmid: 18584656
references.citation: Bioorg Med Chem Lett. 2010 Jan 15;20(2):508-12
references.pmid: 20004575
references.citation: N Biotechnol. 2010 Feb 28;27(1):53-8
references.pmid: 20004754
references.citation: Biotechnol Bioeng. 1990 Jan 20;35(2):132-7
references.pmid: 18592502
references.citation: Mutat Res. 2008 Dec 8;657(2):98-104
references.pmid: 18775795
references.citation: Int J Mol Med. 1999 Oct;4(4):377-80
references.pmid: 10493978
references.citation: J Agric Food Chem. 2011 Oct 12;59(19):10693-8
references.pmid: 21854055
references.citation: Appl Microbiol Biotechnol. 2011 Mar;89(5):1435-42
references.pmid: 21038097
references.citation: 3 Biotech. 2013 Feb;3(1):1-9
references.pmid: 28324347
references.citation: AAPS PharmSciTech. 2005 Sep 30;6(2):E209-22
references.pmid: 16353980
references.citation: J Biotechnol. 2005 Nov 21;120(3):273-83
references.pmid: 16087266
references.citation: Yeast. 1998 Sep 15;14(12):1069-87
references.pmid: 9778794
references.citation: J Agric Food Chem. 2005 Apr 6;53(7):2746-50
references.pmid: 15796620
references.citation: Plant Cell Physiol. 2003 Jan;44(1):96-101
references.pmid: 12552153
references.citation: J Agric Food Chem. 2010 Aug 11;58(15):8651-7
references.pmid: 20681655
references.citation: Int J Mol Sci. 2013 Feb 26;14(3):4613-28
references.pmid: 23443161
references.citation: J Nanobiotechnology. 2005 Jan 20;3(1):1
references.pmid: 15661076
references.citation: Eur J Biochem. 1994 Jun 1;222(2):461-6
references.pmid: 8020484
references.citation: Appl Biochem Biotechnol. 2012 Apr;166(8):1927-40
references.pmid: 22383051
references.citation: BMB Rep. 2011 Feb;44(2):87-95
references.pmid: 21345306
references.citation: Science. 1983 Feb 11;219(4585):722-7
references.pmid: 17814033
references.citation: Biochim Biophys Acta. 2011 Aug;1807(8):977-88
references.pmid: 21167811
references.citation: Biochim Biophys Acta. 2000 Dec 29;1543(2):223-238
references.pmid: 11150608
references.citation: Antimicrob Agents Chemother. 2011 Feb;55(2):478-86
references.pmid: 21135183
references.citation: J Mater Chem B. 2013 Apr 28;1(16):2205-2212
references.pmid: 32260854
references.citation: Nat Protoc. 2007;2(5):1022-33
references.pmid: 17546007
references.citation: Cell Immunol. 2011;271(2):308-18
references.pmid: 21855057
references.citation: J Chromatogr B Analyt Technol Biomed Life Sci. 2013 Apr 1;923-924:98-101
references.pmid: 23500353