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Journal of Food Science and Biotechnology

Corresponding Author(s)

张俊(1993—),男,博士,讲师,主要从事酶工程研究。E-mail:2523050416@qq.com

Abstract

[Objective ] This study aims to construct chitosan and chitin-based carriers with hierarchical porous lamellar structures,systematically evaluate the immobilization effects of these carriers for Pseudomonas fluorescens lipase (PFL) and the performance of the immobilized enzyme in catalyzing the hydrolysis of fish oil.[Method ] Chitin/chitosan were modified via the sol-gel method to prepare the carriers for the binding with free PFL.The enzymatic properties of the immobilized PFL were then evaluated,and then analysed the hydrolysate composition of anchovy oil catalyzed by the immobilized PFL.[Result ] The modified carriers exhibited lamellar structures with abundant free amino groups on the surface,which facilitated covalent immobilization of the enzyme.The chitosan-based carrier achieved the immobilization efficiency of 89.26%,the enzyme loading of 2.21 g/mg,and the PFL enzyme activity of 221.33 U/mg (2.54 folds that of the free enzyme ).The chitin-based carrier showed the immobilization efficiency of 71.62%,the enzyme loading of 1.84 g/mg,and the PFL enzyme activity of 185.34 U/mg (2.12 folds that of the free enzyme ).The chitosan and chitin-immobilized PFL exhibited optimal catalytic activity at 55 ℃ and pH 8.0,along with significant tolerance to organic solvents such as methanol,ethanol,and glycerol and storage stability.After 11 days of storage at 4 ℃,the enzyme activity reached 115.11 U/mg and 91.43 U/mg,respectively.In the hydrolysis of anchovy oil,the hydrolysis degrees of the chitosan and chitin-immobilized PFL were 23.1% and 19.7%,respectively.The proportions of EPA and DHA in the chitosan-immobilized PFL increased to 25.34% and 18.51%,respectively,while the proportions of EPA and DHA in the chitin-immobilized PFL increased to 25.96% and 18.89%,respectively.[Conclusion ] This study demonstrates the potential of hierarchical chitosan and chitin-based carriers in enhancing the catalytic performance of lipase and provides a theoretical basis and practical support for the development of immobilized enzymatic systems for the efficient enrichment of ω-3 fatty acids from deep-sea fish oils.

Publication Date

1-15-2026

First Page

141

Last Page

153

DOI

10.12441/spyswjs.20250805003

References

[1] ZHANG J,CHEN X Y,LV P M,et al.Bionic-immobilized recombinant lipase obtained via bio-silicification and its catalytic performance in biodiesel production [J].Fuel,2021,304:121594.
[2] ISMAIL A R,BAEK K H.Lipase immobilization with support materials,preparation techniques,and applications:present and future aspects [J].International Journal of Biological Macromolecules,2020,163:1624 -1639.
[3] BISHT M,THAYALLATH S K,BHARADWAJ P,et al.Biomass-derived functional materials as carriers for enzymes:towards sustainable and robust biocatalysts [J].Green Chemistry,2023,25(12):4591 -4624.
[4] 张灿,姜国芳,杨江楠,等.多孔材料固定化脂肪酶的研究进展[J].分子催化,2020,34(4):378-396.ZHANG C,JIANG G F,YANG J N,et al.Research progress of immobilized lipase on porous materials [J].Journal of Molecular Catalysis (China),2020,34(4):378-396.(in Chinese )
[5] EL KNIDRI H,BELAABED R,ADDAOU A,et al.Extraction,chemical modification and characterization of chitin and chitosan [J].International Journal of Biological Macromolecules,2018,120:1181 -1189.
[6] KUMARI S,RATH P,SRI HARI KUMAR A,et al.Extraction and characterization of chitin and chitosan from fishery waste by chemical method [J].Environmental Technology & Innovation,2015,3:77-85.
[7] HOU F R,GONG Z Q,JIA F J,et al.Insights into the relationships of modifying methods,structure,functional properties and applications of chitin:a review[J].Food Chemistry,2023,409:135336.[8] ZHANG J,LEI J C,LI K T,et al.Sustainable design of hollow macroporous chitin carrier with enhanced loading capacity for high-performance lipase bioreactor [J].International Journal of Biological Macromolecules,2025,310:143494.
[9] MAERVOET V E T,DE MEY M,BEAUPREZ J,et al.Enhancing the microbial conversion of glycerol to 1,3-propanediol using metabolic engineering [J].Organic Process Research & Development,2011,15(1):189-202.
[10] SETH S,CHAKRAVORTY D,DUBEY V K,et al.An insight into plant lipase research:challenges encountered[J].Protein Expression and Purification,2014,95:13-21.
[11] MANGOS T J,JONES K C,FOGLIA T A.Lipase-catalyzed synthesis of structured low-calorie triacylglycerols [J].Journal of the American Oil Chemists ’ Society,1999,76(10):1127 -1132.
[12] 张惠君,王兴国,金青哲.3种海洋鱼油脂肪酸组成及其位置分布 [J].食品与机械,2017,33(9):59-63.ZHANG H J,WANG X G,JIN Q Z.Composition and positional distribution of fatty acids in triacylglycerols of three marine fish oils [J].Food and Machinery,2017,33(9):59-63.(in Chinese )
[13] 李艳阳,王春艳,罗婷予,等.脂肪酶水解金枪鱼油的工艺优化及脂质成分分析 [J].广东海洋大学学报,2024,44(6):109-117.LI Y Y,WANG C Y,LUO T Y,et al.Process optimization and lipid composition analysis of tuna oil hydrolyzed by lipase [J].Journal of Guangdong Ocean University,2024,44(6):109-117.(in Chinese )
[14] DWIVEDEE B P,SONI S,LAHA J K,et al.Facile immobilization of Pseudomonas fluorescens lipase on polyaniline nanofibers (PANFs-PFL ):a route to develop robust nanobiocatalyst [J].International Journal of Biological Macromolecules,2018,119:8-14.
[15] SRIWONG C,SUKYAI P.Simulated elephant colon for cellulose extraction from sugarcane bagasse:an effective pretreatment to reduce chemical use [J].Science of the Total Environment,2022,835:155281.
[16] ALMULAIKY Y Q,ALKABLI J,EL-SHISHTAWY R M.Improving enzyme immobilization:a new carrier-based magnetic polymer for enhanced covalent binding of laccase enzyme [J].International Journal of Biological Macromolecules,2024,282:137362.
[17] RASMUSSEN H Ø,WOLLENBERG D T W,WANG H B,et al.The changing face of SDS denaturation:complexes of Thermomyces lanuginosus lipase with SDS at pH 4.0,6.0 and 8.0[J].Journal of Colloid and Interface Science,2022,614:214-232.
[18] DA ROCHA T N,CARBALLARES D,GUIMARÃES J R,et al.Determination of immobilized lipase stability depends on the substrate and activity determination condition:stress inactivations and optimal temperature as biocatalysts stability indicators [J].Sustainable Chemistry and Pharmacy,2022,29:100823.
[19] ZHA F C,SHI M,LI H,et al.Biomimetic mineralization of lipase@MOF biocatalyst for ease of biodiesel synthesis:structural insights into the catalytic behavior[J].Fuel,2024,357:129854.
[20] STEMLER C D,SCHERF K A.Comparative characterization of baking lipase substrate specificities using emulsions and the p-nitrophenyl assay [J].LWT-Food Science and Technology,2022,168:113914.
[21] 李佥,曾祥冰,孙西同,等.酚氨基改性 MCM- 41固定化脂肪酶的工艺优化及应用 [J].食品工业科技,2025,46(5):168-177.LI Q,ZENG X B,SUN X T,et al.Process optimization and application of lipase immobilized on phenol amino modified MCM- 41[J].Science and Technology of Food Industry,2025,46(5):168-177.(in Chinese )
[22] ATİROĞLU V.Lipase immobilization on synthesized hyaluronic acid-coated magnetic nanoparticle-functionalized graphene oxide composites as new biocatalysts:improved reusability,stability,and activity[J].International Journal of Biological Macromolecules,2020,145:456-465.
[23] WANG S H,MENG X H,ZHOU H,et al.Enzyme stability and activity in non-aqueous reaction systems:a mini review [J].Catalysts,2016,6(2):32.
[24] 覃红玉,张俊,李昆太,等.木质纤维甘蔗渣载体固定化嗜热脂肪酶及其催化特性研究 [J].食品工业科技,2025,46(21):228-237.QIN H Y,ZHANG J,LI K T,et al.Immobilization of thermophilic lipase on lignocellulosic sugarcane bagasse carrier and its biocatalytic properties [J].Science and Technology of Food Industry,2025,46(21):228-237.(in Chinese)
[25] XU J C,LI Y Y,YANG J W,et al.Plasma etching effect on the molecular structure of chitosan-based hydrogels and its biological properties [J].International Journal of Biological Macromolecules,2023,230:123257.
[26] AFSHARI O,SALEHI Z.β-Cyclodextrin and APTES functionalized magnetic iron oxide nanoparticles for improved lipase immobilization,transesterification efficiency,and reusability [J].Journal of Industrial and Engineering Chemistry,2025,144:476-486.
[27] MU L X,WU L Q,WU S Q,et al.Progress in chitin/chitosan and their derivatives for biomedical applications:where we stand [J].Carbohydrate Polymers,2024,343:122233.
[28] ENGELMANN J I,SILVA P P,IGANSI A V,et al.Structured lipids by swine lard interesterification with oil and esters from common carp viscera [J].Journal of Food Process Engineering,2018,41(4):e12679.

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