It is a feasible way to effectively control the content of ethyl carbamate in Huangjiu (Chinese rice wine) by using new immobilized neutral ureases to degrade urea in Huangjiu.In recent years, in-situ synthesis of immobilized enzymes based on metal-organic frameworks (MOFs) has attracted researchers' attention.In this study, by comparing the two MOF materials, ZIF-8 and MIL-100(Fe), MIL-100(Fe) was selected as the carrier according to the characteristics of Huangjiu to synthesize in-situ the immobilized neutral urease [NU@MIL-100(Fe)] at room temperature and in water environment.It was characterized by X-ray diffractometer (XRD) and scanning electron microscopy (SEM), and its enzymatic properties were investigated.The results showed that the crystal structure and morphology of MIL-100(Fe) were not affected by the immobilization of the enzyme when it was used as the carrier with good acid stability and alcohol tolerance.Compared with free neutral urease, the optimal pH, optimal temperature and the effect of alcohol on activity of NU@MIL-100(Fe) did not change significantly, but NU@MIL-100(Fe) showed higher acid activity and better alcohol tolerance, and its acid stability was also improved.When NU@MIL-100(Fe) was used to degrade urea in Huangjiu, the urea degradation reached to 79.96% after 4 h treatment and 93.83% after 24 h treatment, even 48.29% after 4 h treatment in the 3rd reuse, which showed a good urea degradation effect and reusability.The results provide a new reference for the practical application of immobilized neutral urease in Huangjiu.
[1] WEBER J V, SHARYPOV V I.Ethyl carbamate in foods and beverages:A review[J].Environmental Chemistry Letters, 2009, 7(3):233-247.
[2] JIAO Z H, DONG Y C, CHEN Q H.Ethyl carbamate in fermented beverages:Presence, analytical chemistry, formation mechanism, and mitigation proposals[J].Comprehensive Reviews in Food Science and Food Safety, 2014, 13(4):611-626.
[3] 李童, 钱斌, 周建弟, 等.中性脲酶固定化降解黄酒中尿素[J].食品与发酵工业, 2021, 47(9):70-75.
LI T, QIAN B, ZHOU J D, et al.Degradation of urea in Huangjiu by immobilized neutral urease[J].Food and Fermentation Industries, 2021, 47(9):70-75.
[4] 范文来, 王栋.近10年我国传统饮料酒白酒和黄酒品质安全研究现状与展望[J].食品安全质量检测学报, 2019, 10(15):4811-4829.
FAN W L, WANG D.Current practice and future trends of alcoholic beverages safety of China traditional Baijiu and Huangjiu in recent decades[J].Journal of Food Safety & Quality, 2019, 10(15):4811-4829.
[5] GARCIA-GALAN C, BERENGUER-MURCIA Á, FERNANDEZ-LAFUENTE R, et al.Potential of different enzyme immobilization strategies to improve enzyme performance[J].Advanced Synthesis & Catalysis, 2011, 353(16):2885-2904.
[6] MAGHRABY Y R, EL-SHABASY R M, IBRAHIM A H, et al.Enzyme immobilization technologies and industrial applications[J].ACS Omega, 2023, 8(6):5184-5196.
[7] DUAN W J, ZHAO Z F, AN H D, et al.State-of-the-art and prospects of biomolecules:Incorporation in functional metal-organic frameworks[J].Topics in Current Chemistry (Cham), 2019, 377(6):34.
[8] HU Y L, DAI L M, LIU D H, et al.Progress & prospect of metal-organic frameworks (MOFs) for enzyme immobilization (enzyme/MOFs)[J].Renewable and Sustainable Energy Reviews, 2018, 91:793-801.
[9] LIU W L, YANG N S, CHEN Y T, et al.Lipase-supported metal-organic framework bioreactor catalyzes warfarin synthesis[J].Chemistry, 2015, 21(1):115-119.
[10] CAO S L, YUE D M, LI X H, et al.Novel nano-/micro-biocatalyst:Soybean epoxide hydrolase immobilized on UiO-66-NH2 MOF for efficient biosynthesis of enantiopure (R)-1, 2-octanediol in deep eutectic solvents[J].ACS Sustainable Chemistry & Engineering, 2016, 4(6):3586-3595.
[11] ZHENG H Q, ZHANG Y N, LIU L F, et al.One-pot synthesis of metal-organic frameworks with encapsulated target molecules and their applications for controlled drug delivery[J].Journal of the American Chemical Society, 2016, 138(3):962-968.
[12] 苏伟, 李青云, 刘幽燕, 等.ZIF-8原位合成法固定化脂肪酶CRL[J].中国油脂, 2021, 46(2):107-113.
SU W, LI Q Y, LIU Y Y, et al.Immobilization of lipase CRL on ZIF-8 by in situ synthesis[J].China Oils and Fats, 2021, 46(2):107-113.
[13] LIU J, LIANG J Y, XUE J Y, et al.Metal-organic frameworks as a versatile materials platform for unlocking new potentials in biocatalysis[J].Small, 2021, 17(32):e2100300.
[14] LIANG X, LI Q, SHI Z Y, et al.Immobilization of urease in metal-organic frameworks via biomimetic mineralization and its application in urea degradation[J].Chinese Journal of Chemical Engineering, 2020, 28(8):2173-2180.
[15] BUNZEN H.Chemical stability of metal-organic frameworks for applications in drug delivery[J].ChemNanoMat, 2021, 7(9):998-1007.
[16] HORCAJADA P, SURBLÉ S, SERRE C, et al.Synthesis and catalytic properties of MIL-100(Fe), an iron(III) carboxylate with large pores[J].Chemical Communications, 2007(27):2820-2822.
[17] QUIJIA C R, LIMA C, SILVA C, et al.Application of MIL-100(Fe) in drug delivery and biomedicine[J].Journal of Drug Delivery Science and Technology, 2021, 61:102217.
[18] MA X, LEPOITEVIN M, SERRE C.Metal-organic frameworks towards bio-medical applications[J].Materials Chemistry Frontiers, 2021, 5(15):5573-5594.
[19] ZHANG F M, SHI J, JIN Y, et al.Facile synthesis of MIL-100(Fe) under HF-free conditions and its application in the acetalization of aldehydes with diols[J].Chemical Engineering Journal, 2015, 259:183-190.
[20] GUESH K, CAIUBY C A D, SANCHEZ-SANCHEZ M.Sustainable preparation of MIL-100(Fe) and its photocatalytic behavior in the degradation of methyl orange in water[J].Crystal Growth & Design, 2017, 17(4):1806-1813.
[21] MEIJERS C A M, RUTTEN J C J M.A simple rapid method for the determination of total nitrogen in urine[J].Clinica Chimica Acta, 1969, 24(2):308-310.
[22] 周霞. 解淀粉芽孢杆菌脲酶基因在枯草芽孢杆菌中的重组表达及尿素降解应用研究[D].无锡:江南大学, 2021.
ZHOU X.Heterologous expression of urease-encoded gene from Bacillus amyloliquefaciens in recombinant Bacillus subtilis and its application in urea degradation[D].Wuxi:Jiangnan University, 2021.
[23] ZHUANG J L, LIU X Y, MAO H L, et al.Hollow carbon polyhedra derived from room temperature synthesized iron-based metal-organic frameworks for supercapacitors[J].Journal of Power Sources, 2019, 429:9-16.
[24] BARJASTEH M, VOSSOUGHI M, BAGHERZADEH M, et al.MIL-100(Fe) a potent adsorbent of dacarbazine:Experimental and molecular docking simulation[J].Chemical Engineering Journal, 2023, 452:138987.
[25] DAIF M A, EL-DIN M M.Studies on urea hydrolysis.Part 2.Effects of some heavy metals on urease activity[J].Beitrage Zur Tropischen Landwirtschaft und Veterinarmedizin, 1979, 17(3):261-266.
