Comparison of the influence of pH on the selectivity of free and immobilized trypsin for β-lactoglobulin hydrolysis

被引:23
作者
Mao, Yuhong [1 ]
Krischke, Maria [1 ]
Hengst, Claudia [1 ]
Kulozik, Ulrich [1 ,2 ]
机构
[1] Tech Univ Munich, Chair Food & Bioproc Engn, Weihenstephaner Berg 1, D-85354 Freising Weihenstephan, Germany
[2] Tech Univ Munich, Inst Food & Hlth ZIEL, Technol Unit, Weihenstephaner Berg 1, D-85354 Freising Weihenstephan, Weihenstephan, Germany
关键词
beta-lactoglobulin; Tryptic hydrolysis; Immobilization; pH; Selectivity; TRYPTIC HYDROLYSIS; ENZYME SELECTIVITY; DIGESTION; PROTEINS; KINETICS; RELEASE;
D O I
10.1016/j.foodchem.2018.01.151
中图分类号
O69 [应用化学];
学科分类号
081704 ;
摘要
Although immobilized trypsin is a viable alternative to the free one in solution for producing protein hydrolysates, the change of selectivity introduced by immobilization is unclear. In this study, we compared the selectivity of free and immobilized trypsin towards different cleavage sites of beta-lactoglobulin (beta-Lg) with a focus on the impact of environmental pH. Both free and immobilized trypsin exhibited greater accessibility to native beta-Lg at elevated pH (from pH 7.2 to 8.7). Additionally, free trypsin preferred to attack cleavage sites located at the C-terminus at pH 7.8, whereas an opposite preference for the N-terminus was observed at pH 8.7. Regarding the immobilized trypsin, the pH did not significantly influence its preference for the C-or N-terminus. Generally, immobilization of trypsin resulted in more focused cleavage within its specificity during the initial stage of hydrolysis, and some peptides were formed more rapidly by the immobilized trypsin.
引用
收藏
页码:194 / 202
页数:9
相关论文
共 23 条
  • [1] ENZYMATIC-HYDROLYSIS OF PROTEINS FOR INCREASED SOLUBILITY
    ADLERNISSEN, J
    [J]. JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 1976, 24 (06) : 1090 - 1093
  • [2] [Anonymous], ELECTROPHORESIS
  • [3] Improvement of the stability and activity of immobilized trypsin on modified Fe3O4 magnetic nanoparticles for hydrolysis of bovine serum albumin and its application in the bovine milk
    Atacan, Keziban
    Cakiroglu, Bekir
    Ozacar, Mahmut
    [J]. FOOD CHEMISTRY, 2016, 212 : 460 - 468
  • [4] Determination of the influence of the pH of hydrolysis on enzyme selectivity of Bacillus licheniformis protease towards whey protein isolate
    Butre, Claire I.
    Sforza, Stefano
    Wierenga, Peter A.
    Gruppen, Harry
    [J]. INTERNATIONAL DAIRY JOURNAL, 2015, 44 : 44 - 53
  • [5] Introducing enzyme selectivity: a quantitative parameter to describe enzymatic protein hydrolysis
    Butre, Claire I.
    Sforza, Stefano
    Gruppen, Harry
    Wierenga, Peter A.
    [J]. ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 2014, 406 (24) : 5827 - 5841
  • [6] Impact of the environmental conditions and substrate pre-treatment on whey protein hydrolysis: A review
    Cheison, Seronei Chelulei
    Kulozik, Ulrich
    [J]. CRITICAL REVIEWS IN FOOD SCIENCE AND NUTRITION, 2017, 57 (02) : 418 - 453
  • [7] Analysis of the Effect of Temperature Changes Combined with Different Alkaline pH on the β-Lactoglobulin Trypsin Hydrolysis Pattern Using MALDI-TOF-MS/MS
    Cheison, Seronei Chelulei
    Brand, Janina
    Leeb, Elena
    Kulozik, Ulrich
    [J]. JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 2011, 59 (05) : 1572 - 1581
  • [8] Effect of genetic variation on the tryptic hydrolysis of bovine β-lactoglobulin A, B, and C
    Creamer, LK
    Nilsson, HC
    Paulsson, MA
    Coker, CJ
    Hill, JP
    Jiménez-Flores, R
    [J]. JOURNAL OF DAIRY SCIENCE, 2004, 87 (12) : 4023 - 4032
  • [9] EFFECTS OF IMMOBILIZATION ON INTRINSIC KINETICS AND SELECTIVITY OF TRYPSIN
    DUGGAL, SK
    BUCHHOLZ, K
    [J]. EUROPEAN JOURNAL OF APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 1982, 16 (2-3): : 81 - 87
  • [10] β-Lactoglobulin tryptic digestion: A model approach for peptide release
    Fernandez, Ayoa
    Riera, Francisco
    [J]. BIOCHEMICAL ENGINEERING JOURNAL, 2013, 70 : 88 - 96