Research and Application of Antifreeze Peptides


   The market sales of low-temperature cold chain food in developed countries account for 60% to 70% of total food sales, and in my country it is 25% to 30%. There is still huge room for growth. Food-derived antifreeze peptides that have the functions of reducing the growth rate of ice crystals and inhibiting recrystallization are used as food supplements (Figure 1 shows the mode of action of antifreeze peptides and ice crystal molecules). They have significant effects in the preservation and quality maintenance of low-temperature cold chain foods. The advantages of safety, health, and nutrition meet the needs of the big health industry in the new era.

  Figure 1 Diagram of the interaction between antifreeze peptides and ice crystal molecules

  What is an antifreeze peptide?

   Antifreeze peptides are a class of small molecule proteins or protein hydrolysates that have low-temperature protection under icing or sub-icing conditions and reduce frost damage. Antifreeze peptides not only have good antifreeze activity, but also have relatively stable physical and chemical properties. They directly interact with ice crystal molecules without the need for embedding (Figure 2 is the dynamic process diagram of antifreeze peptides and ice crystals). Food-derived antifreeze peptides have sufficient raw materials and controllable production technology, which is convenient for industrialized production and industrialized large-scale applications. Compared with natural isolated and purified antifreeze proteins, antifreeze polypeptides are more suitable for industrial production and applications.

  Figure 2 Kinetic process of antifreeze peptides and ice crystals

  The research background of antifreeze peptides

   In a low temperature environment, most organisms will be injured by freezing, because the body fluid of the organism will freeze at the temperature below the freezing point, which will cause fatal damage to the organism’s cells. In order to adapt to the cold environment, some special organisms can survive. After long-term evolution, a class of proteins with antifreeze physiological functions are produced in the body.

This protein was first introduced in Antarctic fish by animal physiologist DeVries in 1969. It is found in the serum in the body and is separated and purified, which is called antifreeze protein (AFP). AFP is a special protein that inhibits the growth and recrystallization of ice crystals. It has three basic characteristics: heat hysteresis activity, recrystallization inhibition effect and ice nucleus isomerization effect. Studies have found that the antifreeze activity of AFP mainly depends on the local specific polypeptide chain domains.

Obtaining specific polypeptide chain domain fragments has become a new research direction of AFP. The use of biological enzymatic hydrolysis technology to screen high-activity antifreeze peptides from food-derived proteins is a key breakthrough that restricts the industrial application of AFP.

  The function of antifreeze peptide

The functions of    antifreeze peptides can be summarized as three functions: thermal hysteresis, recrystallization inhibition, and ice nucleation isomerization (Figure 3 is a schematic diagram of the cryogenic protection function and molecular mechanism of antifreeze peptides).

  Figure 3 Schematic diagram of the low-temperature protection function and molecular mechanism of antifreeze peptides

   Application of antifreeze peptides

   Antifreeze peptides, as highly active natural antifreeze agents, have played a unique role in food, agriculture, and medicine. In the field of low-temperature cold-chain foods, such as meat slices, minced meat products, ice cream, noodle products, etc.

added with anti-freeze peptides, the size of ice crystals, recrystallization and low-temperature deterioration can be significantly reduced in the production formula, processing, storage and distribution conditions. , Significantly improve product quality and extend product shelf life. In the agricultural field, antifreeze polypeptides cloned and expressed through transgenic technology have achieved the effect of protecting plants against freezing and cold and improving the environment for seed preservation. In the medical field, it is mainly used as a cryoprotectant for freezing and ultra-low temperature storage of organisms, organs, tissues and cell lines or as a chemical adjuvant to protect target tissues in cancer cryosurgery.

  Book Features

The book “Research and Application of Antifreeze Polypeptides” systematically summarizes the research progress of antifreeze proteins and antifreeze peptides.

The author has co-authored with Dalian University of Technology, Shanghai Jiaotong University, Anjing Food Co., Ltd., Anhui Guopeptide Biotechnology Co., Ltd., etc. Related professional teams cooperated, focusing on the source, structural characteristics and classification of antifreeze proteins and antifreeze peptides, activity detection and evaluation methods, preparation methods, mechanism of action, and practical applications. In addition, the research on antifreeze proteins and antifreeze peptides Prospects for new technologies and applications.

   “Research and Application of Antifreeze Peptides”

  ISBN: 9787030666246

  Wang Shaoyun is waiting

   “Thirteenth Five-Year” National Key Publications Publication Planning Project

   Chief Editor: Jia Chao

About the Author

  Wang Shaoyun, Ph.D., second-level professor, doctoral supervisor. From 2006 to 2009, he was engaged in post-doctoral research at the University of Wisconsin (UW-Madison) and the University of California Davis (UC-Davis), and was selected as a leader in science and technology innovation in the National “Ten Thousand Talents Program” and a young and middle-aged leader in science and technology innovation by the Ministry of Science and Technology , Provincial high-level innovative talents, provincial scientific and technological innovation leaders, provincial colleges and universities outstanding talents in the new century.

Served as the head of the national food science and engineering “first-class specialty”, the leader of the Fujian food and bioengineering “first-class discipline”, the head of the Fujian Marine Functional Product Collaborative Innovation Center, the dean (executive) of the School of Biological Science and Engineering, Fuzhou University, Dean of the Institute of Marine Science and Technology, Fuzhou University.

Concurrently serves as the editorial board member of Food Science and Human Wellness, Hans Journal of Food and Nutrition Science, Food Science, Food Industry Science and Technology, and Journal of Fuzhou University (Natural Science Edition), member of the Council of China Food Science and Technology Association, and Chinese and Foreign Food Technology” the first batch of translation experts.

He has presided over more than 30 projects at or above the provincial and ministerial level, compiled 4 books, obtained 53 authorized invention patents, published more than 230 papers, and included more than 180 papers in SCI/EI.

Won the International Food Functional Factors (ICOFF) Academic Conference Award, the first prize of China Industry-University-Research Cooperation Innovation Achievement Award, the first prize of National Food Industry-University-Research Outstanding Scientific Research Achievements, the first prize of the 3rd China Food Science and Technology Achievement Award, China Chemical Industry Federation Science and Technology The first prize of progress, the first prize of Fujian Science and Technology Progress, the second prize of Fujian Natural Science, the second prize of Fujian Science and Technology Progress.

Won the Science and Technology Innovation-Outstanding Youth Award of China Food Science and Technology Society, Baosteel Outstanding Teacher Award, Lu Jiaxi Outstanding Mentor Award, Provincial Outstanding Teacher Award, Provincial Outstanding Tech Honorary titles such as “Best Tech Worker” and “New Long March Assaulter”.

brief introduction

The book “Research and Application of Antifreeze Peptides” systematically summarizes the research progress of antifreeze proteins and antifreeze peptides, focusing on the sources, structural characteristics and classification, activity detection and evaluation methods, preparation of antifreeze proteins and antifreeze peptides.

Methods, mechanism of action and practical applications, etc. In addition, new technologies and applications in the research of antifreeze proteins and antifreeze peptides are prospected. This book is comprehensive and detailed, with practical, practical and maneuverable characteristics. It is suitable for scientific research personnel and management personnel engaged in food science, agricultural science, and food biotechnology. It can also be used as a reference for postgraduates and teachers of related majors in colleges and universities. book.

  Book list

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table of Contents
Chapter 1 Introduction 1
1.1 Introduction to antifreeze protein/peptide 1
1.2 The source of antifreeze protein/peptide 2
1.2.1 The source of antifreeze protein 2
1.2.2 The source of antifreeze peptides 3
1.3 The function of antifreeze protein/peptide 4
1.3.1 Thermal hysteresis activity 4
1.3.2 Inhibition of recrystallization 5
1.3.3 Modification of ice crystal morphology 5
1.3.4 Lowering the supercooling point 6
1.3.5 Protect cell membrane 6
1.3.6 Dual functional characteristics 7
1.4 Safety evaluation of antifreeze protein/peptide 8
1.4.1 Assessment of sensitization 9
1.4.2 Toxicity assessment 12
References 14


Chapter 2 Structural Features and Classification of Antifreeze Proteins/Peptides 20
2.1 Overview of the structural features and classification of antifreeze proteins/peptides 20
2.2 Structural characteristics and classification of antifreeze proteins 23
2.2.1 Antifreeze protein in fish 23
2.2.2 Antifreeze proteins in insects 32
2.2.3 Antifreeze proteins in plants 36
2.2.4 Antifreeze proteins in bacteria 39
2.2.5 Antifreeze proteins in fungi 40
2.3 Structural characteristics and classification of antifreeze peptides 41
2.3.1 Structural characteristics of antifreeze peptides 41
2.3.2 Classification of antifreeze peptides 44


References 50
Chapter 3 Antifreeze Protein/Peptide Activity Detection and Evaluation Method 57
3.1 Overview of liveness detection and evaluation methods 57
3.2 Antifreeze protein/peptide detection and evaluation methods 58
3.2.1 Detection and evaluation method of antifreeze activity based on thermal hysteresis principle 58
3.2.2 Evaluation method of antifreeze activity based on the principle of inhibition of ice crystal recrystallization 63
3.2.3 Low temperature protection of organisms 66
3.2.4 Food system cryogenic protection law 66
3.2.5 Other methods 66
3.3 Detection and evaluation of antifreeze activity of marine fish AFP 67
3.4 Detection and evaluation of antifreeze activity of insect AFP 67
3.5 Detection and evaluation of antifreeze activity of plant AFP 68
3.6 Detection and evaluation of antifreeze activity of bacterial and fungal AFP 70


References 70
Chapter 4 Preparation of Antifreeze Protein 74
4.1 Overview of antifreeze protein preparation 74
4.1.1 Natural purification method 74
4.1.2 Genetically modified method 77
4.1.3 Separation and purification of antifreeze protein 82
4.1.4 The preparation of antifreeze protein from complex to simple 86
4.2 Preparation of antifreeze protein by natural purification method 87
4.2.1 Preparation of fish-derived antifreeze protein 87
4.2.2 Preparation of insect-derived antifreeze protein 90
4.2.3 Preparation of plant-derived antifreeze protein 94
4.2.4 Preparation of antifreeze protein from microbial sources 100
4.3 Preparation of antifreeze protein by transgenic method 102
4.3.1 Preparation of antifreeze protein by transgenic method 102
4.3.2 Example of preparation of antifreeze protein transgenic method 109

References 122
Chapter 5 Preparation, Separation and Purification of Antifreeze Peptides 130
5.1 Overview of antifreeze peptide preparation 130
5.1.1 Proteolysis method 130
5.1.2 Biological extraction method 131
5.1.3 Synthesis method 132
5.1.4 Genetic engineering method 132
5.1.5 Chemical modification method 132
5.1.6 Crude Antifreeze Peptides Extracted by Ice Affinity Purification 133
5.1.7 Separation, purification and analysis methods of peptides 134
5.2 Preparation of antifreeze peptides by biological enzymatic method 147
5.3 Examples of preparation of antifreeze peptides 157
5.3.1 Preparation by biological enzymatic method 157
5.3.2 Examples of preparation of genetic engineering methods 164


References 169
Chapter 6 Mechanism of Action of Antifreeze Proteins/Peptides 174
6.1 Overview of antifreeze protein/peptide mechanism of action 174
6.2 Adsorption inhibition model 178
6.3 Surface complementary model 182
6.4 Hydrogen atom bonding model 184
6.5 Affinity interaction coupling agglomeration model 184
6.6 Inclusion compound anchoring model 185
6.7 The dipole-dipole model 188
6.8 Crystal Occupation-Lattice Matching Model 189
6.9 Rigid body energy model 191


References 192
Chapter 7 Application of Antifreeze Protein/Peptide 195
7.1 Overview of the application of antifreeze proteins/peptides 195
7.1.1 The application status of antifreeze proteins/peptides in the food industry 196
7.1.2 The role in medicine 202
7.1.3 Application prospects 205
7.2 Application in meat and its products 207
7.2.1 The concept of freezing and freezing 208
7.2.2 The quality changes of meat during freezing and freezing storage 208
7.2.3 Antifreeze protection agent 210
7.2.4 Application of antifreeze protein/polypeptide in meat and meat products 214
7.3 Application in fish and its products 217


7.3.1 Frozen processing of fish and fish meat products 217
7.3.2 The freezing and denaturation mechanism of fish meat 218
7.3.3 The mechanism of action of antifreeze protection agents 219
7.4 Application in noodles and their products 226
7.4.1 Problems in frozen dough processing 226
7.4.2 Application of antifreeze protein/peptide in noodles and noodle products 227
7.5 Application in ice cream products 237
7.5.1 Overview of ice cream and challenges in the manufacturing process 237
7.5.2 Application of antifreeze protein/peptide in ice cream 238
7.6 Application in fruits, vegetables and their products 242
7.6.1 Methods of Keeping Fruits and Vegetables Fresh 242
7.6.2 Antifreeze activity of antifreeze protein on fruit and vegetable processing 244
7.6.3 Antifreeze activity of antifreeze protein on storage of fruits and vegetables 245
7.6.4 The application prospects of antifreeze protein in fruit and vegetable preservation 247
7.7 Application in bacterial cells and organisms 248
7.7.1 Antifreeze activity of antifreeze protein against probiotics 250
7.7.2 Antifreeze activity of antifreeze protein against yeast 252
7.7.3 Antifreeze activity of antifreeze proteins on organisms 254


References 256
Chapter 8 New Technology and Application Prospects of Antifreeze Protein/Peptide Research 273
8.1 Proteomics Technology 273
8.1.1 Overview of Proteomics 273
8.1.2 The application of proteomics in antifreeze research 274
8.2 Peptomics Technology 275
8.2.1 Overview of Peptomics 275
8.2.2 Application of Peptidomics in the Research of Bioactive Peptides 276
8.3 Genetic engineering technology 279
8.3.1 Overview of genetic engineering 279
8.3.2 Overview of antifreeze protein genes 279
8.3.3 Application of genetic engineering technology in frost resistance research 279
8.4 Protein Engineering Technology 283
8.4.1 Overview of protein engineering technology 283
8.4.2 Application of protein engineering in antifreeze protein 283
8.5 Bioinformatics Technology 284
8.5.1 Overview of Bioinformatics Technology 284
8.5.2 Application of bioinformatics technology in antifreeze protein 285

References 286
Appendix Chinese and English abbreviations 292
Index 297