Synergistic combination of cryoprotectants improved freeze-dried survival rate and viable counts of Lactiplantibacillus plantarum
Wenlong Ma
College of Food Science and Engineering, Yangzhou University, Yangzhou, 225127 China
Contribution: Conceptualization, Methodology, Validation, Writing - review & editing, Funding acquisition, Visualization, Data curation, Software
Search for more papers by this authorYinghua Li
College of Food Science and Engineering, Yangzhou University, Yangzhou, 225127 China
Contribution: Data curation, Investigation, Validation, Writing - original draft, Software, Visualization
Search for more papers by this authorWenli Kang
Ausnutria Institute of Food and Nutrition, Ausnutria Dairy (China) Co., Ltd., Changsha, 410083 China
Human Engineering Research Center of Human Microecological Products, Changsha, 410083 China
Contribution: Conceptualization, Validation, Methodology, Visualization
Search for more papers by this authorYifeng Han
Jiangsu College of Tourism, Yangzhou, 225127 China
Contribution: Methodology, Formal analysis
Search for more papers by this authorBoxing Yin
Jiangsu Diary Biotechnology Engineering Research Center, Kang Yuan Diary Co. Ltd, Yangzhou, 225127 China
Contribution: Project administration, Supervision
Search for more papers by this authorRenqin Yang
Jiangsu Diary Biotechnology Engineering Research Center, Kang Yuan Diary Co. Ltd, Yangzhou, 225127 China
Contribution: Project administration, Funding acquisition
Search for more papers by this authorRongxue Tang
Ausnutria Institute of Food and Nutrition, Ausnutria Dairy (China) Co., Ltd., Changsha, 410083 China
Contribution: Validation, Formal analysis, Investigation
Search for more papers by this authorCorresponding Author
Lina Pan
Ausnutria Institute of Food and Nutrition, Ausnutria Dairy (China) Co., Ltd., Changsha, 410083 China
Human Engineering Research Center of Human Microecological Products, Changsha, 410083 China
School of minerals Processing and Bioengineering, Central South University, Changsha, 410083 China
Author for correspondence. E-mail: [email protected]; [email protected]
Contribution: Project administration, Supervision
Search for more papers by this authorJiaqi Wang
Ausnutria Institute of Food and Nutrition, Ausnutria Dairy (China) Co., Ltd., Changsha, 410083 China
Human Engineering Research Center of Human Microecological Products, Changsha, 410083 China
Contribution: Supervision, Project administration
Search for more papers by this authorWei Li
Ausnutria Institute of Food and Nutrition, Ausnutria Dairy (China) Co., Ltd., Changsha, 410083 China
Contribution: Supervision, Project administration
Search for more papers by this authorCorresponding Author
Yujun Huang
College of Food Science and Engineering, Yangzhou University, Yangzhou, 225127 China
Author for correspondence. E-mail: [email protected]; [email protected]
Contribution: Conceptualization, Funding acquisition, Project administration, Writing - review & editing
Search for more papers by this authorRuixia Gu
College of Food Science and Engineering, Yangzhou University, Yangzhou, 225127 China
Human Engineering Research Center of Human Microecological Products, Changsha, 410083 China
Contribution: Project administration, Funding acquisition, Supervision
Search for more papers by this authorWenlong Ma
College of Food Science and Engineering, Yangzhou University, Yangzhou, 225127 China
Contribution: Conceptualization, Methodology, Validation, Writing - review & editing, Funding acquisition, Visualization, Data curation, Software
Search for more papers by this authorYinghua Li
College of Food Science and Engineering, Yangzhou University, Yangzhou, 225127 China
Contribution: Data curation, Investigation, Validation, Writing - original draft, Software, Visualization
Search for more papers by this authorWenli Kang
Ausnutria Institute of Food and Nutrition, Ausnutria Dairy (China) Co., Ltd., Changsha, 410083 China
Human Engineering Research Center of Human Microecological Products, Changsha, 410083 China
Contribution: Conceptualization, Validation, Methodology, Visualization
Search for more papers by this authorYifeng Han
Jiangsu College of Tourism, Yangzhou, 225127 China
Contribution: Methodology, Formal analysis
Search for more papers by this authorBoxing Yin
Jiangsu Diary Biotechnology Engineering Research Center, Kang Yuan Diary Co. Ltd, Yangzhou, 225127 China
Contribution: Project administration, Supervision
Search for more papers by this authorRenqin Yang
Jiangsu Diary Biotechnology Engineering Research Center, Kang Yuan Diary Co. Ltd, Yangzhou, 225127 China
Contribution: Project administration, Funding acquisition
Search for more papers by this authorRongxue Tang
Ausnutria Institute of Food and Nutrition, Ausnutria Dairy (China) Co., Ltd., Changsha, 410083 China
Contribution: Validation, Formal analysis, Investigation
Search for more papers by this authorCorresponding Author
Lina Pan
Ausnutria Institute of Food and Nutrition, Ausnutria Dairy (China) Co., Ltd., Changsha, 410083 China
Human Engineering Research Center of Human Microecological Products, Changsha, 410083 China
School of minerals Processing and Bioengineering, Central South University, Changsha, 410083 China
Author for correspondence. E-mail: [email protected]; [email protected]
Contribution: Project administration, Supervision
Search for more papers by this authorJiaqi Wang
Ausnutria Institute of Food and Nutrition, Ausnutria Dairy (China) Co., Ltd., Changsha, 410083 China
Human Engineering Research Center of Human Microecological Products, Changsha, 410083 China
Contribution: Supervision, Project administration
Search for more papers by this authorWei Li
Ausnutria Institute of Food and Nutrition, Ausnutria Dairy (China) Co., Ltd., Changsha, 410083 China
Contribution: Supervision, Project administration
Search for more papers by this authorCorresponding Author
Yujun Huang
College of Food Science and Engineering, Yangzhou University, Yangzhou, 225127 China
Author for correspondence. E-mail: [email protected]; [email protected]
Contribution: Conceptualization, Funding acquisition, Project administration, Writing - review & editing
Search for more papers by this authorRuixia Gu
College of Food Science and Engineering, Yangzhou University, Yangzhou, 225127 China
Human Engineering Research Center of Human Microecological Products, Changsha, 410083 China
Contribution: Project administration, Funding acquisition, Supervision
Search for more papers by this authorAbstract
Well-designed cryoprotectants are crucial for successful probiotic application. In this study, based on literature analysis, characteristics and economic analysis of cryoprotectants, the sucrose (8.2%)–skim milk (8.0%)–inulin (8.2%) combination was proposed and optimised to enhance freeze-drying resistance of probiotics. Survival, viable cell counts and inactivation rate during storage at 4°C of Lactiplantibacillus plantarum grx16 protected by the combination was 94.9%, 1.89 × 1011 cfu/g and 0.0116. The optimised sucrose–skim milk–polymer prebiotics combination could also effectively protect several other probiotics, with the survival rate being 89.2–95.6%. These results will aid in the rational development of synergistic cryoprotectant formulations for probiotics.
CONFLICT OF INTEREST
The authors declare no competing interests.
Open Research
DATA AVAILABILITY STATEMENT
The data that support the findings of this study are available from the corresponding author upon reasonable request.
Supporting Information
| Filename | Description |
|---|---|
| idt13035-sup-0001-TablesS1-S3.xlsxExcel 2007 spreadsheet , 40.9 KB | Table S1 The 123 references and the composition of cryoprotectants used. Table S2 Cryoprotectants usage frequency and concentration used analysis for 171 lactic acid bacteria. Table S3 Cryoprotectants usage frequency and concentration used analysis for 46 Lactiplantibacillus plantarum strains. |
| idt13035-sup-0002-TableS4.docxWord 2007 document , 20.3 KB | Table S4 Analysis of variance (ANOVA) results of the quadratic model. |
| idt13035-sup-0003-FigureS1.docxWord 2007 document , 374.6 KB | Figure S1 Commonly used cryoprotectants analysis for lactobacillus based on literature |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
REFERENCES
- Cao R D, Sogabe T, Mikajiri S and Kawai K (2022) Effects of sucrose, carnosine, and their mixture on the glass transition behavior and storage stability of freeze-dried lactic acid bacteria at various water activities. Cryobiology 106 131–138.
- Carvalho A S, Silva J, Ho P, Teixeira P, Malcata F X and Gibbs P (2003) Protective effect of sorbitol and monosodium glutamate during storage of freeze-dried lactic acid bacteria. Le Lait 83 203–210.
- Carvalho A S, Silva J, Ho P, Teixeira P, Malcata F X and Gibbs P (2004) Relevant factors for the preparation of freeze-dried lactic acid bacteria. International Dairy Journal 14 835–847.
- Chen B Y, Wang X Y, Li P Z et al. (2023) Exploring the protective effects of freeze-dried lactobacillus rhamnosus under optimized cryoprotectants formulation. LWT–Food Science and Technology 173 114295.
- Chin Y W, Lee S, Yu H H, Yang S J and Kim T W (2021) Combinatorial effects of protective agents on survival rate of the yeast starter, Saccharomyces cerevisiae 88-4, after freeze-drying. Microorganisms 9 13.
- Cui S M, Hu M Y, Sun Y Y et al. (2023) Effect of trehalose and lactose treatments on the freeze-drying resistance of lactic acid bacteria in high-density culture. Microorganisms 11 48.
- Deshwal G K, Singh A K, Kumar D and Sharma H (2020) Effect of spray and freeze drying on physico-chemical, functional, moisture sorption and morphological characteristics of camel milk powder. LWT–Food Science and Technology 134 110117.
- Di L J, Ma W L, Kang W L et al. (2022) Synergistic combination of cryoprotectants for high freeze-dried survival rate and viable cell counts of Streptococcus thermophilus. Drying Technology 41 1444–1453.
- Gao X W, Kong J, Zhu H K, Mao B Y, Cui S M and Zhao J X (2022) Lactobacillus, Bifidobacterium and Lactococcus response to environmental stress: Mechanisms and application of cross-protection to improve resistance against freeze-drying. Journal of Applied Microbiology 132 802–821.
- Jingjing E, Jingya Z, Rongze M, Zichao C, Caiqing Y, Ruixue W, Qiaoling Z, Ying Y, Jing L and Junguo W (2023) Study of the internal mechanism of l-glutamate for improving the survival rate of Lactiplantibacillus plantarum LIP-1 after freeze-drying. Innovative Food Science & Emerging Technologies 84 103253.
- Kurtmann L, Carlsen C U, Skibsted L H and Risbo J (2009) Water activity-temperature state diagrams of freeze-dried lactobacillus acidophilus (La-5): Influence of physical state on bacterial survival during storage. Biotechnology Progress 25 265–270.
- Li B K, Tian F W, Liu X M, Zhao J X, Zhang H and Chen W (2011) Effects of cryoprotectants on viability of lactobacillus reuteri CICC6226. Applied Microbiology and Biotechnology 92 609–616.
- Lu Y J, Huang L J, Yang T B, Lv F X and Lu Z X (2017) Optimization of a cryoprotective medium to increase the viability of freeze-dried Streptococcus thermophilus by response surface methodology. LWT–Food Science and Technology 80 92–97.
- Mensink M A, Van Bockstal P J, Pieters S et al. (2015) In-line near infrared spectroscopy during freeze-drying as a tool to measure efficiency of hydrogen bond formation between protein and sugar, predictive of protein storage stability. International Journal of Pharmaceutics 496 792–800.
- Ming L C, Rahim R A, Wan H Y and Ariff A B (2009) Formulation of protective agents for improvement of lactobacillus salivarius I 24 survival rate subjected to freeze drying for production of live cells in powderized form. Food and Bioprocess Technology 2 431–436.
- Misra S, Mandliya S, Pandey P, Panigrahi C, Dalbhagat C G and Mishra H N (2023) Effect of spray- and freeze-dried microcapsules containing probiotics and and γ-aminobutyric acid on nutritional, physicochemical, textural, pasting, rheological, and microstructural characteristics of composite dough. Food and Bioprocess Technology 17 1–15.
- Morsy M K, Morsy O M, Abdelmonem M A and Elsabagh R (2022) Anthocyanin-colored microencapsulation effects on survival rate of lactobacillus rhamnosus GG, color stability, and sensory parameters in strawberry nectar model. Food and Bioprocess Technology 15 352–367.
- Nguyen T H, Kim J S, Kwon H J and Kang C H (2023) The effect of a glutathione (GSH)-containing Cryo-protectant on the viability of probiotic cells using a freeze-drying process. Fermentation-Basel 9 9.
- Pyar H and Peh K K (2011) Effect of cryoprotective agents on survival and stability of lactobacillus acidophilus cultured in food-grade medium. International Journal of Dairy Technology 64 578–584.
- Qu H X, Zhang L F, Liu X X, et al. (2023) Positive effect of Limosilactobacillus plantarum grx16 on nonalcoholic fatty liver disease in rats. International Journal of Dairy Technology 76 710–716.
- Savedboworn W, Teawsomboonkit K, Surichay S, Riansa-ngawong W, Rittisak S, Charoen R and Phattayakorn K (2019) Impact of protectants on the storage stability of freeze-dried probiotic lactobacillus plantarum. Food Science and Biotechnology 28 795–805.
- Ta L P, Bujna E, Antal O, Ladányi M, Juhász R, Szécsi A, Kun S, Sudheer S, Gupta V K and Nguyen Q D (2021) Effects of various polysaccharides (alginate, carrageenan, gums, chitosan) and their combination with prebiotic saccharides (resistant starch, lactosucrose, lactulose) on the encapsulation of probiotic bacteria Lactobacillus casei 01 strain. International Journal of Biological Macromolecules 183 1136–1144.
- Teng D, Kawai K, Mikajiri S and Hagura Y (2017) Stabilization of freeze-dried Lactobacillus paracasei subsp paracasei JCM 8130(T) with the addition of disaccharides, polymers, and their mixtures. Bioscience Biotechnology and Biochemistry 81 768–773.
- Tonnis W F, Mensink M A, De Jager A, Maarschalk K V, Frijlink H W and Hinrichs W L J (2015) Size and molecular flexibility of sugars determine the storage stability of freeze-dried proteins. Molecular Pharmaceutics 12 684–694.
- Wang A Y, Lin J and Zhong Q X (2020a) Synergistic effects of whey protein isolate and amorphous sucrose on improving the viability and stability of powdered lactobacillus salivarius NRRL B-30514. LWT–Food Science and Technology 118 108722.
- Wang G Q, Pu J, Yu X Q, Xia Y J and Ai L Z (2020b) Influence of freezing temperature before freeze-drying on the viability of various lactobacillus plantarum strains. Journal of Dairy Science 103 3066–3075.
- Wang G Q, Luo L Y, Dong C et al. (2021) Polysaccharides can improve the survival of Lactiplantibacillus plantarum subjected to freeze-drying. Journal of Dairy Science 104 2606–2614.
- Wang R X, Sun R Y, Yang Y et al. (2022) Effects of salt stress on the freeze-drying survival rate of Lactiplantibacillus plantarum LIP-1. Food Microbiology 105 104009.
- Wolkers W F and Oldenhof H (2021) Principles underlying cryopreservation and freeze-drying of cells and tissues. Methods in molecular biology (Clifton, N.J.) 2180 3–25.
- Yoha K S, Anukiruthika T, Anila W, Moses J A and Anandharamakrishnan C (2021) 3D printing of encapsulated probiotics: Effect of different post-processing methods on the stability of Lactiplantibacillus plantarum (NCIM 2083) under static in vitro digestion conditions and during storage. LWT–Food Science and Technology 146 111461.
- Zeng Z D, Zeng X Q, Guo Y X, Wu Z, Cai Z D and Pan D D (2022) Determining the role of UTP-Glucose-1-phosphate uridylyltransferase (GalU) in improving the resistance of lactobacillus acidophilus NCFM to freeze-drying. Food 11 1719.
- Zhang J, Liu Q, Chen W, Du G and Chen J (2016) Short communication: Protection of lyophilized milk starter lactobacillus casei Zhang by glutathione. Journal of Dairy Science 99 1846–1852.
- Zhou T, Kang W L, Han Y F et al. (2023) Combinedly increased viability of Lactiplantibacillus plantarum grx16 by co-encapsulation of cryoprotectants and porous starch within calcium alginate capsules. International Journal of Food Science and Technology 58 5291–5298.
