Our website uses cookies. By continuing to use this site, you agree to our use of cookies in accordance with this notice and the Terms of Use.
Journals KemSU
Scientific journal "Bulletin of KemSU. Series: Humanities and Social Sciences Scientific journal "Bulletin of KemSU. Series: Political, Sociological and Economic sciences Scientific journal "Bulletin of KemSU. Series: Biological, Engineering and Earth Sciences SibScript (until February 17, 2023 – Bulletin of KemSU Strategizing: Theory and Practice Foods and Raw Materials Virtual Communication and Social Networks Food Processing: Techniques and Technology Dairy industry Cheese- and buttermaking Science Evolution
Submit manuscript
Food Processing: Techniques and Technology
UDC 63

LACTULOSE BIOSYNTHESIS WITH ENZYMES OF LACTOSE-FERMENTING MICROMYCETES AND BACTERIA Lactulose Biosynthesis with Enzymes of Lactose-Fermenting Micromycetes and Bacteria

Published in Food Processing: Techniques and Technology · Volume 54, Issue 4 · Pages 645–657 · Rubric: ORIGINAL ARTICLE
DOI: https://doi.org/10.21603/2074-9414-2024-4-2533
Received: 14.03.2024 Accepted: 07.05.2024 Published: 24.12.2024 Language of publication: RUS
Authors
SV
Svetlana Ryabtseva
ORCID
MA
Maria Shpack
ORCID
AL
Alexei Lodygin
ORCID SPIN
AL
Alexander Serov
SPIN
et al.
SE
Serafima Sazanova
ORCID
MA
Marina Skorokhodova
ORCID
VE
Vera Romakhova
ORCID
ABSTRACT
Lactulose is a well-studied prebiotic popular in medicine and the food industry. β-Galactosidases offer an alternative to traditional chemical methods of lactulose production. The article describes lactulose biosynthesis with crude enzyme preparations of β-galactosidases obtained by separate and joint cultivation of lactose-fermenting yeasts and lactic acid bacteria in whey permeate. The research featured the following producers of β-galactosidases: lactose-fermenting yeasts of Kluyveromyces lactis VKM Y-1333 and Y-1339, Kluyveromyces marxianus VKM Y-459 and Y-1338, and viscous strains of lactic acid bacteria of Lactobacillus acidophilus and Streptococcus thermophilis. The β-galactosidase activity was measured using the method of o-nitrophenyl-β-D-galactopyranoside hydrolysis. The method of high-performance liquid chromatography made it possible to reveal the carbohydrate composition. The lactulose formation by yeast enzymes demonstrated the following pattern: its concentration increased rapidly during the first 30 min of reaction with subsequent decrease or stabilization. Some combined unpurified enzyme preparations of yeasts and lactic acid microorganisms were more active than β-galactosidases obtained by separate cultivation. The patterns of lactulose formation depended on both the yeast strain and the type of lactic acid bacteria. The highest yields of lactulose belonged to the samples that combined β-galactosidases of all yeast strains with Streptococcus thermophilis. Co-cultivation of yeast and lactic acid bacteria in some combinations produced combined crude enzyme preparations with higher activity and greater lactulose yields than separate yeast cultivation. These results can help to obtain purified enzyme preparations of β-galactosidases and lactulose-containing products from secondary dairy raw materials.
KEYWORDS
Lactulose, lactose, transglycosylation, β-galactosidases, Kluyveromyces marxianus, Kluyveromyces lactis, Lactobacillus acidophilus, Streptococcus thermophilus
Funding
The research was supported by t he Ministry of Science and Higher Education of the Russian Federation as part of a comprehensive high-tech project of First Russian high-tech lactulose prebiotic and functional dairy ingredients for import substitution in medicine, veterinary practice, baby food, and medical preventive products for humans and animals (Agreement No. 075-11-2022-021, April 07, 2022, on federal budget subsidies for cooperation between state scientific institutions and the real sector of the economy in the sphere of high-tech production); Decree No. 218, April 9, 2010, of the Government of the Russian Federation to the North Caucasian Federal University .
Text References
Text
Text (PDF)
Read Download

1. Vera C, Guerrero C, Illanes A. Trends in lactose-derived bioactives: synthesis and purification. Systems Microbiology and Biomanufacturing. 2022;2:393–412. http://doi.org/10.1007/s43393-021-00068-2

2. Nooshkam M, Babazadeh A, Jooyandeh H. Lactulose: Properties, technofunctional food applications, and food grade delivery system. Trends in Food Science and Technology. 2018;80:23–34. https://doi.org/10.1016/j.tifs.2018.07.028

3. Ryabtseva SA, Khramtsov AG, Budkevich RO, Anisimov GS, Chuklo AO, Shpak MA. Physiological effects, mechanisms of action and application of lactulose. Problems of Nutrition. 2020;89(2):5–20. (In Russ.). https://doi.org/10.24411/0042-8833-2020-10012; https://www.elibrary.ru/TNXHMW

4. Ryabtseva SA. Lactulose technology. Moscow: DeLi print; 2003. 232 p. (In Russ.).

5. Sitanggang AB, Drews A, Kraume M. Recent advances on prebiotic lactulose production. World Journal of Microbiology and Biotechnology. 2016;32(9):154. https://doi.org/10.1007/s11274-016-2103-7

6. Silvério SC, de Macedo EA, Teixeira JA, Rodrigues LR. Biocatalytic approaches using lactulose: end product compared with substrate. Comprehensive Reviews in Food Science and Food Safety. 2016;15(5):878–896. https://doi.org/10.1111/1541-4337.12215

7. Vera C, Illanes A. Lactose-derived nondigestible oligosaccharides and other high added-value products. In: Illanes A, Guerrero C, Vera C, Wilson L, Conejeros R, Scott F, editors. Lactose-derived prebiotics. A process perspective. Academic Press; 2016. pp. 87–110. https://doi.org/10.1016/B978-0-12-802724-0.00003-2

8. Wang M, Wang L, Lyu X, Hua X, Goddard JM, Yang R. Lactulose production from lactose isomerization by chemo-catalysts and enzymes: Current status and future perspectives. Biotechnology Advances. 2022;60:108021. https://doi.org/10.1016/j.biotechadv.2022.108021

9. Ryabtseva SA, Khramtsov AG, Shpak MA, Lodygin AD, Anisimov GS, Sazanova SN, et al. Biotechnology of Lactulose Production: Progress, Challenges, and Prospects. Food Processing: Techniques and Technology. 2023;53(1):97–122c. https://doi.org/10.21603/2074-9414-2023-1-2419; https://www.elibrary.ru/DVBEZS

10. de Albuquerque TL, de Sousa M, Gomes e Silva NC, Girão Neto CAC, Gonçalves LRB, Fernandez-Lafuente R, et.al. β-Galactosidase from Kluyveromyces lactis: Characterization, production, immobilization and applications - A review. International Journal of Biological Macromolecules. 2021;191:881–898. https://doi.org/10.1016/j.ijbiomac.2021.09.133

11. Lyutova LV, Naumov GI, Shnyreva AV, Naumova ES. Molecular polymorphism of β-galactosidase LAC4 genes in dairy and natural strains of Kluyveromyces yeasts. Molecular Biology. 2021;55(1):75–85. (In Russ.). https://doi.org/10.31857/ S0026898421010109; https://www.elibrary.ru/DXUCCI

12. Lyutova LV, Naumova ES. Inter-strain hybridization of Kluyveromyces lactis for creating efficient lactose-fermenting yeast. Biotekhnologiya. 2021;37(4):43–50. (In Russ.). https://doi.org/10.21519/0234-2758-2021-37-4-43-50; https://www.elibrary.ru/XZBKCB

13. Lyutova LV, Naumova ES. Comparative analysis of fermentation of lactose and its components, glucose and galactose, by interstrain hybrids of dairy yeast Kluyveromyces lactis. Biotechnology. 2023;39(1):3–11. (In Russ.). https://doi.org/10.56304/S0234275823010064; https://www.elibrary.ru/BMMPOR

14. Ruiz-Ramírez S, Jiménez-Flores R. Properties of β-Galactosidases derived from Lactobacillaceae species and its capacity for galacto-oligosaccharides production. Journal of Dairy Science. 2023;106(12):8193–8206. https://doi.org/10.3168/jds.2023-23392

15. Dorau R, Jensen PR, Solem C. Purified lactases versus whole-cell lactases-the winner takes it all. Applied Microbiology and Biotechnology. 2021;105:4943–4955. https://link.springer.com/article/10.1007/s00253-021-11388-7

16. Wang Q, Lillevang SK, Rydtoft SM, Xiao H, Fan M-T, Solem C, et.al. No more cleaning up - Efficient lactic acid bacteria cell catalysts as a cost-efficient alternative to purified lactase enzymes. Applied Microbiology and Biotechnology. 2020;104:6315–6323. https://link.springer.com/article/10.1007/s00253-020-10655-3

17. Hashem AM, El-Azeem Ismail SA, Helmy WA, El-Mohamady Y, Abou-Romia R. Factors affecting the production of lactulose by Lactobacillus acidophilus NRRL 4495 b-galactosidase and its biological activity. Malaysian Journal of Microbiology. 2013;9():1–6. http://doi.org/10.21161/mjm.43612

18. Ryabtseva SA, Kotova AA, Skripnyuk AA. Yeast in the processing of dairy raw materials. St. Petersburg: Lan; 2019. 120 p. (In Russ.).

19. Botvynko A, Bednářová A, Henke S, Shakhno N, Čurda L. Production of galactooligosaccharides using various combinations of the commercial β-galactosidases. Biochemical and Biophysical Research Communications. 2019;517(4):762–766. https://doi.org/10.1016/j.bbrc.2019.08.001

20. Inchaurrondo VA, Yantorno OM, Voget CE. Yeast growth and beta-galactosidase production during aerobic batch cultures in lactose-limited synthetic medium. Process biochemistry. 1994;29(1):47–54. https://doi.org/10.1016/0032-9592(94)80058-8

21. Guerrero C, Vera C, Plou F, Illanes A. Influence of reaction conditions on the selectivity of the synthesis of lactulose with microbial b-galactosidases. Journal of Molecular Catalysis B: Enzymatic. 2011;72(3–4):206–212. https://doi.org/10.1016/j.molcatb.2011.06.007

22. Hua X, Yang R, Shen Q, Ye F, Zhang W, Zhao W. Production of 1-lactulose and lactulose using commercial b-galactosidase from Kluyveromyces lactis in the presence of fructose. Food Chemistry. 2013;137(1–4):1–7. https://doi.org/10.1016/j.foodchem.2012.10.003

23. Sitanggang AB, Drews A, Kraume M. Influences of operating conditions on continuous lactulose synthesis in an enzymatic membrane reactor system: A basis prior to long-term operation. Journal of Biotechnology. 2015;203:89–96. https://doi.org/10.1016/j.jbiotec.2015.03.016

24. De Albuquerque TL, Gomes SDL, D’Almeida AP, Fernandez-Lafuente R, Gonçalves LRB, Rocha MVP. Immobilization of β-galactosidase in glutaraldehyde-chitosan and its application to the synthesis of lactulose using cheese whey as feedstock. Process Biochemistry. 2018;73:65–73. https://doi.org/10.1016/j.procbio.2018.08.010

25. Neto CACG, Silva NCGE, de Oliveira Costa T, de Albuquerque TL, Gonçalves LRB, Fernandez-Lafuente R, et.al. The beta-galactosidase immobilization protocol determines its performance as catalysts in the kinetically controlled synthesis of lactulose. International Journal of Biological Macromolecules. 2021;176:468–478. https://doi.org/10.1016/j.ijbiomac.2021.02.078

26. Lee Y-J, Kim CS, Oh D-K. Lactulose production by beta-galactosidase in permeabilized cells of Kluyveromyces lactis. Applied Microbiology and Biotechnology. 2004;64:787–793. http://doi.org/10.1007/s00253-003-1506-1

27. Guerrero C, Vera C, Conejeros R, Illanes A. Transgalactosylation and hydrolytic activities of commercial preparations of β-galactosidase for the synthesis of prebiotic carbohydrates. Enzyme and Microbial Technology. 2015;70:9–17. https://doi.org/10.1016/j.enzmictec.2014.12.006

28. Ryabtseva SA, Skripnyuk AA, Kotova AA, Khramtsov AG, Rodnaya AB, Lodygin AD, et al. Method for combined enzyme beta-galactosidase production. Russia Patent Ru 2622078. 2017.

29. Zhao R, Chen Z, Liang J, Dou J, Guo F, Xu Z, et.al. Advances in Genetic Tools and Their Application in Streptococcus thermophilus. Foods. 2023;12(16):3119. https://doi.org/10.3390/foods12163119

This work is licensed under Creative Commons Attribution 4.0 International
PDF Text JATS XML
The contact persons
Alexander Prosekov
Chief editor
Alexander Prosekov
4562
downloads
RINC RINC Science Index Crossref
CC
This work is licensed under Creative Commons Attribution 4.0 International Creative Commons
© КемГУ, 1997–2025
Agreement Personal data protection and processing policy Support Powered by Editorum, 2026 Instructions
Login or Create
* Forgot your password?