Production of β-galactosidase from new actinobacterial species

Elif Gülşen Karabacak; A. Osman Adıguzel; Hayrettin Saygin; Ahmet Hilmi Çon

doi:10.21448/ijsm.1620801

Production of β-galactosidase from new actinobacterial species

Abstract

Streptomyces himalayensis 3K401, Jiangella aurantiaca 8K307T, and Jiangella ureilytica KC603T isolates were identified as potential β-galactosidase producers. The specific activities of the partially purified enzyme preparations were measured as 3.588 ± 0.00 U/mg protein, 1.320 ± 0.09 U/mg protein, and 1.262 ± 0.02 U/mg protein, respectively. The optimal operating conditions for β-galactosidase derived from S. himalayensis 3K401, J. aurantiaca 8K307T, and J. ureilytica KC603T were determined to be 40°C at pH 7.0, 50°C at pH 7.0, and 50°C at pH 9.0, respectively. Notably, all enzyme solutions retained activity at elevated temperatures (60–70°C). Comparative analysis revealed that β-galactosidases from Jiangella isolates exhibited greater thermal stability at 50°C and 60°C than the enzyme from S. himalayensis 3K401. This suggests that these enzymes could remain active following pasteurization at relatively high temperatures. Furthermore, enzyme solutions from Jiangella isolates demonstrated activity across a broad pH range, including acidic conditions (pH 4.0 and 5.0), which aligns with their potential application in the production of drinking milk. Additionally, the operational pH profiles of these enzymes, particularly their activity under neutral and alkaline conditions, highlight their potential utility in pharmacological applications beyond the food industry.

Keywords

References

Adıgüzel, A.O. (2020). Production and characterization of thermo-, halo-and solvent-stable esterase from Bacillus mojavensis TH309. Biocataylsis and Biotransformation, 38(3), 1-17. https://doi.org/10.1080/10242422.2020.1715370
Bradford, M.M. (1976). A rapid sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72(1-2), 248–254. https://doi.org/10.1016/0003-2697(76)90527-3
Braga, A.R.C, Gomes, P.A., & Kalil, S.J. (2012). Formulation of culture medium with agroindustrial waste for β-galactosidase production from Kluyveromyces marxianus ATCC 16045. Food and Bioprocess Technology, 5(5), 1653- 1663. https://doi.org/10.1007/s11947-011-0511-0
Ceresino, E.B., Melo, R.R., Kuktaite, R., Hedenqvist, M.S., Zucchi, T.D., & Johansson, E. (2018). Transglutaminase from newly isolated Streptomyces sp. CBMAI 1617: Production optimization, characterization and evaluation in wheat proteinand dough systems. Food Chemistry, 241, 403-410. https://doi.org/10.1016/j.foodchem.2017.09.010
Chanalia P., Gandhi, D., Attri, P., & Dhanda, S. (2018). Purification and characterization of β-galactosidase from probiotic Pediococcus acidilactici and its use in milk lactose hydrolysis and galactooligosaccharide synthesis. Bioorganic Chemistry, 77, 176 189. https://doi.org/10.1016/j.bioorg.2018.01.006
Chandler, V., Donovan, S., Goodwin, S., Sprague, K., & Stiefbold, C. (1998). Enzyme Kinetics. In J. Karcher (Ed.), Proceedings of the 19th Workshop/Conference of the Association for Biology Laboratory Education (ABLE) (PP 81-97). Oregon.
Collinge, A.E., Neubeck, C., & Udinsky, J. (2023, Dec 13). Thermostable Lactase. (US patent no: 3,816,259). https://patents.google.com/patent/US3816259A/en Accessed 13 Dec 2023
Cui, L., Du, G., Zhang, D., Liu, H., Chen, J. (2007). Purification and characterization of transglutaminase from a newly isolated Streptomyces hygroscopicus. Food Chemistry, 105(2), 612-618. https://doi.org/10.1016/j.foodchem.2007.04.020

De-Macias, M.E., De Nadra, M.C.M., & de Saad, A.M.S. (1983). Isolation and purification of beta galactosidase of Lactobacillus murinus CNRZ 313. Current Microbiology, 9, 99-103. https://doi.org/10.1007/BF01568916
Domurcuk, G. (2018). Farklı inkübasyon koşulları ve stres faktörlerinin mikrobiyal transglutaminaz üretimine etkisi, enzimin kısmi saflaştırılması ve karakterizasyonu [The effects of different incubation conditions and stress factors on microbial transglutaminase production, partial purification, and characterization of the enzyme] [Unpublished type of Master’s thesis]. Gazi Osmanpaşa University.
Duan, X., Hu, S., Qi, X., & Gu, Z. (2017). Optimal extracellular production of recombinant Bacillus circulans β-galactosidase in Escherichia coli BL21(DE3). Process Biochemistry, 53, 17-24. https://doi.org/10.1016/j.procbio.2016.11.008
Ekren, G.S. (2013). Fitaz üreten fungusdan enzimin üretimi, saflaştırılması ve karakterizasyonu [Production, purification, and characterization of the enzyme from a phytase-producing fungus] [Unpublished type of Master’s thesis]. Adnan Menderes University.
Fu, L., Wang, Y., Ju, J., Cheng, L., Xu, Y., Yu, B., et al. (2020). Extracellular production of active-form Streptomyces mobaraensis transglutaminase in Bacillus subtilis. Applied Microbiology and Biotechnology, 104(2), 623–631. https://doi.org/10.1007/s00253-019-10256-9
Groep, M.E., Gregory, M.E., Kershenbaum, L.S., & Bogle, I.D.L. (2000). Performance modeling and simulation of biochemical process sequences with interacting unit operations. Biotechnology and Bioengineering, 67(3), 300–311. https://doi.org/10.1002/(SICI)1097-0290(20000205)67:3<300::AID-BIT6>3.0.CO;2-P
Gupte, A.M., & Nair, J.S. (2010). β-Galactosidase production and ethanol fermentation from whey using Kluyveromyces marxianus NCIM 3551. NISCAIR-CSIR, 69(11), 855–859.
Hildebrandt, P., Wanarska, M., & Kur, J. (2009). A new cold-adapted β-D-galactosidase from the Antarctic Arthrobacter sp. 32c – Gene cloning, overexpression, purification and properties. BMC Microbiology, 9(1), 151. https://doi.org/10.1186/1471-2180-9-151
Işık, B. (2012). Çinko eksik koşullarda yetiştirilmiş Hordeum spontaneum C.Koch gövdesinden kurulmuş baskılayıcı çıkarım hibridizasyon kütüphanesinden seçilen cDNA’ların moleküler ve biyoinformatik analizleri. [Molecular and bioinformatics analyses of cDNAs selected from a suppressive subtractive hybridization library constructed from the stem of Hordeum spontaneum C. Koch grown under zinc-deficient conditions] [Unpublished type of Master’s thesis]. Yıldız Technical University.
Jin, M., Huang, J., Pei, Z., Gao, H., & Chang, Z. (2016). Purification and characterization of a high-salt-resistant microbial transglutaminase from Streptomyces mobaraensis. Journal of Molecular Catalysis B: Enzymatic, 133, 6 11. https://doi.org/10.1016/j.molcatb.2016.07.005 Karabacak, E.G., Adıgüzel, A.O., Saygın, H., & Çon, A.H. (2022). Aktinobakteri izolatlarının transglutaminaz, levansukraz ve beta galaktozidaz üretim yetenekleri [Transglutaminase, levansucrase, and beta-galactosidase production abilities of Actinobacteria isolates]. Akademik Gıda, 20(1), 30–39. https://doi.org/10.24323/akademik-gida.1097824
Karagöz, F.P., Demir, Y., Kotan, M.Ş., Dursun, A., Beydemir, Ş., & Dikbaş, N. (2021). Purification of the phytase enzyme from Lactobacillus plantarum: The effect on pansy growth and macro-micro element content. Biotechnology and Applied Biochemistry, 68, 1067–1075. https://doi.org/10.1002/bab.2026
Kaur, H., & Gill, P.K. (2019). Microbial enzymes in food and beverages processing. In A. M. Grumezescu & A.M. Holban (Eds.), Engineering tools in beverage industry (PP. 255–282). Woodhead Publishing.
Kılıç, Y., Yüksekdağ, Z., & Yüksekdağ, H. (2014). Lactobacillus ve Bifidobacterium cinsi bakterilerin beta galaktozidaz enzim aktivitelerinin belirlenmesi [Determination of beta-galactosidase enzyme activities of Lactobacillus and Bifidobacterium bacteria]. Gıda, 39(4), 211–218.
Kim, C.S., Ji, E.S., & Oh, D.K. (2004). Characterization of a thermostable recombinant β-galactosidase from Thermotoga maritima. Journal of Applied Microbiology, 97(5), 1006–1014. https://doi.org/10.1111/j.1365-2672.2004.02377.x
Kosciow, K., & Deppenmeier, U. (2020). Characterization of three novel β-galactosidases from Akkermansia muciniphila involved in mucin degradation. International Journal of Biological Macromolecules, 149, 331–340. https://doi.org/10.1016/j.ijbiomac.2020.01.246
Laemmli, U.K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 22, 680–685. https://doi.org/10.1038/227680a0
Maity, M., Sanyal, S., Bhowal, J., & Bhattacharyya, D.K. (2013). Studies on isolation and characterization of lactase produced from soil bacteria. International Journal of Recent Science Research, 2(8), 92–94.
Nagy, V., & Szakacs, G. (2008). Production of transglutaminase by Streptomyces isolates in solid fermentation. Letters in Applied Microbiology, 25(8), 122 127. https://doi.org/10.1111/j.1472-765X.2008.02395.x
Nakao, M., Harada, M., Kodama, Y., Nakayama, T., Shibano, Y., & Amachi, T. (1994). Purification and characterization of a thermostable β-galactosidase with high transgalactosylation activity from Saccharopolyspora rectivirgula. Applied Microbiology and Biotechnology, 40(5), 657-663.
Özarslan, S. (2018). Kefir danesinden izole edilen Kluyveromyces marxianus’dan üretilen β-galaktosidaz enziminin kısmi saflaştırılması ve karakterizasyonu [Partial purification and characterization of β-galactosidase enzyme produced by Kluyveromyces marxianus isolated from kefir grains] [Unpublished type of dissertation]. Süleyman Demirel University.
Pawlak-Szukalska, A., Wanarska, M., Popinigis, A.T., & Kur, J.A. (2014). Novel cold-active β-d-galactosidase with transglycosylation activity from the Antarctic Arthrobacter sp. 32cB: Gene cloning, purification, and characterization. Process Biochemistry, 49(12), 2122-2133. https://doi.org/10.1016/j.procbio.2014.09.018
Princely, S., Basha, N.S., Kirubakaran, J.J., & Dhanaraju, M.D. (2013). Biochemical characterization, partial purification, and production of an intracellular β-galactosidase from Streptococcus thermophilus grown in whey. European Journal of Experimental Biology, 3(2), 242-251.
Sanchez, J., & Hardisson, C. (1979). Induction of β-galactosidase in Streptomyces violaceus. Canadian Journal of Microbiology, 25(7), 833-840. https://doi.org/10.1139/m79-123
Sarrouh, B., Santos, T.M., Miyoshi, A., Dias, R., & Azevedo, V. (2012). Up-to-date insight on industrial enzymes applications and global market. Journal of Bioprocess Biotechniques, 4(2), 1-10. https://doi.org/10.4172/2155-9821.S4-002
Shirling, E.B., & Gottlieb, D. (1966). Methods for characterization of Streptomyces species. International Journal of Systematic Bacteriology, 16(3), 313 340. https://doi.org/10.1099/00207713-16-3-313
Stockinger Lab. (2020, Jul 26). Dialysis tubing lab protocols. https://stockingerlab.osu.edu/sites/stockinger/files/imce/PDFs/Protocols/DialysisTubing.pdf
Topal, Ş. (1985). Enzimler, mikrobiyolojik yolla enzim üretimi ve bu teknolojide renninin yeri [Enzymes, microbial production of enzymes, and the role of rennin in this technology]. Gıda, 10(1), 25-37.
Türkan, F., Huyut, Z., Demir, Y., Ertaş, F., & Beydemir, Ş. (2019). The effects of some cephalosporins on acetylcholinesterase and glutathione S-transferase: An in vivo and in vitro study. Archives of Physiology and Biochemistry, 125(3), 235 243. https://doi.org/10.1080/13813455.2018.1452037
Uyanık, A. (2008). Beta-galaktozidaz enziminin mikrobiyal hücrelerden izolasyonu ve karakterizasyonu [Isolation and characterization of beta-galactosidase enzyme from microbial cells] [Unpublished type of Master’s thesis]. Ankara University.
Yamada, M., Chiba, S., Endo, Y., & Isobe, K. (2017). New alkalophilic β-galactosidase with high activity in the alkaline pH region from Teratosphaeria acidotherma AIU BGA-1. Journal of Bioscience and Bioengineering, 123(1), 15 19. https://doi.org/10.1016/j.jbiosc.2013.03.012

Details

Primary Language

English

Subjects

Enzymes

Journal Section

Research Article

Authors

Elif Gülşen Karabacak ^*
0000-0002-3513-0986
Türkiye

A. Osman Adıguzel
0000-0002-5602-5886
Türkiye

Hayrettin Saygin
0000-0002-8642-5872
Türkiye

Ahmet Hilmi Çon This is me
0000-0002-1225-0133
Türkiye

Early Pub Date

September 1, 2025

Publication Date

December 5, 2025

Submission Date

January 16, 2025

Acceptance Date

July 5, 2025

Published in Issue

Year 2025 Volume: 12 Number: 4

DOI

https://doi.org/10.21448/ijsm.1620801

IZ

https://izlik.org/JA93LZ59GA

Cite

RIS / Bibtex

APA

Karabacak, E. G., Adıguzel, A. O., Saygin, H., & Çon, A. H. (2025). Production of β-galactosidase from new actinobacterial species. International Journal of Secondary Metabolite, 12(4), 868-876. https://doi.org/10.21448/ijsm.1620801

AMA

1.Karabacak EG, Adıguzel AO, Saygin H, Çon AH. Production of β-galactosidase from new actinobacterial species. Int. J. Sec. Metabolite. 2025;12(4):868-876. doi:10.21448/ijsm.1620801

Chicago

Karabacak, Elif Gülşen, A. Osman Adıguzel, Hayrettin Saygin, and Ahmet Hilmi Çon. 2025. “Production of β-Galactosidase from New Actinobacterial Species”. International Journal of Secondary Metabolite 12 (4): 868-76. https://doi.org/10.21448/ijsm.1620801.

EndNote

Karabacak EG, Adıguzel AO, Saygin H, Çon AH (December 1, 2025) Production of β-galactosidase from new actinobacterial species. International Journal of Secondary Metabolite 12 4 868–876.

IEEE

[1]E. G. Karabacak, A. O. Adıguzel, H. Saygin, and A. H. Çon, “Production of β-galactosidase from new actinobacterial species”, Int. J. Sec. Metabolite, vol. 12, no. 4, pp. 868–876, Dec. 2025, doi: 10.21448/ijsm.1620801.

ISNAD

Karabacak, Elif Gülşen - Adıguzel, A. Osman - Saygin, Hayrettin - Çon, Ahmet Hilmi. “Production of β-Galactosidase from New Actinobacterial Species”. International Journal of Secondary Metabolite 12/4 (December 1, 2025): 868-876. https://doi.org/10.21448/ijsm.1620801.

JAMA

1.Karabacak EG, Adıguzel AO, Saygin H, Çon AH. Production of β-galactosidase from new actinobacterial species. Int. J. Sec. Metabolite. 2025;12:868–876.

MLA

Karabacak, Elif Gülşen, et al. “Production of β-Galactosidase from New Actinobacterial Species”. International Journal of Secondary Metabolite, vol. 12, no. 4, Dec. 2025, pp. 868-76, doi:10.21448/ijsm.1620801.

Vancouver

1.Elif Gülşen Karabacak, A. Osman Adıguzel, Hayrettin Saygin, Ahmet Hilmi Çon. Production of β-galactosidase from new actinobacterial species. Int. J. Sec. Metabolite. 2025 Dec. 1;12(4):868-76. doi:10.21448/ijsm.1620801