Evaluation of Inhibitory Activity of α-Amylase and α-Glucosidase Enzymes and Aantioxidant Capacity of Metabolites Obtained from of Lactobacillus delbrueckii ssp bulgaricus PTCC1900 Culture Media

Authors
Faezeh shirkhan 1
Saeed Mirdamadi 2
Mahta Mahta Mirzaei 3
Behrouz Akbari-Adergani 4
nikoo nasoohi 5
1 Department of Food Science and Technology, Faculty of Pharmacy, Tehran Medical Sience, Islamic Azad,Tehran, Iran
2 Professor, Department of Biotechnology, Iranian Research Organization for Science & Technology (IROST), Tehran, Iran
3 Assistant Professor, Department of Food Science and Technology, Shahr-e-Qods Branch, Islamic Azad University, Tehran, Iran
4 Professor of food and drug laboratory research center, food and drug administration, ministry of health and medical education, Tehran, Iran
5 Assistant Professor, Department of Biochemistry and Biophysics, Faculty of Advanced Science and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran
10.22034/FSCT.19.125.47
Abstract
Today, the use of Lactobacill strains and their metabolites is the new strategies in control of diabetes and oxidative stress. Therefore, the present study aimed to investigate the antidiabetic and antioxidant properties of metabolites obtained from of Lactobacillus delbrueckii ssp. bulgaricus culture media. The samples were evaluated including cell free supernatant (CFS), cell free extracts (CFE), cell free supernatant of fermented milk (CFS-FM) and bioactive peptides separated from ultrafiltration (3,5,10 kDa). The ability of bacteria in proteolysis of protein of fermented milk (FM) was evaluated by O-phthaldialdehyde method compared with non-fermented milk (NFM). The antidiabetic activity was measured based on inhibition of α-amylase and α-glucosidase. The antioxidant activity was evaluated by inhibition of 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS). The results showed that the sample of CFS-FM inhibited α-amylase (25%) and α-glucosidase (26.6%) enzymes. Among the peptide fractions, fraction of >10 kDa with higher protein content (13.816 mg (had the highest inhibition of α-amylase (80.64%) and α-glucosidase enzyme (54%) (p<0.05). The inhibitory efficiency ratio (IER) showed that the 3-5 kDa fraction had more inhibitory than other peptide fractions (p<0.05). Also, the CFS and CFE inhibited α-glucosidase enzyme by 50% and 25%, respectively. In case of antioxidant properties, it was observed that the free radicals scavenging of CFS-FM increased compared with NFM (p<0.05). Also, the peptide fractions of 3-5 kDa (80.85%) and >10 kDa (43.63%) had the highest DPPH and ABTS free radicals scavenging. The antioxidant capacity of trolox equivalent from peptide fractions (µmTE/mg protein) showed that the <3 kDa fraction had the highest antioxidant activity (p<0.05). The ability to free radicals scavenging in CFS and CFE was observed. Results indicate the importance of Lactobacillus delbrueckii as a starter culture and its functional role.
Keywords
Diabetes
α-amylase
α-glucosidase
Antioxidant
Metabolite
Lactobacillus delbrueckii

Subjects

[1] Zeng, Z., Luo, JY., Zuo, FL., Yu, R., Zhang, Y., Ma, HQ.& Chen, SW. 2016. Bifidobacteria possess inhibitory activity against dipeptidyl peptidase‐IV. Letters in Applied Microbiology, 62(3), 250-255.
[2] Rajoka, MS., Shi, J., Mehwish, HM., Zhu, J., Li, Q., Shao, D., Huang, Q. & Yang, H. 2017. Interaction between diet composition and gut microbiota and its impact on gastrointestinal tract health. Food Science and Human Wellness, 6(3), 121-130.
[3] Cabello-Olmo, M., Oneca, M., Torre, P., Sainz, N., Moreno-Aliaga, MJ., Guruceaga, E., Díaz, JV., Encio, IJ., Barajas, M. & Araña, M. 2019. A fermented food product containing lactic acid bacteria protects ZDF rats from the development of type 2 diabetes. Nutrients, 11(10), 25-30.
[4] Graham, K., Rea, R., Simpson, P. & Stack, H. 2019. Enterococcus faecalis milk fermentates display antioxidant properties and inhibitory activity towards key enzymes linked to hypertension and hyperglycaemia. Journal of Functional Foods, 58, 292-300.
[5] Chen, P., Zhang, Q., Dang, H., Liu, X., Tian, F., Zhao, J., Chen, Y., Zhang, H. & Chen, W. 2014. Screening for potential new probiotic based on probiotic properties and α-glucosidase inhibitory activity. Food Control, 35(1), 65-72.
[6] Qian, B., Xing, M., Cui, L., Deng, Y., Xu, Y., Huang, M. & Zhang, S. 2011. Antioxidant, antihypertensive and immunomodulatory activities of peptide fractions from fermented skim milk with Lactobacillus delbrueckii ssp. bulgaricus LB340.
Journal of Dairy Research, 78(1), 72-79.
[7] Moslehishad, M., Ehsani, MR., Salami, M., Mirdamadi, S., Ezzatpanah, H., Naslaji, AN. & Moosavi-Movahedi, AA. 2013. The comparative assessment of ACE-inhibitory and antioxidant activities of peptide fractions obtained from fermented camel and bovine milk by Lactobacillus rhamnosus PTCC 1637. International Dairy Journal, 29(2), 82-87.
[8] Ayyash, M., Al-Nuaimi, AK., Al-Mahadin, S. & Liu, SQ. 2018. In vitro investigation of anticancer and ACE-inhibiting activity, α-amylase and α-glucosidase inhibition, and antioxidant activity of camel milk fermented with camel milk probiotic: A comparative study with fermented bovine milk. Food Chemistry, 15(239), 588-597.
[9] González-Montoya, M., Hernández-Ledesma, B., Mora-Escobedo, R. & Martínez-Villaluenga, C. 2018. Bioactive peptides from germinated soybean with anti-diabetic potential by inhibition of dipeptidyl peptidase-IV, α-amylase, and α-glucosidase enzymes. International Journal of Molecular Sciences, 19(10), 1-14.
[10] Muganga, L., Liu, X., Tian, F., Zhao, J., Zhang, H. & Chen, W. 2015. Screening for lactic acid bacteria based on antihyperglycaemic and probiotic potential and application in synbiotic set yoghurt. Journal of Functional Foods, 1(16), 125-136.
[11] Tagliazucchi, D., Martini, S. & Solieri, L. 2019. Bioprospecting for bioactive peptide production by lactic acid bacteria isolated from fermented dairy food. Fermentation, 5(4), 1-34.
[12] Stratton, I. M., Adler, A. I., Neil, H. A. W., Matthews, D. R., Manley, S. E., Cull, C. A., ... & Holman, R. R. 2000. Association of glycaemia with macrovascular and microvascular complications of type 2 diabetes (UKPDS 35): prospective observational study. British Medical Journal, 321(7258), 405-412.
[13] Dan, T., Ren, W., Liu, Y., Tian, J., Chen, H., Li, T., & Liu, W. 2019. Volatile flavor compounds profile and fermentation characteristics of milk fermented by Lactobacillus delbrueckii subsp. bulgaricus. Frontiers in Microbiology, 10, 1-12.
[14] Soleymanzadeh, N., Mirdamadi, S. & Kianirad M. 2016. Antioxidant activity of camel and bovine milk fermented by lactic acid bacteria isolated from traditional fermented camel milk (Chal). Dairy Science & Technology, 96(4), 443-457.
[15] Hatree, EF. 1972. Determination of protein: a modification of the Lowry method that gives a linear photometric response. Analytical Biochemistry, 48, 422-427.
[16] Church, FC, Swaisgood, HE., Porter, DH., Catignani, GL. 1983. Spectrophotometric assay using o-phthaldialdehyde for determination of proteolysis in milk and isolated milk proteins. Journal of Dairy Science, 66(6), 1219-1227.
[17] Son, S. & Lewis, BA. 2002. Free radical scavenging and antioxidative activity of caffeic acid amide and ester analogues: Structure-activity relationship. Journal of Agricultural and Food Chemistry, 50(3), 468-472.
[18] Mukherjee, S,. Pawar, N., Kulkarni, O., Nagarkar, B., Thopte, S., Bhujbal, A. & Pawar, P. (2011). Evaluation of free-radical quenching properties of standard Ayurvedic formulation Vayasthapana Rasayana. BMC Complementary and Alternative Medicine, 11(1), 1-6.
[19] Al Kanhal, H. A. 2010. Compositional, technological and nutritional aspects of dromedary camel milk. International Dairy Journal, 20(12), 811-821.
[20] Vollet Marson, G., Belleville, M. P., Lacour, S. & Dupas Hubinger, M. 2021. Membrane fractionation of protein hydrolysates from by-Products: recovery of valuable compounds from spent yeasts. Membranes, 11(23), 1-19.
[21] Rubak, YT., Nuraida, L., Iswantini, D. &Prangdimurti, E. 2020. Angiotensin-I-converting enzyme inhibitory peptides in milk fermented by indigenous lactic acid bacteria. Veterinary World, 13(2), 345-353.
[22] Aloğlu, H. Ş., & Öner, Z. 2011. Determination of antioxidant activity of bioactive peptide fractions obtained from yogurt. Journal of Dairy Science, 94(11), 5305-5314.
[23] Soleymanzadeh, N., Mirdamadi, S., Mirzaei, M. &Kianirad, M. 2019. Novel β-casein derived antioxidant and ACE-inhibitory active peptide from camel milk fermented by Leuconostoc lactis PTCC1899: Identification and molecular docking. International Dairy Journal, 97, 201-208.
[24] Ramchandran, L., & Shah, N. P. 2008. Proteolytic profiles and angiotensin‐I converting enzyme and α‐glucosidase inhibitory activities of selected lactic acid bacteria. Journal of Food Science, 73(2), 75-81.
[25] Famuwagun, A. A., Alashi, A. M., Gbadamosi, S. O., Taiwo, K. A., Oyedele, D., Adebooye, O. C., & Aluko, R. E. 2021. Effect of protease type and peptide size on the In vitro antioxidant, antihypertensive and anti-diabetic activities of eggplant leaf Protein hydrolysates. Foods, 10(5), 1-22.
[26] Castañeda-Pérez, E., Jiménez-Morales, K., Quintal-Novelo, C., Moo-Puc, R., Chel-Guerrero, L., & Betancur-Ancona, D. 2019. Enzymatic protein hydrolysates and ultrafiltered peptide fractions from Cowpea Vigna unguiculata L bean with in vitro antidiabetic potential. Journal of the Iranian Chemical Society, 16(8), 1773-1781.
[27] Frediansyah, A., Nurhayati, R., & Sholihah, J. 2019. Lactobacillus pentosus isolated from Muntingia calabura shows inhibition activity toward alpha-glucosidase and alpha-amylase in intra and extracellular level. 2nd International Conference on Natural Products and Bioresource Sciences-2018. In IOP Conference Series: Earth and Environmental Science, 251(1), 1-6.
[28] Son, SH., Jeon, HL., Yang, SJ., Lee, NK. & Paik HD. 2017. In vitro characterization of Lactobacillus brevis KU15006, an isolate from kimchi, reveals anti-adhesion activity against foodborne pathogens and antidiabetic properties. Microbial Pathogenesis, 112, 135-141.
[29] Yusuf, D., Nuraida, L., Dewanti-Hariyadi, R. &Hunaefi, D. 2021. In vitro Antioxidant and α-glucosidase inhibitory activities of Lactobacillus spp. isolated from indonesian kefir grains. Applied Food Biotechnology, 8(1), 39-46.
[30] Aguilar-Toalá, J. E., Santiago-López, L., Peres, C. M., Peres, C., Garcia, H. S., Vallejo-Cordoba, B., ... & Hernández-Mendoza, A. 2017. Assessment of multifunctional activity of bioactive peptides derived from fermented milk by specific Lactobacillus plantarum strains. Journal of Dairy Science, 100(1), 65-75.
[31] Ngoh, YY., & Gan CY. 2016. Enzyme-assisted extraction and identification of antioxidative and α-amylase inhibitory peptides from Pinto beans (Phaseolus vulgaris cv. Pinto). Food Chemistry, 190, 331-337.
[32] Mirzaei, M., Mirdamadi, S., Ehsani, MR., Aminlari, M. & Hosseini, E. 2015. Purification and identification of antioxidant and ACE-inhibitory peptide from Saccharomyces cerevisiae protein hydrolysate. Journal of Functional Foods, 19, 259-68.
[33] Wiriyaphan, C., Chitsomboon, B. & Yongsawadigul, J. 2012. Antioxidant activity of protein hydrolysates derived from threadfin bream surimi by products. Food Chemistry, 132, 104-111.
[34] Sarmadi, B.H., & Ismail, A. 2010. Antioxidative peptides from food proteins: a review. Peptides, 31(10),1949-1956.