Purification of a protein-based α-glucosidase inhibitor from Aspergillus niger VTCC 031
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https://doi.org/10.15625/vjbt-22393Keywords:
Aspergillus niger VTCC 031, α-glucosidase inhibitor, chromatography.Abstract
Alpha-glucosidase inhibitors, such as acarbose, voglibose, and miglitol, constitute a prevalent class of pharmaceuticals employed in the clinical management of diabetes. These inhibitors originate mostly from microorganisms, including Aspergillus sp., Bacillus sp., and Actinoplanes sp., representing a promising natural resource for identifying α-glucosidase inhibitors (AGIs) that offer enhanced therapeutic efficacy with reduced adverse effects. Aspergillus sp. is a fungus that has a rich array of secondary metabolites and produces several AGIs. In this work, the A. niger VTCC 031 demonstrated the capacity to manufacture compounds with significant α-glucosidase inhibitory activity, achieving a rate of 67.24%. The aqueous phase, obtained after ethyl acetate extraction, was purified using a Sephadex G100 column followed by a 30 kDa cut-off column, resulting in the purification of the α-glucosidase inhibitor characterized by low molecular weight protein properties. The SDS-PAGE chromatogram indicates that the active ingredient had a molecular weight of 14 kDa, with α-glucosidase inhibitory activity of 80.2% and a recovery efficiency of 0.02%. This study provides the scientific foundation for ongoing research into optimal conditions for the extraction and purification of the α-glucosidase inhibitor derived from the A. niger strain, a metabolically active fungus found in Vietnam.
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References
American Diabetes Association (2015). Standards of medical care in diabetes. Diabetes Care 2015,38 (Suppl. 1), S1–S2. https://doi.org/10.2337/dc15-S003
American Diabetes Association (2020) Classification and diagnosis of diabetes: Standards of medical care in diabetes-2020. Diabetes Care, 44(Supplement_1), S14-S31. https://doi.org/10.2337/dc20-S002
Bassam B. J., Caetano-Anolles G., and Gresshoff P. M. (1991). Fast and sensitive silver staining of DNA in polyacrylamide gels. Analytical Biochemistry, 196(1), 80-83. https://doi.org/10.1016/0003-2697(91)90120-I
Bradford M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248-254. https://doi.org/10.1016/0003-2697(76)90527-3
Carrascosa M., Pascual F., Aresti S. and (1997). Acarbose-induced acute severe hepatotoxicity. Lancet 349(9053), 698-699. https://doi.org/10.1016/S0140-6736(05)60134-1
Coniff R. F., Shapiro J. A., Seaton T. B., and Bray G. A. (1995). Multicenter, placebo-controlled trial comparing acarbose (BAY g 5421) with placebo, tolbutamide, and tolbutamide-plus-acarbose in non-insulin-dependent diabetes mellitus. American Journal of Medicine, 98(5), 443-451. https://doi.org/10.1016/S0002-9343(99)80 343-X
del Moral S., Barradas‑Dermitz D. M., and Aguilar‑Uscanga M. G. (2018). Production and biochemical characterization of α‑glucosidase from Aspergillus niger ITV‑01 isolated from sugar cane bagasse. 3 Biotech, 8(1), 7. https://doi.org/10.1007/s13205-017-1029-6
Dewi R. T., Tachibana S., Fajriah S., and Hanaf M. (2015). α-Glucosidase inhibitor compounds from Aspergillus terreus RCC1 and their antioxidant activity. Medicinal Chemistry Research, 24, 737–743. https://doi.org/10.1007/S00044-014-1164-0
Dewi R. T, Suparman A, Mulyani H, and Lotulung P. D. N (2016). Identification of a new compound as α-glucosidase inhibitor from Aspergillus aculeatus. Annales bogorienses, 20(1), 18-22
Duong-Ly K. C., and Gabelli S. B. (2014). Gel filtration chromatography (size exclusion chro matography) of proteins. Methods in Enzy mology, 541, 105-114. https://doi.org/10.1016/B978-0-12-420119-4.00009-4
Holt R. I. G., Peveler R. C., and Byrne C. D. (2004). Schizophrenia, the metabolic syndrome and diabetes. Diabetic Medicine, 21(6), 515-523. https://doi.org/10.1111/j.1464-5491.2004.01199.x
International Diabetes Federation (2021). IDF Diabetes Atlas. ISBN, 978-2-930229-98-0
Johnston P. S., Lebovitz H. E., Coniff R. F., Simonson D. C., Raskin P., and Munera C. L. (1998). Advantages of α-glucosidase inhibition as monotherapy in elderly type 2 diabetic patients. Journal of Clinical Endocrinology & Metabolism, 83(5), 1515-1522. https://doi.org/10.1 210/jcem.83.5.4824
Kang M. G., Yi S. H., and Lee J. S. (2013). Production and characterization of a new α-glucosidase inhibitory peptide from Aspergillus oryzae N159-1. Mycobiology, 41(3), 149–154. https://doi.org/10.5941/MYCO.2013.41.3.149
Kim Y., Wang M., and Rhee H. (2004). A novel α-glucosidase inhibitor from pine bark. Carbohydrate Research, 339(3), 715-717. https://doi.org/10.1016/j.carres.2003.11.005
Liu B., Chen N., Chen Y. X., and Shen J. J. (2021). A new benzophenone with biological activities purified from Aspergillus fumigatus SWZ01. Natural Product Research, 35(24), 5710-5719 https://doi.org/10.1080/14786419.2020.1825427
Nguyen T. C., Le T. H., Mai V. H., Hoang T. Y., Nguyen T. T., Dao T. M. A., et al. (2021a). Optimization and purification of α-glucosidase inhibitor from Bacillus subtilis YT20. Vietnam Journal of Science and Technology, 59(2), 179-188. https://doi.org/10.15625/2525-2518/59/2/14928
Nguyen T. H.T ., Doris Y .Y. T., Kit Wayne C., Nguyen T. L., Le T. H., Nguyen T. C., et al. (2021b). Discovery of α glucosidase inhibitors from marine microorganisms: Optimization of culture conditions and medium composition. Molecular Biotechnology, 63(11), 1004-1015. https://doi.org/10.1007/s12033-021-00362-3
Nguyen T. T., Nguyen T. T., Phan T. H. T., Tran T. T., Pham D. N., Le T. H., et al. (2022). Isolation, selection and evaluation of α-glucosidase inhibitory activity from endophytic streptomyces sp. isolated from Citrus Myrtifolia cultivar in Hoa Binh Vietnam. Vietnam Journal of Biotechnology, 20(4), 693-704. https://doi.org/10.15625/1811-4989/17172
Sambrook J., and Russell D. W. (2006). SDS-polyacrylamide gel electrophoresis of proteins. 2006(4):pdb.prot4540. https://doi.org/10.1101/pdb.prot4540
Santiago M., and Strobel S. (2013). Thin layer chromatography. Methods in enzymology, 533, 303-324. https://doi.org/10.1016/b978-0-12-420067-8.00024-6
Singh B., and Kaur A. (2016). Antidiabetic potential of a peptide isolated from an endophytic Aspergillus awamori. Journal of Applied Microbiology, 120(2), 301-311. https://doi.org/10.1111/jam.12998
Sun C. X., Liu X. Y., Sun N., Zhang X., Shah M., Zhang G., et al. (2022). Cytotoxic nitrobenzoyl sesquiterpenoids from an Antarctica sponge-derived Aspergillus insulicola. Journal of Natural Products, 85(4), 987-996. https://doi.org/10.1021/acs.jnatprod.1c01118
Ye G., Huang C., Li J., Chen T., Tang J., Liu W., et al. (2021). Isolation, structural characterization and antidiabetic activity of new diketopiperazine alkaloids from mangrove endophytic fungus Aspergillus sp. 16-5c. Mar Drugs, 19(7), 402. https://doi.org/10.3390/md19070402
Zhao H.Y., Anbuchezhian R., Sun W., Shao C.L., Zhang F.L., Yin Y., et al. (2016). Cytotoxic nitrobenzoyloxy-substituted sesquiter -penes from sponge-derived endozoic fungus Aspergillus insulicola MD10-2. Current Pharmaceutical Biotechnology, 17(3), 271-274. https://doi.org/10.2174/1389201017666151223123424
Zhao W., Zeng Y., Chang W., Chen H., Wang H., Dai H., et al. (2023). Potential α-glucosidase inhibitors from the deep-sea sediment-derived fungus Aspergillus insulicola. Marine Drugs, 21(3), 157. https://doi.org/10.3390/md21030157
