Bioinformatics study of phytase from Aspergillus niger for use as feed additive in livestock feed

Hamdan Maulana1, Yantyati Widyastuti2, Nina Herlina2, Abun Hasbuna1, Aas Syiarudin Hasbi Al-Islahi3, Lita Triratna2, Novi Mayasari1
1Faculty of Animal Husbandry, Department of Nutrition and Feed Technology, Universitas Padjadjaran, Jatinangor, Sumedang, Indonesia
2National Research and Innovation Agency (BRIN), Research Center for Applied Microbiology, Cibinong, Bogor, Indonesia
3PT. Berdikari Persero, Jakarta Pusat, Indonesia

Tóm tắt

Phytase supplementation in rations can reduce their phytic acid composition in order to enhance their nutritional value. Aspergillus niger is a fungus that can encode phytase. This study aims to determine the characteristics of its DNA sequences and amino acid composition that encode the phytase enzyme, as well as to determine the primer designs. This study used gene sequence data and protein-encoding phytase from Aspergillus niger that was collected manually from NCBI and PDB. The data was analyzed using SPDBV and then be aligned using the ClustalW Multiple Alignment features. The phylogenetic tree was built by Mega11 software. Primers were designed from selected candidate sequences that were analyzed. The designed primers were then simulated for PCR using FastPCR and SnapGene software. There are 18 Aspergillus niger phytases in NCBI which is 14.87% of the total Aspergillus. There are 14 Aspergillus niger phytases that have identity above 95%. Aspergillus niger 110. M94550.1 is the closest strain to the PDB template. Candidate sources of phytase genes are Aspergillus niger 110.M94550.1, 48.2.BCMY01000003.1, and 92.JQ654450.1. The primer design has 2 possibilities of self-annealing and high melting temperature on the reverse primer. PCR simulation shows that the primer design can attach completely but still has the possibility of mispriming. This study suggests promising results for the future development of phytase enzyme production from Aspergillus niger as a feed additive using genetic engineering to enhance the quality of livestock feed in Indonesia.

Tài liệu tham khảo

Fatima I, Safdar N, Akhtar W, Munir A, Saqib S, Ayaz A, Bahadur S, Alrefaei AF, Ullah F, Zaman, (2023) Evaluation of potential inhibitory effects on acetylcholinesterase, pancreatic lipase, and cancer cell lines using raw leaves extracts of three fabaceae species. Heliyon 9:2–7. https://doi.org/10.1016/j.heliyon.2023.e15909 Yuan HB, Tang RC, Yu CB (2021) Flame Retardant functionalization of microcrystalline cellulose by phosphorylation reaction with phytic acid. Int J Mol Sci 22(17):1–10. https://doi.org/10.3390/ijms22179631 Kumar V, Sinha AK, Kajbaf K (2019) Phytic acid and phytase enzyme. In: Johnson J, Wallace TC (eds) Whole Grains and Their Bioactives: Composition and Health, 1st edn. John Wiley & Sons Ltd., Hoboken, pp 467–483 Elkhateeb YAM, Fadel M (2021) Bioinformatic studies, experimental validation of phytase production and optimization of fermentation condition for enhancing phytatse enzyme production by different microorganisms under solid-state fermentation. Open Microbiol J 16:1–11. https://doi.org/10.2174/18742858-v16-e2202160 Duliński R, Zdaniewicz M, Pater A, Poniewska D, Żyła K (2020) The impact of phytases on the release of bioactive inositols, the profile of inositol phosphates, and the release of selected minerals in the technology of buckwheat beer production. Biomolecules 10(2):1–10. https://doi.org/10.3390/biom10020166 Choi J, Kim SH (2017) A genome Tree of Life for the fungi kingdom. Proc Natl Acad Sci USA 114:9391–9396. https://doi.org/10.1073/pnas.1711939114 Houshyar M, Saki AA, Alikhani MY, Bedford MR, Soleimani M, Kamareheiy F (2023) Approaches to determine the efficiency of novel 3-phytase from Klebsiella pneumoniae and commercial phytase in broilers from 1 to 14 d of age. Poult Sci 102(11):1–10. https://doi.org/10.1016/j.psj.2023.103014 Mkumbe BS, Sajidan PA, Susilowati A, Greiner R (2022) Production, characterization, and molecular phylogenetic analysis of phytase from Aspergillus niger isolates of an indonesia origin. Hayati J Biosci 29(4):468–477. https://doi.org/10.4308/hjb.29.4.468-478 Corrêa TLR, de Araújo EF (2020) Fungal phytases: from genes to applications. Braz J Microbiol 51:1009–1020. https://doi.org/10.1007/s42770-020-00289-y Cangussu ASR, Almeida DA, Aguiar RWS, Barbosa LCB, Cangussu EWS, Brandi IV, Portella ACF, Santos GF, Sobrinho EM, Lima WJN (2017) Characterization of the Catalytic Structure of Plant Phytase, Protein Tyrosine Phosphatase-Like Phytase, and Histidine Acid Phytases and Their Biotechnological Applications. J mycol 2018:1–8. https://doi.org/10.1155/2018/8240698 Oakley AJ (2010) The structure of Aspergillus niger phytase phyA in complex with a phytate mimetic. Biochem Biophys Res Commun 397:45–749. https://doi.org/10.1016/j.bbrc.2010.06.024 Neira-Vielma AA, Aguilar CN, Ilyina A, Contreras-Esquivel JC, Carbeiro-de-Cunha M, Michelena-Alvarez G, Martinez-Hernandez JL (2018) Purification and biochemical characterization of an Aspergillus niger phytase produced by solid-state fermentation using triticale residues as substrate. Biotechnol Rep 17:49–54. https://doi.org/10.1016/j.btre.2017.12.004 Niño-Gómez DC, Rivera-Hoyos CM, Morales-Álvarez ED, Reyes-Montaño EA, Vargas-Alejo NE, Ramírez-Casallas IN, Türkmen KE, Sáenz-Suárez H, Sáenz-Moreno JA, Poutou-Piñales RA, González-Santos J, Arévalo-Galvis A (2017) (In silico) characterization of 3-phytase A and 3-phytase B from Aspergillus niger. Enzyme Res 2017:1–22. https://doi.org/10.1155/2017/9746191 Mustopa AZ, Meilina L, Irawan S, Ekawati N, Fathurahman AT, Triratna L, Kusumawati A, Prastyowati A, Nurfatwa M, Hertati A, Harmoko R (2022) Construction, expression, and in vitro assembly of virus-like particles of L1 protein of human papillomavirus type 52 in Escherichia coli BL21 DE3. J Genet Eng Biotechnol 20(19):1–14. https://doi.org/10.1186/s43141-021-00281-5 Zaman W, Ye J, Ahmad M, Saqib S, Shinwari ZK, Chen Z (2022) Phylogenetic Exploration of Traditional Chinese Medical Plants: A Case Study on Lamiaceae. Pak J Bot 54(3):1033–1040. https://doi.org/10.30848/PJB2022-3(19) Sadiqi S, Hamza M, Ali F, Alam S, Shakeela Q, Ahmed S, Ayaz A, Ali S, Saqib S, Ullah F, Zaman W (2022) Molecular Characterization of Bacterial Isolates from Soil Samples and Evaluation of their Antibacterial Potential against MDRS. Molecules 27(19):1–12. https://doi.org/10.3390/molecules27196281 Farias N, Almeida I, Meneses C (2018) New bacterial phytase through metagenomic prospection. Molecules 23(2):448. https://doi.org/10.3390/molecules23020448 Parekh P, Tang Z, Turner PC, Moyer RW, Tan W (2010) Aptamers recognizing glycosylated hemagglutinin expressed on the surface of vaccinia virus-infected cells. Anal Chem 82(20):8642–8649. https://doi.org/10.1021/ac101801j Gross E, van Iersel L, Janssen R, Jones M, Long C, Murakami Y (2021) Distinguishing level-1 phylogenetic networks on the basis of data generated by Markov processes. J Math Biol 83(32):1–24. https://doi.org/10.1007/s00285-021-01653-8 Kamel AA (2003) Bioinformatic tools and guideline for PCR primer design. Afr J Biotechnol 2(5):1–5. https://doi.org/10.5897/AJB2003.000-1019 Simkhah M, Dehghan EMJ, Zeinoddini M, Pourmahdi N (2019) Simple and rapid detection of Burkholderia and Variolla using multiplex-PCR. Res Mol Med 7(3):39–46. https://doi.org/10.32598/rmm.7.3.39 Brinkman FSL, Leipe DD (2001) Phylogenetic analysis. In: Baxevanis AD, Oulette BFF (eds) Bioinformatics a Practical Guide to The Analysis of Genes and Proteins, 2nd edn. John Wiley & Sons Ltd, Toronto, pp 323–357 Wang J, Zhang D, Xiao J, Wu X (2022) Effects of Enzymatic Modification and Cross-Linking with Sodium Phytate on the Structure and Physicochemical Properties of Cyperus esculentus Starch. Foods 11(17):1–13. https://doi.org/10.3390/foods11172583 Tavakol M, Vaughan TJ (2020) The structural role of osteocalcin in bone biomechanics and its alteration in Type-2 Diabetes. Sci Rep 10(1):1–10. https://doi.org/10.1038/s41598-020-73141-w Chen C, Cheng K, Ko T, Guo R (2015) Current progresses in phytase research: three-dimensional structure and protein engineering. CHEMBIOENG REV 2(2):76–86. https://doi.org/10.1002/cben.201400026 Kokkinidis M, Glykos NM, Fadouloglou VE (2012) Protein Flexibility and Enzymatic Catalysis. Adv Protein Chem Struct Biol 87:181–218. https://doi.org/10.1016/B978-0-12-398312-1.00007-X Armenteros JJA, Tsirigos K, Sønderby CK, Petersen TN, Winther O, Brunak S, von Heijne G, Nielsen H (2019) SignalP 5.0 improves signal peptide predictions using deep neural networks. Nat Biotechnol 37(4):420–423. https://doi.org/10.1038/s41587-019-0036-z Nelson LD, Cox MM (2017) Amino Acids, Peptides, and Proteins. In, Nelson LD and Cox MM (eds) Lehninger: Principles of Biochemistry Seventh Edition, W. H. Freeman and Company, New York, pp 197–215 Saqib S, Zaman W, Ayaz A, Habib S, Bahadur S, Hussain S, Muhammad S, Ullah F (2020) Postharvest disease inhibition in fruit by synthesis and characterization of chitosan iron oxide nanoparticles. Biocatal Agric Biotechnol 28:1–8. https://doi.org/10.1016/j.bcab.2020.101729 Yin Z, Ramshani Z, Waggoner JJ, Pinsky BA, Senapati S, Chang HC (2020) A non-optical multiplexed PCR diagnostic platform for serotype-specific detection of dengue virus. Sens Actuators B Chem 310:1–8. https://doi.org/10.1016/j.snb.2020.127854 Kumar A, Kaur J (2014) Primer based approach for PCR amplification of high GC content gene: Mycobacterium gene as a model. Mol Biol Int 2014:1–7. https://doi.org/10.1155/2014/937308 Sasmito DEK, Kurniawan R, Muhimmah I (2014) Karakteristik primer pada polymerase chain reaction (PCR) untuk sekuensing DNA: Mini review. Paper presented at Seminar Nasional Informatika Medis (SNIMed), University of Islam Indonesia, Yogyakarta, 6 December 2014.