Purification and biofabrication of 5-aminolevulinic acid for photodynamic therapy against pathogens and cancer cells

Abstract

5-Aminolevulinic acid (5-ALA) is a non-proteinogenic amino acid which has involved in heme metabolism of organisms, and has been widely applied in agriculture, and medical fields nowadays. 5-ALA is used in the elimination of pathogens or cancer cells by photodynamic therapy (PDT) owing to the photosensitizer reaction which releases the reactive oxygen species (ROS). Currently, biofabrication of 5-ALA is regarded as the most efficient and eco-friendly approach, but the complicated ingredient of medium causes the nuisance process of purification, resulting in low recovery and high producing cost. In this study, hydrogen chloride, sodium acetate, and ammonia were examined to maximize the recovery of 5-ALA from ion-exchange chromatography (IEC), thus a 92% recovery in 1 M ammonia at pH 9.5 was obtained. Afterward, the activated carbon was used for decolorization to further remove the pigments from the eluent. Four organic solvents, i.e., diethyl ether, methanol, ethanol, and acetone were compared to extract and form 5-ALA precipitation. The purified 5-ALA was verified to eliminate 74% of A549 human lung cancer and 83% of A375 melanoma skin cancer cell. Moreover, Proteus hauseri, Aeromonas hydrophila, Bacillus cereus, and Staphylococcus aureus were killed via anti-microbial PDT with 1% 5-ALA and reached 100% killing rate at optimal condition. With the addition of 0.05% 5-ALA during the culture, the growth of microalgae Chlorella sorokiniana was improved to against a common aquatic pathogen, A. hydrophila. The broad application of 5-ALA was demonstrated in this study for the first time.

Keywords

5-Aminolevulinic acid / Ion exchange chromatography / Photodynamic therapy / Antimicrobial / Cancer cell / Microalgae

References

[1] Armbruster CE, Mobley HL. Merging mythology and morphology: the multifaceted lifestyle of Proteus mirabilis. Nat Rev Microbiol, 2012, 10: 743-754.

[2] Bunke A, Zerbe O, Schmid H, Burmeister G, Merkle HP, Gander B. Degradation mechanism and stability of 5-aminolevulinic acid. J Pharm Sci, 2000, 89: 1335-1341.

[3] Cai J, Zheng Q, Huang H, Li B. 5-aminolevulinic acid mediated photodynamic therapy inhibits survival activity and promotes apoptosis of A375 and A431 cells. Photodiagnosis Photodyn Ther, 2018, 21: 257-262.

[4] Chen H, Jiang JG. Toxic effects of chemical pesticides (trichlorfon and dimehypo) on Dunaliella salina. Chemosphere, 2011, 84: 664-670.

[5] Di Venosa G, Fukuda H, Perotti C, Batlle A, Casas A. A method for separating ALA from ALA derivatives using ionic exchange extraction. J Photochem Photobiol B, 2004, 75: 7-163.

[6] Din NAS, Lim SJ, Maskat MY, Abd Mutalib S, Zaini NAM. Lactic acid separation and recovery from fermentation broth by ion-exchange resin: a review. Bioresour Bioprocess, 2021, 8: 1-23.

[7] Hotta Y, Tanaka T, Takaoka H, Takeuchi Y, Konnai M. Promotive effects of 5-aminolevulinic acid on the yield of several crops. Plant Growth Regul, 1997, 22: 109-114.

[8] Inoue K. 5-Aminolevulinic acid-mediated photodynamic therapy for bladder cancer. Int J Urol, 2017, 24: 97-101.

[9] Jiao K, Chang J, Zeng X, Ng IS, Xiao Z, Sun Y, Lin L. 5-Aminolevulinic acid promotes arachidonic acid biosynthesis in the red microalga Porphyridium purpureum. Biotechnol Biofuels, 2017, 10: 1-10.

[10] Juzeniene A, Juzenas P, Iani V, Moan J. Topical application of 5-aminolevulinic acid and its methylester, hexylester and octylester derivatives: considerations for dosimetry in mouse skin model¶. Photochem Photobiol, 2002, 76: 329-334.

[11] Kennedy J, Pottier RH, Pross DC. Photodynamic therapy with endogenous protoporphyrin: IX: basic principles and present clinical experience. J Photochem Photobiol B, 1990, 6: 143-148.

[12] Klausen M, Ucuncu M, Bradley M. Design of photosensitizing agents for targeted antimicrobial photodynamic therapy. Molecules, 2020, 25: 5239.

[13] Kussovski V, Mantareva V, Angelov I, Orozova P, Wöhrle D, Schnurpfeil G, Avramov L. Photodynamic inactivation of Aeromonas hydrophila by cationic phthalocyanines with different hydrophobicity. FEMS Microbiol Lett, 2009, 294: 133-140.

[14] Lin JY, Xue C, Tan SI, Ng IS. Pyridoxal kinase PdxY mediated carbon dioxide assimilation to enhance the biomass in Chlamydomonas reinhardtii CC-400. Bioresour Technol, 2021, 322.

[15] Lyu X, Lyu Y, Yu H, Chen W, Ye L, Yang R. Biotechnological advances for improving natural pigment production: a state-of-the-art review. Bioresour Bioprocess, 2022, 9: 1-38.

[16] Matsumura Y, Takeshima YI, Okita H. A convenient method for introducing oxo group into the β-position of cyclic amines and its application to synthesis of δ-aminolevulinic acid. Bull Chem Soc Jpn, 1994, 67: 304-306.

[17] Miyachi N, Tanaka T, Nishikawa S, Takeya H, Hotta Y. Preparation and chemical properties of 5-aminolevulinic acid and its derivatives. Porphyrins, 1998, 7: 342-347.

[18] Moreira MJA, Gando-Ferreira LM. Separation of phenylalanine and tyrosine by ion-exchange using a strong-base anionic resin. II Cyclic Adsorption/desorption Studies. Biochem Eng J, 2012, 67: 241-250.

[19] Okada H, Tanaka T, Nomura T (2016) Method for producing 5-aminolevulinic acid hydrochloride. EP Patent 1,927,586, 14 Apr 2016.

[20] Patrickios CS, Yamasaki EN. Polypeptide amino acid composition and isoelectric point ii. comparison between experiment and theory. Anal Biochem, 1995, 231: 82-91.

[21] Pérez-Laguna V, García-Luque I, Ballesta S, Pérez-Artiaga L, Lampaya-Pérez V, Samper S, Gilaberte Y. Antimicrobial photodynamic activity of Rose Bengal, alone or in combination with Gentamicin, against planktonic and biofilm Staphylococcus aureus. Photodiagnosis Photodyn Ther, 2018, 21: 211-216.

[22] do Prado-Silva L, Alvarenga VO, Braga GÚ, Sant’Ana AS. Inactivation kinetics of Bacillus cereus vegetative cells and spores from different sources by antimicrobial photodynamic treatment (aPDT). LWT-Food Sci Technol, 2021, 142: 111037.

[23] Sarmah P, Dan MM, Adapa D, Sarangi TK. A review on common pathogenic microorganisms and their impact on human health. Electron J Biol, 2018, 14: 50-58.

[24] Sasaki K, Watanabe M, Tanaka T. Biosynthesis, biotechnological production and applications of 5-aminolevulinic acid. Appl Microbiol Biotechnol, 2002, 58: 23-29.

[25] Shukla SK, Sharma AK, Gupta V, Kalonia A, Shaw P. Challenges with wound infection models in drug development. Curr Drug Targets, 2020, 21: 1301-1312.

[26] Simões D, Miguel SP, Ribeiro MP, Coutinho P, Mendonça AG, Correia IJ. Recent advances on antimicrobial wound dressing: A review. Eur J Pharm Biopharm, 2018, 127: 130-141.

[27] Tachiya N (2016) Novel crystal of 5-aminolevulinic acid phosphate and process for production thereof. EP Patent 2,053,039, 16 Mar 2016.

[28] Teijo MJ, Diez BA, Battle A, Fukuda H. Modulation of 5-Aminolevulinic acid mediated photodynamic therapy induced cell death in a human lung adenocarcinoma cell line. Integr Cancer Sci Ther, 2016, 3: 450-459.

[29] Tripetch P, Srzednicki G, Borompichaichartkul C. Separation process of 5-aminolevulinic acid from Rhodobacter spaeroides for increasing value of agricultural product by ion exchange chromatography. Acta Hort, 2013, 1011: 265-271.

[30] Wachowska M, Muchowicz A, Firczuk M, Gabrysiak M, Winiarska M, Wańczyk M, Golab J. Aminolevulinic acid (ALA) as a prodrug in photodynamic therapy of cancer. Molecules, 2011, 16: 4140-4164.

[31] Wang H, Zhang W, Chen L, Wang J, Liu T. The contamination and control of biological pollutants in mass cultivation of microalgae. Bioresour Technol, 2013, 128: 745-750.

[32] Wang J, Zhang J, Li J, Dawuda MM, Ali B, Wu Y, Xie J. Exogenous application of 5-aminolevulinic acid promotes coloration and improves the quality of tomato fruit by regulating carotenoid metabolism. Front Plant Sci, 2021, 12: 683868.

[33] Xue C, Hsu KM, Ting WW, Huang SF, Lin HY, Li SF, Ng IS. Efficient biotransformation of L-lysine into cadaverine by strengthening pyridoxal 5’-phosphate-dependent proteins in Escherichia coli with cold shock treatment. Biochem Eng J, 2020, 161: 107659.

[34] Yi YC, Shih IT, Yu TH, Lee YJ, Ng IS. Challenges and opportunities of bioprocessing 5-aminolevulinic acid using genetic and metabolic engineering: a critical review. Bioresour Bioprocess, 2021, 8: 1-18.

[35] Yi YC, Xue C, Ng IS. Low-carbon-footprint production of high-end 5-aminolevulinic acid via integrative strain engineering and rubisco-equipped Escherichia coli. ACS Sustain Chem Eng, 2021, 9: 15623-15633.

[36] Yu TH, Tan SI, Yi YC, Xue C, Ting WW, Chang JJ, Ng IS. New insight into the codon usage and medium optimization toward stable and high-level 5-aminolevulinic acid production in Escherichia coli. Biochem Eng J, 2022, 177: 108259.

[37] Zhang ZJ, Li HZ, Zhou WJ, Takeuchi Y, Yoneyama K. Effect of 5-aminolevulinic acid on development and salt tolerance of potato (Solanum tuberosum L.) microtubers in vitro. Plant Growth Regul, 2006, 49: 27-34.

[38] Zhu Z, Jiang J, Fa Y. Overcoming the biological contamination in microalgae and cyanobacteria mass cultivations for photosynthetic biofuel production. Molecules, 2020, 25: 5220.

Funding

Ministry of Science and Technology, Taiwan(MOST 108-2221-E-006-004-MY3)