Deteksi Gen DXS dan Penentuan Jalur Biosintesis Karotenoid pada Chlorella pyrenoidosa

*Ramadhebi Monalita  -  Departemen Biologi Fakultas Sains dan Matematika, Universitas Diponegoro, Indonesia
Hermin Kusumaningrum  -  Departemen Biologi Fakultas Sains dan Matematika, Universitas Diponegoro, Indonesia
Anto Budiharjo  -  Departemen Biologi Fakultas Sains dan Matematika, Universitas Diponegoro, Indonesia

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Abstract

Sintesis karotenoid alami belum pernah melebihi produk sintetik pada skala komersial. Kurangnya pemahaman mengenai aspek mikrobiologis dan ekofisiologis isolat penghasil karotenoid menyebabkan terjadinya kesalahan penamaan spesies. Satu isolat lokal alga hijau yang digunakan sebagai pakan alami sumber karotenoid dalam industri makanan yaitu Chlorella pyrenoidosa. Akumulasi karotenoid jalur non- MVA pada alga hijau ditentukan oleh enzim D-1-deoksixilulosa 5-fosfat sintase, yang disandi oleh gen D- 1-deoksixilulosa 5-fosfat sintase (DXS). Tujuan utama penelitian ini adalah untuk mendeteksi gen DXS sebagai penyandi enzim kunci biosintesis karotenoid pada C. pyrenoidosa apakah juga mengikuti jalur non- MVA untuk biosintesis karotenoidnya atau tidak. Metode karakterisasi morfologis dan ekofisiologis dilakukan berdasarkan pengamatan secara berkala dan deteksi gen DXS menggunakan panduan Kuzuyama (2000). Hasil analisis keserupaan gen DXS C. pyrenoidosa pada daerah lestari memperlihatkan dapat mendeteksi parsial gen DXS dari Chlamydomonas reinhardtii. Tidak adanya hambatan pertumbuhan pada C. pyrenoidosa dengan lovastatin menunjukkan jalur non-MVA adalah jalur yang digunakan dalam biosintesis karotenoidnya.

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  1. Amaya, D. B. R. 2016. Natural Food Pigments And Colorants. Current Opinion in Food Science 5(7): 20 – 26
  2. Bacon, K. 2001. Photosynthesis: Photobiochemistry and Photobiophysics. Kluwer Academics Publisher, Netherland
  3. Bai, C., T. Capell, J. Berman, V. Medina, G. Sandmann, P. Christou and Zhu. 2015. Bottlenecks In Carotenoid Biosynthesis And Accumulation In Rice Endosperm Are Influenced By The Precursor–Product Balance. Plant Biotechnology Journal 14(1): 195-205
  4. Beltra´ n, J., B. Kloss, J.P. Hosler, J. Geng, A. Liu, A Modi, J.H. Dawson, M. Sono, M. Shumskaya and C. Ampomah-Dwamena. 2015. Control Of Carotenoid Biosynthesis Through A Heme- Based Cis-Trans Isomerase. Nature Chemical Biology 11(8): 598–605
  5. Bishop, W.M. and H.M. Zubeck. 2012. Evaluation Of Microalgae For Use As Nutraceuticals And Nutritional Supplements. Journal of Nutrion and Food Science 2(5): doi: 10.4172/2155-9600.1000147
  6. Brown, M. S and J.G. Hay. 2009. Essentials of Medical Genomics. New Jersey, USA
  7. Carretero-Paulet, L., A. Cairó, D. Talavera, A. Saura,
  8. S. Imperial, M. Rodríguez-Concepción, N. Campos and A. Boronat. 2013. Functional And Evolutionary Analysis Of DXL1, A NonEssential Gene Encoding A 1- Deoxy-D- Xylulose 5-Phosphate Synthase Like Protein
  9. In Arabidopsis Thaliana . Gene 524(1): 40–53. Cazzonelli, C.I. and B.J. Pogson. 2010. Source to Sink: Regulation Of Carotenoid Biosynthesis In Plants. Trends in Plant Science 15(5): 266–274
  10. Chacon-Lee T.L. and G.E. Gonzalez-Marino. 2010. Microalgae For “Healthy” Foods Possibilities And Challenges. Comprehensive Reviews in Food Science and Food Safety 9(6): 655–675
  11. Cunningham, F.X., H. Lee and E. Gantt. 2007. Carotenod Biosynthesis In The Primitive Red Alga Cyanidioschyzon Merolae. Eukaryotic Cell 6(3): 533-545
  12. del Campo, A.J., M. Garcia-Gonzảles, and M.G Guerrero. 2007. Outdoor Cultivation Of Microalgae For Carotenoid Production: Current State And Perspectives. Applied Microbiolgy and Biotechnology 74(6): 1163-1174
  13. Eisenreich, W., A. Bacher, D. Arigoni and F. Rohdich. 2004. Biosynthesis Of Isoprenoids Via The NonMevalonate Pathway. Cellular and Molecular Life Science 61(12): 1401–1426
  14. Estevez, J. M., A. Cantero, A. Reindl, S. Reichler and P. Leon. 2001. 1-Deoxy-D-Xylulose-5-Phosphate Synthase, A Limiting Enzyme For Plastidic Isoprenoid Biosynthesis In Plants. The Journal of Biology Chemistry 276(25): 22901–22909
  15. Gill, S. R., M. Pop, R. T. Deboy, P.B Eckburg, P.J. Turnbaugh, B.S. Samuel, J.I. Gordon, D.A. Relman, C.M. Fraser-Liggett and K.E. Nelson. 2006. Metagenomic Analysis Of The Human Distal Gut Microbiome. Science 312(5778): 1355–1359
  16. Gonzalez-Jorge, S., S.H. Ha, M. Magallanes- Lundback, L.U. Gilliland, A. Zhou, A.E. Lipka, Y.N. Nguyen, R. Angelovici, H. Lin and J. Cepela
  17. Carotenoid Cleavage Dioxygenase4 is A Negative Regulator Of Β-Carotene Content In Arabidopsis Seeds. The Plant Cell 25(12):4812– 4826
  18. Hendry, G.A.F. and Grime, J.P. 1993. Methods on Comparative Plant Ecology, A Laboratory Manual. London: Chapman and Hill
  19. Iwamoto, H. 2004. Industrial Production Of Microalgal Cell-mass and Secondary Products-Major Industrial Species: Chlorella. In : A. Richmond (Ed.). Handbook of Microalgal Culture. Biotechnology and Applied Phycology. Blackwell Publishing
  20. Jahns, P. and A.R. Holzwarth. 2012. The Role Of The Xanthophyll Cycle And Of Lutein In Photoprotection Of Photosystem II. Biochimica et Biophysica Acta-Bioenergetics 1817(1): 182–193
  21. Jones, OG. 2016. Recent Advances In The Functionality Of Non-Animal-Sourced Proteins Contributing To Their Use In Meat Analogs. Current Opinion in Food Science 9(7): 7–13
  22. Kiokias, S., C. Proestos and Varzakas. 2016. A Review of the Structure, Biosynthesis, Absorption of Carotenoids-Analysis and Properties of their Common Natural Extracts. Current Research in Nutrition and Food Science 2(4): 25-37
  23. Kong, W., S. Hua, H. Cao, Y. Mu, H. Yang, H. Song and C. Xia. 2012. Optimization Of Mixotrophic Medium Components For Biomass Production And Biochemical Composition Biosynthesis By Chlorella Vulgaris Using Response Surface Methodology. Journal of the Taiwan Institute of Chemical Engineers. 43(3):360-367
  24. Krinsky N.I. and E.J. Johnson. 2005. Carotenoid Actions And Their Relation To Health And Disease. Molecular Aspects of Medicine. 26(6):459–516
  25. Kusumaningrum, H.P., 2003. Karakterisasi Alga Hijau Dunaliella sp. dan Isolat Sianobakteria serta Deteksi Gen DXS Penyandi Enzim Kunci Biosintesis Karotenoid. Disertasi. Yogyakarta, Universitas Gadjah Mada
  26. Kusumaningrum, H.P. and M. Zainuri. 2013. Aplikasi Pakan Alami Kaya Karotenoid Untuk Post Larvae Penaeus Monodon Fab. Ilmu Kelautan: Indonesian Journal of Marine Sciences. 3(18): 143 – 149
  27. Kusmiati, Agustini, S.R. Tamat and M. Irawati. 2010. Ekstraksi Danpurifikasi Senyawa Lutein Dari Mikroalga Chlorella Pyrenoidosa Galur Lokal Ink. Jurnal Kimia Indonesia 1(5): 30-34
  28. Li, L. and H. Yuan. 2013. Chromoplast Biogenesis And Carotenoid Accumulation. Archives of Biochemistry and Biophysics 539(2): 102–109
  29. Lichtenthaler, H.K. 2000. Non-mevalonate Isoprenoid Biosynthesis: Enzymes, Genes and Inhibitors. Biochemical Society Transactions 28(6):785-9
  30. Liu, G., H. Gerken, J. Huang and F. Chen. 2013. Engineering Of An Endogenous Phytoene Desaturase Gene As A Dominant Selectable Marker For Chlamydomonas reinhardtii Transformation And Enhanced Biosynthesis Of Carotenoids. Process Biochemistry 48(5-6): 788795
  31. Liu, J., Y. Xu, L. Liang and J. Wei. 2015. Molecular Cloning, Characterization And Expression Analysis Of The Gene Encoding 1-Deoxy- DXylulose 5-Phosphate Reductoisomerase From Aquilaria Sinensis (Lour.). Gilg. Journal Of Genetics 94(2): 239–249
  32. Lohr, M., C. Im and A.R. Grossman. 2005. Genomebased Examination of Chlorophyll and Carotenoid Biosynthesis in Chlamydomonas reinhardtii. Plant physiology 138(1):490-515
  33. Marchal, L., M. Mojaat-Guemir, A. Foucalt And J. Pruvast. 2013. Centrifugal Partition Extraction Of B-Carotene From Dunaliella Salina For Efficient And Biocompatible Recovery Of Metabolites. Bioresource Technology 1(134): 396-400
  34. Mikami, K and M. Hosokawa. 2013. Biosynthetic Pathway and Health Benefits of Fucoxanthin, an Algae-Specific Xanthophyll in Brown Seaweeds. Internationa Journal of Molecular Science 14(7): 13763-13781
  35. Merchant, R. and C.A. Andre. 2000. A Review Of Recent Clinical Trials Of The Nutritional Supplement Chlorella Pyrenoidosa In The Treatment Of Fibromyalgia, Hypertension, And Ulcerative Colitis. Alternative therapies in health and medicine 7(3): 79-91
  36. Ndiha, B. and L. Limantara. 2009. Karotenoid pada Bahan Makanan. Prosiding Seminar Nasional Biologi, Lingkungan dan Pembelajarannya. Jurdik Biologi. FMIPA Universitas Negeri Yogyakarta. P. 75-84
  37. Neilson, J.A.D. and D.G. Durnford. 2010. Structural And Functional Diversification Of The LightHarvesting Complexes In Photosynthetic Eukaryotes. Photosynthesis Research 106(1-2): 57–71
  38. Neuman, H., N. Galpaz, F.X. Cunningham, D. Zamir and J. Hirschberg. 2014. The Tomato Mutation Nxd1 Reveals A Gene Necessary For Neoxanthin Biosynthesis And Demonstrates That Violaxanthin Is A Sufficient Precursor For Abscisic Acid Biosynthesis. The Plant Journal 78(1): 80–93
  39. Nisar N., L. Li, S. Lu, N.C. Khin and B.J. Pogson. 2015. Carotenoid metabolism in plants. Molecular Plant 8(1):68–82
  40. Nugraheni, A. S., M. Majid, K. Lia, W. Yustin and O.K. Radjasa. 2010. Characterization of Carotenoid Pigmen from Bacterial Symbionts of Seagrass Thallasia hemprichii. J. Coast. Dev 14 (1): 51-60
  41. Pankratov, I., R. McQuinn, J. Schwartz, E. Bar, Z. Fei, E. Lewinsohn, D. Zamir, J.J. Giovannoni and J. Hirschberg. 2016. Fruit Carotenoid-Deficient Mutants In Tomato Reveal A Function Of The Plastidial Isopentenyl Diphosphate Isomerase (IDI1) In Carotenoid Biosynthesis. The Plant Journal 88(1): 82–94
  42. Pattanaik, B. and P. Lindberg. 2015. Terpenoids And Their Biosynthesis In Cyanobacteria. Life 5 (1): 269–293
  43. Prabowo, D. A. 2009. Optimasi Penembangan Media Untuk Pertumbuhan Chlorella sp. Pada Skala Laboratorium. Skripsi. Bogor, Institut Pertanian Bogor
  44. Prayoga, W. dan A.K. Wardani. 2015. Polymerase Chain Reaction untuk Deteksi Salmonella sp. Kajian Pustaka. Jurnal Pangan dan Agroindustri 3(2):483- 488
  45. Qian, C., E.A. Decker, H. Xiao, D.J. McClements
  46. Physical And Chemical Stability Of ΒCarotene- Enriched Nanoemulsions: Influence Of Ph, Ionic Strength, Temperature, And Emulsifier Type. Journal of Food Chemistry 132(3): 12211229
  47. Rodrigo-Baños M., I. Garbayo, C. Vílchez, M.J. Bonete and R.M. Martínez-Espinosa. 2015. Carotenoids From Haloarchaea And Their Potential In Bioatechnology. Marine Drugs 13(9): 5508–5532
  48. Rodriguez-Concepcion, M. and A. Boronat. 2002. Elucidation Of The Methylerythritol Phosphate Pathway For Isoprenoid Biosynthesis In Bacteria And Plastids. A Metabolic Milestone Achieved Through Genomic. Plant Physiology 130(3): 1079–1089
  49. Ruiz-Sola, M.A., D. Coman, G. Beck, M.V. Barja, M. Colinas, A. Graf, R. Welsch, P. Rutimann, P. Buhlmann and L. Bigler. 2016. Arabidopsis Geranylgeranyl Diphosphate Synthase 11 Is A Hub Isozyme Required For The Production Of Most Photosynthesis-Related Isoprenoids. New Phytologist 209(1): 252–264
  50. Safi, C, D.Z. Liu, B.H.J. Yap, G.J.O. Martin, C. VacaGarcia and P.Y. Pontalier. 2014. A Two-Stage Ultrafiltration Process For Separating Multiple Components Of Tetraselmis Suecica After Cell Disruption
  51. Journal of Applied Phycology 26(6): 2379–2387. Sambrook, J dan Russel D. W. 2001. Molecular Cloning: A Laboratory Manual 3rd edition. Cold Spring Harbor, New York
  52. Sari, L. 2011. Rekayasa Proses Pembuatan Kurva Standart dari Larutan Betakoroten. Banjar baru, Teknologi Industri Pertanian
  53. Schubert, N., E. García-Mendoza and I. Pacheco- Ruiz. 2006. Carotenoid Composition Of Marine Red Algae. Journal of Phycology 42(6): 1208– 1216
  54. Shumskaya, Μ. and E.T. Wurtzel. 2013. The Carotenoid biosynthetic pathway: Thinking in all dimensions. Plant Science 59(208): 58-63
  55. Simamora, L.A., Sudarno and T. Istirokhatun. 2017. Kultivasi mikroalga sebagai metode pengolahan dalam menyisihkan kadar COD dan amonium pada limbah cair tahu. Jurnal Teknik Lingkungan 6(1): 1-14
  56. Stafsnes, M. H., D.J. Kjell, K.A. Geir, V. Svein, E.E. Trond and B. Per. 2010. Isolation and Characterization of Marine Pigmented Bacteria from Norwegian Coastal Waters and Screening for Carotenoidswith UVA-Blue Light Absorbing Properties. Journal Microbiology 48(1): 16-23
  57. Takekoshi, H., G. Suzuki and H. Chubachi. 2005. Effect Of Chlorella Pyrenoidosa On Fecal Excretion And Liver Accumulation Of Polychlorinated Dibenzo-P-Dioxin In Mice. Chemosphere 59(2): 297–304
  58. Tong, Y., P. Su, Y. Zhao, M. Zhang, X. Wang, Y. Liu, X. Zhang, W. Gao and L. Huang. 2015. Molecular Cloning And Characterization of DXS and DXR Genes In The Terpenoid Biosynthetic Pathway of Tripterygium Wilfordii. International Journal of Molecular Science. 16(10):25516–25535
  59. Venkataraman, G .S . 1969. The Cultivation of Algae. Indian Council of Agriciltural Research. New Delhi
  60. Viegas C.V., I. Hachemi, P. Maki-Arvela, A. Smeds, A. Aho, D.Y. Murzin. 2015. Algal Products Beyond Lipids: Comprehensive Characterization Of Different Products In Direct Saponification Of Green Alga Chlorella sp. Algal Research 2(11): 156-164
  61. Vilchez, C., E. Forjan, M. Cuaresma, F. Bedmar, I. Garbayo and J.M. Vega. 2011. Marine Carotenoids: Biological Function and Commercial Applications. Marine Drugs 9(3): 319-333
  62. Waghmare, A. and K. Salve. 2016. Concentration And Characterization Of Microalgae Proteins from Chlorella pyrenoidosa. Bioresources and Bioprocessing 3(1) : 2-9
  63. Wijihastuti. 2011. Optimasi lingkungan Tumbuh Mikroalga dari Kawah Ratu Sukabumi yang Berpotensi sebagai Sumber biodiesel. Bogor, ITB. Xiang, S., G. Usunow, G. Lange, M. Busch and L. Tong. 2007. Crystal Structure of 1-Deoxy-Dxylulose 5- Phosphate Synthase, a Crucial Enzyme for Isoprenoids Biosynthesis. Journal of Biology Chemistry 282(4): 2670-2681
  64. Yoon, S.H., A. Das, H.K. Ryu, H.J. Jang, J.Y. Kim, D.K. Oh, J.D. Keasling and S.W. Kim. 2009. Combinatorial Expression Of Bacterial Whole Mevalonate Pathway For The Production Of Beta-Carotene In E. Coli . J. Biotechnol 140(3- 4): 218-222

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