استخراج، خالص سازی و ارزیابی خواص ضد میکروبی و آنتی اکسیدانی رنگدانه فیکواریترین سیانوباکتری خاکزی Nostoc sp. FA1
محورهای موضوعی : میکروب شناسی کاربردی
بهاره نوروزی
1
(گروه زیست شناسی ، دانشگاه آزاد اسلامی واحد علوم و تحقیقات، تهران، ایران)
امیر علی انوار
2
(گروه بهداشت مواد غذایی، دانشکده دامپزشکی، دانشگاه آزاد اسلامی واحد علوم و تحقیقات، تهران، ایران)
حامد اهری
3
(گروه علوم و صنایع غذایی واحد علوم و تحقیقات، دانشگاه آزاد اسلامی تهران.ایران)
کلید واژه: ویژگی زیست فعالی, نوستوک, فیکواریترین,
چکیده مقاله :
سابقه و هدف:رنگ های طبیعی فیکواریترین سیانوباکتری ها، دارای ویژگی های درمانی بالقوه ای هستند. امروزه جایگزینی آنتی اکسیدان های مصنوعی با آنتی اکسیدان های طبیعی بی ضرر مورد توجه می باشد. هدف ازاین تحقیق، جداسازی و خالص سازی رنگدانه ارغوانی خوراکی فیکواریترین و ارزیابی فعالیت آنتی اکسیدانی و ضد میکروبی سیانوباکتری خاکزی FA1Nostoc sp. می باشد.مواد و روش ها: استخراج و خالص سازی رنگدانه فیکواریترین در چهار مرحله آماده سازی عصاره خام، رسوب گیری با آمونیوم سولفات 65 درصد، دیالیز و کروماتوگرافی تبادل آنیونی انجام گردید. سپس فعالیت آنتی اکسیدانی با روش نیتریک اکساید و فعالیت ضد میکروبی با روش انتشار در دیسک مورد بررسی قرار گرفت.یافته ها: غلظت و خلوص رنگدانه فیکواریترین پس از هر مرحله خالص سازی تا 4 برابر (از 5/1 تا 22/6) افزایش یافت. بیشترین فعالیت ممانعت کنندگی رنگدانه فیکواریترین در مقابل باسیلوس سوبتلیس (5/10± 28/0) و کاندیدا آلبیکنس (98/10±006/0) یافت گردید. علاوه بر آن، نتایج حاصل از بررسی فعالیت آنتی اکسیدانی، افزایش معناداری را با افزایش میزان غلظت رنگدانه نشان داد.نتیجه گیری:نتایج این تحقیق می تواند زمینه ساز معرفی رنگدانه های طبیعی خوراکی سیانوباکتری های خاکزی دارای ویژگی های آنتی اکسیدانی و آنتی میکروبی با قابلیت استفاده در صنایع غذایی و دارویی باشد.
Background & Objectives:Phycoerythrin (PE( found in cyanobacteria are natural dyes which have shown potential therapeutic properties. Nowadays, substitution of artificial oxidants with harmless natural antioxidants is crucial. Hence, the present study was focused on purification process of PE from a soil strain of Nostoc sp.FA1 and evaluating its antioxidant and antimicrobial activity. Materials and Methods: Extraction and purification of PE was performed in four stages of preparation of crude extract, 65% ammonium sulfate precipitation, dialysis and anion exchange chromatography. Subsequently, antioxidant and antimicrobial activity was determined by nitric oxide scavenging and paper disk diffusion method respectively. Results:It was found that concentration and purity of PE were increased after each purification step by almost four times (from 1.5 to 6.22). PE had the strongest inhibitory activity against Bacillus subtilis(10.5 ± 0.28) and Candida albilancs (10.98 ± 0.006). In addition, the results of antioxidant surveys showed that PE is a potent free radical scavenger and this activity increased significantly with the increasing pigment concentration. Conclusion: The results of this study can be the basis for the introduction of natural edible pigments of soil cyanobacteria with antioxidant and antimicrobial properties that can be used in food and pharmaceutical industries.
rich source of novel bioactive compounds with pharmaceutical potential. Journal of
Pharmaceutical Health Services Research. 2018 Mar;9(1(:5-12.
2. Nowruzi B, Sarvari , Blanco S. Applications of cyanobacteria in biomedicine. InHandb.
Algal Sci. Microbiol. Technol. Med 2020. Elsevier Amsterdam. 441-454.
3. Mishra S , Shrivastav A, Pancha I, Jain D, Mishra S. Effect of preservatives for food grade
C-Phycoerythrin, isolated from marine cyanobacteria seudanabaena sp. International ournal
of biological macromolecules. 2010 Dec 1;47(5(:597-602.
4. Stadnichuk IN. Phycobiliproteins: Determination of chromophore composition and content.
Phytochemical Analysis. 1995 Nov;6(6(:281-8.
5. Afreen S, Fatma T. Extraction, purification and characterization of phycoerythrin from
Michrochaete and its biological activities. Biocatalysis and agricultural biotechnology. 2018
Jan 1;13:84-9.
6. Arun N, upta S, Singh DP. Antimicrobial and antioxidant property of commonly found
microalgae irulina latensis, Nostoc uscoru and Chlorella yrenoidosa against some
pathogenic bacteria and fungi. International Journal of Pharmaceutical Sciences and Research.
2012 Dec 1;3(12(:4866.
7. Basheva D, Moten D, Stoyanov P, Belkinova D, Mladenov R, Teneva I. Content of
phycoerythrin, phycocyanin, alophycocyanin and phycoerythrocyanin in some cyanobacterial
strains: Applications. Engineering in Life Sciences. 2018 Nov;18(11(:861-6.
8. Nowruzi B, Blanco S. In silico identification and evolutionary analysis of candidate genes
involved in the biosynthesis methylproline genes in cyanobacteria strains of Iran.
Phytochemistry Letters. 2019 Feb 1;29:199-211.
9. Chakdar H, Pabbi S. Extraction and purification of phycoerythrin from nabaena variabilis
(CCC421(. Phykos. 2012;42(1(:25-31.
10. Román RB, Alvarez-Pez JM, Fernández FA, rima EM. Recovery of pure B-phycoerythrin
from the microalga or hyridiu cruentu . Journal of Biotechnology. 2002 Jan 31;93(1(:
73-85.
11. amble SP, aikar RB, Padalia RB, Shinde D. Extraction and purification of
C-phycocyanin from dry irulina powder and evaluating its antioxidant, anticoagulation and
prevention of DNA damage activity. Journal of Applied Pharmaceutical Science. 2013 Aug
1;3(8(:149.
12. Nowruzi B, Blanco S, Ne adsattari T. Chemical and molecular evidences for the poisoning of
a duck by anatoxin-a, nodularin and cryptophycin at the coast of Lake Shoormast (Mazandaran
province, Iran(. Int J Algae. 2018;20(4(.
13. Nowruzi B, Wahlsten M, Jokela J. A report on finding a new peptide aldehyde from
cyanobacterium Nostoc sp. Bahar M by LC-MS and Marfey’s analysis. Iranian ournal of
biotechnology. 2019 Apr;17(2(.
14. Nowruzi B, havari-Ne ad RA, Sivonen , azemi B, Na afi F, Ne adsattari T. Identification
and toxigenic potential of a Nostoc sp. Algae. 2012 Dec 1;27(4(:303.
15. Hastie CJ, Borthwick EB, Morrison LF, Codd A, Cohen PT. Inhibition of several protein
phosphatases by a non-covalently interacting microcystin and a novel cyanobacterial peptide,
nostocyclin. Biochimica et Biophysica Acta (BBA(- eneral Sub ects. 2005 Nov 15;1726(2(:
187-93.
16. Becher P , Jüttner F. Insecticidal compounds of the biofilm‐forming cyanobacterium
ischerella sp.(ATCC 43239(. Environmental Toxicology: An International Journal. 2005
Jun;20(3(:363-72.
17. Dahms HU, Ying X, Pfeiffer C. Antifouling potential of cyanobacteria: a mini-review.
Biofouling. 2006 Jan 1;22(5(:317-27.
18. Jaiswal P, Singh P , Prasanna R. Cyanobacterial bioactive molecules—an overview of their
toxic properties. Canadian Journal of Microbiology. 2008 Sep;54(9(:701-17.
19. Prasanna R, Nain L, Tripathi R, upta , Chaudhary , Middha S, Joshi M, Ancha R,
aushik BD. Evaluation of fungicidal activity of extracellular filtrates of
cyanobacteria–possible role of hydrolytic enzymes. Journal of basic microbiology. 2008 Jun;48
(3(:186-94.
20. Asthana R , Srivastava A, Singh AP, Singh SP, Nath , Srivastava R, Srivastava BS.
Identification of an antimicrobial entity from the cyanobacterium ischerella sp. isolated from
bark of adirachta indica (Neem( tree. Journal of applied phycology. 2006 Feb 1;18(1(:33-9.
21. awsar SM, Fu ii Y, Matsumoto R, Yasumitsu H, Ozeki Y. Protein R-phycoerythrin from
marine red alga hiroa ance s: extraction, purification and characterization. Phytologia
Balcanica. 2011;17(3(:347-54.
22. hao M, Sun L, Sun S, ong X, Fu X, Chen M. Phycoerythrins in phycobilisomes from the
marine red alga olysi honia urceolata. International ournal of biological macromolecules.
2015 Feb 1;73:58-64.
23. Wang L, Wang S, Fu X, Sun L. Characteristics of an R-Phycoerythrin with two γ subunits
prepared from red macroalga olysi honia urceolata. PLoS One. 2015 Mar 17;10
(3(:e0120333.
24. Parmar A, Singh N , aushal A, Sonawala S, Madamwar D. Purification, characterization
and comparison of phycoerythrins from three different marine cyanobacterial cultures.
Bioresource technology. 2011 Jan 1;102(2(:1795-802.
25. Wang L, Qu Y, Fu X, hao M, Wang S, Sun L. Isolation, purification and properties of an
R-phycocyanin from the phycobilisomes of a marine red macroalga olysi honia urceolata.
PloS one. 2014 Feb 4;9(2(:e87833.
26. Fu X, Sun L, Yang , Lu W, hao M, ong X. The subunits analysis of R-phycoerythrin from
marine red algae by isoelectric focusing. African Journal of Biotechnology. 2011;10(39(:
7640-9.
27. Soni B, isavadiya NP, Dalwadi N, Madamwar D, Winder C, halil C. Purified
c‐phycoerythrin: safety studies in rats and protective role against permanganate‐mediated
fibroblast‐DNA damage. Journal of Applied Toxicology. 2010 Aug;30(6(:542-50.
28. Tan H, ao S, huang Y, Dong Y, uan W, hang , Xu J, Cui J. R-Phycoerythrin induces
S C-7901 apoptosis by arresting cell cycle at S phase. Marine drugs. 2016 Sep;14(9(:166.
29. Ran itha , aushik BD. Purification of phycobiliproteins from Nostoc uscoru .
30. ásquez-Suárez A, Lobos- onzález F, Cronshaw A, Sepúlveda-Ugarte J, Figueroa M,
Dagnino-Leone J, Bunster M, Martínez-Oyanedel J. The γ subunit of R-phycoerythrin from
racilaria chilensis has a typical double linked phycourobilin similar to γ subunit. PloS one.
2018 Apr 10;13(4(:e0195656.
31. Tripathi SN, apoor S, Shrivastava A. Extraction and purification of an unusual phycoerythrin
in a terrestrial desiccation tolerant cyanobacterium yngbya arboricola. Journal of Applied
Phycology. 2007 Oct 1;19(5(:441-7.
32. im JJ, Jeon YM, Noh JH, Lee MY. Isolation and characterization of a new phycoerythrin
from the cyanobacterium ynechococcus sp. ECS-18. Journal of Applied Phycology. 2011 Feb
1;23(1(:137-42.
33. hao M, Sun L, Sun S, ong X, Fu X, Chen M. The 42.1 and 53.7 kDa bands in SDS-PA E
of R-phycoerythrin from olysi honia urceolata. International ournal of biological
macromolecules. 2013 Sep 1;60:405-11.
34. Na denski HM, igova L , Iliev II, Pilarski PS, Lukavský J, Tsvetkova I , Ninova MS,
ussovski . Antibacterial and antifungal activities of selected microalgae and cyanobacteria.
International ournal of food science & technology. 2013 Jul;48(7(:1533-40.
35. Sonani RR, Rastogi RP, Madamwar D. Antioxidant potential of phycobiliproteins: Role in
anti-aging research. Biochem Anal Biochem. 2015;4(172(:2161-1009.
36. Wo cik M, Burzynska-Pedziwiatr I, Wozniak LA. A review of natural and synthetic
antioxidants important for health and longevity. Current Medicinal Chemistry. 2010 Sep 1;17
(28(:3262-88.
37. Nowruzi B, Sarvari , Blanco S. The cosmetic application of cyanobacterial secondary
metabolites. Algal Research. 2020 Aug 1;49:101959
_||_
rich source of novel bioactive compounds with pharmaceutical potential. Journal of
Pharmaceutical Health Services Research. 2018 Mar;9(1(:5-12.
2. Nowruzi B, Sarvari , Blanco S. Applications of cyanobacteria in biomedicine. InHandb.
Algal Sci. Microbiol. Technol. Med 2020. Elsevier Amsterdam. 441-454.
3. Mishra S , Shrivastav A, Pancha I, Jain D, Mishra S. Effect of preservatives for food grade
C-Phycoerythrin, isolated from marine cyanobacteria seudanabaena sp. International ournal
of biological macromolecules. 2010 Dec 1;47(5(:597-602.
4. Stadnichuk IN. Phycobiliproteins: Determination of chromophore composition and content.
Phytochemical Analysis. 1995 Nov;6(6(:281-8.
5. Afreen S, Fatma T. Extraction, purification and characterization of phycoerythrin from
Michrochaete and its biological activities. Biocatalysis and agricultural biotechnology. 2018
Jan 1;13:84-9.
6. Arun N, upta S, Singh DP. Antimicrobial and antioxidant property of commonly found
microalgae irulina latensis, Nostoc uscoru and Chlorella yrenoidosa against some
pathogenic bacteria and fungi. International Journal of Pharmaceutical Sciences and Research.
2012 Dec 1;3(12(:4866.
7. Basheva D, Moten D, Stoyanov P, Belkinova D, Mladenov R, Teneva I. Content of
phycoerythrin, phycocyanin, alophycocyanin and phycoerythrocyanin in some cyanobacterial
strains: Applications. Engineering in Life Sciences. 2018 Nov;18(11(:861-6.
8. Nowruzi B, Blanco S. In silico identification and evolutionary analysis of candidate genes
involved in the biosynthesis methylproline genes in cyanobacteria strains of Iran.
Phytochemistry Letters. 2019 Feb 1;29:199-211.
9. Chakdar H, Pabbi S. Extraction and purification of phycoerythrin from nabaena variabilis
(CCC421(. Phykos. 2012;42(1(:25-31.
10. Román RB, Alvarez-Pez JM, Fernández FA, rima EM. Recovery of pure B-phycoerythrin
from the microalga or hyridiu cruentu . Journal of Biotechnology. 2002 Jan 31;93(1(:
73-85.
11. amble SP, aikar RB, Padalia RB, Shinde D. Extraction and purification of
C-phycocyanin from dry irulina powder and evaluating its antioxidant, anticoagulation and
prevention of DNA damage activity. Journal of Applied Pharmaceutical Science. 2013 Aug
1;3(8(:149.
12. Nowruzi B, Blanco S, Ne adsattari T. Chemical and molecular evidences for the poisoning of
a duck by anatoxin-a, nodularin and cryptophycin at the coast of Lake Shoormast (Mazandaran
province, Iran(. Int J Algae. 2018;20(4(.
13. Nowruzi B, Wahlsten M, Jokela J. A report on finding a new peptide aldehyde from
cyanobacterium Nostoc sp. Bahar M by LC-MS and Marfey’s analysis. Iranian ournal of
biotechnology. 2019 Apr;17(2(.
14. Nowruzi B, havari-Ne ad RA, Sivonen , azemi B, Na afi F, Ne adsattari T. Identification
and toxigenic potential of a Nostoc sp. Algae. 2012 Dec 1;27(4(:303.
15. Hastie CJ, Borthwick EB, Morrison LF, Codd A, Cohen PT. Inhibition of several protein
phosphatases by a non-covalently interacting microcystin and a novel cyanobacterial peptide,
nostocyclin. Biochimica et Biophysica Acta (BBA(- eneral Sub ects. 2005 Nov 15;1726(2(:
187-93.
16. Becher P , Jüttner F. Insecticidal compounds of the biofilm‐forming cyanobacterium
ischerella sp.(ATCC 43239(. Environmental Toxicology: An International Journal. 2005
Jun;20(3(:363-72.
17. Dahms HU, Ying X, Pfeiffer C. Antifouling potential of cyanobacteria: a mini-review.
Biofouling. 2006 Jan 1;22(5(:317-27.
18. Jaiswal P, Singh P , Prasanna R. Cyanobacterial bioactive molecules—an overview of their
toxic properties. Canadian Journal of Microbiology. 2008 Sep;54(9(:701-17.
19. Prasanna R, Nain L, Tripathi R, upta , Chaudhary , Middha S, Joshi M, Ancha R,
aushik BD. Evaluation of fungicidal activity of extracellular filtrates of
cyanobacteria–possible role of hydrolytic enzymes. Journal of basic microbiology. 2008 Jun;48
(3(:186-94.
20. Asthana R , Srivastava A, Singh AP, Singh SP, Nath , Srivastava R, Srivastava BS.
Identification of an antimicrobial entity from the cyanobacterium ischerella sp. isolated from
bark of adirachta indica (Neem( tree. Journal of applied phycology. 2006 Feb 1;18(1(:33-9.
21. awsar SM, Fu ii Y, Matsumoto R, Yasumitsu H, Ozeki Y. Protein R-phycoerythrin from
marine red alga hiroa ance s: extraction, purification and characterization. Phytologia
Balcanica. 2011;17(3(:347-54.
22. hao M, Sun L, Sun S, ong X, Fu X, Chen M. Phycoerythrins in phycobilisomes from the
marine red alga olysi honia urceolata. International ournal of biological macromolecules.
2015 Feb 1;73:58-64.
23. Wang L, Wang S, Fu X, Sun L. Characteristics of an R-Phycoerythrin with two γ subunits
prepared from red macroalga olysi honia urceolata. PLoS One. 2015 Mar 17;10
(3(:e0120333.
24. Parmar A, Singh N , aushal A, Sonawala S, Madamwar D. Purification, characterization
and comparison of phycoerythrins from three different marine cyanobacterial cultures.
Bioresource technology. 2011 Jan 1;102(2(:1795-802.
25. Wang L, Qu Y, Fu X, hao M, Wang S, Sun L. Isolation, purification and properties of an
R-phycocyanin from the phycobilisomes of a marine red macroalga olysi honia urceolata.
PloS one. 2014 Feb 4;9(2(:e87833.
26. Fu X, Sun L, Yang , Lu W, hao M, ong X. The subunits analysis of R-phycoerythrin from
marine red algae by isoelectric focusing. African Journal of Biotechnology. 2011;10(39(:
7640-9.
27. Soni B, isavadiya NP, Dalwadi N, Madamwar D, Winder C, halil C. Purified
c‐phycoerythrin: safety studies in rats and protective role against permanganate‐mediated
fibroblast‐DNA damage. Journal of Applied Toxicology. 2010 Aug;30(6(:542-50.
28. Tan H, ao S, huang Y, Dong Y, uan W, hang , Xu J, Cui J. R-Phycoerythrin induces
S C-7901 apoptosis by arresting cell cycle at S phase. Marine drugs. 2016 Sep;14(9(:166.
29. Ran itha , aushik BD. Purification of phycobiliproteins from Nostoc uscoru .
30. ásquez-Suárez A, Lobos- onzález F, Cronshaw A, Sepúlveda-Ugarte J, Figueroa M,
Dagnino-Leone J, Bunster M, Martínez-Oyanedel J. The γ subunit of R-phycoerythrin from
racilaria chilensis has a typical double linked phycourobilin similar to γ subunit. PloS one.
2018 Apr 10;13(4(:e0195656.
31. Tripathi SN, apoor S, Shrivastava A. Extraction and purification of an unusual phycoerythrin
in a terrestrial desiccation tolerant cyanobacterium yngbya arboricola. Journal of Applied
Phycology. 2007 Oct 1;19(5(:441-7.
32. im JJ, Jeon YM, Noh JH, Lee MY. Isolation and characterization of a new phycoerythrin
from the cyanobacterium ynechococcus sp. ECS-18. Journal of Applied Phycology. 2011 Feb
1;23(1(:137-42.
33. hao M, Sun L, Sun S, ong X, Fu X, Chen M. The 42.1 and 53.7 kDa bands in SDS-PA E
of R-phycoerythrin from olysi honia urceolata. International ournal of biological
macromolecules. 2013 Sep 1;60:405-11.
34. Na denski HM, igova L , Iliev II, Pilarski PS, Lukavský J, Tsvetkova I , Ninova MS,
ussovski . Antibacterial and antifungal activities of selected microalgae and cyanobacteria.
International ournal of food science & technology. 2013 Jul;48(7(:1533-40.
35. Sonani RR, Rastogi RP, Madamwar D. Antioxidant potential of phycobiliproteins: Role in
anti-aging research. Biochem Anal Biochem. 2015;4(172(:2161-1009.
36. Wo cik M, Burzynska-Pedziwiatr I, Wozniak LA. A review of natural and synthetic
antioxidants important for health and longevity. Current Medicinal Chemistry. 2010 Sep 1;17
(28(:3262-88.
37. Nowruzi B, Sarvari , Blanco S. The cosmetic application of cyanobacterial secondary
metabolites. Algal Research. 2020 Aug 1;49:101959