Cloning of a C-phycocyanin Alpha Subunit from Thermosynechococcus sp. TUBT-T01 and Prediction of Its Properties

Plaipol DEDVISITSAKUL; Sichon HUADRAKSASAT; Supenya CHITTAPUN; Theppanya CHAROENRAT; Chanitchote PIYAPITTAYANUN

doi:10.48048/wjst.2018.3626

Authors

Plaipol DEDVISITSAKUL School of Science, Mae Fah Luang University, Chiang Rai 57100 http://orcid.org/0000-0002-6124-4012
Sichon HUADRAKSASAT Biotechnology, Faculty of Science and Technology, Thammasat University, Rangsit Campus, Prathum Thani 12120
Supenya CHITTAPUN Biotechnology, Faculty of Science and Technology, Thammasat University, Rangsit Campus, Prathum Thani 12120
Theppanya CHAROENRAT Biotechnology, Faculty of Science and Technology, Thammasat University, Rangsit Campus, Prathum Thani 12120
Chanitchote PIYAPITTAYANUN Biotechnology, Faculty of Science and Technology, Thammasat University, Rangsit Campus, Prathum Thani 12120

DOI:

https://doi.org/10.48048/wjst.2018.3626

Keywords:

Cyanobacteria, hot spring, alpha subunit, C-phycocyanin gene, Thermostability, Thermosynechococcus sp. TUBT-T01

Abstract

C-Phycocyanin, a blue-colored and water soluble protein, is a class of phycobiliproteins that are the major light-harvesting pigments of a photosynthetic system in cyanobacteria. C-phycocyanins are utilized in many industries, including as natural colorants in food and cosmetics and as antioxidant compounds. However, the uses of C-phycocyanins have been limited due to their vulnerability to high temperatures. Therefore, the objective of this study was to identify and analyze the C-phycocyanin gene isolated from Thermosynechococcus sp. TUBT-T01, living in a hot spring in Surat Thani province, in the hope that this C-phycocyanin exhibited thermostable properties and that their applications could be expanded over a wide range of industries. In the present study, the polymerase chain reaction of the gene encoding alpha subunits of C-phycocyanin (cpcA) was performed, using primers designed based upon the sequence alignments of cpcA from Thermosynechococcus sp. available in the GenBank database. The putative cpcA, with an approximate size of 500 base pairs, was detected on an agarose gel. The DNA sequencing analysis indicated that the cpcA was 489 base pairs in length, and its nucleotide sequence was 94 % identical to those of thermophilic Thermosynechococcus sp. NK55, T. elongatus BP-1, and Synechococcus vulcanus. The deduced amino acid sequence was very similar to those of Thermosynechococcus sp. NK55, T. elongatus BP-1, and S. vulcanus. The data derived from the homologous model revealed that the presence of Asp28, Lys32, and Ser72 in the alpha subunit of C-phycocyanin from Thermosynechococcus sp. TUBT-T01 could provide the high thermostability property of this protein.

Downloads

Download data is not yet available.

Metrics

Metrics Loading ...

Author Biography

Plaipol DEDVISITSAKUL, School of Science, Mae Fah Luang University, Chiang Rai 57100

School of Science, Mae Fah Luang University

References

WF Vincent. Cyanobacteria. Laval University, Quebec City, Canada, Elsevier Inc, 2009, p. 226-32.

WR Hess. Cyanobacterial genomics for ecology and biotechnology. Curr. Opin. Microbiol. 2011; 14, 608-14.

GE Fogg, WDP Stewart, P Fay and AE Walsby. The Blue-Green Algae. Academic Press, London and New York, 1973, p. 459

N Thajuddin and G Subramanian. Cyanobacterial biodiversity and potential applications in biotechnology. Curr. Sci. 2005; 89, 47-57.

Govindjee and D Shevela. Adventures with cyanobacteria: A personal perspective. Front Plant Sci. 2011; 6, 2-28.

KM Minkova, AA Tchernov, MI Tchorbadjieva, ST Fournadjieva, RE Antova and MC Busheva. Purification of C-phycocyanin from Spirulina (Arthrospira) fusiformis. J. Biotechnol. 2003; 102, 55-9.

R MacColl. Cyanobacterial phycobilisomes. J. Struct. Biol. 1998; 124, 311-34.

SR Kalla, LK Lind, J Lidholm and P Gustaffson. Transcriptional organization of the phycocyanin subunit gene clusters of the cyanobacerium Anacystis nidulans UTEX 625. J. Bacteriol. 1998; 170, 2961-70.

Y Nakamura, T Kaneko, S Sato, M Ikeuchi, H Katoh, S Sasamoto, A Watanabe, M Iriguchi, K. Kawashima, T Kimura, Y Kishida, C Kiyokawa, M Kohara, M Matsumoto, A Matsuno, N Nakazaki, S Shimpo, M Sugimoto, C Takeuchi, M Yamada and S Tabata. Complete genome structure of the thermophilic cyanobacterium Thermosynechococcus elongatus BP-1. DNA Res. 2002; 9, 123-30.

WR Belknap and R Haselkorn. Cloning and light regulation of expression of the phycocyanin operon of the cyanobacterium Anabaena. Embo J. 1987; 6, 871-84.

M Kuddus, P Singh, G Thomas and A Al-Hazimi. Recent developments in production and biotechnological applications of C-Phycocyanin. BioMed. Res. Int. 2013; 2013, 742859.

T Kato. Blue pigment from Spirulina. New Food Indust. 1994; 29, 17-24

T Hirata, M Tanaka, M Ooike, T Tsunomura and M Sakaguchi. Antioxidant activities of phycocyanobilin prepared from Spirulina platensis. J. Appl. Phycol. 2000; 12, 435-9.

L Jespersen, LD Strømdahl and K Olsen. Heat and light stability of three natural blue colorants for use in confectionery and beverages. Eur. Food Res. Tech. 2005; 220, 261-6.

KS Mishra, A Shrivastav and S Mishra. Effect of preservatives for food grade C-PC from Spirulina platensis. Process Biochem. 2008; 43, 339-45.

C Romay, R Delgado, D Remirez, R González and A Rojas. Effects of phycocyanin extract on tumor necrosis factor-alpha and nitrite levels in serum of mice treated with endotoxin. Arzneimittelforschung 2001; 51, 733-6.

CM Shih, SN Cheng, CS Wong, YL Kuo and TC Chou. Antiinflammatory and antihyperalgesic activity of C-phycocyanin. Anesth Analg. 2009; 108, 1303-10.

C Romay, R Gonzalez, N Ledon, D Remirez and Rimbau. C-phycocyanin: a biliprotein with antioxidant, anti-inflammatory and neuroprotective effects. Curr. Protein Pept. Sci. 2003; 4, 207-16.

K Bagchi and S Puri. Free radicals and antioxidants in health and disease: A review. Eastern Mediterr. Health J. 1998; 4, 350-60.

A Nunomura, R Castellani, X Zhu, P Moreira, G Perry and M Smith. Involvement of oxidative stress in Alzheimer disease. J. Neuropathol. Exp. Neurol. 2006; 65, 631-41.

RM Tamimi, SE Hankinson, H Campos, D Spiegelman, S Zhang, GA Colditz, WC Willett and DJ Hunter. Plasma carotenoids, retinol, and tocopherols and risk of breast cancer. Am. J. Epidemiol. 2005; 161, 153-60.

G Davì, A Falco and C Patrono. Lipid peroxidation in diabetes mellitus. Antioxid. Redox. Sig. 2005; 7, 256-68.

C Hitchon and H El-Gabalawy. Oxidation in rheumatoid arthritis. Arthritis Res. Ther. 2004; 6, 265-78.

Richa, VK Kannaujiya, M Kesheri, G Singh and RP Sinha. Biotechnological potentials of phycobiliproteins. Int. J. Pharm. Bio. Sci. 2011; 2, B446-B454.

JY Leu, TH Lin, MJP Selvamani, HC Chen, JZ Liang and KM Pan. Characterization of a novel thermophilic cyanobacterial strain from Taian hot springs in Taiwan for high CO2 mitigation and C-phycocyanin extraction. Process Biochem. 2013; 48, 41-8.

MR Edwards, C Hauer, RF Stack, LE Eisele and R MacColl. Thermophilic C- phycocyanin: Effect of temperature, monomer stability, and structure. Biochim. Biophys. Acta 1997; 1321, 157-64.

N Adir, Y Dobrovetsky and N Lerner. Structure of c-phycocyanin from the thermophilic cyanobacterium Synechococcus vulcanus at 2.5 angstrom: Structural implications for thermal stability in phycobilisome assembly. J. Mol. Biol. 2001; 313, 71-81.

K Suwanmanee, T Charoenrat and S Chittapun. Isolation and cultivation of thermophilic blue green algae from hot springs in Surat thani province. In: Proceeding of the 7th National Conference on Algae and Plankton, Narai Hotel, Bangkok, Thailand, 2015. p. 1-12.

A Untergasser, I Cutcutache, T Koressaar, J Ye, BC Faircloth, M Remm and SG Rozen. Primer3-new capabilities and interfaces. Nucleic. Acids Res. 2012; 40, e115.

SF Altschul, W Gish, W Miller, EW Myers and D J Lipman. Basic local alignment search tool. J. Mol. Biol. 1990; 215, 403-10.

SF Altschul, TL Madden, AA Schäffer, J Zhang, Z Zhang, W Miller and DJ Lipman. Gapped BLAST and PSI-BLAST: A new generation of protein database search programs. Nucleic. Acids Res. 1997; 25, 3389-402.

S Kumar, G Stecher and K Tamura. MEGA7: Molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol. Biol. Evol. 2016; 33, 1870-4.

E Gasteiger, C Hoogland, A Gattiker, S Duvaud, MR Wilkins, RD Appel and A Bairoch. Protein Identification and Analysis Tools on the ExPASy Server. The Proteomics Protocols Handbook, Humana Press, 2005, p. 571-607.

J Garnier, JF Gibrat and B Robson. GOR method for predicting protein secondary structure from amino acid sequence. Meth. Enzymol. 1996; 266, 540-53.

F Sievers, A Wilm, D Dineen, TJ Gibson, K Karplus, W Li, R Lopez, H McWilliam, M Remmert, J Söding, JD Thompson and DG Higgins. Fast scalable generation of high-quality protein multiple sequence alignments using Clustal Omega. Mol Syst Biol. 2011; 7, 539.

A Marx and N Adir. Allophycocyanin and phycocyanin crystal structures reveal facets of phycobilisome assembly. Biochim. Biophys. Acta 2013; 1827, 311-8.

M Duerring, GB Schmidt and R Huber. Isolation, crystallization, crystal structure analysis and refinement of constitutive C-phycocyanin from the chromatically adapting cyanobacterium Fremyella diplosiphon at 1.66 A resolution. J. Mol. Biol. 1991; 217, 577-92.

K Brejc, R Ficner, R Huber and S Steinbacher. Isolation, crystallization, crystal structure analysis and refinement of allophycocyanin from the cyanobacterium Spirulina platensis at 2.3 A resolution. J. Mol. Biol. 1995; 249, 424-40.

T Schirmer, W Bode and R Huber. Refined three-dimensional structures of two cyanobacterial C-phycocyanins at 2.1 and 2.5 A resolution: A common principle of phycobilin-protein Interaction. J. Mol. Biol. 1987; 196, 677-95.

C Ohto, C Ishida, H Nakane, M Muramatsu, T Nishino and S Obata. A thermophilic cyanobacterium Synechococcus elongatus has three different class I prenyltransferase genes. Plant Mol. Biol. 1999; 40, 307-21.

R Prabha, DP Singh and A Rai. Looking into the genome of Thermosynechococcus elongatus (thermophilic cyanobacteria) with codon selection and usage perspective. Int. J. Bioinform. Res. Appl. 2015; 11, 130-41.

S Stolyar, Z Liu, V Thiel, LP Tomsho, N Pinel, WC Nelson and JK Fredrickson. Genome sequence of the thermophilic cyanobacterium Thermosynechococcus sp. strain NK55a. Genome Announc. 2014; 2, e01060-e010113.

K Guruprasad, BV Reddy and MW Pandit. Correlation between stability of a protein and its dipeptide composition: A novel approach for predicting in vivo stability of a protein from its primary sequence. Protein Eng. 1990; 4, 155-61.

J Kyte and RF Doolittle. A simple method for displaying the hydropathic character of a protein. J. Mol. Biol. 1982; 157, 105-32.

VK Kannaujiya, A Rahman, Adinath, S Sundaram and RP Sinha. Structural and functional dynamics of tyrosine amino acid in phycocyanin of hot-spring cyanobacteria: A possible pathway for internal energy transfer. Gene Rep. 2016; 5, 83-91

N Inoue, T Emi, Y Yamane, Y Kashino, H Koike and K Satoh. Effect of hight-temperature treatments on a thermophilic cyanobacterium Synechococcus vulcanus. Plant Cell Physiol. 2000; 41, 515-22.

P Turner, G Mamo and EN Karlsson. Potential and utilization of thermophiles and thermostable enzymes in biorefining. Microb. Cell Fact. 2007; 6, 9.

XY Guan, WJ Zhang, XW Zhang, YX Li, JF Wang, HZ Lin and S Qin. A potent anti‐oxidant property: Fluorescent recombinant α‐phycocyanin of Spirulina. J. Appl. Microbial. 2009; 106, 1093-100.

S Qin, Z Tang, F Lin, LA Sung and CK Tseng. Genomic cloning, expression and recombinant protein purification of α and β subunits of the allophycocyanin gene (apc) from the cyanobacterium Anacystis nidulans UTEX 625. J. Appl. Phycol. 2004; 16, 483-7.

Z Shoja, HR Memari and MR Ardakani. Cloning and expression of Beta Subunit Gene of Phycocyanin from Spirulina platensis in Escherichia coli. Jundishapur J. Microbiol. 2015; 8, e17809.

T Cherdkiatikul and Y Suwanwong. Production of the α and β subunits of Spirulina allophycocyanin and C-phycocyanin in Escherichia coli: A comparative study of their antioxidant activities. J. Biomol. Screen 2014; 19, 959-65.

MF Hossain, PR Ratnayake, K Meerajini and KL Wasantha Kumara. Antioxidant properties in some selected cyanobacteria isolated from fresh water bodies of Sri Lanka. Food Sci. Nutr. 2016; 4, 753-8.

A Patel, S Mishra and PK Ghosh. Antioxidant potential of C-phycocyanin isolated from cyanobacterial species Lyngbya, Phormidium and Spirulina spp. Indian J. Biochem. Biol. 2006; 43, 25-31.

B Soni, U Trivedi and D Madamwar. A novel method of single step hydrophobic interaction chromatography for the purification of phycocyanin from Phormidium fragile and its characterization for antioxidant property. Bioresource Tech. 2008; 99, 188-94.