Production Of Phycobiliproteins Research Articles

Evaluation of growth and phycobiliprotein composition of cyanobacteria isolates cultivated in different nitrogen sources

Phycobiliproteins, light-harvesting pigments found in cyanobacteria and in some eukaryotic algae, have numerous commercial applications in food, cosmetic, and pharmaceutical industries. Colorant production from cyanobacteria offers advantages over their production from higher plants, as cyanobacteria have fast growth rate and high photosynthetic efficiency and require less space. In this study, three cyanobacteria strains were studied for phycobiliprotein production and the influence of sodium nitrate, potassium nitrate and ammonium chloride on the growth and phycobiliprotein composition of the strains were evaluated. In the batch culture period of 12 days, Phormidium sp. and Pseudoscillatoria sp. were able to utilize all tested nitrogen sources; however, ammonium chloride was the best nitrogen source for both strains to achieve maximum growth rate μ = 0.284 ± 0.03 and μ = 0.274 ± 0.13 day−1, chlorophyll a 16.2 ± 0.5 and 12.2 ± 0.2 mg L−1, and phycobiliprotein contents 19.38 ± 0.09 and 19.99 ± 0.14% of dry weight, whereas, for Arthrospira platensis, the highest growth rate of μ = 0.304 ± 0.0 day−1, chlorophyll a 19.1 ± 0.5 mg L−1, and phycobiliprotein content of 22.27 ± 0.21% of dry weight were achieved with sodium nitrate. The phycocyanin from the lyophilized cyanobacterial biomass was extracted using calcium chloride and food grade purity (A620/A280 ratio > 0.7) was achieved. Furthermore, phycocyanin was purified using two-step chromatographic method and the analytical grade purity (A620/A280 ratio > 4) was attained. SDS-PAGE demonstrated the purity and presence of two bands corresponding to α- and β-subunits of the C-phycocyanin. The results showed that Phormidium sp. and Pseudoscillatoria sp. could be good candidates for phycocyanin production.

Effects of different light source and media on growth and production of phycobiliprotein from freshwater cyanobacteria

The aim of this study was to determine the effect of different light sources and media (wastewater and BBM) on the growth of Pseudanabaena mucicola and its phycobiliprotein production. Results showed that P. mucicola grown in white light using wastewater as medium attributed higher biomass (0.55 g L−1) and when extracted with water, also showed significantly higher (P < .05) production (237.01 mg g−1) and purity (1.14) of phycobiliprotein. This study validated that phycobiliprotein extracted from P. mucicola using water can be food grade natural blue pigment. Moreover, cyanobacteria grown in wastewater could cut down the production cost of phycobiliprotein.

Phycobiliprotein production by a novel cold desert cyanobacterium Nodularia sphaerocarpa PUPCCC 420.1

The present study focuses on finding a good source of phycobiliproteins (PBP). We report a new cyanobacterium Nodularia sphaerocarpa PUPCCC 420.1 as a good producer of PBP. The organism produced 445.6 μg PBP mg−1 dry biomass. The growth and PBP production of organism were optimized by varying pH of growth medium, nitrogen sources, light quality and sugars. The optimized conditions for PBP production were as follows: pH 8.0, 5 mmol L−1 KNO3, 10 mmol L−1 NaNO2, 0.5% sucrose and green light. The PBP production under these conditions ranged from 486 to 676.3 μg mg−1 dry biomass. The PBP were more stable when stored in alkaline pH at 4 °C under dark. As per survey of literature, except for Anabaena fertilissima, the amount of PBP is significantly higher than the amount of PBP produced by other cyanobacteria. Thus, this organism is a good candidate for the PBP production at commercial level.

Economic feasibility of microalgal bacterial floc production for wastewater treatment and biomass valorization: A detailed up-to-date analysis of up-scaled pilot results

The economic potential of outdoor microalgal bacterial floc (MaB-floc) raceway ponds as wastewater treatment technology and bioresource of biomass for fertilizer, shrimp feed, phycobiliproteins and biogas in Northwest Europe is assessed. This assessment is based on cost data provided by industry experts, on experimental data obtained from pilot-scale outdoor MaB-floc ponds treating aquaculture and food-industry effluents, and from different biomass valorization tests. MaB-floc ponds exhibit a cost-performance of EUR 0.25–0.50m−3 wastewater which is similar to conventional wastewater treatment technologies. The production cost of MaB-flocs in aquaculture and food industry effluent is EUR 5.29 and 8.07kg−1TSS, respectively. Capital costs and pond mixing costs are the major expenses. Commercializing MaB-flocs as aquaculture feed generates substantial revenues, but the largest profit potential lies in production of high-purity phycobiliproteins from MaB-flocs. These results highlight the large economic potential of MaB-floc technology, and justify its further development.

Food-industry-effluent-grown microalgal bacterial flocs as a bioresource for high-value phycochemicals and biogas

Microalgal bacterial floc (MaB-floc) raceway ponds are a novel sunlight-based technology to grow biomass on food-industry effluent and flue gas (33.9kgvolatilesolids(VS)hapond−1d−1). The MaB-floc biorefinery concept of high-value phycochemicals and biogas was screened to find a suitable valorization strategy for this novel biomass. Freezing and aqueous extraction of MaB-flocs followed by size exclusion chromatography yielded 22.4gC-phycocyanin(C-PC)kg−1 VS with a purity of 1.32 (24.5% recovery) and 9.5gC-phycoerythrin(C-PE)kg−1 VS with a purity of 1.06 (20.9% recovery). Anaerobic digestion of the extracted MaB-flocs resulted in 272NLCH4g−1 VS. Moreover, increasing the suspended solid (SS) loading of food industry effluent for one day, significantly reduced the biochemical methane yield by 13.6%, and the C-PC and C-PE yield of total crude extracts by 74.5% and 65.5%, respectively. In contrast, it increased the neophytadiene yield by 45.1%. This study highlights the large potential of these MaB-flocs as a bioresource for production of phycobiliproteins, biogas and neophytadiene. Further research is needed to improve the phycochemical extraction and purification processes, and to confirm a huge economic potential.

Recent advances in production, purification and applications of phycobiliproteins.

An obligatory sunlight requirement for photosynthesis has exposed cyanobacteria to different quantity and quality of light. Cyanobacteria can exhibit efficient photosynthesis over broad region (450 to 650 nm) of solar spectrum with the help of brilliantly coloured pigment proteins called phycobiliproteins (PBPs). Besides light-harvesting, PBPs are found to involve in several life sustaining phenomena including photoprotection in cyanobacteria. The unique spectral features (like strong absorbance and fluorescence), proteineous nature and, some imperative properties like hepato-protective, anti-oxidants, anti-inflammatory and anti-aging activity of PBPs enable their use in food, cosmetics, pharmaceutical and biomedical industries. PBPs have been also noted to show beneficial effect in therapeutics of some disease like Alzheimer and cancer. Such large range of applications increases the demand of PBPs in commodity market. Therefore, the large-scale and coast effective production of PBPs is the real need of time. To fulfil this need, many researchers have been working to find the potential producer of PBPs for the production and purification of PBPs. Results of these efforts have caused the inventions of some novel techniques like mixotrophic and heterotrophic strategies for production and aqueous two phase separation for purification purpose. Overall, the present review summarises the recent findings and identifies gaps in the field of production, purification and applications of this biological and economically important proteins.

Effect of gibberellic acid on the cyanobacterium Nostoc linckia

We investigated the influence of gibberellic acid (GA3; 0, 1, 10, and 100 μM) on Nostoc linckia culture at 7, 14, and 21 days. The fresh and dry weight of N. linckia was increased considerably by the 10 and 100 μM GA3 treatments. A reduction in heterocyst frequency was observed in cultures treated with 1 and 10 μM GA3. Adding GA3 to N. linckia culture had a little effect on cell size. The amount of chlorophyll a and carotenoids decreased at all concentrations of GA3. The amount of phycocyanin increased up to twofold in 7-day-old culture treated with 1 μM GA3, and similar changes were observed for allophycocyanin and phycoerythrin content after 7 days. The effect of GA3 on reducing sugar content was different and was dependent on the growth period. A reduction in soluble sugar content was detected after GA3 application in 7- and 14-day-old cyanobacteria. Cultures treated with GA3 had a higher protein content after 14 days and a lower protein content after 7 and 21 days, and reduced nitrogenase activity after 7, 14, and 21 days. Our data show that GA3 application can be a suitable and inexpensive way to increase N. linckia biomass and phycobiliprotein production.

Hyperproduction of phycobiliproteins by the cyanobacterium Anabaena fertilissima PUPCCC 410.5 under optimized culture conditions

This study was targeted to find new sources of phycobiliproteins (PBP) as these are important as natural colours, antioxidants and fluorescent markers. Here we report Anabaena fertilissima PUPCCC 410.5 as a good producer of PBP. Control cultures of the cyanobacterium produced 383μg PBP mg−1 dry biomass on day 8. The pH of the medium, supplementation of nitrogen source, sugar and illumination of cultures with different colours of light were optimized for PBP production. Optimized individual parameters were: pH of the medium 7.5; supplementation of 2mmol nitrite L−1 or 0.5% sucrose; and illumination of cultures with blue light. Total PBP production under these conditions ranged between 627 and 696μgmg−1 dry biomass, resulting in nearly 1.6 fold increase, with a significant increase in phycocyanin content. Incubation of 2mmol nitrite L−1 supplemented cultures in blue light resulted in nearly 4.5 fold increase in phycoerythrin compared to the control cultures. The PBP/phycocyanin/phycoerythrin production under the optimized conditions by Anabaena fertilissima PUPCCC 410.5 is significantly higher than the PBP production by the other cyanobacterial strains reported in literature. Thus this cyanobacterial strain is a promising candidate for PBP production at commercial level.

Production of phycobiliproteins by Arthrospira platensis under different lightconditions for application in food products

There has been an increase in investment in research on new sources of natural pigments for food application. Some cyanobacteria can change the structures responsible for light harvesting and cellular processes according to the wavelength and light intensity. This phenomenon has been described as complementary chromatic adaptation. The present study aimed to investigate the growth of Arthrospira platensis using different light qualities, irradiance, and wavelength by evaluating the production of biomass, proteins, and phycobiliproteins. The occurrence of the chromatic adaptation phenomenon in this cyanobacterium was also investigated. The microorganism used in this study, A. platensis, was grown in a Zarrouk medium under three irradiance levels, 50, 100, and 150 μmol fotons.m–2.s–1 with illumination provided by white and green fluorescent lamps. The condition of 150 µmol fotons.m–2.s–1 white light was the one that promoted the highest biomass production of A. platensis cultures (2115.24 mg.L–1). There was no difference in the production of total protein and total phycobiliproteins under the studied conditions. It is likely that the large supply of nitrogen in the Zarrouk medium was sufficient for cell growth and maintenance, and it supplied the production of accessory pigments composed of protein. Finally, there was no evidence of the complementary chromatic adaptation phenomenon in A. platensis cultivated under green light. Moreover, this condition did not increase phycocyanin production.

Towards the biorefinery concept: Interaction of light, temperature and nitrogen for optimizing the co-production of high-value compounds in Porphyridium purpureum

The interactive effects of light, temperature and nitrogen regime on phycobiliprotein (PB) production and other compounds such as fatty acids, pigments and carbohydrates, were studied during batch-cultivation of Porphyridium purpureum, a red microalga, containing multiple compounds of commercial interest. Results indicate that nitrogen-replete modes, such as semi-continuous or continuous regime represent the most suitable culture strategy for PB, carbohydrate, total fatty acid (TFA) and eicosapentaenoic acid (EPA) production in P. purpureum. Nitrate-deficiency causes a strong decrease in growth performance, as well as in its PE, TFA and EPA contents which may be related to membrane degradation; but induces carbohydrate accumulation. Nitrate-starved cells of P. purpureum had the ability to restore PB and TFA contents, and specifically phycoerythrin (PE) and EPA levels, after medium refreshment, suggesting an almost complete regeneration of the plastidic membranes and phycobilisomes. Using response surface methodology (RSM), our results highlight for the first time the optimally combined light and temperature conditions necessary to promote growth and compound production, in particular PB, in P. purpureum batch-cultivated in nitrogen-replete medium. A simultaneous increase in light and temperature causes a strong decrease in cellular PB, TFA, EPA and pigment contents, suggesting a severe damage and possible disruption of thylakoid membranes. The highest PB content (~2.9% d.w.) was reached under combined low light (30μmolm−2s−1) and low temperature (10°C). Despite of this, maximal PB productivity was obtained at 20°C and under low light intensity, reaching up to 33.3mgL−1 (~2%d.w.). Under such specific growth conditions, P. purpureum biomass also contained substantial amounts of other valuable products (i.e., carbohydrates, EPA, Chl. a, zeaxanthin, β-carotene) which could therefore be co-extracted, with PB, by applying a biorefinery approach.

Modulation of phycobiliprotein production in Nostoc muscorum through culture manipulation

Nostoc sp. BTA87 was procured from National fresh water cyanobacterial and microalgal repository of IBSDDBT, Imphal, Manipur, India. This was identified as Nostoc muscorum based upon morphological characteristics and 16S rRNA gene sequence analysis. Cultivation of organism under two cultural conditions indicated that phycobiliproteins enhanced under dialyzed conditions in comparison to non-dialyzed conditions. Red light was more effective to green and blue light for enhanced production of phycocyanin and allophycocyanin whereas phycoerythrin was produced more under green light in dialyzed condition. The optimum pH for maximum production of all the three components of phycobiliproteins was 8.0. The results clearly indicated that the production of phycobiliproteins can be enhanced by cultivating cyanobacterium in dialyzed conditions at pH-8.0 under optimum light quality and Nostoc muscorum can be a potential candidate for the commercial production of phycobiliproteins.

Effect of external pH on cyanobacterial phycobiliproteins production and ammonium excretion

The response of phycobiliproteins to light qualities in Anabaena circinalis S. K. Ojit, Th. Indrama, O. Gunapati, S. O. Avijeet, S. A. Subhalaxmi, Ch. Silvia, D. W. Indira, Kh. Romi, Sh. Minerva, D. A. Thadoi, O. N. Tiwari, G. D. Sharma

Influence of light on phycobiliprotein production in three marine cyanobacterial cultures

Complementary chromatic adaptation, a photomorphogenetic response, known to occur in many cyanobacteria, enables them to efficiently absorb prevalent wavelengths of light in the environment. In the present study, we have described the influence of light on phycobiliprotein production in three marine phycoerythrin producing cyanobacterial cultures, namely, Lyngbya sp. A09DM, Phormidium sp. A27DM and Halomicronema sp. A32DM. A comparative study (UV-visible overlay spectra and SDS-PAGE analyses) of phycobiliproteins purified from all the three cultures grown in white, yellow, red and green lights has been confirmed. White light was taken as control. Red and green lights were taken to check their effect on phycocyanin and phycoerythrin production, respectively. Yellow light was studied as its wavelength falls in between green and red light. Lyngbya sp. A09DM was found to be the best chromatically adapting cyanobacterium followed by Halomicronema sp. A32DM. These two cultures can be placed in group III chromatic adaptors. Phormidium sp. A27DM was the least chromatically adapting culture and can be placed in group II chromatic adaptors. The study signifies that even light plays an important role along with nutrient availability in adapting cultures to changing environmental conditions.

Isolation, identification and characterization of novel thermotolerant Oscillatoria sp. N9DM: Change in pigmentation profile in response to temperature

Two thermotolerant cyanobacterial strains N9DM I and N9DM II were collected from Tuwa geothermal spring, Panchmahal, Gujarat, India, from different wells having temperature 30±2°C and 55±2°C respectively. Analysis of 16S rRNA gene sequence revealed that both the strains were nearest neighbour of Oscillatoria sp. In addition to high temperature tolerance Oscillatoria N9DM sp. changes phycobilisome rod pigmentation as exposed to various temperatures. Oscillatoria N9DM sp. produces phycoerythrin (PE) and phycocyanin (PC) at 30±2°C and 55±2°C, respectively whereas at intermediate temperature (42±2°C) it produced phycoerythrocyanin (PEC). Moreover reverted cultures (normally grown at 55±2°C but now sub-cultured and grown at 30±2°C) started producing PE. Furthermore, Oscillatoria N9DM sp. produced PE, PC and PEC in white, red and yellow light respectively and it showed direct correlation to different proteins produced at various temperatures. To best of our knowledge, this is the first report describing correlation of temperature and its effect on the phycobiliprotein production. This study for the first time also tried to explain that thermo-adaptation might have been evolved along with complementary chromatic adaptation (CCA) in certain thermotolerant cyanobacteria where fluctuation of temperature was prevalent throughout the year.

Application of molecular genetic and microbiological techniques in ecology and biotechnology of cyanobacteria

The review discusses the advances and problems in biotechnology and ecology of cyanobacteria and considers the possibilities of molecular genetic and microbiological techniques in this field. Due to the ease of cultivation, high growth rate, availability of synchronous cultures, and existence of numerous molecular genetic and microbiological techniques for various cyanobacterial strains, cyanobacteria—prokaryotic organisms that are ancient relatives of the chloroplasts—are model organisms in the studies of photosynthesis, dinitrogen fixation, cell division, hydrogen production, and in a number of other areas of basic and applied science. These techniques make possible deeper understanding of the role of cyanobacteria in various ecosystems and utilization of their potential in numerous applied projects, including production of molecular hydrogen, phycobiliproteins, and cyanophycin; formation of nanoparticles; removal of heavy metals from the environment; substrate biodegradation; manufacture of products for medicine and food industry; and solution of the problem of cyanobacterial toxins in freshwater and marine environments.

Screening of Cyanobacteria for Phycobiliproteins and Effect of Different Environmental Stress on Its Yield

Among 18 strains, cyanobacterium Anabaena NCCU-9 contained the highest amount of phycobiliprotein (91 mg/g dry cell weights). Therefore, the effects of various environmental stresses were investigated on its phycobiliprotein production potential. The ideal conditions observed were 30 degrees C, 25 micromol photons/m2/s, white light, pH-8, 16:8 light and dark regimes, nitrogen free medium and 10 mM sodium chloride. Among three pesticides studied malathion showed highest toxicity. Under heavy metal stress the order of toxicity was chromium > cadmium > lead > nickel > copper > zinc.

Statistical optimization of culture media for production of phycobiliprotein by Synechocystis sp. PCC 6701

Synechocystis sp. PCC 6701 has a brilliantly colored pigment, phycobiliprotein containing phycoerythrin. Culture medium was optimized by sequential designs in order to maximize phycobiliprotein production. The observed fresh weights after 6 days were 0.58 g/L in BG-11, 0.83 g/L in medium for Scenedesmus sp. and 0.03∼0.52 g/L in the other tested media. Medium for Scenedesmus sp. was selected to be optimized by fractional factorial design and central composite design since the medium maintained a more stable pH within a desirable range due to higher contents of phosphate. The fractional factorial design had seven factors with two levels: KNO3, NaNO3, NaH2PO4, Na2HPO4, Ca(NO3)2, FeEDTA, and MgSO4. From the result of fractional factorial design, nitrate and phosphate were identified as significant factors. A central composite design was then applied with four variables at five levels each: nitrate, phosphate, pH, and light intensity. Parameters such as fresh weight and phycobiliprotein contents were used to determine the optimum value of the four variables. The proposed optimum media contains 0.88 g/L of nitrate, 0.32 g/L of phosphate under 25 μE·m−2·s−1 of light intensity. The maximum phycobiliprotein contents have been increased over 400%, from 4.9 to 25.9 mg/L after optimization.

Crescimento mixotrófico de &lt;em&gt;Nostoc&lt;/em&gt; sp. Glucose, sacarose e melaço de cana-de-açúcar foram testados como substratos para produção de biomassa e ficobiliproteinas

Glucose, sacarose, and sugarcane molasses were tested as substrates for production of biomass and phycobiliproteins by Nostoc sp., varying their concentrations in relation to a mineral medium, BG11. All substrates increased the biomass and phycobiliproteins when compared with the control. Sugarcane molasses showed to be the best substrate for production of both biomass and phycobiliproteins. Greater biomass production occurred in sugarcane molasses 1.0 g L-1 and it was 5.7 times greater than the control. With glucose, it was in 2.5 g L-1 and sucrose, in 1.5 g L-1, reaching 2.5 and 4.8 times greater than the control, respectively. For phycobiliproteins, the major production was in sugarcane molasses 1.0 g L-1, 12.5 times greater than the control. With glucose, it was in 1.0 g L-1 and sucrose, in 0,5 g L-1, reaching 3.0 and 4.5 times greater than the control, respectively. The Nostoc sp. assayed can grow mixotrophically, using glucose, sucrose, and sugarcane molasses as organic substrates, and a greater production of biomass and phycobiliproteins can be reached when compared with the autotrophic growth.

Phycobiliproteins as a commodity: trends in applied research, patents and commercialization

Phycobiliproteins are a group of colored proteins commonly present in cyanobacteria and red algae possessing a spectrum of applications. They are extensively commercialized for fluorescent applications in clinical and immunological analysis. They are also used as a colorant, and their therapeutic value has also been categorically demonstrated. However, a comprehensive knowledge and technological base for augmenting their commercial utilities is lacking. Hence, this work is focused towards this objective by means of analyzing global patents and commercial activities with application oriented research. Strategic mining of patents was performed from global patent databases resulting in the identification of 297 patents on phycobiliproteins. The majority of the patents are from USA, Japan and Europe. Patents are grouped into fluorescent applications, general applications and production aspects of phycobiliproteins and the features of each group are discussed. Commercial and applied research activities are compared in parallel. It revealed that US patents are mostly related to fluorescent applications while Japanese are on the production, purification and application for therapeutic and diagnostic purposes. Fluorescent applications are well represented in research, patents and commercial sectors. Biomedical properties documented in research and patents are not ventured commercially. Several novel applications are reported only in patents. The paper further pinpoints the plethora of techniques used for cell breakage and for extraction and purification of phycobiliproteins. The analysis identifies the lacuna and suggests means for improvements in the application and production of phycobiliproteins.

Enhancing pigment accumulation inAnabaenastrains using sugars

The pigment accumulating potential of two strains of Anabaena was evaluated in the presence of various sugars (glucose, sucrose and jaggery), when incubated under light/darkness. Both strains exhibited excellent potential for phycocyanin (PC), phycoerythrin (PE) and total phycobiliproteins (PBS) production in light, in the presence of sugars. In terms of accumulation of % phycobiliproteins / proteins, Anabaena cylindrica (ATCC 29414) was observed to be more responsive to jaggery while sucrose and glucose were more stimulatory for Anabaena sp. (cyanobiont isolated from Azolla mexicana ). Significant enhancement in PC and PE content by supplementation with sugars emphasises the need for in-depth analysis of the interactions between assimilation of sugars and pigment production pathways in cyanobacteria.