Phenylalanine ammonia lyase (PAL) enzyme activity and antioxidant properties of some cyanobacteria isolates
Highlights
► Antioxidant properties and PAL activity of cyanobacterial isolates evaluated together. ► Microalgae have the possibility of usage as natural antioxidant and PAL sources. ► l-phe induces the total phenolic accumulation in cyanobacteria. ► Cyanobacteria may have the potential to be used in the treatment of phenylketonuria.
Introduction
Phenylalanine ammonia lyase (PAL) catalyses the non-oxidative deamination of phenylalanine to trans-cinnamate and directs the carbon flow from the shikimate pathway to the branches of phenylpropanoid metabolism. It is a secondary metabolic pathway that is operative in higher plants and some other organisms which are mainly involved in defense mechanisms (MacDonald & D’ Cunha, 2007). The properties, regulation, expression, and cellular distributions of PAL have been extensively studied, and the crystal structures of PAL from parsley and the yeast Rhodosporidium toruloides have been determined (Reichert, He, & Dixon, 2009). PAL is an inducible enzyme, and the incorporation of l-phe in the fermentation medium has been indicated as significantly inducing PAL activity (MacDonald & D’ Cunha, 2007).
The first described PALs from cyanobacteria were Anabaena variabilis and Nostoc punctiforme. In comparison to eukaryotic homologues, the cyanobacterial PALs are 20% smaller in size, but it is indicated that they share similar substrate selectivity and kinetic activity toward l-phe compared with l-tyrosine. Recombinant PAL is currently being evaluated for use in humans as enzyme substitution therapy for the treatment of the inherited metabolic disease phenylketonuria (PKU) resulting from the loss or impairment of phenylalanine hydoxilase, an enzyme that metabolizes l-phe (Moffitt et al., 2007).
The general phenylpropanoid metabolism generates various common known secondary metabolites such as lignin or flavonoid and other aromatic metabolites such as coumarins, phenolic volatiles, or hydrolyzable tannins. The genes of the shikimate pathway have partly originated from the prokaryotic ancestor of cyanobacterial origin (Vogt, 2010).
The chemical activities of polyphenols in terms of their reducing properties as hydrogen or electron donating agents predict their potential for action as free-radical scavengers (antioxidants). The activity of an antioxidant is determined by its reactivity as a hydrogen or electron donating agent (which is related to its reduction potential). Polyphenols possess ideal structural chemistry for free radical-scavenging activities and have been shown to be more effective antioxidants in vitro than vitamins E and C on a molar basis (Rice-Evans, Miller, & Paganga, 1997).
Cyanobacteria are prokaryotic organisms that have served as important model organisms for studying oxygenic photosynthesis and have played a significant role in the Earth’s history as primary producers of atmospheric oxygen. Synechocystis sp. PCC 6803 was the first cyanobacteria that had its genome sequenced in 1996 (Nakao et al, 2010). In contrast, limited screening programs have been conducted on cyanobacteria due to the scarcity of expertise in physiological research and industrial production (Natrah, Yusoff, Shariff, Abas, & Mariana, 2007).
Plants possess many compounds, but their rapid production and usage for industrial and medical purposes is not easy, because plant cultivation is dependent on local soil and climatic conditions, and their attainment of optimal growth takes a longer time. In this study, we aimed at investigating some cyanobacteria isolates for the relationship between their PAL enzyme activity, total phenolics, and antioxidant activities for the first time. Determination of a cyanobacteria isolate that has a high and inducable PAL activity would create new evaluation areas in its industrial usage, and also cyanobacteria can serve as a potential resource in rapid production of the PAL enzyme.
Section snippets
Culture conditions and microorganisms
The isolates representing filamentous and non-filamentous cyanobacteria forms from different water sources were obtained from the Gazi University Faculty of Science, Biotechnology Laboratory culture collection. A confirmatory sequence analysis also was achieved for the study.
Isolation and purification of the isolates were performed by dilution and plating water samples. The isolates were cultured in BG11 medium (Rippka, Deruelles, Waterbury, Herdman, & Stanier, 1979) at 25 °C with a light/dark
Results and discussion
Current enzyme replacement therapies for PKU are focused on PAL, which has many advantages compared with phenylalanine hydroxylase (PAH) enzyme that is produced in the body to degrade l-phe, and recently, cyanobacterial PALs are evaluated for the design and construction of an effective protein-based treatment for PKU (Moffitt et al. 2007).
In the present study, PALs from the cyanobacteria are particular to Synechocystis, Leptolyngbya, and Oscillatoria species and are described for the first
Conclusions
Antioxidant and PAL activity of cyanobacterial isolates was reported for the first time together. The results of the study also showed that the microalgae have the possibility of usage as natural antioxidant sources. PALs isolated from different organisms so far exhibit problems related to administration, immunogenecity, and stability. With such screen studies, we may have the chance to find a candidate cyanobacteria that has the potential to be used as a PAL source in the tratment of
References (30)
Rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding
Analytical Biochemistry
(1976)- et al.
Action of phenolic derivates (acetoaminophen, salicylate and 5-aminosalicylate) as inhibitors of membrane lipid peroxidation and as peroxyl radical scavengers
Archives of Biochemistry and Biophysics
(1994) - et al.
Cloning and expression of a phenylalanine ammonia-lyase gene (BoPAL2) from Bambusa oldhamii in Escherichia coli and Pichia pastoris
Protein Expression and Purification
(2010) - et al.
Evaluation of antioxidant capacity and total phenolic content of different fractions of selected microalgae
Food Chemistry
(2007) - et al.
Antioxidant capacity of some edible and wound healing plants in Oman
Food Chemistry
(2007) - et al.
Antioxidant properties of phenolic compounds
Trends in plant science
(1997) - et al.
Homeostasis between lipid peroxidation and antioxidant enzyme activities in healthy human aging
Mechanisms of Ageing and Development
(1992) - et al.
Dietary oxidative stress and antioxidant defense with an emphasis on plant extract administration
Cell Stress and Chaperones
(2012) Phenylpropanoid biosynthesis
Molecular Plant
(2010)- et al.
Antimicrobial and antioxidant activities of Russula delica Fr
Food and Chemical Toxicology
(2009)
Production of phenolic compounds from Spirulina maxima microalgae and its protective effects
African Journal of Biotechnology
Chemical, antioxidant and antimicrobial investigations of Pinus cembra L. bark and needles
Molecules
Antioxidant determinations by the use of a stable free radical
Nature
Nitric Oxide alleviates oxidative damage in the green algae Chlorella pyrenoidosa caused by UV-B radiation
Folia Microbiologica
Ethanol synthesis by genetic engineering in cyanobacteria
Applied and Environmental Microbiology
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