Computational Approach for Enzymes Present in Capsicum Annuum

Review Paper

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ch permitsch Phosphate 2012), blood k is properlyk is cited. All rights reserved Allrights et al., , , Enzymes, ExPAsy www.ijddr.in Int. J. Drug Dev. & Res & Dev. Drug J. Int. et al., Open Access Contents Access Open Copyright © 2013 By IYPF By 2013 © Copyright 2012a), antibacterial effect of e helpful for application these for of helpful e Capsicum annuum: Capsicum peroxidase, polyphenol oxida viz. et al., Capasicum annuum developing degenerative macular diseases degeneration in women such (Moller et 2006). al., It as also inhibit carcinogenic activity (Nishino et al., 2002) and posterior reduce subcapsular (50% cataracts reduced in subjects rate) with higher plasma lutein concentration (Gale et al., 2003). An inclusion complex of red bell pigments with pepper β-cyclodextrin is used a as natural colorant in yogurt (Lidiane et al., 2013). antimicrobial antimicrobial effect of (Shivendu garlic and cinnamon marigold leaves (Nandita computational approach approach. should be biological applied Computational properties to and understand its mechanism trends of research is continuously shifting towards nanotechnology (Kingsley coagulation property of marigold leaves or sweet bell pepper is one of the more economical

Int.Drug J. Dev. Res.,& October -December 2013, 5(4): her her medicinal properties. This review highlights the isher and licensee and isher IYPF. This is Access Open an article whi uniprot, swissprot and Brenda enzyme db and ExPAsy

sing conditions for e.g. shelf life of fruits and vegetables can bstract: bstract: eywords: Epimerase contribute to its properties by various molecular mechanisms. Capasicum annuum asparaginase, Polygalactouronase, Capsanthin and Ribulose- 3- mechanisms these Understandingwillb of enzymes in food processing and in the production of food ingredients. increasing sophistication of processing industries creates a demand for a broad variety of proces enzymes with characteristics decreasingincreased by peroxidaselevels of the polyphenoloxidase. and compatible with food K DB. protein A and agriculturally viable vegetable grown all over the world owing to antioxidant i and ot essential enzymes present and its online mode tools of action using bioinformatics protein databases. The enzymes tyrosinase, catecholase, Pectin esterase, Catalase, 9-lipoxygenase, L

genera and all are widely f f Biosciences & Capsicum © 2013 Ramalinagam Chidambaram ntroduction cultivated cultivated throughout world. studied have higher specific activity Most and protein enzymes content in green than in the red pepper (Mateos et al., 2003). properties are It’s the main attributes medicinal to be focused and research on purposes. Further, bellpepper issweet antioxidant also a good source of bioactive compounds, such as the carotenoids (Kim substances can et substantially al., reduce the 2004). risk of These Capsicum annuum (Sweet bell common ingredient of staple food. In Brazil and in pepper) is a some other countries, the sweet bell considered pepper is one commodities in terms of of production volume and the commercially (De et al., 2009). There more are various species from important I

CoveredEmbase,&Scopus in Elsevier Technology Vellore-14 University, VIT Tamilnadu,India email: [email protected], School of Bio Sciences and Sciences Bio of School Technology, University, VIT Tamilnadu,India CorrespondingAuthors: RamalingamChidambaram Professor, School o ShivenduRanjan Dasgupta Nandita Arkadyuti RoyChakraborty Melvin SamuelS Pradeep SinghJadon Chidambaram* Ramalinagam

A reviewA

Computational Computational forapproach enzymespresent in

Page 88 (Shivendu et al., 2012b). This review provides some hydrogen peroxide as the electron acceptor to

essential information about the enzymes present in catalyse a number of oxidative reactions. Most capsicum annuum retrieved from online haem peroxidases follow the given reaction database viz. swissprot, uniprot, ExPAsy, and scheme:

BRENDA ENZYME DB. Fe 3+ + H 2O2 [Fe 4+ =O]R' (Compound I) + H 2O UniProt (launched on 15 December 2003) [Fe 4+ =O]R' + substrate [Fe 4+ =O]R (Compound II) comprises of three components: Firstly, the UniProt + oxidised substrate

Knowledgebase which will continue the work of [Fe 4+ =O]R + substrate Fe 3+ + H 2O + oxidised Swiss-Prot, TrEMBL and PIR by providing an expertly substrate Capsicumannuum curated database; Secondly, the UniProt Archive The enzyme reacts with one equivalent of H 2O2 to (UniParc) which is updated with novel sequences give [Fe 4+ =O]R' (compound I). This is a daily; Thirdly, the UniProt non-redundant reference oxidation/reduction reaction involves transfer of

databases (UniProt NREF), which provide concise two electrons where H 2O2 is reduced to water and views on top of the UniProt Knowledgebase and the enzyme is oxidised. One oxidising equivalent UniParc (Rolf et al., 2004). Swiss-Prot is the leading resides on iron, giving the oxyferryl intermediate universal curated protein sequence database, (Nelson et al., 1994), while in many peroxidases ymes present in which contained as of November 2003 (release the porphyrin (R) is oxidised to the porphyrin pi-

42.6) contains 140,000 curated sequence entries cation radical (R'). Compound I then oxidises an 89 Page from over 8300 different species. Being non- organic substrate to give a substrate radical (Li redundant, means that all reports for a given and Poulos, 1994). protein are merged into a single entry, and is POD mainly oxidises phenols to phenoxy radicals, highly integrated with other databases (Gasteiger which participate in reactions where polymers et al., 2001). ExPASy is the SIB Bioinformatics and oligomers are produced which are further Resource Portal which provides access to responsible for enzymatic browning and off-flavor scientific databases and software tools in different development (Morimoto et al., 2000). It is involved areas of life sciences including proteomics, in photorespiration and assimilation of genomics, phylogeny, systems biology, population symbiotically induced nitrogen in plant-specific genetics, transcriptomics etc (Panu et al., 2012). functions. Recent postgenetic approaches in BRENDA (BRaunschweig ENzyme DAtabase) was transcriptome and proteome showed that plant created in 1987 at the German National Research peroxisomes have more important roles in various Center for Biotechnology at Braunschweig and is metabolic processes including biosynthesis of currently being maintained by the University of plant hormones (Mano and Nishimura, 2005). Cologne (Priti et al., 2003). POD can be found in plants, animals and microbes. It is one of the most thermostable Peroxidase (EC number: 1.11.1) enzymes performing single electron oxidation on a It belongs to a large Family of Class III Plant wide variety of compounds in the presence of

Peroxidases i.e. Oxidoreductases. It acts on hydrogen peroxide. POD reduces H 2O2 to water peroxide as an acceptor. Further peroxidases while oxidizing a variety of other substrates. The (POD) are haem-containing enzymes that use catalytic process of POD occurs in a multi step

ShivenduRanjan et al; Computational approach for enz Covered in Scopus & Embase, Elsevier Int. J. Drug Dev. & Res., October -December 2013, 5 (4): 88-97 © 2013 Ramalinagam Chidambaram et al, publisher and licensee IYPF. This is an Open Access article which permits unrestricted noncommercial use, provided the original work is properly cited.

reaction. This is shown in the following scheme, reaction of which is shown in Figure 1 (Prontipa et where S stands for substrate and S • represents the al., 2010).

corresponding radical. AH 2 and AH • represent a The browning process can be caused by both reducing substrate and its radical product, enzymatic and non-enzymatic biochemical respectively (Lengauer and Rarey, 1996). reactions. Polyphenol oxidase (PPO) and peroxidase (POD) are two well known enzymes POD + H 2O2 Compound I + H 2O involved in the browning process (Sapers, 1993; Compound I+S Compound II + S • Compound II + S • POD+S • Chen et al., 2000; Deepa et al., 2007). PPO A simplified equation for peroxidase activity may catalyzes the conversion of phenolic compounds be given as: H2O2+ 2AH 2 2H 2O + AH • to quinones and assists their products’

polymerization in the presence of oxygen leading Furthermore, the related activity of PPO and POD to the formation of undesirable brown pigments is due to the generation of hydrogen peroxide and off-flavored products. The browning of during the oxidation of phenolic compounds in injured, peeled or diseased fruit tissues can causes PPO-catalyzed reactions (Francisco and Juan, undesirable changes in quality during handling, 2001; Khan and Robinson, 1994; Sugai and Tadini, processing or storage (Yemenicioglu et al., 1999).

2006). Peroxidise is the enzyme responsible for off- Review Different forms of PPO are found in Capsicum flavour development, so its inactivation will inhibit annuum , such as tyrosinase (Nisit et al., 2007), Page 90 Page any such off-flavour development (Yemenicioglu cathecolase (Deepa et al., 2007). et al., 1999). Tyrosinase/Cresolase/Phenolase (EC 1.14.18.1) Polyphenol Oxidase (EC number: 1.14.18.1) Generally known as Tyrosinase but can also be Paper It also belongs to a large family of Class III Plant named as monophenol monooxygenase, Peroxidases i.e. Oxidoreductases enzyme family so phenolase, monophenol oxidase, cresolase, its mode of action is similar to that of POD i.e. acts monophenolase, tyrosine-dopa oxidase, on paired donors with O 2 as the oxidant and monophenol monooxidase, incorporation or reduction of oxygen. The monophenoldihydroxyphenylalanine oxygen incorporated oxygen need not be derived from oxidoreductase, N-acetyl-6-hydroxytryptophan O2; or with another compound as a single donor, oxidase, monophenol, dihydroxy-L-phenylalanine and incorporation of one oxygen atom into the oxygen oxidoreductase, o-diphenol oxygen other donor (Francisco and Juan, 2001; Khan and oxidoreductase, phenol oxidase. Its synthetic Robinson, 1994). Polyphenol oxidase (PPO) is a name is L-tyrosine,L-dopa:oxygen oxidoreductase dicopper-containing enzyme which is involved in (BRENDA, 2013; ExPAsy, 2013). the oxidation of the polyphenols from plants. PPO It is type III copper protein found in a variety of shows a higher activity with diphenols (Onsa et al., bacteria, fungi, plants, insects, crustaceans, and 2000). Two kinds of reactions catalyzed by PPO mammals, which is involved in the synthesis of are the hydroxylation of monophenols to o- betalains and melanin. The enzyme, which is diphenol and the oxidation of o-diphenol to o- activated upon binding molecular oxygen, can quinone (Francisco and Juan, 2001) schematic catalyse both a monophenolase reaction cycle (reaction 1) or a diphenolase reaction cycle Covered in Scopus & Embase, Elsevier Int. J. Drug Dev. & Res., October -December 2013, 5 (4): 88-97 © 2013 Ramalinagam Chidambaram et al, publisher and licensee IYPF. This is an Open Access article which permits unrestricted noncommercial use, provided the original work is properly cited. (reaction 2). During the monophenolase cycle, oxidation of catechols (Mayer and Harel, 1979;

one of the bound oxygen atoms is transferred to a Gerdemann et al., 2002).

monophenol (such as L-tyrosine), generating an o- 2 catechol + O 2 = 2 1,2-benzoquinone + 2 H2O diphenol intermediate, which is subsequently Pectinesterase (EC 3.1.1.11) oxidized to an o-quinone and released along with Pectinesterase which can also be called as pectin a water molecule. The enzyme remains inactive in demethoxylase, pectin methoxylase, pectin its deoxy state and is further restored to the active methylesterase, pectase, pectin methyl esterase oxy state by the binding of a new oxygen and pectinoesterase. The Systematic name is molecule. During the diphenolase cycle the pectin pectylhydrolase (BRENDA, 2013; ExPAsy, Capsicumannuum enzyme binds to an external diphenol molecule 2013). Pectin in presence of water is converted (such as L-dopa) and oxidizes it to an o-quinone into n methanol and pectate with the help of that is released along with a water molecule, enzyme pectin esterase (Figure 2).

leaving the enzyme in the intermediate met state. pectin + n H2O = n methanol + pectate The enzyme then binds to a second diphenol It is involved in the regulation of the cell wall molecule and repeats the process, thereby rigidity and also plays a key role in process of fruit ending in a deoxy state (Rolff et al., 2011; Steiner ripening (Castro et al., 2004). Temperature stability ymes present in et al., 1996). is indicated by presence of 75% enzyme activity at

Reaction (1) 55°C, 10% activity activity activity at 75°C and 5% 91 Page L-tyrosine + O 2 = dopaquinone + H 2O (overall at 85°C, maximum stability of it is found upto a reaction) (1a) L-tyrosine + ½ O2 = L-dopa maximum temperature of 95°C (Mejia-Cordova et (1b) L-dopa + ½ O2 = dopaquinone + H 2O al., 2005). Reaction (2) 2 L-dopa + O 2 = 2 dopaquinone + 2 H 2O 7. Catalase (Ec 1.11.1.6)

The enzyme Catalase is also referred as equilase, Catechol oxidase/ Catecholase ( EC 1.10.3.1 ) caperase, optidase, catalase-peroxidase, CAT Generally known as Catechol oxidase but can and its systematic name is hydrogen- additionally called as diphenol oxidase, o- peroxide:hydrogen-peroxide – oxidoreductase diphenolase, polyphenol oxidase, pyrocatechol (BRENDA, 2013; ExPAsy, 2013). oxidase, dopa oxidase, catecholase, o-diphenol It is an oxidoreductase by classification and its oxygen oxidoreductase and o-diphenol mode of action is the same as others mentioned oxidoreductase, but the synthetic name is 1,2- previously i.e. it reduces H 2O2 to water ( 2 H2O2 = O 2 benzenediol:oxygen oxidoreductas (BRENDA, + 2 H2O) (Dounce, 1983). This enzyme can also act 2013; ExPAsy, 2013). as a peroxidase for which several organic It is also a type 3 copper protein that catalyses the substances like ethanol can act as a hydrogen oxidation of catechol (i.e., o-diphenol) to the donor. A manganese protein containing Mn III in corresponding o-quinone exclusively. The enzyme the resting state, which also belongs here, is often also acts on a variety of substituted catechols. called pseudocatalase (Kono and Fridovich, Unlike tyrosinase which can catalyse both the 1983). It is found in stems, leaves, roots and monooxygenation of monophenols and the developing fruits of capsicum annuum and serves to protect cells from the toxic effects of hydrogen

peroxide by reducing it to water. There are 492 al., 2007). It is responsible for asparagine amino acids present in it (Kwon and An, 2001). degradation (Yao et al., 2005) and is involved in the interconversion of aspartate and asparagines Linoleate 9S-lipoxygenase /9-lipoxygenase (EC (Falzone et al., 1988). 1.13.11.58) It is found in Capsicum annuum, Datura inoxia, The above enzyme can also be termed as 9- Pisum sativum, Solanum lycopersicum, Tamarindus lipoxygenase, 9 S-lipoxygenase, linoleate 9- indica, Vigna unguiculata, Withania somnifera etc lipoxygenase, LOX1 (gene name), 9S-LOX, and with the function of asparatate degradation and the synthetic names is linoleate:oxygen 9 S- is also involved in the interconversion of aspartate oxidoreductase (BRENDA, 2013; ExPAsy, 2013). It and asparagines (Oza et al., 2009). converts linolate to 9-hydroperoxy-10,12- Polygalactouronase/PG (EC 3.2.1.15) octadecadinoate in presence of oxygen. (Gigot It can also be called as pectin depolymerise, et al., 2010) pectinase, endopolygalacturonase, pectolase, Overall Reaction: pectin hydrolase, pectin polygalacturonase,

Linoleate + O 2 = (9 S,10 E,12 Z)-9-hydroperoxy-10,12- endo-polygalacturonase, poly-α-1,4- octadecadienoate galacturonide glycanohydrolase, Review It contains a nonheme iron and a common plant endogalacturonase and endo-D-galacturonase. lipoxygenase that oxidizes linoleate and alpha- Its synthetic name is poly (1, 4-α-D-galacturonide) Page 92 Page linolenate, the two most common glycanohydrolase (BRENDA, 2013; ExPAsy, 2013). It polyunsaturated fatty acids in plants by inserting is involved in the random hydrolysis of (1→4)-α-D-

molecular oxygen at the C9 position with (S)- galactosiduronic linkages in pectate and other Paper configuration. The enzyme plays a physiological galacturonans. It splits pectin chain by adding a role during the early stages of seedling growth molecule of water and breaks the linkage (Bannenberg et al., 2009). between two galacturonan units. These enzymes

In capsicum annuum it regulates defense and cell catalyze reactions that break α- 1,4-glycosidic death response to microbial pathogens. The bonds (Huang et al., 1990). optimum temperature for its activity is 25°C with PG activity increases during ripening. Ripening an optimum pH of 6 ranging upto a mximum of pH specific pectin methylesterase, active in vivo, 6.5 (Hwang and Hwang, 2010). appears to enhance PG-mediated pectin ultra- degradation, resulting in cell wall dissolution and L-Asparaginase/ L-asparaginase (EC 3.5.1.1) emergence of deciduous fruit traits. Some pectin Its accepted name is asparaginase and can be methylesterase isozymes are apparently inactive also called as asparaginase II, L-asparaginase, in vivo, particularly in green fruit and throughout colaspase, elspar, leunase, crasnitin and α- ripening in the ‘Hard Pick’ line, thereby limiting PG- asparaginase. The synthetic name is L-asparagine mediated pectin depolymerization which results in amidohydrolase (BRENDA, 2013; ExPAsy, 2013). It moderately difficult fruit separation from the calyx catalyzes the reaction in which asparaginase is (Rancibia and Motsenbocker, 2006). It is also involved in cells (Figure 3) overall reaction is as; L- involved in homogalacturonan degradation

asparagine + H 2O = L-aspartate + NH 3 (Narta et (Willats et al., 2001). Exo-poly-α-D- Covered in Scopus & Embase, Elsevier Int. J. Drug Dev. & Res., October -December 2013, 5 (4): 88-97 © 2013 Ramalinagam Chidambaram et al , publisher and licensee IYPF. This is an Open Access article which permits unrestricted noncommercial use, provided the original work is properly cited. galacturonosidase (EC 3.2.1.82) (exoPG) phosphoketopentose epimerase, ribulose 5-

hydrolyzes peptic acid from the non-reducing phosphate 3-epimerase, D-ribulose phosphate-3- end, further releasing digalacturonate (He and epimerase, D-ribulose 5-phosphate epimerase, D- Collmer, 1990). It is found in Capsicum frutescens ribulose-5-P 3-epimerase, D-xylulose-5-phosphate

annuum and capsicum annuum with the similar functions 3-epimerase, pentose-5-phosphate 3-epimerase

of ripening (Gong et al., 2008). and Systematic name is D-ribulose-5-phosphate 3- epimerase (BRENDA, 2013; ExPAsy, 2013). Enzyme Capsanthin (EC 5.3.99.8) ribulose-phosphate 3-epimerase is involved in the Its accepted name is capsanthin/capsorubin conversion of D-ribulose 5-phosphate into D- Capsicum synthase and some other names are as CCS, xylulose 5-phosphate as shown in Figure 4 (Thom ketoxanthophyll synthase, capsanthin-capsorubin et al., 1998) synthase and Systematic name is violaxanthin- In capsicum it is mainly involved in the pentose- capsorubin isomerise (BRENDA, 2013; ExPAsy, phosphate pathway, although it has many 2013). Capsanthin catalyses two reactions from functions like the Bifidobacterium shunt, Calvin- substrate antheraxanthin and violaxanthin which Benson-Bassham cycle, D-arabinose degradation produce capsanthin and capsorbin respectively II, formaldehyde assimilation II (RuMP Cycle), ymes present in (Bouvier et al., 1994). formaldehyde assimilation III (dihydroxyacetone This multifunctional enzyme is induced during the cycle), heterolactic fermentation, pentose Page 93 Page chromoplast differentiation in plants; the gene is phosphate pathway (non-oxidative branch) and specifically expressed during chromoplast the ribitol degradation (Thom et al., 1998). development in fruits thereby accumulating ketocarotenoids but not in mutants impaired in this biosynthetic step (Bouvier et al., 1994). In capsicum annuum it is involved in apsanthin and capsorubin biosynthesis (Al-Babili et al., 2000). It Figure 1 : Hydroxylation of monophenols to diphenols and to o-quinonne has relation with molecular genetics as y locus in

pepper is responsible for capsanthin-capsorubin synthase which ultimately relates to the color of capsicum annuum (Popovsky and Paran, 2000). The capsanthin-capsorubin synthase gene is

activated specifically during the final stages of Capsicum fruit ripening (Lefebvre et al., 1998). Figure 2: Overall reaction in which enzyme asparaginase is involved

Ribulose-Phosphate 3-Epimerase (EC 5.1.3.1) Its accepted name is ribulose-phosphate 3- epimerase but follows some other names such as phosphoribulose epimerase, erythrose-4- phosphate isomerise, phosphoketopentose 3- epimerase, xylulose phosphate 3-epimerase,

Figure 3: Overall reaction in which ribulose- 10) Falzone CJ, Karsten WE, Conley JD, Viola RE (1988) L- phosphate 3-epimerase is involved aspartase from Escherichia coli : substrate specificity

and role of divalent metal ions. Biochemistry 27(26): References 9089-93. 11) Francisco ATB, Juan Carlos E (2001) Phenolic 1) Al-Babili S, Hugueney P, Schledz M, Welsch R, compounds and related enzymes as determinants Frohnmeyer H, Laule O, Beyer P (2000) Identification of quality in fruits and vegetables. J Sci Food Agric of a novel gene coding for neoxanthin synthase 81: 853–876. from Solanum tuberosum. FEBS Lett 485(2-3):168-72. 12) Gasteiger E, Jung E, Bairoch A (2001) SWISS-PROT: 2) Bannenberg G, Martinez M, Hamberg M, connecting biomolecular knowledge via a protein Castresana C (2009) Diversity of the enzymatic database. Curr Issues Mol Biol 3 (3); 47-55. activity in the lipoxygenase gene family of 13) Gerdemann C, Eicken C, Krebs B (2002) The crystal Arabidopsis thaliana . Lipids 44 :85–95. structure of catechol oxidase: new insight into the 3) Bouvier F, Hugueney P, d'Harlingue A, Kuntz M, function of type-3 copper proteins. Acc Chem Res Camara B (1994) Xanthophyll biosynthesis in 35: 183–191. chromoplasts: isolation and molecular cloning of an 14) Gong Z, Yang R, Li D (2008) Specific expression gene enzyme catalyzing the conversion of 5,6- from Capsicum annuum after inoculated with epoxycarotenoid into ketocarotenoid. Plant J 6: 45– Phytophthora capsici.; Submitted (DEC-2008) to the 54. EMBL/GenBank/DDBJ databases. 4) Gale CR, Hall NF, Phillips DIW, Martyn CN (2003) Review 15) He SY, Collmer A (1990) Molecular cloning, Lutein and zeaxanthin status and risk of age-related nucleotide sequence, and marker exchange

Page 94 Page macular degeneration. Investigative mutagenesis of the exo-poly-alpha-D- Ophthalmology Visual Science 44: 2461–2465. galacturonosidase-encoding pehX gene of Erwinia 5) Castro SM. Van Loey A, Saraiva JA, Smout C, chrysanthemi EC16. J Bacteriol 172: 4988-4995.

Hendrickx M (2004) Activity and process stability of 16) Huang JH, Schell MA (1990) DNA sequence analysis Paper purified green pepper ( Capsicum annuum ) pectin of pglA and mechanism of export of its methylesterase. J Agric Food Chem: 52: 5724-5729. polygalacturonase product from Pseudomonas 6) Cédric Gigot, Marc Ongena, Marie-Laure solanacearum . J Bacteriol 172: 3879-3887. Fauconnier, Jean-PaulWathelet, Patrick Du Jardin,

17) Hwang IS, Hwang BK (2010) The pepper 9- Philippe Thonart (2010) The lipoxygenase metabolic lipoxygenase gene CaLOX1 functions in defense pathway in plants: potential for industrial production and cell death responses to microbial pathogens. of natural green leaf volatiles. Biotechnol Agron Soc Plant Physiol 152: 948-967. Environ 14(3): 451-460. 18) Kengo Morimoto, Kenji Tatsumi, Ken-Ichi Kuroda 7) Chen L, Mehta A, Berenbaum M, Zangerl AR, (2000) Peroxidase catalyzed co-polymerization of Engeseth NJ (2000) Honeys from different floral pentachlorophenol and a potential humic sources as inhibitors of enzymatic browning in fruit precursor. Soil Biology & Biochemistry 32: 1071-1077. and vegetable homogenates. J Agric Food Chem 19) Khan AA, Robinson DS (1994) Hydrogen donor 48(10): 4997-5000. specificity of mango isoperoxidases. Food Chem. 8) De Azevedo-Meleiro CH, Rodriguez-Amaya DB 49: 407–410. (2009) Qualitative and quantitative differences in 20) Kono Y, Fridovich I (1983) Isolation and the carotenoid composition of yellow and red characterization of the pseudocatalase of peppers determined by HPLC-DAD-MS. J Sep Sci Lactobacillus plantarum . J Biol Chem 258: 6015– 32(21): 3652-3658. 6019. 9) Dounce AL (1983) A proposed mechanism for the 21) Kwon SI, An CS (2001) Molecular cloning, catalatic action of catalase. J Theor Biol 105(4): 553- characterization and expression analysis of a 67.

Covered in Scopus & Embase, Elsevier Int. J. Drug Dev. & Res., October -December 2013, 5 (4): 88-97 © 2013 Ramalinagam Chidambaram et al, publisher and licensee IYPF. This is an Open Access article which permits unrestricted noncommercial use, provided the original work is properly cited. catalase cDNA from hot pepper ( Capsicum 32) N Deepa, Charanjit Kaur, Binoy George, Balraj

annuum L .). Plant Sci 160(5): 961-969. Singh, Kapoor HC (2007) Antioxidant constituents in 22) Lefebvre V, Kuntz M, Camara B, Palloix A (1998) The some sweet pepper ( Capsicum annuum L .) capsanthin-capsorubin synthase gene: a candidate genotypes during maturity. LWT - Food Science and gene for the y-locus controlling the red fruit colour in Technology 40 (1): 121–129. pepper. Plant Mol Biol 36: 785-789. 33) Kingsley JD, Shivendu R, Nandita D, Proud S (2013) 23) Lengauer T, Rarey M (1996) Review Computational Nanotechnology for tissue engineering: Need, methods for biomolecular docking. Curr Opin Struct techniques and applications. Journal of pharmacy Biol 6(3): 402-406. research 7: 200-204. 24) Li H, Poulos TL (1994) Structural variation in heme 34) Nandita D, Shivendu R, Proud S, Rahul J, Swati M,

Capsicumannuum enzymes: a comparative analysis of peroxidase and Arabi MSMA (2012) Antibacterial activity of leaf P450 crystal structures. Structure 2(6): 461-464. extract of Mexican marigold (Tagetes erecta) 25) Lidiane Martins MG, Nicolly P, Valéria GC, Deborah against different gram positive and gram negative QF, Kátia G, de Lima A (2013) Inclusion complexes bacterial strains. Journal of Pharmacy Research of red bell pepper pigments with β-cyclodextrin: 5(8): 4201-4203. Preparation, characterisation and application as 35) Nelson RE, Fessler LI, Takagi Y, Blumberg B, Keene natural colorant in yogurt. Food Chemistry. doi: DR, Olson PF, Parker CG, Fessler JH (1994) 10.1016/j.foodchem.2012.09.065. Peroxidasin: a novel enzyme-matrix protein of ymes present in 26) Mano S, Nishimura M (2005) Plant peroxisomes. Drosophila development. EMBO J 13(15): 3438-3447. Vitam Horm 72: 111-54. 36) Nishino H, Murakoshi M, Ii T, Takemura M, Kuchide M, 27) Mateos RM, León AM, Sandalio LM, Gómez M, del Kanazawa M, Mou XY, Wad S, Masuda M, Ohsaka Y, Page 95 Page Río LA, Palma JM (2003) Peroxisomes from pepper yogosawa S, Satomi Y, Jinno K (2002) Carotenoids in fruits ( Capsicum annuum L .): purification, cancer chemoprevention. Cancer Metastasis Rev characterisation and antioxidant activity. J Plant 21: 257-264. Physiol 160(12): 1507-16. 37) Nisit Kittipongpatana, Piyamart Maneerat, 28) Mayer AM, Harel E (1979) Polyphenol oxidases in Patchanee Pattanakitkosol, Ornanong S plants. Phytochemistry 18(2): 193–215. Kittipongpatana (2007) Effect of Some Factors on 29) Mejia-Cordova SM, Montanez JC, Aguilar CN, de la the Growth of Capsicum annuum L . Cell Suspension Luz Reyes-Vega M, de la Garza H, Hours RA, Culture and Biotransformationof Hydroquinone to Contreras-Esquivel (2005) Extraction of Arbutin. CMU J Nat Sci 6(2): 207-218. pectinesterase from Jalapeno chili pepper 38) Onsa GH, bin Saari N, Selamat J, Bakar J (2000) (Capsicum annuum ) and its thermal stability. Food Latent polyphenol oxidases from sago log Sci. Biotechnol. 14: 185-189. (Metroxylon sagu): partial purification, activation, 30) Moeller SM, Parekh N, Tinker L, Ritenbaugh C, Blodi and some properties. J Agric Food Chem 48(10): B, Wallace RB, Mares JA, CAREDS Research Study 5041-5045. Group (2006) Associations between intermediate 39) Oza VP, Trivedi SD, Parmar PP, Subramanian RB age-related macular degeneration and lutein and (2009) Withania somnifera (Ashwagandha): a novel zeaxanthin in the Carotenoids in Age-related Eye source of L-asparaginase. J Integr Plant Biol 51: 201- Disease Study (CAREDS): ancillary study of the 206. Women's Health Initiative. Arch Ophthalmol. 124(8): 40) Panu Artimo, Manohar Jonnalagedda, Konstantin 1151-1162. Arnold, Delphine Baratin, Gabor Csardi, Edouard de 31) Narta UK, Kanwar SS, Azmi W (2007) Castro, Séverine Duvaud, Volker Flegel, Arnaud Pharmacological and clinical evaluation of Fortier, Elisabeth Gasteiger, Aurélien Grosdidier, lasparaginase in the treatment of leukemia. Crit Rev Céline Hernandez, Vassilios Ioannidis, Dmitry Oncol Hematol 61 (3): 208-221. Kuznetsov, Robin Liechti, Sébastien Moretti, Khaled

Mostaguir, Nicole Redaschi, Grégoire Rossier, Ioannis 49) S Kim, J Park, IK Hwang (2004) Composition of main Xenarios, Heinz Stockinger (2012) ExPASy: SIB carotenoids in Korean red pepper ( Capsicum bioinformatics resource portal. Nucleic Acids Res annuum, L. ) and changes of pigment stability during doi: 10.1093/nar/gks400. the drying and storage process. Journal of Food 41) Priti Pharkya, Evgeni V Nikolaev, Costas D Maranas Science 69: 39 - 44. (2003) Review of the BRENDA Database. Metabolic 50) Sapers GM (1993) Browning of foods: Control by Engineering 5(2): 71–73. sulfites, antioxidants, and other means. Food 42) Prontipa Nokthai, Vannajan Sanghiran Lee, Lalida Technol 68:5–84. Shank (2010) Molecular Modeling of Peroxidase and 51) Steiner U, Schliemann W, Strack D (1996) Assay for Polyphenol Oxidase: Substrate Specificity and tyrosine hydroxylation activity of tyrosinase from Active Site Comparison. Int J Mol Sci 11 (9): 3266– betalain-forming plants and cell cultures. Anal 3276. Biochem 238: 72–75. 43) Popovsky S, Paran I (2000) Molecular genetics of the 52) Sugai AY, Tadini CC (2006) Thermal inactivation of y locus in pepper: its relation to capsanthin- mango ( Mangifera indica variety Palmer) puree capsorubin synthase and to fruit color. Theor Appl peroxidase. Proceedings of 2006 CIGR Section VI Genet 101: 86-89. International Symposium on Future of Food 44) Rancibia RA, Motsenbocker CE (2006) Pectin Engineering. Warsaw, Poland. 2006 methylesterase activity in vivo differs from activity in 53) The BRENDA enzyme database, Available online at

vitro and enhances polygalacturonase-mediated http://www.brenda-enzymes.org/ (Retrieved date: Review pectin degradation in tabasco pepper. J Plant 16 Feb 2013). Physiol 163: 488-496. 54) The ExPASy Enzyme database, Available online at Page 96 Page 45) Rolf Apweiler, Amos Bairoch, Cathy H Wu, Winona C http://www.enzyme.expasy.org/ (Retrieved date: 25 Barker, Brigitte Boeckmann, Serenella Ferro, Feb 2013) Elisabeth Gasteiger, Hongzhan Huang, Rodrigo 55) Thom E, Möhlmann T, Quick WP, Camara B,

Lopez, Michele Magrane, Maria J Martin, Darren A Neuhaus HE (1998) Sweet pepper plastids: enzymic Paper Natale, Claire O’Donovan, Nicole Redaschi, Lai-Su L equipment, characterisation of the plastidic Yeh (2004) UniProt: the Universal Protein oxidative pentose-phosphate pathway, and knowledgebase. Nucleic Acids Res doi: transport of phosphorylated intermediates across

10.1093/nar/gkh131. the envelope membrane. Planta 204: 226-233. 46) Rolff M, Schottenheim J, Decker H, Tuczek F (2011) 56) Willats WG, McCartney L, Mackie W, Knox JP (2001)

Copper-O2 reactivity of tyrosinase models towards Pectin: cell biology and prospects for functional external monophenolic substrates: molecular analysis. Plant Mol Biol 47(1-2): 9-27. mechanism and comparison with the enzyme. 57) Yao M, Yasutake Y, Morita H, Tanaka I (2005) Chem Soc Rev 40: 4077–4098. Structure of the type I L-asparaginase from the 47) Shivendu R, Nandita D, Proud S, Madhumita R, hyperthermophilic archaeon Pyrococcus horikoshii Ramalingam C (2012a) Comparative study of at 2.16 angstroms resolution. Acta Crystallogr D Biol antibacterial activity of garlic and cinnamon at Crystallogr 61(3): 294-301. different temperature and its application on 58) Yemenicioglu A, Özkan M, Cemeroglu B (1999) preservation of fish. Advances in Applied Science Some characteristics of polyphenol oxidase and Research 3(1): 495-501. peroxidase from taro ( Colocasia antiquorum ). Tr J 48) Shivendu R, Nandita D, Proud S (2012b) Time Agric Forestry 23: 425–430. dependent and spectrophotometric analysis of blood clotting property of marigold leaf extract and comparative study of its antibacterial activity. J Biotechnol Biomater 2(6): 276

Covered in Scopus & Embase, Elsevier Int. J. Drug Dev. & Res., October -December 2013, 5 (4): 88-97 © 2013 Ramalinagam Chidambaram et al, publisher and licensee IYPF. This is an Open Access article w hich permits unrestricted noncommercial use, provided the original work is properly cited.

Article History:------

Date of Submission: 22-04-2013

Date of Acceptance: 29-04-2013

Conflict of Interest: NIL

Source of Support: NONE

Capsicumannuum

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