Biochemical characterisation of chlorophyllase from leaves of selected Prunus species - A comparative study
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Despite senescence-induced chlorophyll depletion in plants has been widely studied, the enzymatic background of this physiologically regulated process still remains highly unclear. The purpose of this study was to determine selected biochemical properties of partially purified fractions of chlorophyllase (Chlase, chlorophyll chlorophyllido-hydrolase, EC 126.96.36.199) from leaves of three Prunus species: bird cherry (Prunus padus L.), European plum (Prunus domestica L.), and sour cherry (Prunus cerasus L.). Secondarily, this report was aimed at comparing seasonal dynamics of Chlase activity and chlorophyll a (Chl a) content within investigated plant systems. Molecular weight of native Chlase F1 has been estimated at 90 kDa (bird cherry) and approximately 100 kDa (European plum and sour cherry), whereas molecular mass of Chlase F2 varied from 35 kDa (European plum) to 60 kDa (sour cherry). Furthermore, enzyme fractions possessed similar optimal pH values ranging from 7.6 to 8.0. It was found that among a broad panel of tested metal ions, Hg+2, Fe+2, and Cu+2 cations showed the most pronounced inhibitory effect on the activity of Chlase. In contrast, the presence of Mg+2 ions influenced a subtle stimulation of the enzymatic activity. Importantly, although Chlase activity was negatively correlated with the amount of Chl a in leaves of examined Prunus species, detailed comparative analyses revealed an incidental decrement of enzymatic activity in early or moderately senescing leaves. It provides evidence that foliar Chlase is not the only enzyme involved in autumnal chlorophyll breakdown and further in-depth studies elucidating this catabolic process are required.
- Arkus KA, Cahoon EB, Jez JM (2005) Mechanistic analysis of wheat chlorophyllase. Arch Biochem Biophys 438: 146-155.
- Arkus KA, Jez JM (2006) Development of a high-throughput purification method and a continuous assay system for chlorophyllase. Anal Biochem 353: 93-98.
- Arriagada-Strodthoff P, Karboune S, Neufeld RJ, Kermasha S (2007) Optimization of chlorophyllase-catalyzed hydrolysis of chlorophyll in monophasic organic solvent media. Appl Biochem Biotechnol 142: 263-275.
- Azoulay-Shemer T, Harpaz-Saad S, Belausov E, Lovat N, Krokhin O, Spicer V, Standing KG, Goldschmidt EE, Eyal Y (2008) Citrus chlorophyllase dynamics at ethylene-induced fruit color-break: a study of chlorophyllase expression, posttranslational processing kinetics, and in situ intracellular localization. Plant Physiol 148: 108-118.
- Azoulay-Shemer T, Harpaz-Saad S, Cohen-Peer R, Mett A, Spicer V, Lovat N, Krokhin O, Brand A, Gidoni D, Standing KG, Goldschmidt EE, Eyal Y (2011) Dual N- and C-terminal processing of citrus chlorophyllase precursor within the plastid membranes leads to the mature enzyme. Plant Cell Physiol 52: 70-83.
- Banaś AK, Łabuz J, Sztatelman O, Gabryś H, Fiedor L (2011) Expression of enzymes involved in chlorophyll catabolism in Arabidopsis is light controlled. Plant Physiol 157: 1497-1504.
- Barry CS (2009) The stay-green revolution: Recent progress in deciphering the mechanisms of chlorophyll degradation in higher plants. Plant Sci 176: 325-333.
- Beisel KG, Jahnke S, Hofmann D, Köppchen S, Schurr U, Matsubara S (2010) Continuous turnover of carotenes and chlorophyll a in mature leaves of Arabidopsis revealed by 14CO2 pulse-chase labeling. Plant Physiol 152: 2188-2199.
- Ben-Yaakov E, Harpaz-Saad S, Galili D, Eyal Y, Goldschmidt E (2006) The relationship between chlorophyllase activity and chlorophyll degradation during the course of leaf senescence in various plant species. Isr J Plant Sci 54: 129-135.
- Büchert AM, Civello PM, Martínez GA (2011) Chlorophyllase versus pheophytinase as candidates for chlorophyll dephytilation during senescence of broccoli. J Plant Physiol 168: 337-343.
- Büchert AM, Civello PM, Martínez GA (2011) Effect of hot air, UV-C, white light and modified atmosphere treatments on expression of chlorophyll degrading genes in postharvest broccoli (Brassica oleracea L.) florets. Sci Hortic 127: 214-219.
- Chen L-FO, Lin C-H, Kelkar SM, Chang Y-M, Shaw J-F (2008) Transgenic broccoli (Brassica oleracea var. italica) with antisense chlorophyllase (BoCLH1) delays postharvest yellowing. Plant Sci 174: 25-31.
- Ciepiela AP, Sytykiewicz H, Czerniewicz P (2005) Changes in the activity of chlorophyllase and Mg-dechelatase in bird cherry leaves induced by Rhopalosiphum padi /L./ feeding. Aphids & Other Hemipterous Insects 11: 25-31.
- Cowan AK (2009) Plant growth promotion by 18:0-lyso-phosphatidylethanolamine involves senescence delay. Plant Signal Behav 4: 324-327.
- Criado MN, Motilva MJ, Ramo T, Romero MP (2006) Chlorophyll and carotenoid profile and enzymatic activities (chlorophyllase and lipoxygenase) in olive drupes from the fruit-setting period to harvest time. J Amer Soc Hort Sci 131: 593-600.
- Distefano G, Las Casas G, Caruso M, Todaro A, Rapisarda P, La Malfa S, Gentile A, Tribulato E (2009) Physiological and molecular analysis of the maturation process in fruits of clementine mandarin and one of its late-ripening mutants. J Agric Food Chem 57: 7974-4982.
- Fang Z, Bouwkamp JC, Solomon T (1987) Chlorophyllase activities and chlorophyll degradation during leaf senescence in non-yellowing mutant and wild type of Phaseolus vulgaris. J Exp Bot 49: 503-510.
- Fernandez-Lopez JA, Almela L, Almansa MS, Lopez-Roca JM (1992) Partial purification and properties of chlorophyllase from chlorotic Citrus limon leaves. Phytochemistry 31: 447-449.
- Gaffar R, Kermasha S, Bisakowski B (1999) Biocatalysis of immobilized chlorophyllase in a ternary micellar system. J Biotechnol 75: 45-55.
- Gómez-Lobato ME, Hasperué JH, Civello PM, Chaves AR, Martínez GA (2012) Effect of 1-MCP on the expression of chlorophyll degrading genes during senescence of broccoli (Brassica oleracea L.). Sci Hortic 144: 208-211.
- Gupta S, Gupta SM, Kumar N (2011) Role of chlorophyllase in chlorophyll homeostasis and post-harvest breakdown in Piper betle L. leaf. Indian J Biochem Biophys 48: 353-360.
- Gupta S, Gupta SM, Sane AP, Kumar N (2012) Chlorophyllase in Piper betle L. has a role in chlorophyll homeostasis and senescence dependent chlorophyll breakdown. Mol Biol Rep 39: 7133-7142.
- Harpaz-Saad S, Azoulay T, Arazi T, Ben-Yaakov E, Mett A, Shiboleth YM, Hörtensteiner S, Gidoni D, Gal-On A, Goldschmidt EE, Eyal Y (2007) Chlorophyllase is a rate-limiting enzyme in chlorophyll catabolism and is posttranslationally regulated. Plant Cell 19: 1007-1022.
- Hornero-Méndez D, Mínguez-Mosquera MI (2001) Properties of chlorophyllase from Capsicum annuum L. fruits. Z Naturforsch C 56: 1015-1021.
- Hörtensteiner S (2006) Chlorophyll degradation during senescence. Annu Rev Plant Biol 57: 55-77.
- Ihl M, Martin AS, Bifani V (2000) Chlorophyllase and quality of 'Granny Smith' apples during storage. Gartenbauwissenschaft 65: 266-271.
- Ihl M, Monsalves M, Bifani V (1998) Chlorophyllase inactivation as a measure of blanching efficacy and colour retention of artichokes (Cynara scylomus L.). Lebensm Wiss Technol 31: 50-56.
- Johnson-Flanagan AM, McLachlan G (1990) The role of chlorophyllase in degreening canola (Brassica napus) seeds and its activation by sublethal freezing. Physiol Plantarum 80: 460-466.
- Karboune S, Neufeld R, Kermasha S (2005) Immobilization and biocatalysis of chlorophyllase in selected organic solvent systems. J Biotechnol 120: 273-283.
- Kariola T, Brader G, Li J, Palva ET (2005) Chlorophyllase 1, a damage control enzyme, affects the balance between defense pathways in plants. Plant Cell 17: 282-294.
- Kuroki M, Shioi Y, Sasa T (1981) Purification and properties of soluble chlorophyllase from tea leaf sprouts. Plant Cell Physiol 22: 717-725.
- Laemmli UK (1970) Cleavage of structural protein during the assembly of the head of bacteriophage T4. Nature 227: 680-685.
- Layne E (1957) Spectrophotometric and turbidimetric methods for measuring proteins. Methods Enzymol 3: 447-455.
- Lee GC, Chepyshko H, Chen HH, Chu CC, Chou YF, Akoh CC, Shaw JF (2010) Genes and biochemical characterization of three novel chlorophyllase isozymes from Brassica oleracea. J Agric Food Chem 58: 8651-8657.
- Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265-275.
- Majumdar S, Ghosh S, Glick BR, Dumbroff EB (1991) Activities of chlorophyllase, phosphoenolpyruvate carboxylase and ribulose-1,5-bisphosphate carboxylase in the primary leaves of soybean during senescence and drought. Physiol Plantarum 8: 473-480.
- McFeeters RF, Chichester CO, Whitaker JR (1971) Purification and properties of chlorophyllase from Ailanthus altissima (tree-of-heaven). Plant Physiol 47: 609-618.
- Mihailovic N, Drazic G, Vucinic Z (2008) Effects of aluminium on photosynthetic performance in Al-sensitive and Al-tolerant maize inbred lines. Photosynthetica 46: 476-480.
- Okazawa A, Tang L, Itoh Y, Fukusaki E, Kobayashi A (2006) Characterization and subcellular localization of chlorophyllase from Ginkgo biloba. Z Naturforsch C 61: 111-117.
- Pshibytko N, Kalitukho NL, Zhavoronkova NB, Kabashnikova LF (2004) The pool of chlorophyllous pigments in barley seedlings of different ages under heat shock and water deficit. Russ J Plant Physiol 51: 15-20.
- Saidi MN, Jbir R, Ghorbel I, Namsi A, Drira N, Gargouri-Bouzid R (2012) Brittle leaf disease induces an oxidative stress and decreases the expression of manganese-related genes in date palm (Phoenix dactylifera L.). Plant Physiol Biochem 50: 1-7.
- Schelbert S, Aubry S, Burla B, Agne B, Kessler F, Krupinska K, Hörtensteiner S (2009) Pheophytin pheophorbide hydrolase (pheophytinase) is involved in chlorophyll breakdown during leaf senescence in Arabidopsis. Plant Cell 21: 767-785.
- Schenk N, Schelbert S, Kanwischer M, Goldschmidt EE, Dörmann P, Hörtensteiner S (2007) The chlorophyllases AtCLH1 and AtCLH2 are not essential for senescence-related chlorophyll breakdown in Arabidopsis thaliana. FEBS Lett 581: 5517-5525.
- Stangarlin JR, Pascholati SF (2000) Activities of ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco), chlorophyllase, β-1,3 glucanase and chitinase and chlorophyll content in bean cultivars (Phaseolus vulgaris) infected with Uromyces appendiculatus. Summa Phytopathol 26: 34-42.
- Sytykiewicz H, Czerniewicz P, Leszczyński B (2008) Molecular characteristics of sucrose synthase isolated from bird cherry leaves. Herba Pol 54: 41-49.
- Sytykiewicz H, Czerniewicz P, Sprawka I, Krzyzanowski R (2013) Assessment of chlorophyll amount in aphid-injured seedling leaves of different maize genotypes. Acta Biol Cracov Bot (in press).
- Tang L, Okazawa A, Itoh Y, Fukusaki E, Kobayashi A (2004) Expression of chlorophyllase is not induced during autumnal yellowing in Ginkgo biloba. Z Naturforsch C 59: 415-420.
- Terpstra W, Lambers JWJ (1983) Interactions between chlorophyllase, chlorophyll a, plant lipids and Mg2+. Biochim Biophys Acta 746: 23-31.
- Todorov DT, Karanov EN, Smith AR, Hall MA (2003a) Chlorophyllase activity and chlorophyll content in wild and mutant Arabidopsis thaliana. Biol Plant 46: 125-127.
- Todorov DT, Karanov EN, Smith AR, Hall MA (2003b) Chlorophyllase activity and chlorophyll content in wild type and eti5 mutant Arabidopsis thaliana subjected to low and high temperatures. Biol Plant 46: 633-636.
- Trebitsh T, Goldschmidt EE, Riov J (1993) Ethylene induces de novo synthesis of chlorophyllase, a chlorophyll degrading enzyme, in Citrus fruit peel. Proc Natl Acad Sci 90: 9441-9445.
- Tsuchiya T, Ohta H, Masuda T, Mikami B, Kita N, Shoi Y, Takamiya K (1997) Purification and characterization of two isozymes of chlorophyllase from mature leaves of Chenopodium album. Plant Cell Physiol 38: 1026-1031.
- Tsuchiya T, Suzuki T, Yamada T, Shimada H, Masuda T, Ohta H, Takamiya K (2003) Chlorophyllase as a serine hydrolase: identification of a putative catalytic triad. Plant Cell Physiol 44: 96-101.
- Wang H-C, Huang X-M, Hu G-B, Yang Z, Huang H-B (2005) A comparative study of chlorophyll loss and its related mechanism during fruit maturation in the pericarp of fast- and slow-degreening litchi pericarp. Sci Hortic 106: 247-257.
- Yang CM, Chang IF, Lin SJ, Chou CH (2004) Effects of three allelopathic phenolics on chlorophyll accumulation of rice (Oryza sativa) seedlings: II. Stimulation of consumption-orientation. Bot Bull Acad Sin 45: 119-125.
- Yi Y, Neufeld R, Kermasha S (2007) Controlling sol-gel properties enhancing entrapped membrane protein activity through doping additives. J Sol-Gel Sci Technol 43: 161-170.
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