J . Sci. Food Agric.


The Peroxidase Enzyme Activity of some Vegetables and its Resistance to Heat
Nezih Muftugil
TUBITAK, Marmara Research Institute, Nutrition and Food Technology Department, Gebze, Kocaeli, Turkey (Manuscript received 12 April 1985)

The peroxidase enzyme activities of some fresh vegetables (cabbage, leeks, carrot, spinach, celery, squash, potatoes, onions and green beans) were determined. The peroxidase activities of cabbage and green beans were high. Onions showed very little peroxidase activity. The vegetables were blanched in hot water at 75",85" and 95°C. Peroxidase inactivation was faster at the higher temperature blanch. Blanching of green beans, potatoes and squash at 75°C for 30 min was not sufficient for complete inactivation of peroxidase enzyme. Peroxidase enzyme inactivation was also affected by the type of vegetable and the size of vegetable pieces. Keywords: Peroxidase; vegetables; blanching; heat treatment; enzyme activity.

1. Introduction
Blanching is an important process in the preparation of vegetables for canning, freezing and dehydration. It consists of a mild heat treatment accomplished by exposing the vegetables to hot water or steam at 60-100°C for several minutes. This treatment is quite acceptable since most vegetables are cooked prior to consumption. Raw and underblanched vegetables discolour, lose flavour and develop off-odours during frozen and dried storage. It is now widely recognised that the loss of quality that unblanched vegetables experience during frozen and dried storage is caused by enzymic action.'-3 Despite the efforts of many investigators over the past 30 years the enzymes responsible for quality deterioration have not been positively identified.4 There is a general agreement that where flavour is concerned several enzymic systems may be involved, working in sequence or simultaneously. Peroxidase, a member of a large group of enzymes called the oxidoreductases, is considered to have an empirical relationship to off-flavours and off-colours in raw and unblanched vegetables, although the reactions involved have not been conclusively identified.5 It has been found that enzyme peroxidase present in all vegetables is the most heat stable enzyme in plank5 For this reason peroxidase activity is widely used as an index of blanching. In this study the peroxidase enzyme activity of some vegetables was determined and the effect of blanching temperature on the inactivation of peroxidase enzyme was investigated.

2. Experimental
2.1 Materials In this study nine different kinds of vegetables were used as the raw materials (Table 1). Vegetables were size graded, trimmed, washed and some of them were peeled. Cabbage (cut 5mm), leek (cut lOmm), carrot (cut lOmm), spinach (leaves), celery (cut lOmm), squash (cut lOmm), potato (10mm cubes), onion (cut 5mm) and green beans (cut 20mm) were used, one commercial variety of each blanched in water at three different temperatures (75,85 and 95°C).

14 20. 3. Temperature of the blanching water was controlled by an immersed thermocouple connected to a digital thermometer.10 4.41 11.6).50 22.0 32.43 6.20 3.95 30.61 62.56 14.07 64.7ml of the same buffer was added to the reference cuvette.63 45. A known weight of puree (approximately 50mg) was weighed into each of two spectrophotometer cuvettes.96 68.08 51.70 57.80 37. The peroxidase activity of some fresh vegetables Veeetable Cabbage Leeks Carrots Spinach Celery Squash Potatoes Onions Green beans Botanical name Brassica oleracea Allium porrum Daucus carota Spinacia oleracea Apium graveolens Cucurbito pep0 Solanum tuberosum Allium cepa Phaseolus vulgaris Peroxidase enzyme activity (unit 100n") 56.17 41.20 58.2 Method The quantitative peroxidase activities of raw and blanched vegetables were assayed according to the method described by Harvey et aL6 Vegetable purees were made by blending 50g of the sample with 50ml of 0 .40 45. After blanching the vegetables were cooled in cold water.46 42. 2 ~ acetate buffer (pH 5. Table 2. Results and discussion Peroxidase was found in all samples of fresh vegetables investigated (Table 1).46 4.7ml of acetate buffer was added to the sample cuvette whilst 2.45 1.88 64.41 0.46 Blanching was performed in a blanching kettle with 8 parts of water to 1 part vegetable. 0. Both cuvette were then shaken prior to determining the rate of formation of brown colour in the sample by measuring the absorbance of light at a wavelength of 420nm in a Varian Techtron Model 635 spectrophotometer. 2.70 61.37 79.40 1.90 68.13 47.02 0.5% guaicol and 0.44 39.61 27. Miiftugil Table 1.32 55.15 510 2. 2ml of acetate buffer was added to the sample cuvette whilst 2.1% hydrogen peroxidase was added to the sample cuvette.80 51.75 - 53. After homogenisation the suspended vegetable tissue was returned to the cuvettes.10 600 900 1200 1500 18M 100 100 100 100 100 100 100 100 100 66.60 31.24 19.50 59.05 36.10 86.10 27.72 73. This rate was generally linear with time in the early stages of the reaction and the gradient of the line was taken to be proportional to the concentration of peroxidase in the sample.31 .87 59.73 1. The inactivation of peroxidase during the blanching of some vegetables in water at 75°C Peroxidase activity 01 fresh vegetable (W) Vegetable Cabbage Leeks Carrots Spinach Celery Squash Potatoes Onion Green beans 0 15 30 60 90 120 Blanching: time (s) 180 240 300 26.02 37. The contents of the cuvette were transferred to.878 N.02 12.43 40.8 89. a small tissue grinder. Cabbage and green beans had high enzyme activities whereas in ordon and carrots the peroxidase enzyme activity was low. and homogenised in.7ml of an aqueous mixture (1:l) of 0.62 2.

potatoes and cabbage at 75°C for 30min was not sufficient for complete inactivation of peroxidase.76 100 72.97 53.62 100 39. Conclusions Peroxidase enzyme inactivation time depended on the temperature of the blanching water.45 - 45 Blanching time (s) 60 90 120 2.76 1.88 26.97 33.92 72.Peroxidase enzyme activity of vegetables 879 Table 3. During blanching.95 The effect of blancing temperature on the inactivation of peroxidase enzyme is given in Tables 2.18 1.48 47. Blanching of green beans.15 420 600 900 1080 1200 18.77 0.30 14.8' Table 4. a significant proportion .82 8.77 2. The inactivation of peroxidase during the blanching of some vegetables in water at 85°C ~~ ~ Peroxidase activity of fresh vegetable (%) - Vegetable Cabbage Leeks Carrots Spinach Celery Squash Potatoes Onion Green beans 0 100 15 30 60 90 2.27 11.80 6.25 73.26 3. peroxidase inactivation was also affected by the size of vegetable pieces.05 180 0.18 10.19 100 11. Gelatinisation of starch in potatoes by heat caused formation of a thin film on the surface of the potatoe cubes and thereby decreased the inactivation rate of peroxidase enzyme.76 100 43. Attempts to inhibit all peroxidase activity is not strictly necessary.12 5. In industrial blanching processes the peroxidase enzyme test is commonly used to determine whether a sufficient blanch has been given.92 210 240 100 100 100 100 100 100 100 100 100 4.46 100 94.60 26.46 23.90 35.00 32.88 Blanching time (s) 120 180 240 360 1.13 59.80 3. Peroxidase enzyme was most easily inactivated at the high temperature blanch.72 9.8 Jansen reported that when peroxidase is considered responsible for changes in frozen vegetables.83 1.90 38.02 11.17 16.20 0.20 41.13 2. then the measurement of total peroxidase and of the loss therein could have little meaning.46 1. size of vegetables and the nature of the enzyme present.85 9.25 32.35 40. The enzyme inactivation in spinach and cabbage samples which have large surface to volume ratios and comparably high enzyme activities was faster.50 - 56. the locale and occurrence of isoenzymes should also be considered. A full account of the biochemistry of this enzyme has been given by Saunders et aL7 It has been stated that various types of peroxidase exist and the properties depend to some extent on its source.75 100 40.02 2.42 17.70 100 37.50 84.57 2.93 27.62 6.59 13.The intensity of heat treatment applied during blanching caused differences in enzyme activity of the vegetables.94 6.72 14. 4.' If one or more isoenzyme is responsible for adverse changes in frozen vegetables.82 21.17 23.40 7. 3 and 4.79 26.85 0. The inactivation of peroxidase during the blanching of some vegetables in water at 95°C Peroxidase activity of fresh vegetable (%) Vegetable Cabbage Leeks Carrots Spinach Celery Squash Potatoes Onion Green beans 0 15 16.91 0.17 30 4.14 26.17 100 69.10 24. Fawpenetration of heat to the centres could explain fast enzyme inactivation during blanching.57 7.

. 8. Food Res. E. F.. M." Some observations support the view that the enzyme lipoxygenase in green beans.J. Origin of off-odors in frozen green beans.S. 1978. B. pp.. for some vegetables complete inactivation of peroxidase enzyme is necessary in order to obtain good quality frozen stored products.S. Adv. H. Food Technol. Butlenvorths. R. 11S114. Miiftiigil of active peroxidase can be left in many vegetables and a long shelf life in frozen storage can still be achieved.O.3. Lipid oxidation in frozen vegetables in relation to flavour change. 11. 1&11.L. 5. Chow. 1952 17. 1-5. No. 5 . Adams. pp. Hartzler.S. D. C. Peroxidase. 31. Olson. Watts. Paris 1981.M. 10. Holmes-Siedle. However. 1964.B. 1969. Ongley. Watts.T.. S.271-282. Food Technol. 17S179. 15-29.. Rhee. 19-42. A.R. J ." There is some concern that the heat treatment required to give complete inactivation may represent an excessive blanch in many cases. spinach. 3. 1951. New York. Stark. M. 1964.B. J.E. Food Res. 1. pp. Nahrung.. B. P. (Van Arsdel. Eds).. Lindquist.H. R. The enzyme content and the quality of frozen vegetables.F.. 42. Thermal requirements for blanching of fruits and vegetables to be frozen. Food Chem. 657-679. Effect of enzyme inactivation on quality retention in frozen Brussel Sprouts. peas and cauliflowers is responsible for the development of off-flavour during frozen storage12 but its low measurable activity in the fresh products makes this enzyme less suitable as a blanching parameter. M.P. Subjective and Objective Methods Used in Blanching Research in 1976177.S. Masure. 2. 1966.P. Kent..R. Wiley. Guerrant. Technical Memorandum. Quality related chemical and physical changes in frozen foods.. Adams. Boggs. 91. Burnette. Dietrich.C.880 N. Residual peroxidase activity. 12-32. I. 4. Proceedings of Internationa[ Meeting Day of 'Blanching of Fruits and Vegetables to be Fro Zen-Present Knowledge and Future Perspectives'.C. The Campden Food Preservation Research Association. Aylward.G.. References 1. B. Peroxidase and its relationship to food flavour and quality. F. 17. E. . N. Gloucestershire. Harvey. Effect of blanching and of frozen storage of vegetables on ascorbic acid retention and the concomitant activity of certain enzymes. 7. Jansen. 1975. M.. 1977. Butler.. L. A review. K. In: Quality and Stabiliry of Frozen Foods. 1969..J. Food Sci.M. 19. 9. Science for Stability Programme to whom thanks are due. W.B. Baardseth. 23.. Food Sci. Saunders. Haisman. Oxidation systems in fruits and vegetables. F.C. 178.M. P. Acknowledgement This work forms part of a research project sponsored by the Scientific Affairs of NATO. 198-199 12. Copley. 1-76. Powel. B. 6 . 2-32. Bottcher. Quality changes of frozen vegetables. W. 1977.. M.