Effects of temperature,solid content and pH on the stability of black carrot anthocyanins

Anthocyanin stability of black carrots was studied at various solid contents (11, 30, 45 and 64° Brix) and pHs (4.3 and 6.0) during both heating, at 70–90 °C, and storage at 4–37 °C. Monomeric anthocyanin degradation fitted a first-order reaction model. Degradation of monomeric anthocyanins increased with increasing solid content during heating, while it decreased during storage. For example, at pH 4.3, half-life periods for anthocyanins at 30, 45 and 64° Brix were, respectively, 8.4, 6.9 and 5.2 h during heating at 80 °C and 18.7, 30.8 and 35.9 weeks during storage at 20 °C. At 30–64° Brix, increasing pH from 4.3 to 6.0 enhanced the degradation of anthocyanins during heating. The effect of pH on thermal stability of anthocyanins was also studied at six different pHs (2.5–7.0) in citrate-phosphate buffer solutions and significant decrease in anthocyanin stability was observed at pHs above 5.0. Higher activation energies (E_a) were obtained during heating than during storage with increasing solid contents. At 30–64° Brix, E_a values ranged from 68.8 to 95.1 kJ mol⁻¹ during heating and from 62.1 to 86.2 kJ mol⁻¹ during storage. Q₁₀ values at 20–37 °C were as high as 3.1 at 45° Brix and 3.6 at 64° Brix.     

Consumer acceptance of roasted peanuts affected by storage temperature and humidity conditions

Consumer acceptance and intensity ratings of roasted peanuts stored at temperatures of 23, 30, 35, and 40 °C, and water activities (a_w) of 0.33, 0.44, 0.54, 0.67 and 0.75 were determined over time. Consumer acceptance ratings, including overall, appearance, color, and texture were affected by storage water activity and time, but not by storage temperature. Consumer intensity ratings of crunchiness were affected by storage water activity and time, but not storage temperature. Aroma acceptance, flavor acceptance, and roasted peanutty and stale/oxidized/rancid intensity ratings of roasted peanuts were dependent on storage temperature, water activity and time.Shelf-life of roasted peanuts was predicted by all consumer attributes (R²>0.60) and was best predicted by aroma acceptance (R²=0.75). Using contour plots with ratings >5.0 for all acceptance attributes, the shelf-life of roasted peanuts stored at 23 °C and between 0.33 and 0.75 a_w was limited by overall acceptance and decreased by approximately 50% with each 0.1 a_w increase. At accelerated temperatures of 30, 35 and 40 °C, shelf-life of roasted peanuts was predominantly limited by flavor acceptance (>5.0), and to a lesser extent by overall acceptance (>5.0). Shelf-life of roasted peanuts stored at accelerated temperatures decreased by 50% or more with each 0.1 a_w increase.     

Immobilization of naringinase from Aspergillus niger CECT 2088 in poly(vinyl alcohol) cryogels for the debittering of juices

Naringinase, induced from Aspergillus niger CECT 2088 cultures, was immobilized into a polymeric matrix consisting of poly(vinyl alcohol) (PVA) hydrogel, cryostructured in liquid nitrogen, to obtain biocatalytically active beads. The effects of matrix concentration, enzyme load and pH on immobilization efficiency were studied. Between 95% and 108% of the added naringinase was actively entrapped in PVA cryogel, depending on the conditions of immobilization used. The optimal conditions were: 8% (w/v) PVA at pH 7 and 1.6–3.7 U ml⁻¹ of enzyme load. The pH/activity profiles revealed no change in terms of shape or optimum pH (4.5) upon immobilization of naringinase. However, the optimum temperature was shifted from 60 °C to 70 °C and the activation energy of reaction, E_a, was decreased from 8.09 kJ mol⁻¹ to 6.36 kJ mol⁻¹ by immobilization. The entrapped naringinase could be reused through six cycles (runs of 24 h at 20 °C), retaining 36% efficacy for the hydrolysis of naringin in simulated juice.     

Effect of water activity, temperature and incubation time on growth and ochratoxin A production by Aspergillus niger and Aspergillus carbonarius on maize kernels

The aim of this study was to determine the effects of water activity (a_w) (0.92-0.98), temperature (5-45 °C) and incubation time (5-60 days) on growth and ochratoxin A (OTA) production by Aspergillus niger and Aspergillus carbonarius on maize kernels using a simple method. Colony diameters of both strains at 0.92 a_w were significantly lower than those at 0.96 and 0.98 a_w levels. The optimum growth temperature range for A. niger was 25-40 °C and for A. carbonarius 20-35 °C. A. niger produced OTA from 15 to 40 °C, and the highest OTA level was recorded at 15 °C. The concentration of OTA produced at 0.92 a_w was significantly lower than those at 0.96 and 0.98 a_w. A. carbonarius produced OTA from 15 to 35 °C and the maximum concentration was achieved at 15 °C, although not differing statistically from the concentration detected at 20 °C. At 0.98 a_w the OTA concentration was significantly higher than at 0.96 and 0.92 a_w. Our results show that maize supports both growth and OTA production by A. niger and A. carbonarius. The studied strains were able to produce OTA in maize kernels from the fifth day of incubation over a wide range of temperatures and water availabilities. Although the limit of quantification of our method was higher than that required for the analysis of OTA in food commodities, it has proved to be a useful and rapid way to detect OTA production by fungi inoculated onto natural substrates, in a similar way as for pure culture. Both species could be a source of OTA in this cereal in temperate and tropical zones of the world.     

Water activity and temperature effects on mycotoxin production by Alternaria alternata on a synthetic tomato medium

Alternaria spp. have been reported to be the most frequent fungal species invading tomatoes. Certain species, in particular the most common one, A. alternata, are capable of producing several mycotoxins in infected plants and in agricultural commodities. Alternariol (AOH), alternariol monomethyl ether (AME), and tenuazonic acid (TA) are some of the main Alternaria mycotoxins that can be found as contaminants of food. The objective of this study was to determine the effect of water activity (a_w, 0.904, 0.922, 0.954, and 0.982) and temperature (6, 15, 21 and 35 °C) on mycotoxin production on a synthetic tomato medium of a cocktail inoculum of five strains of A. alternata isolated from tomato fruits affected by Blackmould. The optimum AOH production occurred at 0.954 a_w after 28 days of incubation at 21 °C. A temperature of 21 °C was the most favourable for AOH synthesis at all a_w levels. The maximum concentration of AME was determined at 0.954 a_w and 35 °C. The optimum conditions for TA accumulation were 0.982 a_w and 21 °C. At the 0.904 a_w no growth or germination was registered at 6 °C and 15 °C over the whole incubation period. At 21 °C and 35 °C growth occurred slowly but none of the toxins were detected at this a_w level. In general, high a_w levels were favourable for mycotoxin production. None of the other toxins was detected at quantifiable levels at 6 °C after the whole incubation period. A storage temperature of 6 °C or below could be considered as safe for tomato fruits and high moisture tomato products (a_w > 0.95), in relation with Alternaria toxins. The results obtained here could be extrapolated to evaluate the risk of spoilage in tomato fruits and tomato products caused by this pathogen.     

Purification and characterization of olive (Olea europaea L.) peroxidase – Evidence for the occurrence of a pectin binding peroxidase

Peroxidase from olive fruit (Olea europaea L., cv Douro) in a black ripening stage was purified to electrophoretic homogeneity, resulting in four cationic and four anionic fractions. The anionic fractions accounted for 92% of recovered activity and showed molecular masses of 18–20 kDa. The anionic fraction PODa4, the predominant fraction that comprised about 70% of total recovered activity, showed an isoelectric point of 4.4 and optimum pH and temperature of, respectively, 7.0 and 34.7 °C, and apparent K_m values of 41.0 and 0.53 mM, for phenol and H₂O₂, respectively. From the activity-temperature profile, the denaturation temperature and the changes in enthalpy and heat capacity for unfolding of PODa4 were estimated as being, respectively, 36.5 °C, 411.2 and −13.6 kJ mol⁻¹ K⁻¹. The activation energy for phenol oxidation by PODa4 was 99.1 kJ mol⁻¹, corresponding to a calculated temperature coefficient (Q₁₀) of 4. The arabinose (39 mol%) and galacturonic acid (38 mol%) content of the carbohydrate moiety indicated the existence of pectic material in the purified PODa4 fraction. Co-migration of the carbohydrate with the protein band in the isoelectric focusing electrophoresis, points to PODa4 fraction as being a pectin type binding peroxidase.     

Influence of water activity and temperature on growth and mycotoxin production by Alternaria alternata on irradiated soya beans

The aim of this study was to determine the effects of water activity (a_w) (0.99-0.90), temperature (15, 25 and 30 °C) and their interactions on growth and alternariol (AOH) and alternariol monomethyl ether (AME) production by Alternaria alternata on irradiated soya beans. Maximum growth rates were obtained at 0.980 a_w and 25 °C. Minimum a_w level for growth was dependent on temperature. Both strains were able to grow at the lowest a_w assayed (0.90). Maximum amount of AOH was produced at 0.98 a_w but at different temperatures, 15 and 25 °C, for the strains RC 21 and RC 39 respectively. Maximum AME production was obtained at 0.98 a_w and 30 °C for both strains. The concentration range of both toxins varied considerably depending on a_w and temperature interactions. The two metabolites were produced over the temperature range 15 to 30 °C and a_w range 0.99 to 0.96. The limiting a_w for detectable mycotoxin production is slightly greater than that for growth. Two-dimensional profiles of a_w × temperature were developed from these data to identify areas where conditions indicate a significant risk from AOH and AME accumulation on soya bean. Knowledge of AOH and AME production under marginal or sub-optimal temperature and a_w conditions for growth can be important since improper storage conditions accompanied by elevated temperature and moisture content in the grain can favour further mycotoxin production and lead to reduction in grain quality. This could present a hazard if the grain is used for human consumption or animal feedstuff.     

Effect of fenpropimorph, prochloraz and tebuconazole on growth and production of T-2 and HT-2 toxins by Fusarium langsethiae in oat-based medium

Fusarium langsethiae has been isolated from infected cereals in central and northern Europe where it has been identified in the last decade as the main species involved in the occurrence of high levels of T-2 and HT-2 toxins, mainly in oats. The efficacy of three fungicides (prochloraz, tebuconazole, fenpropimorph) for controlling growth of two strains of F. langsethiae isolated from oats was examined at 0.96 and 0.98 a_w at 15, 20 and 25 °C on oat-based media. The concentrations necessary for 50 and 90% growth inhibition (ED₅₀ and ED₉₀ values) were determined. The effect on the trichothecene type A mycotoxins T-2 and HT-2 was also determined. Without fungicides both strains grew faster at 0.98 than at 0.96 a_w and the influence of temperature on growth rates was 25 > 20 > 15 °C. Prochloraz and tebuconazole were more effective than fenpropimorph against F. langsethiae. Strain, temperature and type of fungicide significantly influenced the ED₅₀ and ED₉₀ values for growth. The concentration ranges under different environmental conditions were: prochloraz (0.03-0.1 and 0.3-1.5), tebuconazole (0.06-0.9 and 1.3-8.2), and fenpropimorph (22-59 and 125-215 mg l⁻¹). Production of T-2 and HT-2 toxins was influenced by temperature, a_w, type of fungicide and dose. Levels of T-2 were usually higher than those of HT-2 under the same conditions. The biosynthesis of T-2 toxin increased after 10 day incubation, but was reduced with decreasing temperature and increasing fungicide dose. At 0.98 a_w T-2 levels increased in cultures containing fenpropimorph while at 0.96 a_w the toxin concentrations increased in response to the other two fungicides. Low doses of prochloraz or tebuconazole enhanced toxin production when compared with untreated cultures for strain 2004-59 at 0.96 a_w and 20-25 °C. HT-2 was hardly detectable in the treatments with prochloraz or tebuconazole at 0.98 a_w. This is the first study on the effect of these anti-fungal compounds on control of growth of F. langsethiae and on production of T-2 and HT-2 toxins in oat-based media.     

Evaluation of ability of ferulic acid to control growth and fumonisin production of Fusarium verticillioides and Fusarium proliferatum on maize based media

The aim of this study was to evaluate the efficacy of ferulic acid (1, 10, 20 and 25 mM) at different water activity (a_w) values (0.99, 0.98, 0.96 and 0.93) at 25 °C on growth and fumonisin production by Fusarium verticillioides and Fusarium proliferatum on maize based media. For both Fusarium species, the lag phase significantly decreased (p ≤ 0.001), and the growth rates increased (p ≤ 0.001) at the lowest ferulic acid concentration used (1 mM), regardless of the a_w. However, high doses of ferulic acid (10 to 25 mM) significantly reduced (p ≤ 0.001) the growth rate of both Fusarium species, regardless of the a_w. In general, growth rate inhibition increased as ferulic acid doses increased and as media a_w decreased. Fumonisin production profiles of both Fusarium species showed that low ferulic acid concentrations (1–10 mM) significantly increased (p ≤ 0.001) toxin production, regardless of the a_w. High doses of ferulic acid (20–25 mM) reduced fumonisin production, in comparison with the controls, by both Fusarium species but they were not statistically significant in most cases. The results show that the use of ferulic acid as a post-harvest strategy to reduce mycotoxin accumulation on maize needs to be discussed.     

Distribution of zearalenone in malted barley fractions dependent on Fusarium graminearum growing conditions

Zearalenone (ZON) distribution was measured in main fractions of malted barley, dependent on incubation time (17, 26, 34 days), water activity (0.95 and 0.98) and temperature (20 °C and 30 °C). Malted samples were sterilised and inoculated with Fusarium graminearum. ZON levels were higher (p < 0.01) in bran and germ than flour under almost all growing conditions. Incubation at 30 °C resulted in generally lower ZON levels in germ and bran, regardless of a_w and incubation time. After 34 days, ZON levels in flour from samples that were incubated at the higher temperature rose significantly. At 20 °C ZON concentration showed a bell-shaped concentration profile with increasing incubation time in bran and germ, whilst ZON levels in flour increased at a_w 0.98 and dropped at the lower a_w. The results indicate the importance of storage conditions for ZON levels in commercially relevant grain fractions of malted barley and help predict existing mycotoxin levels or manipulate storage conditions to reduce ZON content.     

Properties of polyphenol oxidase from Anamur banana (Musa cavendishii)

Polyphenol oxidase (PPO) was extracted from Anamur banana, grown in Turkey, and its characteristics were studied. The optimum temperature for banana PPO activity was found to be 30 °C. The pH-activity optimum was 7.0. From the thermal inactivation studies, in the range 60–75 °C, the half-life values of the enzyme ranged from 7.3 to 85.6 min. The activation energy (E_a) and Z values were calculated to be 155 kJ mol⁻¹ and 14.2 °C, respectively. K_m and V_max values were 8.5 mM and 0.754 OD₄₁₀ min⁻¹, respectively. Of the inhibitors tested, ascorbic acid and sodium metabisulphite were the most effective.     

Thermal inactivation kinetics of Rabdosia serra (Maxim.) Hara leaf peroxidase and polyphenol oxidase and comparative evaluation of drying methods on leaf phenolic profile and bioactivities

Inactivation kinetics of peroxidase and polyphenol oxidase in fresh Rabdosia serra leaf were determined by hot water and steam blanching. Activation energy (52.30 kJ mol⁻¹) of polyphenol oxidase inactivation was higher than that (20.15 kJ mol⁻¹) of peroxidase. Water blanching at 90 °C or steam blanching at 100 °C for 90 s was recommended as the preliminary treatment for the retention of phenolics. Moreover, comparative evaluation of drying methods on the phenolics profiles and bioactivities of R. serra leaf were conducted. The results indicated that only intact leaf after freeze drying retained the initial quality. The sun- and air-dried leaves possessed identical phenolic profiles. The homogenised leaf (after freeze-drying) possessed a lower level of phenolics due to enzymatic degradation. Good antioxidant activities were detected for the sun- and air-dried leaves. There was insignificant difference in anti-tyrosinase and anti-α-glucosidase activities among sun-, air-, and freeze-dried leaves.     

Release kinetics of nisin from biodegradable poly(butylene adipate-co-terephthalate) films into water

The release kinetics of nisin from poly(butylene adipate-co-terephthalate) (PBAT) to distilled water was studied at of 5.6, 22 and 40 °C. The release kinetics of nisin from PBAT film was described using Fick’s second law of diffusion, partition coefficient, and Weibull model. The diffusion coefficients (D) determined were 0.93, 2.29, and 5.78 × 10⁻¹⁰ cm²/s at 5.6, 22, and 40 °C, respectively. The partition coefficients (K) calculated were 0.84, 3.89, and 5.2 × 10³ at 5.6, 22, and 40 °C, respectively. The nisin release data at selected temperatures were fitted with the Weibull model (R² > 0.97) with b and n values ranging from 0.02 to 0.98 and from 0.28 to 0.45, respectively. The temperature dependence of D, K, and Weibull model parameter b was modeled using the Arrhenius equation giving values of activation energy (E_a) of 38.3 kJ mol⁻¹ (for D), 38.5 kJ mol⁻¹ (for K), and 79.5 kJ mol⁻¹ (for b).     

Degradation kinetics of anthocyanins in acerola pulp: Comparison between ohmic and conventional heat treatment

Degradation kinetics of monomeric anthocyanins in acerola pulp during thermal treatment by ohmic and conventional heating was evaluated at different temperatures (75–90 °C). Anthocyanin degradation fitted a first-order reaction model and the rate constants ranged from 5.9 to 19.7 × 10⁻³ min⁻¹. There were no significant differences between the rate constants of the ohmic and the conventional heating processes at all evaluated temperatures. D-Values ranged from 116.7 to 374.5 for ohmic heating and from 134.9 to 390.4 for conventional heating. Values of the free energy of inactivation were within the range of 100.19 and 101.35 kJ mol⁻¹. The enthalpy of activation presented values between 71.79 and 71.94 kJ mol⁻¹ and the entropy of activation ranged from −80.15 to −82.63 J mol⁻¹ K⁻¹. Both heating technologies showed activation energy of 74.8 kJ mol⁻¹ and close values for all thermodynamic parameters, indicating similar mechanisms of degradation.     

Kinetics of the formation of radicals in meat during high pressure processing

The kinetics of the formation of radicals in meat by high pressure processing (HPP) has been described for the first time. A threshold for the radicals to form at 400 MPa at 25 °C and at 500 MPa at 5 °C has been found. Above this threshold, an increased formation of radicals was observed with increasing pressure (400–800 MPa), temperature (5–40 °C) and time (0–60 min). The volume of activation (ΔV^#) was found to have the value −17 ml mol⁻¹. The energy of activation (E_a) was calculated to be 25–29 kJ mol⁻¹ within the pressure range (500–800 MPa) indicating high independence on the temperature at high pressures whereas the reaction was strongly dependent at atmospheric pressure (E_a = 181 kJ mol⁻¹). According to the effect of the processing conditions on the reaction rate, three groups of increasing order of radical formation were established: (1) 55 °C at 0.1 MPa, (2) 500 and 600 MPa at 25 °C and 65 °C at 0.1 MPa, and (3) 700 MPa at 25 °C and 75 °C at 0.1 MPa. The implication of the formation of radicals as initiators of lipid oxidation under HPP is discussed.     

Is intraspecific variability of growth and mycotoxin production dependent on environmental conditions? A study with Aspergillus carbonarius isolates

The aim of this study was to assess the impact of suboptimal environmental conditions on the intraspecific variability of A. carbonarius growth and OTA production using thirty isolates of A. carbonarius.Three a_w/temperature conditions were tested, one optimal (0.98a_w/25 °C) and two suboptimal: 0.90a_w/25 °C and 0.98a_w/37 °C as suboptimal water activity and temperature, respectively, which might take place through over ripening and dehydration of grapes. For each condition, 12 Petri dishes were inoculated, and colony growth and OTA production were measured over time.ANOVA revealed significant differences among μ and λ within the 30 assayed isolates. Coefficients of variation (CV%) revealed a wider dispersion of growth rates at 0.90a_w/25 °C compared to 0.98a_w/25 °C, and a more than 4-fold higher CV at 0.98a_w/37 °C compared to 0.98a_w/25 °C. However, dispersion of lag phases was similar at 0.98a_w/25 °C and 0.90a_w/25 °C and wider at 0.98a_w/37 °C.There were significant differences (p < 0.05) among OTA levels (ng/mm²) for the different conditions, values being lower under marginal conditions, and particularly at 0.98a_w/37 °C. Coefficients of variation (CV%) revealed a wider dispersion of OTA production at 0.90a_w/25 °C compared to 0.98a_w/25 °C, while CV at 0.98a_w/37 °C was similar to that at 0.98a_w/25 °C.In order to address the strain variability in growth initiation and prove the well-established notion of reducing OTA in foods by preventing fungal growth, a greater number of strains should be included when developing models for conditions that are suboptimal both for a_w for OTA production and temperature levels for growth.     

Pre-processed spice mix formulation and changes in its quality during storage

Wet and dry spice and condiments of optimized quantities were pre-processed and mixed to prepare a spice mix formulation. The formulation was unit packed in 100 g quantities in glass bottles and held separately at 27±2 or 37±2 °C for 6 months. Freshly prepared spice mix contained (g/100 g) 6.8 moisture, 21.4 fat, 5.1 protein, 7.2 ash and 14.2 NaCl. Gradual decrease in pH from 4.0 to 3.7 and 4.0 to 3.6, increase in a_w from 0.52 to 0.54 and 0.52 to 0.57, increase in free fatty acid (FFA) values (as % oleic acid) from 0.14 to 0.56 and 0.14 to 0.58 and increase in thiobarbituric acid (TBA) values (mg malonaldehyde/kg sample) from 2.5 to 2.9 and 2.5 to 2.9 were noticed during storage at 27±2 or 37±2 °C respectively. Similarly decrease in Hunter L^* (lightness), a^* (redness) and b^* (yellowness) values were noted during the same period. Standard plate count (SPC), spore counts and staphylococci counts (log  cfu/g) were low, 4.0-4.7, 1.2-2.0 and 1.4-2.0, respectively, and yeast and moulds and coliforms could not be detected in all the samples throughout the storage period of 6 months. Two ready-to-serve products, viz., dhal fry and chicken fry prepared by using spice-mix stored for up to 6 months were judged acceptable as indicated by sensory quality scores of 6.8-7.9 (on a 9-point hedonic scale). The spice mix formulation prepared is useful in the preparation of varieties of vegetarian and non-vegetarian ready-to-serve products such as aloo mattar, gobi masala, mattar mushroom, kheema mattar, Mutton chilly fry, etc.     

TBARS predictive models of pork sausages stored at different temperatures

2-Thiobarbituric acid reactive substances (TBARS) is an important quality index for pork sausages. To study this in pork sausages during storage, kinetic models were developed to predict TBARS content changes of pork sausages at different temperatures. The predictive models of TBARS content with respect to storage time and temperature were developed based on primary and Arrhenius equations. The regression coefficients (R² > 0.95) indicated the acceptability of the primary reaction and Arrhenius model for predicting TBARS content changes of pork sausages. The activation energy (E_A) of TBARS is 14.12 kJ mol^− 1, and the corresponding rate constant (k₀) is 9.262 × 10¹⁰. Relative errors between predicted and measured values of TBARS content are all within ± 8%. Thus, the established model could effectively predict the TBARS content of pork sausages between 5 and 35 °C during storage.     

A β-galactosidase from chick pea (Cicer arietinum) seeds: Its purification,biochemical properties and industrial applications

A β-galactosidase from Cicer arietinum seeds has been purified to apparent electrophoretic homogeneity using a combination of various fractionation and chromatographic techniques, giving a final specific activity of 220 units mg⁻¹, with approximately 1840 fold purification. Analysis of the protein by SDS–PAGE revealed two subunits with molecular masses of 48 and 38 kDa, respectively. These bands were further confirmed with LC–MS/MS, indicating that Chick pea β-galactosidase (CpGAL) is a heterodimer. Molecular mass was determined to be 85 kDa by Superose-12 FPLC column, which is in agreement with the molecular mass suggested by mass spectroscopy to be 83 kDa. The optimum pH of the enzyme was 2.8 and it hydrolysed o-nitrophenyl β-d galactopyranoside (ONPG) with a K_m value of 1.73 mM at 37 °C. The energy of activation (E_a) calculated in the range of 35 to 60 °C, using Arrhenius equation, was determined to be 11.32 kcal mol⁻¹. The enzyme could also hydrolyse lactose, with an optimum pH of 4.0 at 40 °C. K_m and E_a for lactose hydrolysis was found to be 10 mM and 10.57 kcal mol⁻¹, respectively. The enzyme was found to be comparatively thermostable showing maximum activity at 60 °C for both ONPG and lactose. Galactose was found to be the competitive inhibitor. β-Galactosidase also exhibited glycoproteineous properties when applied on Con-A Sepharose column. The enzyme was localised in germinated seeds with X-gal activity staining and shown to be expressed prominently at grown radical tip and seed coat. Sequence alignment of CpGAL with other known plant β-galactosidase showed high amino acid sequence homology.