Background: Consistency is one of the main traits that define
commercial quality and price of tomato paste. Pectins are partially
responsible for consistency in tomato paste, therefore enzymatic pectin
modification could be used to increase paste consistency. Results: This
work reports the effects of a commercial enzymatic preparation of
pectin-methyl-esterase (PME) (NovoShape\u2122) on tomato paste
consistency taking into account variables as enzyme/substrate ratio
(0,1% w/w - 1% w/w), reaction time (0 hr - 3 hrs) and reaction
temperature (40\ubaC-60\ubaC). The results indicate that
NovoShape\u2122 increased consistency when reaction temperature ranged
from 40 to 50\ubaC with an enzyme/substrate ratio of 0.5 to 1 (l PME
solution/g tomato paste on dry base). On the other hand, enzymatic
treatment was not effective at 60\ubaC with an enzyme/substrate ratio
of 0.1%. Conclusions: Based on these results, addition of
NovoShape\u2122 is a good technological approach to increase tomato
paste consistency

Caballero-Vald\ue9s, Eduardo

Fern\ue1ndez-Parodi, Jos\ue9

P\ue9rez-Torres, Eduardo

Z\ufa\uf1iga-Hansen, Mar\ueda Elvira

Bioline International

Electronic Journal of Biotechnology ISSN: 0717-3458 Electron. J. Biotechnol. http://www.ejbiotechnology.info DOI: 10.2225/vol15-issue3-fulltext-5                                                                                                             SHORT COMMUNICATION Effect of a commercial pectinmethylesterase on tomato paste consistency María Elvira Zúñiga-Hansen1,2   · Eduardo Pérez-Torres1 · Eduardo Caballero-Valdés1,2 ·  José Fernández-Parodi2 1 Centro Regional de Estudios en Alimentos Saludables (CREAS), CONICYT-Regional GORE R06I1004, Valparaíso, Chile 2 Pontificia Universidad Católica de Valparaíso, Facultad de Ingeniería, Escuela de Ingeniería Bioquímica, Valparaíso, Chile  Corresponding author: direccion@creas.cl; mzuniga@ucv.cl Received November 30, 2011 / Accepted March 27, 2012 Published online: May 15, 2012 © 2012 by Pontificia Universidad Católica de Valparaíso, Chile Abstract Background: Consistency is one of the main traits that define commercial quality and price of tomato paste. Pectins are partially responsible for consistency in tomato paste, therefore enzymatic pectin modification could be used to increase paste consistency. Results: This work reports the effects of a commercial enzymatic preparation of pectin-methyl-esterase (PME) (NovoShape™) on tomato paste consistency taking into account variables as enzyme/substrate ratio (0,1% w/w - 1% w/w), reaction time (0 hr - 3 hrs) and reaction temperature (40ºC-60ºC). The results indicate that NovoShape™ increased consistency when reaction temperature ranged from 40 to 50ºC with an enzyme/substrate ratio of 0.5 to 1 (l PME solution/g tomato paste on dry base). On the other hand, enzymatic treatment was not effective at 60ºC with an enzyme/substrate ratio of 0.1%. Conclusions: Based on these results, addition of NovoShape™ is a good technological approach to increase tomato paste consistency. Keywords: pectin methyl esterase, tomato paste, viscosity     INTRODUCTION Tomato paste is obtained by concentrating tomato juice after removal of skin and seeds. Its commercial quality is associated with colour, lycopene concentration and consistency, therefore, improvement of any of these traits determines a better competitiveness and commercial value in the market (Haley and Smith, 2003; Colle et al. 2010).  Consistency of tomato products is strongly affected by the composition of pectins, which are high molecular weight polysaccharides of partially methylated units of D-galacturonic acid (Anthon et al. 2002; Van Buggenhout et al. 2009). Removal of methyl groups from methoxyl pectin by pectin methyl esterase (PME) has been used to increase consistency in some foods by allowing cross linking of divalent cations with pectin molecules (Anthon et al. 2005; Plaza et al. 2008).  Based on the above mentioned, enzymes involved in pectin modification could have important applications for tomato industry (Van Buggenhout et al. 2009). The present study proposes the use of a commercial preparation of PME, NovoShape™, as a strategy to increase tomato paste consistency and, therefore, its commercial value. The main variables to carry out the study are enzyme/substrate Zúñiga-Hansen et al. DOI: 10.2225/vol15-issue3-fulltext-5                                                                                                                                                    2 ratio and reaction time and temperature, which are reported in order to determine the best conditions to use PME preparation in tomato paste industry. MATERIALS AND METHODS  Materials Tomato pastes were purchased from commercial brands Aconcagua Foods and Dos Caballos. A commercial preparation of pectin methyl transferase (EC. 3.2.1.11) from Aspergillus aculeatus, was used in the assays (NovoShape™, Novo Nordisk, Bagsvaerd, Denmark). Sample preparation Commercial tomato pastes were adjusted to 12ºBrix by addition of distilled water. NovoShape™ was added to previously homogenized samples at 0.1, 0.5 and 1% w/w and incubated at 40, 50 and 60ºC for 3 hrs in water bath with temperature control. Enzymatic reaction was stopped by thermal shock at 100ºC during 5 min and the paste obtained was reconstituted to reach 12ºBrix with distilled water. Samples were measured with a Bronswick consistometer (CSC Scientific, USA). Consistency measurement This method determines consistency of almost any viscous substance by measuring the distance a material flows under its own weight during a time interval of 30 sec in a Bronswick consistometer at 20ºC. Made of stainless steel, unit has spirit level and two levelling screws. Trigger-held gate is spring-operated to permit instantaneous release. Trough is graduated in 0.5 cm divisions and requires 75 mL sample. The details of the present procedure are mentioned in ASTM F1080-93. Statistical analysis Samples were measured in duplicate and significant differences were determined by ANOVA (P < 0.05) using the software GraphPad Prism version 5.01.  RESULTS AND DISCUSSION  Consistency is highly dependent on crosslinking of pectin with divalent cations such as calcium (Slavov et al. 2009; Wei et al. 2009; Yoo et al. 2009). However, the fact that the treatments with 0.5% w/w and 1% w/w of NovoShape™ increased tomato paste consistency throughout 3 hrs of assay (P < 0.05) without addition of divalent cations suggests that consistency is not limited by these elements in tomato paste, which can be observed in Figure 1a and c. It must be taking into account that the lower the Bronswick value of consistency, the higher the sample consistency. A Bronswick consistency value decreased from 6 cm/30 sec to 4 cm/30 sec, which could increase the commercial value of tomato paste in about 50%, and therefore was considered the value to be reached in these experiments. Samples treated with 0.1% w/w NovoShape™ showed similar consistencies compared to control conditions (Figure 1a).  Incubation for longer times would probably result in a higher paste consistency; however, those treatments could not be considered because of the delay involved in processing several thousand tons of tomato per season, taking into account the productivity as criteria. Treatment with 1% w/w NovoShape™ increased consistency to 4 cm/30 sec in half time compared to the same preparation at 0.5% w/w (Figure 1b and c). Despite the shorter time necessary to reach the target consistency with NovoShape™ at 1% w/w, this amount of enzyme preparation represents a high cost to be used in practical applications in tomato paste industry. Therefore, treatment with NovoShape™ at 0.5% w/w is recommended when a compromise among consistency, productivity and enzyme cost is considered.  Effect of a commercial pectinmethylesterase on tomato paste consistency DOI: 10.2225/vol15-issue3-fulltext-5                                                                                                                                                    3 A sudden change in consistency was observed at 0.5% w/w NovoShape™ during the second hour of treatment compared to the first one (Figure 1b). This is probably due to a threshold of pectin demethylation necessary to sustain the cross linking with divalent cations in the paste to increase consistency. It has been shown that PME from Aspergillus aculeatus cleaves methoxyl groups in a random way and the determinant factor in gel formation by calcium/pectin interactions is the number of successive carboxyl groups along the pectin backbone and not only their total number (Axelos et al. 1996). Based on the latter, the higher change in consistency observed during the second hour of treatment might be related to the time necessary to have enough successive carboxyl groups free to interact with divalent cations in tomato pectin treated with NovoShape™.  Fig. 1 Increasing of sample consistency of tomato paste using three different amounts of PME (NovoShape™) and temperatures (control at room temperature, 40ºC, 50ºC and 60ºC). (a) 0.1% NovoShapeTM; (b) 0.5% NovoShapeTM and (c) 1% NovoShapeTM Zúñiga-Hansen et al. DOI: 10.2225/vol15-issue3-fulltext-5                                                                                                                                                    4 Treatment with 0.5% w/w NovoShape™ at 40ºC resulted in an increased consistency from 20 min onwards, meanwhile it took more than 1 hr to observe significant changes in paste consistency at 50ºC (P < 0.05) (Figure 1b). PME from A. aculeatus has been shown to follow a first order model of inactivation in the range of 46-56ºC (Duvetter et al. 2005). Therefore, a higher degree of enzymatic inactivation at 50ºC may explain why longer treatments were necessary to increase consistency at this temperature. Differences between 40ºC and 50ºC incubation treatments were not observed at 1% w/w NovoShape™ (Figure 1c), suggesting that despite the inactivation by temperature, remaining PME activity at this concentration was more than enough to saturate the system. CONCLUDING REMARKS  Our results suggest that NovoShape™ added at 0.5% w/w could have important applications in tomato paste industry by increasing consistency and therefore its commercial value. Availability of a PME less susceptible to high temperature and a complete economic evaluation may help to determine the feasibility of its use in tomato paste industry Financial support: Proyecto FONDECYT Nº 1070258 and Centro Regional de Estudios en Alimentos Saludables (CREAS), CONICYT-Regional GORE Valparaíso R06I1004. REFERENCES ANTHON, G.E.; SEKINE, Y.; WATANABE, N. and BARRETT, D.M. (2002). Thermal inactivation of pectin methylesterase, polygalacturonase, and peroxidase in tomato juice. Journal of Agricultural and Food Chemistry, vol. 50, no. 21, p. 6153-6159. [CrossRef] ANTHON, G.E.; BLOT, L. and BARRETT, D.M. (2005). Improved firmness in calcified diced tomatoes by temperature activation of pectin methylesterase. Journal of Food Science, vol. 70, no. 5, p. C342-C347. [CrossRef] AXELOS, M.A.V.; GARNIER, C.; RENARD, C.M.G.C. and THIBAULT, J.F. (1996). Interactions of pectins with multivalent cations: Phase diagrams and structural aspects. Progress in Biotechnology, vol. 14, p. 35-45. [CrossRef] COLLE, I.; VAN BUGGENHOUT, S.; VAN LOEY, A. and HENDRICKX, M. (2010). High pressure homogenization followed by thermal processing of tomato pulp: Influence on microstructure and lycopene in vitro bioaccessibility. Food Research International, vol. 43, no. 8, p. 2193-2200. [CrossRef] DUVETTER, T.; LOEY, A.V.; SMOUT, C.; VERLENT, I.; NGUYEN, B.L. and HENDRICKX, M. (2005). Aspergillus aculeatus pectin methylesterase: Study of the inactivation by temperature and pressure and the inhibition by pectin methylesterase inhibitor. Enzyme and Microbial Technology, vol. 36, no. 4, p. 385-390. [CrossRef] HALEY, T.A. and SMITH, R.S. (2003). Evaluation of in-line obsorption photometry to predict consistency of concentrated tomato products. LWT-Food Science and Technology, vol. 36, no. 2, p. 159-164. [CrossRef] PLAZA, L.; DUVETTER, T.; PLANCKEN, I.V.D.; MEERSMAN, F.; LOEY, A.V. and HENDRICKX, M. (2008). Influence of environmental conditions on thermal stability of recombinant Aspergillus aculeatus pectinmethylesterase. Food Chemistry, vol. 111, no. 4, p. 912-920. [CrossRef] SLAVOV, A.; GARNIER, C.; CRÉPEAU, M.-J.; DURAND, S.; THIBAULT, J.F. and BONNIN, E. (2009). Gelation of high methoxy pectin in the presence of pectin methylesterases and calcium. Carbohydrate Polymers, vol. 77, no. 4, p. 876-884. [CrossRef] VAN BUGGENHOUT, S.; SILA, D.N.; DUVETTER, T.; VAN LOEY, A. and HENDRICKX, M. (2009). Pectins in processed fruits and vegetables: Part III-texture engineering. Comprehensive Reviews in Food Science and Food Safety, vol. 8, no. 2, p. 105-117. [CrossRef]  WEI, X.; CHEN, Z.; LU, Y.; XU, H.; CHEN, G. and WU, W. (2009). Physicochemical characterization of a pectin/calcium matrix containing a large fraction of calcium chloride: Implications for sigmoidal release characteristics. Journal of Applied Polymer Science, vol. 113, no. 4, p. 2418-2428. [CrossRef] YOO, S.H.; LEE, B.H.; SAVARY, B.J.; LEE, S.; LEE, H.G. and HOTCHKISS, A.T. (2009). Characteristics of enzymatically-deesterified pectin gels produced in the presence of monovalent ionic salts. Food Hydrocolloids, vol. 23, no. 7, p. 1926-1929. [CrossRef]       Effect of a commercial pectinmethylesterase on tomato paste consistency DOI: 10.2225/vol15-issue3-fulltext-5                                                                                                                                                    5       How to reference this article: ZÚÑIGA-HANSEN, M.E.; PÉREZ-TORRES, E.; CABALLERO-VALDÉS, E. and FERNÁNDEZ-PARODI, J. (2012). Effect of a commercial pectinmethylesterase on tomato paste consistency. Electronic Journal of Biotechnology, vol. 15, no. 3. http://dx.doi.org/10.2225/vol15-issue3-fulltext-5  Note: Electronic Journal of Biotechnology is not responsible if on-line references cited on manuscripts are not available any more after the date of publication. Supported by UNESCO / MIRCEN network.  

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