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Trypsin inhibitors in xoconostle seeds (Opuntia joconostle Weber.)

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Abstract

Seed proteins recovered after heating a seed extract from Opuntia joconostle Weber (xoconostle, an acid cactus pear) were screened for different biochemical activities, detecting only trypsin-like inhibitory activity. Two trypsin-like inhibitor forms from seeds were separated by RP-HPLC and partially sequenced and characterized as an enriched mixture. They were evaluated for inhibition on several serine proteinases, but only trypsin-like inhibition was detected by the inhibitor extract. The two isolated forms, OjTI 1 and OjTI 2 showed low molecular weights of 4.26 and 4.17 kDa as determined by mass spectrometry. An enriched inhibitory fraction showed a high thermal stability by retaining the activity after heating the sample for 1 h at 90 °C, as well as after heating for at 120 °C under 1 kg/cm2 for 15 min at different pH values. Partial sequence of the two forms was determined by mass spectrometry indicating that they were similar and after alignments analysis they showed the highest similarity with the trypsin inhibitor from O. streptacantha and to a lesser extent to other trypsin inhibitors of the MEROPS database families. The inhibitory spectrum was evaluated against several digestive enzymes from pests and beneficial insects from several taxonomic orders.

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Abbreviations

PIs:

Proteinase inhibitors

OjTIs:

Opuntia joconostle trypsin inhibitors

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Acknowledgment

We would like to thank PROMEP for economical support to the project PROMEP/103.5/10/7303 for the scholarship for Aguirrezabala-Cámpano María Teresa and financial support of this research. Also we want to thank to the PAICTY-UANL 2010 budget for financial support of project 47. Also, we want to thank Yolanda Rodríguez Aza and Ma. Cristina Elizarraráz Anaya for their invaluable help and technical support during the purification of the inhibitor through HPLC. Finally, we thank to Biotechnology Laboratory from the Faculty of Agronomy from UANL for the facilities provided to this work.

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Authors and Affiliations

  1. Facultad de Agronomía-Laboratorio de Biotecnología, Universidad Autónoma de Nuevo León (UANL), Francisco Villa s/n, ExHacienda El Canadá, General Escobedo, Nuevo León, México

    María Teresa Aguirrezabala-Cámpano, Sugey Ramona Sinagawa-García, Adriana Gutiérrez-Díez & Jorge Ariel Torres-Castillo

  2. Experimental field General Terán del Instituto Nacional de Investigaciones Forestales Agrícolas y Pecuarias, Nuevo León, México

    Reyna Ivonne Torres-Acosta

  3. Centro de Investigación y de Estudios Avanzados del I. P. N. Unidad Irapuato, Guanajuato, México

    Alejandro Blanco-Labra & Ma. Elizabeth Mediola-Olaya

Authors
  1. María Teresa Aguirrezabala-Cámpano
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  2. Reyna Ivonne Torres-Acosta
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  3. Alejandro Blanco-Labra
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  4. Ma. Elizabeth Mediola-Olaya
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  5. Sugey Ramona Sinagawa-García
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  6. Adriana Gutiérrez-Díez
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  7. Jorge Ariel Torres-Castillo
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Corresponding author

Correspondence to Jorge Ariel Torres-Castillo.

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Aguirrezabala-Cámpano, M.T., Torres-Acosta, R.I., Blanco-Labra, A. et al. Trypsin inhibitors in xoconostle seeds (Opuntia joconostle Weber.). J. Plant Biochem. Biotechnol. 22, 261–268 (2013). https://doi.org/10.1007/s13562-012-0152-z

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  • DOI: https://doi.org/10.1007/s13562-012-0152-z

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