TY - JOUR
T1 - Biological protein precipitation
T2 - a green process for the extraction of cucumisin from melon (Cucumis melo L. inodorus) by-products
AU - Gómez-García, Ricardo
AU - Campos, Débora A.
AU - Aguilar, Cristóbal N.
AU - Madureira, Ana R.
AU - Pintado, Manuela
N1 - Funding Information:
The use of melon by-products as new feedstock for proteins recovery by biological precipitation could decrease the environmental impact as well as minimize the costs associated to the traditional extractive processes. This research demonstrated that melon peels have proteolytic (4.24 U/mg protein) and milk-clotting (6300 MCU/mg protein) activities with an MCA/PA ratio of 1485 at 85 °C. Besides, biological precipitation with CRG allows to improve the biological activity of the proteins recovered from melon peels. A yield of 0.17 g of CUC/100 g of melon by-products can be obtained with the best conditions of precipitation at pH 3 with low concentration of CRG (0.003% w/v), improving 17.65 and 2.11-folds the PA (74.86 U/mg protein) and MCA (405 MCU/mg protein), respectively. Furthermore, these findings support the importance to valorise melon by-products to avoid economic and environmental issues through their reincorporation into the industrial chains as rich sources of value-added proteins.The authors would like to thank the National Council of Science and Technology (CONACyT, Mexico) for PhD fellowship support granted to Gómez-García Ricardo and also the authors would like to thank to the project MultiBiorefinery.: Estratégias multiuso para a valorização de uma gama alargada de subprodutos agroflorestais e das pescas: Um passo em frente na criação de uma biorrefinaria” financiado pelo Programa Operacional Competitividade e Internacionalização (POCI-01-0145-FEDER-016403) e pelo Programa Operacional Regional de Lisboa(LISBOA-01-0145-FEDER-016403), na sua componente FEDER e pela Fundação para a Ciência e Tecnologia, I.P. na componente nacional (SAICTPAC/0040/2015).
Funding Information:
The authors would like to thank the National Council of Science and Technology (CONACyT, Mexico) for PhD fellowship support granted to Gómez-García Ricardo and also the authors would like to thank to the project MultiBiorefinery.: Estratégias multiuso para a valorização de uma gama alargada de subprodutos agroflorestais e das pescas: Um passo em frente na criação de uma biorrefinaria” financiado pelo Programa Operacional Competitividade e Internacionalização (POCI-01-0145-FEDER-016403) e pelo Programa Operacional Regional de Lisboa(LISBOA-01-0145-FEDER-016403), na sua componente FEDER e pela Fundação para a Ciência e Tecnologia, I.P. na componente nacional ( SAICTPAC/0040/2015 ).
Publisher Copyright:
© 2021 Elsevier Ltd
PY - 2021/7
Y1 - 2021/7
N2 - Cucumisin (CUC) from industrial melon by-products was separated for the first time through biological precipitation using carrageenan (CRG). This approach could represent a cost-effective and environmentally friendly process for the industries, avoiding the use of expensive equipment and toxic salts or solvents, such as butanol and ethanol. In this study, biological precipitation of proteins from melon by-products using CRG was studied and compared with conventional precipitation with ammonium sulphate. Different methods were applied for the identification and characterization of isolated proteins, including SDS-PAGE gel, FPLC and proteolytic activity assays. The isolated CUC confirmed a molecular weight of 68 kDa and showed highly stable proteolytic (PA) and milk-clotting (MCA) activities in a wide range of CaCl2 (20–60 mM), pH (5–7) and temperatures (30–85 °C). Melon peel extract demonstrated to possess significant PA (4.24 U/mg protein) and MCA (191.50 MCU/mg protein), but such values were increased by ammonium sulphate precipitation (1.60 and 2.06-folds, respectively), and specially a noticeable increment was observed by biological precipitation with 2.11 and 17.65-folds, respectively, demonstrating the capability to be an effective strategy to isolate and purify CUC, allowing a yield of 0.17 g CUC/100 g of by-products and keeping its biological properties.
AB - Cucumisin (CUC) from industrial melon by-products was separated for the first time through biological precipitation using carrageenan (CRG). This approach could represent a cost-effective and environmentally friendly process for the industries, avoiding the use of expensive equipment and toxic salts or solvents, such as butanol and ethanol. In this study, biological precipitation of proteins from melon by-products using CRG was studied and compared with conventional precipitation with ammonium sulphate. Different methods were applied for the identification and characterization of isolated proteins, including SDS-PAGE gel, FPLC and proteolytic activity assays. The isolated CUC confirmed a molecular weight of 68 kDa and showed highly stable proteolytic (PA) and milk-clotting (MCA) activities in a wide range of CaCl2 (20–60 mM), pH (5–7) and temperatures (30–85 °C). Melon peel extract demonstrated to possess significant PA (4.24 U/mg protein) and MCA (191.50 MCU/mg protein), but such values were increased by ammonium sulphate precipitation (1.60 and 2.06-folds, respectively), and specially a noticeable increment was observed by biological precipitation with 2.11 and 17.65-folds, respectively, demonstrating the capability to be an effective strategy to isolate and purify CUC, allowing a yield of 0.17 g CUC/100 g of by-products and keeping its biological properties.
KW - Biological precipitation
KW - Cucumisin
KW - Melon by-products
KW - Milk-clotting and proteolytic enzyme
KW - Polyelectrolyte
UR - http://www.scopus.com/inward/record.url?scp=85101311182&partnerID=8YFLogxK
U2 - 10.1016/j.foodhyd.2021.106650
DO - 10.1016/j.foodhyd.2021.106650
M3 - Article
AN - SCOPUS:85101311182
SN - 0268-005X
VL - 116
SP - 1
EP - 9
JO - Food Hydrocolloids
JF - Food Hydrocolloids
M1 - 106650
ER -