The necessities of a sustainable agriculture require the use of fertilizing solutions and methods capable to respond to the
needs of agricultural crops and to the requirements of the environment protection. In this regard, extra-radicular
fertilizers and liquid fertilizers with application or incorporation into the soil respond to these priorities. In the past 20
years, the range of fertilizers has greatly diversified, currently being developed the compositions containing minerals
together with natural or synthetic organic substances, with application to a wide range of crops. Organic substances
added are intended to stimulate the activity of photosynthesis, absorption of nutrients, to provide a balanced nutrition, to
combat nutritional deficiencies and reduce the effects of different stressors. The most frequently used organic
substances are protein hydrolysates of plant or animal origin, and various amino-acids. Complex matrices resulted by
including into NPK structures organic substances led to the development of new types of bio-fertilizers whose
agrochemical effects were tested with positive effects on the nutrition of different crops in the conditions present in
Romania. Through a careful selection of raw materials and technological processes, certain formulas of bio-fertilizers
can be used in organic farming, where the imposed requirements are defined by European regulations and verified by
certification bodies. In my own researches the concerns are directed towards producing these fertilizers and their testing
in the vegetation/green house and in field. This paper shows the physical and chemical characteristics for some of the
complex matrices with organic substances used in agriculture

Cioroianu, Traian Mihai

Grigore, Adriana Elena

Mihalache, Daniela

Sîrbu, Carmen Eugenia

Sîrbu, Carmen-Eugenia

Grigore, Adriana-Elena

Cioroianu, Traian-Mihai

ALSE Repository

Lucrări Ştiinţifice – vol. 57 (2) 2014, seria Agronomie 47   PROTEIN HYDROLYSATES AND AMINO-ACIDS FERTILIZERS - PHYSICOCHEMICAL CHARACTERISTICS  Daniela MIHALACHE1, Carmen Eugenia SÎRBU1,  Adriana Elena GRIGORE1, Traian Mihai CIOROIANU1  e-mail: ddobrinescu@yahoo.com   Abstract  The necessities of a sustainable agriculture require the use of fertilizing solutions and methods capable to respond to the needs of agricultural crops and to the requirements of the environment protection. In this regard, extra-radicular fertilizers and liquid fertilizers with application or incorporation into the soil respond to these priorities. In the past 20 years, the range of fertilizers has greatly diversified, currently being developed the compositions containing minerals together with natural or synthetic organic substances, with application to a wide range of crops. Organic substances added are intended to stimulate the activity of photosynthesis, absorption of nutrients, to provide a balanced nutrition, to combat nutritional deficiencies and reduce the effects of different stressors. The most frequently used organic substances are protein hydrolysates of plant or animal origin, and various amino-acids. Complex matrices resulted by including into NPK structures organic substances led to the development of new types of bio-fertilizers whose agrochemical effects were tested with positive effects on the nutrition of different crops in the conditions present in Romania. Through a careful selection of raw materials and technological processes, certain formulas of bio-fertilizers can be used in organic farming, where the imposed requirements are defined by European regulations and verified by certification bodies. In my own researches the concerns are directed towards producing these fertilizers and their testing in the vegetation/green house and in field. This paper shows the physical and chemical characteristics for some of the complex matrices with organic substances used in agriculture.  Key words: fertilizer, protein hydrolysate, ferric chlorosis                                                  1 Institutul National de Cercetare-Dezvoltare pentru Pedologie, Agrochimie si Protectia Mediului - ICPA Bucuresti The analysis of the data existing in the specialized literature regarding the conventional fertilizers of extra-radicular type or those with substances having a stimulating effect of growth indicates the fact that the exclusive use of certain substances in the category of bio-stimulators for the treatment of agricultural crops often does not lease to obtaining significant effects (Cioroianu T.M. et al, 2011; Cioroianu T.M. et al, 2009; Cioroianu T.M. et al, 2011; Cioroianu T.M. et al, 2011). In such cases the “explosive” vegetative development of the plant is not supported by an additional contribution, fast, of macro and micro-elements necessary for the nutrition (Iovi A. et al, 2000). This phenomenon strongly occurs in case of a poor basic fertilization, on degraded soils, as well as in case of unbalanced macronutrient fertilization.   In practice, it is known a wide range of liquid fertilizers, complex solutions with extra-radicular application, having as nutrients nitrogen, phosphorus, potassium, meso and micro-elements that may also contain synthetic organic substances, humic substances, fulvic substances, plant extracts, peptides or protein hydrolysates of animal origin or glycoproteins of plant origin, naphthenates, introduced in order to stimulate the metabolization of nutrients, or to facilitate the absorption of ionic species or molecules.    It is well known that the use of microelements like iron, copper, zinc, calcium, magnesium and manganese chelated with natural organic substances are more easily absorbed by plants, and their presence may destroy or reduce bacteria, viruses, fungi or other pathogens when applied as fertilizing substances to plants.  In the field of agriculture it is recommended the use of extra-radicular fertilizers not only in treating certain nutrition diseases of plants, but also to prevent them, for increased yields, for increased quality of products and to reduce the negative impact on the environment of conventional fertilizers (Dorneanu A. et al., 2003). Also, the plants treated with fertilizers containing natural organic substances with chelating role are more resistant to frost, drought, to biotic and abiotic Universitatea de Ştiinţe Agricole şi Medicină Veterinară Iaşi 48 stress factors (Sîrbu C. et al, 2010; Sîrbu C. et al, 2007; Sîrbu C. et al, 2008; Sîrbu C. et al, 2009; Trandafir V. et al, 2007; Trandafir V. et al, 2003).  MATERIAL AND METHOD  The experiments conducted in the last 5 years by I.N.C.D.P.A.P.M – I.C.P.A. Bucharest (National Research and Development Institute for Soil Science Agro-Chemistry and Environment) to obtain fertilizers with organic substance having a stimulating role of growth have used a protein hydrolysate of animal origin, obtained through neutral hydrolysis, having in its composition proteins, protides and amino-acids free in structure: glycine 30 -38%, alanine 10 – 15%, proline 10 – 15%, glutamic acid – 10%, hydroxyproline 7 – 9%, aspartic acid 4 – 6%,  arginine 4 – 6%, serine 3 – 5 %, threonine 1 – 3%, and as essential amino-acids in significant proportions: lysine 2 – 4%, valine     2 – 4% leucine 2 – 3%, phenylalanine 1.5 – 2%, isoleucine 1 – 1.5 %, histidine 0.7 – 1.5%, methionine 0.2 – 0.5%. The fertilizers of extra-radicular type obtained consist in a matrix of NPK type to which micro and meso-elements are added: Fe, Cu, Zn, Mn, Mg, B, Mo, Co and protein hydrolyzate. The main types of chemical reactions used in the process of obtaining extra-radicular fertilizers were:  - neutralization reaction;  - proton exchange reaction (ionization reaction, neutralization, ion displacement, hydrolysis);  - reactions forming chelates and metal complexes;  The technological process for obtaining fertilizers of extra-radicular type included the following steps:  - obtaining the solution of NPK macro-elements; - obtaining the solution of chelated/complexed microelements;  - obtaining the solution of protein hydrolyzate;  - mixing, homogenization of the three solutions;  The general scheme of processes used to obtain most of the extra-radicular fertilizers having also in their composition protein hydolysates introduced as substances with a bio-stimulator role is shown in the figure 1.    Figure 1 Scheme of general processes used to obtain extra-radicular fertilizers with protein hydrolysates RESULTS AND DISCUSSIONS  Using the scheme of processes shown in the Figure 1, there were realized several versions of extra-radicular fertilizers, with the introduction into a matrix of macro, meso and microelements of the protein hydrolysate of animal origin.  The experimental fertilizers were characterized physically and chemically in the Laboratory for Testing and Control of Fertilizers Quality, accredited by the Romanian Accreditation Association (RENAR), belonging to ICPA Potassium source  Demi water  Phosphorus source  Neutralization process Nitrogen sources  Process of dissolution of nitrogen sources  Chelating/ Complexing agents  Microelements Fe, Cu, Mn, Mg, Zn, Co, Mo, B Chelating – Complexing Process  Mixing – Homogenization Process  Fertilizer  Protein hydrolysate  Lucrări Ştiinţifice – vol. 57 (2) 2014, seria Agronomie 49 Bucharest and agro-chemically tested in the National Network for the authorization of fertilizers to be used in agriculture. The chemical composition of the fertilizers of foliar type experimentally obtained and agro-chemically tested in order to be authorized is presented in the tables 1, 4 and 8 (Regulation (EC) No. 2003/2003 the European Parliament and of the Council of 13 October 2003 on fertilizers, Regulation (EC) No. 834/2007 of the Council of 28 June 2007 on organic production and labeling of organic products, and for repealing the Regulation (EEC) No. 2092/91)  The results of the agro-chemical tests for the experimental fertilizers are shown in the tables 2, 3, 5 – 7.   Table 1 Chemical composition of fertilizers AMINO-FERT N, NK and NPK Chemical composition  AMINO-FERT NK AMINO-FERT N AMINO-FERT NPK  (g/l) (g/l) (g/l) total nitrogen (N) 17,0 18,0 125,0 Phosphorus (P2O5) 0,01 0,01 70,0 Potassium (K2O) 8,5 0,01 60,0 Copper (Cu) 1,0 1,0 0,22 Zinc (Zn) 1,0 1,0 0,18 Iron (Fe) 3,0 2,75 0,45 Manganese (Mn) 0,6 0,55 0,3 Boron (B) 0,5 0,5 0,3 Magnesium (MgO) 3,5 2,5 0,55 Molybdenum (Mo) - - 0,01 Cobalt (Co) - - 0,01 Sulfur (SO3) 28,0 19,0 3,5 Organic substances, of which  95,0 97,0 26,0 Protein substances  95,0 97,0 10,0  Table 2  The effectiveness of AMINOFERT products, applied to sunflower, cultivated on cambic chernozem Unit: TELEORMAN Agricultural Research and Development Station Basic soil fertilization with N-80, P2O5, - 80 Kg/ha Version No .   Treatment  Number of  treatments  Soil concentration %  Quantity of fertilizers used  liters/ha   Production of seeds (kg/ha) Increase  For a treatment  For all the treatment   kg/ha  %  kg/liter  fertilizer used  1 Non-fertilized - - - - 2742 - 100,0 - 2 AMINOFERT N 2 0,5 5,0 10,0 3082 340*** 112,3 34,0 3 AMINOFERT NK 2 0,5 5,0 10,0 3193 451*** 116,4 45,1 4 AMINOFERT NPK 2 0,5 5,0 10,0 3216 474*** 117,3 47,4  Table 3 The effectiveness of AMINO-FERT products, applied to grape vine, variety Chasselas Doré,  cultivated on cambic chernozem. Unit: Institute of Biological Research, Iasi, Experimental Field of the University of Agricultural Sciences and Veterinary Medicine, Iaşi Without basic fertilization in soil Version No .   Treatment  Number of treatments  Soil concentration  %  Quantity of fertilizers used  liters/ha    Production of grapes  (kg/ha) Increase  For a treatment  For all the treatment   kg/ha  %  kg/liter  fertilizer used   1 Non-fertilized  - - - - 11213 - 100.0 - 2 AMINOFERT N 3 0.5 5.0 15.0 15357 4144*** 136.9 276.3 3 AMINOFERT NK 3 0.5 5.0 15.0 15545 4332*** 138.6 288.8 4 AMINOFERT NPK 3 0.5 5.0 15.0 15607 4394*** 139.2 292.9  Universitatea de Ştiinţe Agricole şi Medicină Veterinară Iaşi 50 Table 4 Chemical composition of the fertilizer AMINO-FERT 1H No. Composition (minimum values) AMINO-FERT 1H (g/l) 1 Total nitrogen (N), of which: 130 - Nitric nitrogen  5 - Ammonium nitrogen  5 - Amide nitrogen  120 2 Phosphorus (P2O5) 50 3 Potassium (K2O) 45 4 Copper (Cu) 0.07 5 Zinc (Zn) 0.05 6 Iron (Fe) 0.25 7 Manganese (Mn) 0.1 8 Boron (B) 0.1 9 Magnesium (MgO) 0.1 10 Sulfur (SO3) 0.5 11 Organic substances, of which: 12.0 - Protein substances, hydrolysate 7.5  Table 5  The effectiveness of the product AMINOFERT 1 H applied to H-Olt corn, cultivated on psamosol (sandy soil) Research and Development Center for Agricultural Plants on Sands – DABULENI, Without basic fertilization Version No .   Treatment  Number of treatments  Soil concentration %  Quantity of fertilizers used  liters/ha    Production of grains (kg/ha) Increase  For a treatment  For all the treatment   kg/ha  %  kg/liter  foliar fertilizer   1 Control - - - - 1644 - 100.0 - 2 AMINOFERT 1H  2.0 1.0 5.0 10.0 4522 2878*** 275.0 287.8  Table 6  The effectiveness of the product AMINOFERT 1 H applied to grape vine, variety Sauvignon,  cultivated on faeziom argic. Unit: Institute of Biological Research, Iasi, Experimental Field of the University of Agricultural Sciences and Veterinary Medicine, Iaşi. Basic fertilization with:  Nt-100, P2O5 – 100, K2O – 100 kg/ha Version No .   Treatment  Number of treatments  Soil concentration %  Quantity of fertilizers used  liters/ha    Production of grapes (kg/ha) Increase  For a treatment  For all the treatment   kg/ha  %  kg/liter  foliar fertilizer   1 Control - - - - 5580 - 100.0 - 2 AMINOFERT 1H  3 1.0 10.0 30.0 8030 2450*** 144.0 81.7  Table 7  The effectiveness of the product AMINOFERT 1 H applied to sunflower, variety Favorit, cultivated on cambic chernozem. Unit: TELEORMAN Agricultural Research and Development Station Basic fertilization with N-80, P2O5, - 80 Kg/ha Version No .   Treatment  Number of treatments  Soil concentration %  Quantity of fertilizers used  liters/ha    Production of seeds (kg/ha) Increase  For a treatment  For all the treatment   kg/ha  %  kg/liter  fertilizer   1 Control - - - - 2118 - 100.0 - 2 AMINOFERT 1H  2.0 1.0 5.0 10.0 3134 1016*** 147.9 101.6  Lucrări Ştiinţifice – vol. 57 (2) 2014, seria Agronomie 51 Table 8 Chemical composition of the fertilizer FERT - IRON Chemical composition  FERT – IRON (g/l) Nitrogen (Nt) 60 Phosphorus (P2O5) 23 Potassium (K2O) 27 Copper (Cu) 0.1 Zinc (Zn) 0.5 Iron (Fe) 27 Manganese (Mn) 0.1 Magnesium (Mg) 2.5 Boron (B) 0.5 Sulfur (S) 20 Protein hydrolysate  10  Means were compared by using Student test at the level of significance a = 0.05. According to the data in Tables 2,3,5,6,7,  it can be seen that there are significant differences for the two fertilizers compared to control and they are statistically insured. The performances achieved after the extra-radicular application on the soil Chardonnay grafted on rootstock Berlandiaeri x Riparia SO4-4 (1% solution, three treatments) of the fertilizer FERT - IRON can be summarized as follows:  - it has brought a very significant production increase of 50.7%, compared to the untreated control;  - it has brought an increase in grain dry matter content, statistically highly significant of 9.9% compared to the untreated control;  - it has brought an increase in sugar content from must (of grapes) / unfermented wine, very significant from 254 g/l (untreated control) to 272.0 g/l.  - it has prevented and treated ferric chlorosis of grapevine.       CONCLUSIONS  It has been developed a technology for obtaining extra-radicular fertilizers of the type NPK with macro, meso, micronutrients and protein hydrolysates. There were obtained and characterized physically and chemically five fertilizers with extra-radicular application, agro-chemically tested in the National Network for fertilizers authorization. An increased production obtained by applying extra-radicular fertilizers, compared to the foliar unfertilized control, was significant and ranged between 12% for the sunflower and 50% for the grapevine. There have been formulated and obtained two fertilizers AMINO-FERT N and NK, tested and certified, compatible for application in organic farming. There has been formulated and obtained a fertilizer for the prevention and treatment of ferric chlorosis of grapevine. Fertilizing compositions have been authorized for use in agriculture.    REFERENCES  Cioroianu T.M., Sîrbu C., 2011 - Unconventional Fertilizers – Extra-radicular fertilizers with protein substances, Est Falia Publishing House, Bucharest.  Cioroianu T.M., Sîrbu C., Dumitru M., Dorneanu A., Stefanescu D., 2009 - Unconventional fertilizers – liquid fertilizers, Est Falia Publishing House, Bucharest. Cioroianu T.M., Sîrbu C., Oprica I., Grigore A., Mihalache D., Dumitrascu M., Anton I., Soare M., 2011 - Fertilizers with protein substances, technology, agro-chemical and physical characteristics, Scientific Papers of the Symposium, Series I, vol. XIII - ISSN: 1453-1410, p. 449-453.  Cioroianu T.M., Sîrbu C., Oprica I., Grigore A., Mihalache D., Dumitrascu M., Anton I., Soare M., 2011 - Fertilizers with protein substances, technology, agro-chemical and physical characteristics paper published in the volume of Scientific Papers of the Symposium, Series I, vol. XIII - ISSN: 1453-1410, p. 449-453. Dorneanu A., Dorneanu E., Sîrbu C., Cioroianu T.M., 2003 - Foliar Fertilization a possible technology to support modern, ecological agriculture – Symposium:  "Diversifying the range of fertilizers and improvement of their quality according to the requirements of sustainable agriculture" Bacau, p. 249-257.  Iovi A., Iovi C., Negrea P., 2000 – Ecologic technologies, Chemistry and technology of fertilizers with microelements, Politehnica Publishing House, ISBN 973-9389-56-2. Sîrbu C., Cioroianu T.M., Dumitrascu M., 2010 - New fertilizers with protein structure with phyto-stimulating role, Organic farming - Priorities and Prospects", Scientific works, Series Agronomy, U.Ş.A.M.V. Iaşi, vol. 52.  Sîrbu C., Cioroianu T.M., Dumitru M., Dorneanu A., Mihalache D., Angelescu L., 2007 – Fertilizers containing chelating protein hydrolyzates with applications in agriculture – International Symposium "Ecological restoration and fertilizers requirements in the area of Gorj County, Tg. Jiu, 4 – 5 October 2007 New Agris Publishing House, p. 467- 474.   Universitatea de Ştiinţe Agricole şi Medicină Veterinară Iaşi 52 Sîrbu C., Cioroianu T.M., Dumitru M., Dorneanu A., Negrila M., Mihalache D., Anghelescu L., 2008 - New types of bio-fertilizers with chelating protein hydrolyzed structures, with bio-stimulating and protective role, with applications in sustainable agriculture, Annals of the University of Agricultural Sciences and Veterinary Medicine of Iasi, series Agronomy, vol. 51 (2), p. 189-194. Sîrbu C., Cioroianu T.M., Cojocaru I., Trandafir V., Albu M.G., 2009 - The fertilizers with the protein chelated structures with bio-stimulating role, Journal of Chemistry, vol. 60 (11). Trandafir V., Popescu G., Albu M.G., Iovu H., Georgescu M., 2007 – Collagen bio-products, ARS DOCENDI Publishing House, Bucharest.  Trandafir V., Cioroianu T.M., Bratulescu V., Cioroianu T., 2003 - Use of collagen hydrolisate as an active ingredient in bio-Fertilizers, Cucio Pel. Mat., vol. 79 (6), p 289-293.  *** - Regulation (EC) No. 2003/2003 the European Parliament and of the Council of 13 October 2003 on fertilizers *** - Regulation (EC) No. 834/2007 of the Council of 28 June 2007 on organic production and labeling of organic products, and for repealing the Regulation (EEC) No. 2092/91    

Protein hydrolysates and amino-acids fertilizers – physicochemical characteristics

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Protein hydrolysates and amino-acids fertilizers – physicochemical characteristics

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