Method of processing synanthropic fly larva

FIELD: chemistry, biochemistry.

SUBSTANCE: invention relates to biotechnology. The proposed method comprises thermal processing of larva, separated from a substratum upon termination of their cultivation, for 2 to 15 min at 75 to 100°C. Thermal processing over, larvae are crushed, chitine-containing fraction is separated from hemolymph. Now, the chitine-containing precipitate as-washed and crushed is subjected to enzymic hydrolysis at temperature 35 to 46°C with pH of 6.5 to 8.5 till achieving amine nitrogen content not below 0.8 G/%. Then, the separated chitine-containing precipitate is subjected to alkaline hydrolysis at continuous mixing for 60 to 90 min at 55 to 65°C. Finally, the aforesaid chitine-containing precipitate is subjected to demineralisation by 1.5 to 3.0 percent solution of hydrochloric or nitric acid for 30 to 40 min at 60 to 70°C. After that, the precipitate containing chitine is rinsed with water till pH of 6.5 to 7.5 and dried.

EFFECT: increase in biological value of target products with simultaneous increase in their number and simplification of process method.

4 cl, 1 ex

 

The invention relates to biotechnology, and more specifically to integrated waste processing larvae of synanthropic flies cultured on organic waste from pig and poultry farms.

From the levels known in the art is a method of processing larvae of synanthropic flies, including the cultivation of larvae of synanthropic flies on organic waste, separation of larvae from the substrate at the end of their cultivation, grinding raw larvae until a pasty mass, which first stariway in airtight metal containers, and then sterilized by stepwise increasing the temperature to 150°With (see author's certificate SU - A1 - No. 1517904, 1989).

The disadvantage of this method is that it does not provide deep processing of biomass of larvae that, on the one hand, does not provide the whole range of valuable products (chitin, chitosan, protein hydrolysates), with a wide range of applications, and, on the other hand, leads to lower quality in this way feed additives, due to the presence of components, which in the native form is not fully digested by the body of the animal.

Also known is a method of processing larvae of synanthropic flies, which is taken as a prototype and according to which larvae of synanthropic flies, separated tsubstrate after cultivation for swine manure or poultry litter, freeze, then using a press-fit larvae crush and separate heterosocial fraction from the first target product haemolymph (liquid protein weight), then washed with water heterosocial fraction is subjected to a first demineralization 1,5-5,0% solution of a strong inorganic acid in a ratio of solid and liquid phases, is equal to 1:(3-4) by weight, with constant stirring for 1.5 to 2.0 hours and the temperature of 18-30°and then after washing heterotetramer sediment water heterostasis residue is subjected to alkaline cleaning (alkaline hydrolysis) of 2.0 to 5.0 percent solution of alkali in the ratio of solid and liquid phases, is equal to 1:(3-5) by weight, with constant stirring for 1.5 to 3.0 hours and a temperature of 90-100°With conditions precluding contact of the reaction medium with air. Then separated from the alkaline hydrolysate (which is the next target product), and the precipitate is washed successively with water, ethanol and chloroform, resulting receive the third target product - chitin (see patent RU - A1 No. 2139887, 1999). The main disadvantage of the prototype is that single-stage alkaline hydrolysis held to the same with a hard modes, does not provide effective cleaning of chitin from proteins, leads to the degradation of chitin, reduce biological the practical value of protein, obtained by alkaline hydrolysis, as well as to the necessity of using organic solvents at wash obtained from chitin soluble fractions. However, it has been found that the use in the production of chitin organic solvents, including chloroform, does not allow the use of chitin for food, in particular, biologically active additives. In addition, the implementation of the known method of processing larvae of synanthropic flies deals with the necessity of use of the refrigerator, as well as special tools and protective environments to prevent contact of the reaction medium with air. The complication of the technical realization of this method inevitably leads to an increase in the value of the target products.

The present invention is directed to the solution of the technical problem to provide to increase the biological value of target products, while increasing the number of received target products and simplifying the technical implementation of the method of processing larvae of synanthropic flies. Achievable technical result is to preserve to the greatest extent native properties of chitin, protein feed, obtained from the alkaline hydrolysate, in providing more effective cleaning of chitin from protein at the expense of additional product - farms is ntatives protein hydrolysate, while simplifying the technical implementation of the method of processing larvae of synanthropic flies.

The problem is solved by the fact that in the way pererabotki larvae of synanthropic flies, including the crushing of the larvae and the Department hitinsoderjath fraction from the haemolymph, the demineralization solution of a strong inorganic acid and alkaline cleaning heterotetramer sludge separation of the alkaline hydrolysate according to the invention before the Department hitinsoderjath fraction from the hemolymph of the larvae, which are separated from the substrate at the end of their cultivation, subjected to heat treatment for 2 to 15 minutes at a temperature of 75-100°C, and after crushing cooked larvae separated, washed with water and crushed heterosocial fraction is subjected to enzymatic hydrolysis carried out an extract of the pancreas of proteolytic activity not less than 0.3 PE/G, taken in the ratio of 0.9 to 1.1 liters per kilogram hitinsoderjath fraction at a pH of 6.5-8.5 and a temperature of 35-48°before reaching the content of amino nitrogen is not lower than 0.8 G/%, after enzymatic hydrolysis stop, separating the solution of protein hydrolysate from heterotetramer residue which is subjected to alkaline cleaning from 1.5 to 3.0 percent alkali solution in the ratio of liquid and solid time of 1:(3-4) on the Assos, under continuous stirring for 60 to 90 minutes and a temperature of 55-65°after the separation of heterotetramer precipitate from the alkaline hydrolysate he, after washing with water, is subjected to demineralization 1,5-3,0% solution of hydrochloric or nitric acid in the ratio of solid and liquid phases, is equal to 1:(3-4) by weight, with continuous stirring for 30-40 minutes and a temperature of 60-70°C, after which containing chitin precipitate is washed to a pH of 6.5-7.5, dried or direct the production of chitosan.

In addition, the problem is solved by the fact that:

- obtained solution of the enzymatic protein hydrolysate lighten and dried;

- obtained solution of the alkaline hydrolysate is neutralized to a pH of 6.5-7.5 and dried;

- neutralization of the alkaline hydrolysate is carried out by mixing it with those obtained by demineralization of the liquid fraction.

The advantage of the proposed method for processing larvae of synanthropic flies in the prototype is, first, that the conduct of operations instead of freezing larvae, separated from the substrate at the end of their cultivation, their heat treatment not only provides pasteurization biomass of larvae, but also the suppression tyrosinase reaction that improves the quality of target products while simplifying the technical implementation of the method per the developing larvae of synanthropic flies. Secondly, the proposed carrying out in sequence the two-stage hydrolysis enables you not only to more efficient cleaning of chitin from protein and gain an additional enzymatic protein hydrolysate, which by its biological value than alkaline hydrolysate, but, on the one hand, allows to significantly mitigate the modes of carrying out the alkaline hydrolysis (which provides a higher degree of preservation of the native properties of chitin and increases the biological value of feed protein secreted by alkaline hydrolysis), and on the other hand, eliminates the need for washing the chitin of organic solvents, which expands the scope of its use.

A method of processing larvae of synanthropic flies is as follows. Cultivation of larvae of synanthropic flies carried out on organic waste, preferably in swine manure or poultry litter at a temperature 20-52°C for 5-7 days after depositing in swine manure or poultry litter humidity of 70-85% of eggs synanthropic flies of the calculation, for example, one gram of eggs per kilogram of manure or litter. After culturing larvae of synanthropic flies (which is judged by the temperature decrease of the substrate) larvae separated from the recycled organic waste is in, for example, using a mesh separator.

Larvae of synanthropic flies, separated from the substrate after cultivation for swine manure or poultry litter, exposed for 2 to 15 min heat treatment in water with a temperature of 75-100°that, on the one hand, it provides pasteurization, and on the other hand, provides the inhibition of the enzyme complex involved in the response melanieortega. Thus, improving the quality of target products by suppressing tyrosinase reaction. The duration of heat treatment of larvae in the water is determined primarily by the temperature of the water, namely: the higher the water temperature, the less time for the heat treatment of larvae.

Then the larvae that underwent heat treatment, crush and separate heterosocial fraction from the haemolymph, which is the first target product, for example, using a press cage, the diameter of the drum is preferably of 0.5-0.75 mm. Highlighted the hemolymph (liquid protein fraction of the biomass of larvae) are used in native or dried form (fish meal) for animal feed, for example, in the composition of animal feed or feed additives for making mixes.

Heterosocial fraction washed with water, for example water, ground in a colloid mill, and then subjected to enzymatic what the hydrolysis, carried out an extract of the pancreas of proteolytic activity is not less than 0.3 PE/G per gram of dry substance, for example, cattle or pigs, taken in the ratio of 0.9 to 1.1 liters per kilogram hitinsoderjath fraction at neutral or slightly alkaline conditions (pH 6.5 to 8.5) and temperature 35-48°before reaching the content of amino nitrogen is not lower than 0.8 G/%. After reaching the required content of amino nitrogen spend the acid inactivation of the enzymes contained in the extract of the pancreas, by adding 10% hydrochloric acid up to pH values of 4.5 and 5.2. As a result of enzymatic hydrolysis stops. The resulting solution enzymatic protein hydrolysate is separated from heterotetramer sediment, lighten filter or centrifuge, and then purified enzymatic protein hydrolysate is dried. The resulting powder contains not less than 90 wt.% protein and can be used for preparing microbiological culture media, diet, starter feed, etc.

Washed heterostasis residue is subjected to alkaline cleaning (alkaline hydrolysis), which is carried from 1.5 to 3.0 percent alkali solution in the ratio of solid and liquid phases, is equal to 1:(3-4) by weight at a temperature of 55-65°C for 60-90 minutes and at a constant peremeci the years. The resulting solution of the alkaline hydrolysate is separated from heterotetramer sludge is neutralized, for example, hydrochloric acid to a pH of 6.5-7.5 and dried. Alkaline hydrolysate can be used for livestock feed in liquid and dry form.

Deproteinizing heterostasis the precipitate washed twice with water and subjected to demineralization 1,5-3,0% solution of hydrochloric or nitric acid in the ratio of solid and liquid phases, is equal to 1:(3-4) by weight, with continuous stirring for 30-40 minutes and a temperature of 60-70°C. Separating the liquid fraction, which in the preferred embodiment, the method is mixed with an alkaline hydrolysate, as mentioned above, to a pH of 6.5-7.5 and containing chitin precipitate is washed with water until neutral environment (pH of 6.5-7.5), dried or direct the production of chitosan by deacetylation.

The invention is further explained with a specific example. 10 kg of pig manure contribute 10 g eggs synanthropic flies. Cultured larvae of synanthropic flies in for five days at a temperature of 25°and the moisture content of manure 70-72%. Then the larvae are separated from the substrate using a grid with a cell size of 2×2 mm and subjected to heat treatment at a temperature of 80°within 10 minutes Past thermal processing larvae crush and separate the hemolymph from hitinsoderjath faction. When e is om get 300 grams hitinsoderjath faction, which is washed with water, ground in a colloid mill and subjected to enzymatic hydrolysis. For this heterosocial fraction is placed in the fermenter, the volume was adjusted with water to one liter, petitrenaud pH to 8.0, heated to 45°and poured 300 ml of the extract of the pancreas of cattle. The hydrolysis is carried out at a constant temperature by controlling the content of amino nitrogen. After 12 hours, when the value of the amine nitrogen has reached the value of 0.87 G/%, the hydrolysis is stopped by the addition of 10% hydrochloric acid to a pH of 4.7. The supernatant is separated, lighten on a suction filter and dried in the spray dryer. The obtained dry powder used for preparation of nutrient media under cultivation of microorganisms. Received heterostasis residue is poured 500 ml of 3% alkali solution, heated to 60°; and incubated with constant stirring for 90 minutes Then adosados is separated, neutralized with hydrochloric acid to pH 6.5 and dried. The remaining mass is twice washed with water, pour 500 ml of 3% hydrochloric acid, heated to 60°and with constant stirring withstand 30 minutes After which the contents are moved to filter and separate the liquid fraction. Derived chitin was washed with water to pH 7.0 and dried. The result is 32 grams of chitin is, which meets the technical requirements TU 15-02538-89.

The invention can be used at production facilities, ensuring full utilization of solid organic waste of some species of farm animals and poultry.

1. A method of processing larvae of synanthropic flies, including the crushing of the larvae and the Department hitinsoderjath fraction from the haemolymph, the demineralization solution of a strong inorganic acid and alkaline cleaning heterotetramer sludge separation of the alkaline hydrolysate, characterized in that before the Department hitinsoderjath fraction from the hemolymph of the larvae, which are separated from the substrate at the end of their cultivation, subjected to heat treatment for 2 to 15 minutes at a temperature of 75-100°and after crushing cooked larvae separated, washed with water and crushed heterosocial fraction is subjected to enzymatic hydrolysis carried out an extract of the pancreas of proteolytic activity is not less than 0.3 PE/G taken in the ratio of 0.9-1.1 l / kg hitinsoderjath fraction at a pH of 6.5-8.5 and a temperature of 35-48°before reaching the content of amino nitrogen is not lower than 0.8 G/%, after enzymatic hydrolysis stop, separate the solution of the enzymatic protein hydrolysate from HiTi ostergade sediment, which is subjected to alkaline cleaning 1,5-3,0%alkali solution at a ratio of solid and liquid phases, is equal to 1:(3-4) by weight, with continuous stirring for 60-90 min and a temperature of 55-65°after the separation of heterotetramer precipitate from the alkaline hydrolysate heterostasis residue after cleaning with water is subjected to demineralization 1,5-3,0%solution of hydrochloric or nitric acid in the ratio of solid and liquid phases, is equal to 1:(3-4) by weight, with continuous stirring for 30-40 min and a temperature of 60-70°after which containing chitin precipitate is washed to a pH of 6.5-7.5, dried or direct the production of chitosan.

2. The method according to claim 1, characterized in that the resulting solution of the enzymatic protein hydrolysate lighten and dried.

3. The method according to claim 1, characterized in that the obtained solution of the alkaline hydrolysate is neutralized to a pH of 6.5-7.5 and dried.

4. The method according to claim 3, characterized in that the neutralization of the alkaline hydrolysate is carried out by mixing it with those obtained by demineralization of the liquid fraction.



 

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