CN104195206B - Method for reducing content of volatile basic nitrogen in active peptides of euphausia superba - Google Patents

Abstract

The invention aims at providing a method for reducing the content of volatile basic nitrogen in active peptides of euphausia superba. The method comprises the following steps: enzymatically hydrolyzing euphausia superba powder, and defluorinating to obtain an active peptide aqueous solution; adjusting the solid content of the active peptide aqueous solution of the euphausia superba by diluting or concentrating, and adding a swelling agent into the active peptide aqueous solution of the euphausia superba; heating the active peptide aqueous solution of the euphausia superba in vacuum for a period of time, cooling, and centrifuging to obtain supernatant; desalting the supernatant, and spray-drying to obtain an euphausia superba active peptide powder product with low volatile basic nitrogen content. The method is simple to perform, low in equipment investment, free of environmental pollution, high in efficiency and suitable for industrialized operation; the obtained euphausia superba active peptide powder with low volatile basic nitrogen content has the characteristics of no harmful material residues and high product safety.

Description

A kind of method reducing TVB-N content in krill bioactive peptide

Technical field

The invention belongs to food, medical supplies processing technique field, be specifically related to a kind of method reducing TVB-N content in water-soluble krill bioactive peptide.

Background technology

Total volatile basic nitrogen refers under the effect of bacterium and enzyme, animal food in the process of corruption, the alkaline nitrogenous substancess such as the ammonia that Protein in Food decomposes and produces and amine.Total volatile basic nitrogen is that a class has volatile toxic substance.The content of total volatile basic nitrogen is an important indicator of fishery technology freshness, and freshness and the course of processing of it and raw material have obvious dependency.

Krill is a class marine invertebrate, is distributed in antarctic waters.According to statistics, the standing stock of krill are 6.5-10 hundred million tons, are the huge protein resource treasure-houses of the mankind.In krill dry weight, protein content is about 65-75%, and its amino acid composition is reasonable, essential amino acids content is high, is the important source material of carrying out protein higher value application.

Due to the limitation of processing on ship, krill is after fishing for, and freshness declines rapidly.The decline of raw material freshness, makes the TVB-N content of krill raw material be in a higher initial level.And in the process of preparation krill bioactive peptide, after the process of super-dry, enzymolysis, defluorinate, the link such as concentrated, in sample, TVB-N content rises further.In krill bioactive peptide, the height of TVB-N content directly has influence on the exploitation of its popularization at field of food and krill industry to a certain extent.

Domestic patent CN201020683123.7 and CN201210284607.8 individually discloses a kind of device utilizing total volatile basic nitrogen in vapor removal fish meal, the TVB-N content in fish meal can be reduced within the scope of technic index.But, if under water-soluble krill bioactive peptide is exposed to high-temperature steam, aerobic environment for a long time, the changes such as look change can be caused, causes active peptide powder Quality Down.Therefore, under being necessary to provide a kind of prerequisite not reducing water-soluble krill bioactive peptide nutritive value further, the method for TVB-N content can effectively be reduced.

Summary of the invention

The object of this invention is to provide a kind of method reducing TVB-N content in water-soluble krill bioactive peptide, both the method for TVB-N content in the water-soluble krill bioactive peptide of simple to operate, that cost is low, efficiency is high reduction, thus supplement the deficiencies in the prior art.

Reduce the method for TVB-N content in water-soluble krill bioactive peptide in the present invention, it is characterized in that comprising the steps:

1) in euphausia superba powder, add distilled water, feed liquid mass ratio is 1:6 ~ 15, regulates mixed solution pH value, then in mixed solution, adds proteolytic enzyme be hydrolyzed, and hydrolysis temperature is 40 ~ 60 DEG C, and hydrolysis time is 2 ~ 12h; Hydrolysis terminates post-heating enzymolysis reaction, and centrifugal, obtain supernatant liquor, supernatant liquor, after defluorinate process, obtains defluorinate krill bioactive peptide water solution A;

2) concentration of solid content in regulator solution A, obtains solution B, in solution B, adds raising agent, obtain mixed solution C;

3) by solution C in heated under vacuum, after cooling, centrifuging and taking supernatant liquor, obtains mixed solution D;

4) solution D is through desalination, spraying dry, obtains the krill bioactive peptide powder-product of low volatility alkali nitrogen content.

Wherein, step 1) in proteolytic enzyme be one or more in food grade papoid, bromeline, trypsinase, neutral protease or Sumizyme MP;

Step 2) in the solid content of solution B be 5 ~ 30%;

Step 2) in raising agent be food grade raising agent, comprise potassium alum, tschermigite, secondary calcium phosphate etc., addition is 0.5 ~ 5.0% of solid content in solution B;

Step 3) in vacuum tightness be 0.04 ~ 0.09MPa, Heating temperature is 30 ~ 80 DEG C, and heat-up time is 1 ~ 6h;

Step 4) in the method for desalination comprise in ion exchange resin desalination, macroporous resin desalination, nanofiltration desalination, electrodialytic desalting one or more, during spraying dry, inlet temperature is 120-160 DEG C, and air outlet temperature is 80-120 DEG C;

In the present invention, reached the object removing most of total volatile basic nitrogen in the krill bioactive peptide aqueous solution by the mode of heating under vacuum; In bioactive peptide solution, add raising agent, be more conducive to steaming of total volatile basic nitrogen in solution.In the present invention, under not adding the condition of raising agent, the TVB-N content after vacuum heating treatment in krill active peptide powder can reduce by 50 ~ 70%; Under adding the condition of raising agent, the TVB-N content after vacuum heating treatment in krill active peptide powder can reduce by more than 90%.Method of the present invention is simple to operate, facility investment is few, non-environmental-pollution, efficiency are high, is applicable to industrial operation; The krill active peptide powder of gained low volatility alkali nitrogen content has following feature: unharmful substance is residual, Product Safety is high, TVB-N content is reduced to 10 ~ 35mg/100g by 100 ~ 500mg/100g, ash oontent is lower than 7%, and protein content is higher than 85%.

Embodiment

The measuring method of TVB-N content is with reference to the microdiffusion in GB/T 5009.44-2003 " analytical procedure of GB-meat quail hygienic standard ".Specific as follows:

1) reagent

Unsaturated carbonate potassium solution: take 50g salt of wormwood, adds 50mL water, micro-heating hydrotropy, uses supernatant liquor;

Water-soluble glue: take 10g gum arabic, adds 10mL water, then adds 5mL glycerine and 5g Anhydrous potassium carbonate, grinds well;

Boric acid absorption liquid: 20g/L;

Mixing indicating liquid: configure methyl red-ethanol indicator (2g/L) and methylene blue indicator (1g/L) respectively, facing the used time is mixing indicating liquid by above-mentioned two kinds of indicator balanced mix;

Hydrochloric Standard Titration: 0.010mol/L.

2) analytical procedure

Sample pretreating: take 10g sample, be placed in Erlenmeyer flask, adds 100mL water, frequently jolting, and filter after dipping 30min, filtrate is sample liquid.

Water-soluble glue is applied to the edge of diffusion boat, in ware central authorities, room adds 1mL absorption liquid and 1 mixing indicating liquid.Add 1.00mL sample liquid in ware mistress side, opposite side adds 1mL unsaturated carbonate potassium solution, notes not making two liquid contacts, builds immediately; Rotated gently on desktop by ware after sealing, sample liquid is mixed with alkali lye, then in 37 DEG C of incubators, place 2h, throw off lid, use Hydrochloric Standard Titration titration, terminal is bluish voilet.Do reagent blank test simultaneously.

3) result calculates

In sample, the content of total volatile basic nitrogen calculates according to formula (1).

$X=\frac{(V1-V2)\×c\×14}{m\×1/100}\×100---\left(1\right)$

In formula:

X: the content of total volatile basic nitrogen in sample, unit is the every hectogram of milligram (mg/100g);

V1: mensuration sample liquid consumes the volume of hydrochloric acid standard solution, and unit is milliliter (mL);

V2: sample blank consumes the volume of hydrochloric acid standard solution, and unit is milliliter (mL);

C: the actual concentrations of hydrochloric acid standard solution, unit is mole often liter (mol/L);

14: the quality of the nitrogen suitable with 1.00mL Hydrochloric Standard Titration, unit is milligram (mg);

M: sample mass, unit is gram (g).

Below in conjunction with embodiment, the present invention is described in detail

Embodiment 1

1) euphausia superba powder 5kg is got, 50L distilled water is added in shrimp med, obtain mixed solution, in mixed solution, add 2M HCl or 2M NaOH regulate mixed solution pH value to 8.0, be warming up to 50 DEG C, Sumizyme MP 80g is added in mixed solution, after stirring 10h in a kettle., be heated to 100 DEG C and stop enzyme reaction, centrifuging and taking supernatant liquor, supernatant liquor, after defluorinate process, obtains defluorinate krill bioactive peptide water solution A;

2) make the solid content of solution A reach 15% by vacuum concentration, vacuum tightness is 0.08MPa, and temperature is 60 DEG C, obtains solution B;

4) get solution B 10L, add 15g secondary calcium phosphate wherein, obtain mixed solution C;

5) mixed solution C is heated 3h under vacuum, vacuum tightness is 0.08MPa, and temperature is 60 DEG C, centrifugal after cooling, obtains supernatant liquor D;

6) supernatant liquor D obtains solution E through desalting treatment;

7) solution E is through spray drying treatment (inlet temperature is 135 DEG C, and air outlet temperature is 85 DEG C), obtains the krill bioactive peptide powder-product of low volatility alkali nitrogen content.

In the krill bioactive peptide product of gained low volatility alkali nitrogen content of the present invention, TVB-N content is 10 ~ 35mg/100g, and ash oontent is lower than 7%, and protein content is higher than 85%.And be 100 ~ 500mg/100g without TVB-N content in the krill bioactive peptide powder-product removing total volatile basic nitrogen; Be 280mg/100g without TVB-N content in the krill bioactive peptide powder-product removing total volatile basic nitrogen in the present embodiment.

Embodiment 2

1) get euphausia superba powder 10kg, add 100L95% ethanol, stir 6h in a kettle. in shrimp med, centrifuging and taking precipitates, and vacuum-drying obtains degreasing krill meal;

2) extracting degreasing shrimp med 5kg, 60L distilled water is added in degreasing shrimp med, obtain mixed solution, in mixed solution, add 2M HCl or 2M NaOH regulate mixed solution pH value to 7.5, be warming up to 45 DEG C, neutral protease 100g is added in mixed solution, stir 8h in a kettle., be heated to 100 DEG C and stop enzyme reaction, centrifuging and taking supernatant liquor, supernatant liquor, after defluorinate process, obtains defluorinate krill bioactive peptide water solution A;

4) make the solid content of solution A reach 20% by vacuum concentration, vacuum tightness is 0.07MPa, and temperature is 50 DEG C, obtains solution B;

5) get solution B 10L, add 30g potassium alum wherein, obtain mixed solution C;

6) mixed solution C is heated 4h under vacuum, vacuum tightness is 0.07MPa, and temperature is 50 DEG C, centrifugal after cooling, gets supernatant liquor D;

7) supernatant liquor D obtains solution E through desalting treatment;

8) solution E is through spray drying treatment (inlet temperature is 125 DEG C, and air outlet temperature is 95 DEG C), obtains the krill bioactive peptide powder-product of low volatility alkali nitrogen content.

In the krill bioactive peptide product of gained low volatility alkali nitrogen content of the present invention, TVB-N content is 10 ~ 35mg/100g, and ash oontent is lower than 7%, and protein content is higher than 85%.And be 100 ~ 500mg/100g without TVB-N content in the krill bioactive peptide powder-product removing total volatile basic nitrogen.

The above results shows that method of the present invention can the content of total volatile basic nitrogen in the water-soluble krill bioactive peptide of reduction of significance.

Claims (4)

1. reduce a method for TVB-N content in water-soluble krill bioactive peptide, it is characterized in that, described method comprises the steps:

1) in euphausia superba powder, add distilled water, feed liquid mass ratio is 1:6 ~ 15, regulates mixed solution pH value, then in mixed solution, adds proteolytic enzyme be hydrolyzed, and hydrolysis temperature is 40 ~ 60 DEG C, and hydrolysis time is 2 ~ 12h; Hydrolysis terminates post-heating enzymolysis reaction, and centrifugal, obtain supernatant liquor, supernatant liquor, after defluorinate process, obtains defluorinate krill bioactive peptide water solution A;

2) concentration of solid content in regulator solution A, obtain solution B, the solid content of solution B is 15 ~ 20%; In solution B, add raising agent, obtain mixed solution C; Wherein the addition of raising agent is 0.5 ~ 5.0% of solid content in solution B;

Described raising agent is potassium alum, any one or several in tschermigite, secondary calcium phosphate;

3) by solution C in heated under vacuum, after cooling, centrifuging and taking supernatant liquor, obtains mixed solution D;

Described vacuum condition, wherein vacuum tightness is 0.04 ~ 0.09MPa, and Heating temperature is 50 ~ 60 DEG C, and heat-up time is 1 ~ 6h;

4) solution D is through desalination, spraying dry, obtains the krill bioactive peptide powder-product of low volatility alkali nitrogen content.

2. the method for claim 1, is characterized in that described step 1) in proteolytic enzyme be one or more in food grade papoid, bromeline, trypsinase, neutral protease, Sumizyme MP.

3. the method for claim 1, is characterized in that described step 4) in the method for desalination be any one in ion exchange resin desalination, macroporous resin desalination, nanofiltration desalination, electrodialytic desalting method.

4. the method for claim 1, is characterized in that described step 4) in spraying dry time inlet temperature be 120-160 DEG C, air outlet temperature is 80-120 DEG C.