CN109295146A - Enzyme process abalone enzymatic extract and preparation method thereof - Google Patents
Enzyme process abalone enzymatic extract and preparation method thereof Download PDFInfo
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Abstract
The present invention carries out finishing processing to abalone using the means of bioengineering, by self-dissolving enzymatic hydrolysis, active bacteria fermentation, impulse ultrasound handles and the combination of enzymolysis process, so that the macromolecular substances such as protein in abalone are maximumlly degraded to active small molecular, improve the producing level and nutrition balance of abalone nutritional ingredient, obtained abalone enzymatic extract has good antioxidant activity and anti-fatigue effect, can be used as corresponding efficiency health-care food, drug processes raw material, greatly improve the added value and industrial value of abalone processing.
Description
Technical field:
The invention belongs to bioengineering fields, are related to enzyme engineering and fermentation engineering, and in particular to a kind of abalone Enzymatic Extraction
Object and preparation method thereof, by digesting the abalone enzymatic extract that there is high bioactivity with zymotechnique preparation.
Background technique:
Abalone is a kind of mollusk, is Gastropoda, Probranchia, Archaeogastropoda, Bao Ke, Bao category, is a kind of rare sea
Product.Abalone geographical distribution is wide, and there are Australia, Japan, China, New Zealand, the U.S., Europe and the Middle East one in main place
Band, abalone are traditional high-grade nourishing food, " soft gold " being known as on dining table, dietotherapeutic, according to " Mingyi Bielu " record, Bao
The sweet salty temperature of piscinity, function tonification liver kidney, benefiting shrewd head；It is recorded in Compendium of Material Medica, abalone is mild-natured, and it is sweet in flavor, salty, there is improving eyesight benefit
Void, heat-clearing enriching yin, blood-nourishing benefit stomach, nourishing liver and kidney effect.
With the application of artificial abalone cultural technique, Fujian Province and Guangdong Province become the maximum abalone culture base in China
Ground, wherein Guangdong Province Shanyi City forms one of maximum abalone culture base in the whole nation, produces 900 t of abalone per year, and in lasting
Ascendant trend；And Fujian Province's Partial Sea Area seawater is pure, seaweed is abundant, is very suitable to abalone growth, and abalone yield accounts for the whole nation
Yield 3/4ths or so, while Fujian abalone kind is mostly to hybridize Bao, individual is larger, the growing environment in southern sea area is adapted to,
Form that the time is short, and survival rate is higher, the big abalone Industry Foundation of amount of fine quality brings booming chance, " Zhang to Fujian abalone
State abalone " and " Liaanjiang county abalone " have become the famous brand name of Chinese abalone industry.
However, being still in the junior stage for the processing and utilization of abalone at present, the processing of existing abalone is concentrated mainly on dry
System and instant abalone, the focusing on of product development promote abalone Individual Size, the rehydration technology of dried product, instant abalone
Flavor etc..Processing in relation to abalone at present, nutritional ingredient and physiological activity research very understatement road, few document report mentions
To abalone is processed as abalone sauce or is processed as enzymatic hydrolyzed extract by zymolysis technique, however, for the processing and utilization of abalone
Maximum economic benefit is not yet played, and comprehensive development and utilization degree is inadequate.
Summary of the invention:
Based on the state of development of existing abalone production and processing technology, the present invention is intended to provide a kind of abalone enzymatic extract and its system
Preparation Method, by carrying out the abalone enzymatic extract that high activity is prepared in depth growth working process to abalone, wherein rich
Containing abalone polypeptide, have the function of multinomial physiological hygiene, and excellent flavor, compared to traditional enzymolysis product, bitter taste peptide content is few,
Substantially there is no bitter taste, which can be used for the processing of health food, drug, be with a wide range of applications.
In order to solve the above technical problems, the present invention adopts the following technical scheme:
A kind of abalone enzymatic extract, which is characterized in that be prepared by the following method to obtain:
Pretreatment: step 1 takes fresh abalone, decladding takes abalone meat after removing internal organ, and abalone meat is immersed in ozone water solution
Carry out disinfection processing；
Step 2, the preparation of slurries: the abalone meat that step 1 has been pre-processed is mixed with mixture of ice and water, be placed in colloid mill into
Abalone pretreatment slurries are prepared in row defibrination process；
Step 3, self-dissolving hydrolysis: the abalone pretreatment slurries that step 2 is prepared are irradiated using ultraviolet light, in the process
And it is stirred continuously, self-dissolving hydrolysis；
Step 4, fermentation process: being added oligoisomaltose and glucose in the self-dissolving hydrolysate that step 3 is prepared, and
It is inoculated with saccharomyces cerevisiae and Lactobacillus casei thereto afterwards, carries out fermentation process, obtain tunning；
Impulse ultrasound pretreatment: step 5 carries out impulse ultrasound pretreatment to the tunning that step 4 obtains, it is super to obtain pulse
Sound preconditioning product；
Enzymatic hydrolysis: the pH of impulse ultrasound pretreatment product made from step 5 is adjusted to 7.5-8.2, is added by alkalinity by step 6
Protease, papain, neutral proteinase and compound fertilizer production composition complex enzyme, 40-42 DEG C is heated to, at enzymatic hydrolysis
Reason 3.5-4 hours, obtains enzymolysis liquid, complex enzyme neutral and alkali protease, papain, neutral proteinase and composite flavor albumen
The weight of enzyme is 3:(1-1.5): (1-2): (2-3), the additive amount of complex enzyme are 0.5-0.7%；
Step 7, enzyme deactivation, concentration, as abalone enzymatic extract.
In particular it relates to following specific technical solution:
A kind of abalone enzymatic extract, which is characterized in that be prepared by the following method to obtain:
Pretreatment: step 1 takes fresh abalone, decladding takes abalone meat after removing internal organ, and abalone meat is immersed in ozone water solution
Carry out disinfection processing, and the processing time is 20min, and treatment temperature is 5 DEG C, and ozone concentration is 2-3ppm in ozone water solution；
The preparation of slurries: step 2 abalone meat and mixture of ice and water that step 1 has pre-processed is mixed according to mass ratio 1:2-4
It closes, is placed in colloid mill and carries out defibrination process, cross 30-50 mesh, stir evenly, abalone pretreatment slurries are prepared；
Step 3, self-dissolving hydrolysis: at a temperature of the abalone pretreatment slurries that step 2 is prepared are placed in 2-4 DEG C, using wave
The ultraviolet light of a length of 200-280nm irradiates, and in the process and is stirred continuously, and self-dissolving hydrolyzes 3-4 hours, passes through abalone itself institute
The enzyme contained carries out self-dissolving hydrolysis to macromoleculars such as the protein of its own, obtains self-dissolving hydrolysate；
Step 4, fermentation process: being added oligoisomaltose and glucose in the self-dissolving hydrolysate that step 3 is prepared, from
The quality proportioning of molten hydrolysate, oligoisomaltose and glucose is 100:(0.5-1): (2-3), then inoculation wine brewing thereto
Yeast and Lactobacillus casei, are placed at a temperature of 28-32 DEG C, fermentation process 20-24 hours, obtain tunning；
Impulse ultrasound pretreatment: step 5 carries out impulse ultrasound pretreatment, ultrasonic frequency to the tunning that step 4 obtains
For 20kHz, power 200W, ultrasonic time 10-12 minutes, time impulse ultrasound working clearance ratio was 2s/2s, and it is super to obtain pulse
Sound preconditioning product；
Enzymatic hydrolysis: the pH of impulse ultrasound pretreatment product made from step 5 is adjusted to 7.5-8.2, is added by alkalinity by step 6
Protease, papain, neutral proteinase and compound fertilizer production composition complex enzyme, 40-42 DEG C is heated to, at enzymatic hydrolysis
Reason 3.5-4 hours, obtains enzymolysis liquid, complex enzyme neutral and alkali protease, papain, neutral proteinase and composite flavor albumen
The weight of enzyme is 3:(1-1.5): (1-2): (2-3), the additive amount of complex enzyme are 0.5-0.7%；
Step 7, enzyme deactivation, concentration: by enzymolysis liquid heating water bath made from step 6 to 80 DEG C, destroy the enzyme treatment 10 minutes, so that multiple
Synthase inactivation, is concentrated into the 1/3 of original volume, as abalone enzymatic extract after filtering.
The invention further relates to a kind of preparation methods of abalone enzymatic extract, which is characterized in that this method includes following step
It is rapid:
Pretreatment: step 1 takes fresh abalone, decladding takes abalone meat after removing internal organ, and abalone meat is immersed in ozone water solution
Carry out disinfection processing, and the processing time is 20min, and treatment temperature is 5 DEG C, and ozone concentration is 2-3ppm in ozone water solution；
The preparation of slurries: step 2 abalone meat and mixture of ice and water that step 1 has pre-processed is mixed according to mass ratio 1:2-4
It closes, is placed in colloid mill and carries out defibrination process, cross 30-50 mesh, stir evenly, abalone pretreatment slurries are prepared；
Step 3, self-dissolving hydrolysis: at a temperature of the abalone pretreatment slurries that step 2 is prepared are placed in 2-4 DEG C, using wave
The ultraviolet light of a length of 200-280nm irradiates, and in the process and is stirred continuously, and self-dissolving hydrolyzes 3-4 hours, passes through abalone itself institute
The enzyme contained carries out self-dissolving hydrolysis to macromoleculars such as the protein of its own, obtains self-dissolving hydrolysate；
Step 4, fermentation process: being added oligoisomaltose and glucose in the self-dissolving hydrolysate that step 3 is prepared, from
The quality proportioning of molten hydrolysate, oligoisomaltose and glucose is 100:(0.5-1): (2-3), then inoculation wine brewing thereto
Yeast and Lactobacillus casei, are placed at a temperature of 28-32 DEG C, fermentation process 20-24 hours, obtain tunning；
Impulse ultrasound pretreatment: step 5 carries out impulse ultrasound pretreatment, ultrasonic frequency to the tunning that step 4 obtains
For 20kHz, power 200W, ultrasonic time 10-12 minutes, time impulse ultrasound working clearance ratio was 2s/2s, and it is super to obtain pulse
Sound preconditioning product；
Enzymatic hydrolysis: the pH of impulse ultrasound pretreatment product made from step 5 is adjusted to 7.5-8.2, is added by alkalinity by step 6
Protease, papain, neutral proteinase and compound fertilizer production composition complex enzyme, 40-42 DEG C is heated to, at enzymatic hydrolysis
Reason 3.5-4 hours, obtains enzymolysis liquid, complex enzyme neutral and alkali protease, papain, neutral proteinase and composite flavor albumen
The weight of enzyme is 3:(1-1.5): (1-2): (2-3), the additive amount of complex enzyme are 0.5-0.7%；
Step 7, enzyme deactivation, concentration: by enzymolysis liquid heating water bath made from step 6 to 80 DEG C, destroy the enzyme treatment 10 minutes, so that multiple
Synthase inactivation, is concentrated into the 1/3 of original volume, as abalone enzymatic extract after filtering.
Preferably, in step 4 self-dissolving hydrolysate, oligoisomaltose and glucose quality proportioning be 100:1:2 or
100:0.5:3。
Preferably, complex enzyme neutral and alkali protease, papain, neutral proteinase and composite flavor albumen in step 6
The weight of enzyme is 3:1.5:2:3 or 3:1:2:2.The wherein alkali protease, papain, neutral proteinase
It is commercially available food processing enzyme with compound fertilizer production, such as the relevant enzymes purchased from Novi's letter.
Based on above technical scheme, the invention has the advantages that and the utility model has the advantages that
Firstly, the present invention carries out finishing processing to abalone using the means of bioengineering, sent out by self-dissolving enzymatic hydrolysis, active bacteria
Ferment, impulse ultrasound handle and the combination of enzymolysis process, so that the macromolecular substances such as protein in abalone are maximumlly degraded to
Active small molecular, improves the producing level and nutrition balance of abalone nutritional ingredient, and obtained abalone enzymatic extract has
Good antioxidant activity and anti-fatigue effect, can be used as corresponding efficiency health-care food, drug processes raw material, and greatly mention
The high added value and industrial value of abalone processing.Also, it is directed to the situation of current abalone overproduction, method of the invention
It can be effectively removed the inventory of abalone production, avoid causing abalone price to decline since abalone yield is promoted.
Secondly, the present invention is pioneeringly using self-dissolving hydrolysis, active bacteria fermentation, impulse ultrasound processing and enzymolysis process joint
Treatment process, wherein self-dissolving hydrolysis be using the enzyme abundant contained by abalone itself, after defibrination process, abalone tissue
Be destroyed with cell, the hydrolases such as intracellular protease be discharged into tissue homogenate in, contained by enzyme can be to Bao
Histone, polysaccharide molecule of fish etc. carry out enzymatic hydrolysis discharge active component, in self-dissolving hydrolytic process, using the purple of specific wavelength
Outside line irradiation, the generation that self-dissolving can be excited to digest, while the contaminated bacteria in the ambient enviroments such as air can be also killed, it avoids
The generation of pollution, self-dissolving hydrolysis can also ferment for the active bacteria of next step provides good nutrition, guarantees active bacteria inoculation
After can be proliferated, ferment as quick as thought；Secondly, the present invention uses saccharomyces cerevisiae and Lactobacillus casei in active bacteria fermentation process
In conjunction with mode carry out, and oligoisomaltose and glucose are added in self-dissolving hydrolysate, the special ratios of the two
It ensure that two kinds of bacterium can be proliferated to relative equilibrium, especially oligoisomaltose is only capable of utilizing for Lactobacillus casei, can
Promote the proliferation of Lactobacillus casei, and then the macromolecular nutritional ingredient for abalone of degrading is small-molecule active substance, is saccharomyces cerevisiae
It is utilized, promotes the breeding of saccharomyces cerevisiae, in the breeding of two kinds of bacterium, the macromolecular substances of abalone are further by fermentation
It is degraded to active small molecular substance, and generates certain flavor substance, ensure that product of the invention has good flavor；
Impulse ultrasound processing is creatively utilized in the present invention, passes through pulses ultrasonic frequency, power, processing time and pulse frequency
Adjustment, obtains optimal impulse ultrasound processing mode, and impulse ultrasound processing enables to the histocyte of abalone further
The macromolecular substances such as broken, protein adequately discharge, meanwhile, the active bacteria-that impulse ultrasound processing can be crushed proliferation is dry
Lactobacillus paracasei and saccharomyces cerevisiae discharge the tissue enzyme in the two cell, for the enzymatic hydrolysis of next step, meanwhile, broken yeast
Cell has good delicate flavour, has also sufficiently ensured the excellent flavor of product；Finally, the present invention obtains abalone by enzymatic hydrolysis processing
Enzymatic hydrolyzed extract, inventor have determined complex enzyme by alkali protease, papain, neutral proteinase by largely test
It is formed with compound fertilizer production, wherein the proportion of various enzymes is mutually cooperateed with the result that impulse ultrasound is handled, i.e., in complex enzyme
The enzyme that each enzyme discharges after broken with active bacteria is had an effect jointly, effectively promotes the drop of abalone macromolecular substances
Solution, ensure that macromolecular substances-protein in abalone, polysaccharide molecule are fully hydrolyzed, and excellent flavor, activity is finally made very
High abalone enzymatic hydrolyzed extract.
Specific embodiment:
Embodiment 1: a kind of abalone enzymatic extract, which is characterized in that be prepared by the following method to obtain:
Pretreatment: step 1 takes fresh abalone, decladding takes abalone meat after removing internal organ, and abalone meat is immersed in ozone water solution
Carry out disinfection processing, and the processing time is 20min, and treatment temperature is 5 DEG C, and ozone concentration is 3ppm in ozone water solution；
Step 2, the preparation of slurries: the abalone meat that step 1 has pre-processed is mixed with mixture of ice and water according to mass ratio 1:2,
It is placed in colloid mill and carries out defibrination process, cross 50 meshes, stir evenly, abalone pretreatment slurries are prepared；
Step 3, self-dissolving hydrolysis: at a temperature of the abalone pretreatment slurries that step 2 is prepared are placed in 4 DEG C, using wavelength
It irradiates for the ultraviolet light of 220nm, in the process and is stirred continuously, self-dissolving hydrolyzes 4 hours, passes through enzyme contained by abalone itself
The macromoleculars such as the protein to its own carry out self-dissolving hydrolysis, obtain self-dissolving hydrolysate；
Step 4, fermentation process: being added oligoisomaltose and glucose in the self-dissolving hydrolysate that step 3 is prepared, from
The quality proportioning of molten hydrolysate, oligoisomaltose and glucose is 100:1:2, is then inoculated with saccharomyces cerevisiae and cheese thereto
Lactobacillus is placed at a temperature of 30 DEG C, fermentation process 22 hours, obtains tunning；
Impulse ultrasound pretreatment: step 5 carries out impulse ultrasound pretreatment, ultrasonic frequency to the tunning that step 4 obtains
For 20kHz, power 200W, ultrasonic time 12 minutes, time impulse ultrasound working clearance ratio was 2s/2s, obtained impulse ultrasound
Pretreatment product；
Enzymatic hydrolysis: the pH of impulse ultrasound pretreatment product made from step 5 is adjusted to 8.2, is added by basic protein by step 6
The complex enzyme that enzyme, papain, neutral proteinase and compound fertilizer production form, is heated to 42 DEG C, enzymolysis processing 3.5 is small
When, obtain enzymolysis liquid, the weight of complex enzyme neutral and alkali protease, papain, neutral proteinase and compound fertilizer production
Part proportion is 3:1.5:2:3, and the additive amount of complex enzyme is 0.6%；
Step 7, enzyme deactivation, concentration: by enzymolysis liquid heating water bath made from step 6 to 80 DEG C, destroy the enzyme treatment 10 minutes, so that multiple
Synthase inactivation, is concentrated into the 1/3 of enzymolysis liquid volume, as abalone enzymatic extract after filtering.
Embodiment 2: a kind of abalone enzymatic extract, which is characterized in that be prepared by the following method to obtain:
Pretreatment: step 1 takes fresh abalone, decladding takes abalone meat after removing internal organ, and abalone meat is immersed in ozone water solution
Carry out disinfection processing, and the processing time is 20min, and treatment temperature is 5 DEG C, and ozone concentration is 2ppm in ozone water solution；
Step 2, the preparation of slurries: the abalone meat that step 1 has pre-processed is mixed with mixture of ice and water according to mass ratio 1:4,
It is placed in colloid mill and carries out defibrination process, cross 30 meshes, stir evenly, abalone pretreatment slurries are prepared；
Step 3, self-dissolving hydrolysis: at a temperature of the abalone pretreatment slurries that step 2 is prepared are placed in 2 DEG C, using wavelength
It irradiates for the ultraviolet light of 280nm, in the process and is stirred continuously, self-dissolving hydrolyzes 4 hours, passes through enzyme contained by abalone itself
The macromoleculars such as the protein to its own carry out self-dissolving hydrolysis, obtain self-dissolving hydrolysate；
Step 4, fermentation process: being added oligoisomaltose and glucose in the self-dissolving hydrolysate that step 3 is prepared, from
The quality proportioning of molten hydrolysate, oligoisomaltose and glucose is 100:0.5:3, is then inoculated with saccharomyces cerevisiae thereto and does
Lactobacillus paracasei is placed at a temperature of 28 DEG C, fermentation process 24 hours, obtains tunning；
Impulse ultrasound pretreatment: step 5 carries out impulse ultrasound pretreatment, ultrasonic frequency to the tunning that step 4 obtains
For 20kHz, power 200W, ultrasonic time 12 minutes, time impulse ultrasound working clearance ratio was 2s/2s, obtained impulse ultrasound
Pretreatment product；
Enzymatic hydrolysis: the pH of impulse ultrasound pretreatment product made from step 5 is adjusted to 7.5, is added by basic protein by step 6
The complex enzyme that enzyme, papain, neutral proteinase and compound fertilizer production form, is heated to 40 DEG C, enzymolysis processing 4 is small
When, obtain enzymolysis liquid, the weight of complex enzyme neutral and alkali protease, papain, neutral proteinase and compound fertilizer production
Part proportion is 3:1:2:2, and the additive amount of complex enzyme is 0.7%；
Step 7, enzyme deactivation, concentration: by enzymolysis liquid heating water bath made from step 6 to 80 DEG C, destroy the enzyme treatment 10 minutes, so that multiple
Synthase inactivation, is concentrated into the 1/3 of enzymolysis liquid volume, as abalone enzymatic extract after filtering.
A kind of embodiment 3: preparation method of abalone enzymatic extract, which comprises the steps of:
Pretreatment: step 1 takes fresh abalone, decladding takes abalone meat after removing internal organ, and abalone meat is immersed in ozone water solution
Carry out disinfection processing, and the processing time is 20min, and treatment temperature is 5 DEG C, and ozone concentration is 2ppm in ozone water solution；
The preparation of slurries: step 2 abalone meat and mixture of ice and water that step 1 has pre-processed is mixed according to mass ratio 1:2-4
It closes, is placed in colloid mill and carries out defibrination process, cross 40 meshes, stir evenly, abalone pretreatment slurries are prepared；
Step 3, self-dissolving hydrolysis: at a temperature of the abalone pretreatment slurries that step 2 is prepared are placed in 3 DEG C, using wavelength
It irradiates for the ultraviolet light of 250nm, in the process and is stirred continuously, self-dissolving hydrolyzes 3.5 hours, passes through enzyme contained by abalone itself
Class carries out self-dissolving hydrolysis to macromoleculars such as the protein of its own, obtains self-dissolving hydrolysate；
Step 4, fermentation process: being added oligoisomaltose and glucose in the self-dissolving hydrolysate that step 3 is prepared, from
The quality proportioning of molten hydrolysate, oligoisomaltose and glucose is 100:1:2, is then inoculated with saccharomyces cerevisiae and cheese thereto
Lactobacillus is placed at a temperature of 30 DEG C, fermentation process 22 hours, obtains tunning；
Impulse ultrasound pretreatment: step 5 carries out impulse ultrasound pretreatment, ultrasonic frequency to the tunning that step 4 obtains
For 20kHz, power 200W, ultrasonic time 11 minutes, time impulse ultrasound working clearance ratio was 2s/2s, obtained impulse ultrasound
Pretreatment product；
Enzymatic hydrolysis: the pH of impulse ultrasound pretreatment product made from step 5 is adjusted to 7.8, is added by basic protein by step 6
The complex enzyme that enzyme, papain, neutral proteinase and compound fertilizer production form, is heated to 42 DEG C, enzymolysis processing 3.5 is small
When, obtain enzymolysis liquid, the weight of complex enzyme neutral and alkali protease, papain, neutral proteinase and compound fertilizer production
Part proportion is 3:1.5:2:3, and the additive amount of complex enzyme is 0.5%；
Step 7, enzyme deactivation, concentration: by enzymolysis liquid heating water bath made from step 6 to 80 DEG C, destroy the enzyme treatment 10 minutes, so that multiple
Synthase inactivation, is concentrated into the 1/3 of enzymolysis liquid volume, as abalone enzymatic extract after filtering.
Embodiment 4: in order to separately verify self-dissolving hydrolysis, impulse ultrasound processing and the pairs of abalone enzymatic extract of complex enzyme group
Influence, it is special during present invention test that following three groups of comparative examples are set:
Comparative example A(is used to verify influence of the self-dissolving hydrolysis to enzymatic hydrolyzed extract):
A kind of abalone extract A, which is characterized in that be prepared by the following method to obtain:
Pretreatment: step 1 takes fresh abalone, decladding takes abalone meat after removing internal organ, and abalone meat is immersed in ozone water solution
Carry out disinfection processing, and the processing time is 20min, and treatment temperature is 5 DEG C, and ozone concentration is 3ppm in ozone water solution；
Step 2, the preparation of slurries: the abalone meat that step 1 has pre-processed is mixed with mixture of ice and water according to mass ratio 1:2,
It is placed in colloid mill and carries out defibrination process, cross 50 meshes, stir evenly, abalone pretreatment slurries are prepared；
Step 3, fermentation process: being added oligoisomaltose and glucose in the pretreatment slurries that step 2 is prepared, from
The quality proportioning of molten hydrolysate, oligoisomaltose and glucose is 100:1:2, is then inoculated with saccharomyces cerevisiae and cheese thereto
Lactobacillus is placed at a temperature of 30 DEG C, fermentation process 22 hours, obtains tunning；
Impulse ultrasound pretreatment: step 4 carries out impulse ultrasound pretreatment, ultrasonic frequency to the tunning that step 3 obtains
For 20kHz, power 200W, ultrasonic time 12 minutes, time impulse ultrasound working clearance ratio was 2s/2s, obtained impulse ultrasound
Pretreatment product；
Enzymatic hydrolysis: the pH of impulse ultrasound pretreatment product made from step 4 is adjusted to 8.2, is added by basic protein by step 5
The complex enzyme that enzyme, papain, neutral proteinase and compound fertilizer production form, is heated to 42 DEG C, enzymolysis processing 3.5 is small
When, obtain enzymolysis liquid, the weight of complex enzyme neutral and alkali protease, papain, neutral proteinase and compound fertilizer production
Part proportion is 3:1.5:2:3, and the additive amount of complex enzyme is 0.6%；
Step 6, enzyme deactivation, concentration: by enzymolysis liquid heating water bath made from step 5 to 80 DEG C, destroy the enzyme treatment 10 minutes, so that multiple
Synthase inactivation, is concentrated into the 1/3 of enzymolysis liquid volume, as abalone extract A after filtering.
Comparative example B(is used to verify influence of the impulse ultrasound processing to enzymatic hydrolyzed extract):
A kind of abalone extract B, which is characterized in that be prepared by the following method to obtain:
Pretreatment: step 1 takes fresh abalone, decladding takes abalone meat after removing internal organ, and abalone meat is immersed in ozone water solution
Carry out disinfection processing, and the processing time is 20min, and treatment temperature is 5 DEG C, and ozone concentration is 3ppm in ozone water solution；
Step 2, the preparation of slurries: the abalone meat that step 1 has pre-processed is mixed with mixture of ice and water according to mass ratio 1:2,
It is placed in colloid mill and carries out defibrination process, cross 50 meshes, stir evenly, abalone pretreatment slurries are prepared；
Step 3, self-dissolving hydrolysis: at a temperature of the abalone pretreatment slurries that step 2 is prepared are placed in 4 DEG C, using wavelength
It irradiates for the ultraviolet light of 220nm, in the process and is stirred continuously, self-dissolving hydrolyzes 4 hours, passes through enzyme contained by abalone itself
The macromoleculars such as the protein to its own carry out self-dissolving hydrolysis, obtain self-dissolving hydrolysate；
Step 4, fermentation process: being added oligoisomaltose and glucose in the self-dissolving hydrolysate that step 3 is prepared, from
The quality proportioning of molten hydrolysate, oligoisomaltose and glucose is 100:1:2, is then inoculated with saccharomyces cerevisiae and cheese thereto
Lactobacillus is placed at a temperature of 30 DEG C, fermentation process 22 hours, obtains tunning；
Enzymatic hydrolysis: the pH of impulse ultrasound pretreatment product made from step 4 is adjusted to 8.2, is added by basic protein by step 5
The complex enzyme that enzyme, papain, neutral proteinase and compound fertilizer production form, is heated to 42 DEG C, enzymolysis processing 3.5 is small
When, obtain enzymolysis liquid, the weight of complex enzyme neutral and alkali protease, papain, neutral proteinase and compound fertilizer production
Part proportion is 3:1.5:2:3, and the additive amount of complex enzyme is 0.6%；
Step 6, enzyme deactivation, concentration: by enzymolysis liquid heating water bath made from step 5 to 80 DEG C, destroy the enzyme treatment 10 minutes, so that multiple
Synthase inactivation, is concentrated into the 1/3 of enzymolysis liquid volume, as abalone extract B after filtering.
Comparative example C (for verifying the influence of the pairs of enzymatic hydrolyzed extract of complex enzyme group):
A kind of abalone extract C, which is characterized in that be prepared by the following method to obtain:
Pretreatment: step 1 takes fresh abalone, decladding takes abalone meat after removing internal organ, and abalone meat is immersed in ozone water solution
Carry out disinfection processing, and the processing time is 20min, and treatment temperature is 5 DEG C, and ozone concentration is 3ppm in ozone water solution；
Step 2, the preparation of slurries: the abalone meat that step 1 has pre-processed is mixed with mixture of ice and water according to mass ratio 1:2,
It is placed in colloid mill and carries out defibrination process, cross 50 meshes, stir evenly, abalone pretreatment slurries are prepared；
Step 3, self-dissolving hydrolysis: at a temperature of the abalone pretreatment slurries that step 2 is prepared are placed in 4 DEG C, using wavelength
It irradiates for the ultraviolet light of 220nm, in the process and is stirred continuously, self-dissolving hydrolyzes 4 hours, passes through enzyme contained by abalone itself
The macromoleculars such as the protein to its own carry out self-dissolving hydrolysis, obtain self-dissolving hydrolysate；
Step 4, fermentation process: being added oligoisomaltose and glucose in the self-dissolving hydrolysate that step 3 is prepared, from
The quality proportioning of molten hydrolysate, oligoisomaltose and glucose is 100:1:2, is then inoculated with saccharomyces cerevisiae and cheese thereto
Lactobacillus is placed at a temperature of 30 DEG C, fermentation process 22 hours, obtains tunning；
Impulse ultrasound pretreatment: step 5 carries out impulse ultrasound pretreatment, ultrasonic frequency to the tunning that step 4 obtains
For 20kHz, power 200W, ultrasonic time 12 minutes, time impulse ultrasound working clearance ratio was 2s/2s, obtained impulse ultrasound
Pretreatment product；
Enzymatic hydrolysis: the pH of impulse ultrasound pretreatment product made from step 5 is adjusted to 8.2, is added by basic protein by step 6
The complex enzyme that enzyme, papain, neutral proteinase and compound fertilizer production form, is heated to 42 DEG C, enzymolysis processing 3.5 is small
When, obtain enzymolysis liquid, the weight of complex enzyme neutral and alkali protease and papain is 3:2, the additive amount of complex enzyme
It is 0.5%；
Step 7, enzyme deactivation, concentration: by enzymolysis liquid heating water bath made from step 6 to 80 DEG C, destroy the enzyme treatment 10 minutes, so that multiple
Synthase inactivation, is concentrated into the 1/3 of enzymolysis liquid volume, as abalone enzymatic extract after filtering.
Embodiment 5: safety testing
Acute toxicity test in mice: it is fed using the feed of the product section substitution mouse of 1-3 of the embodiment of the present invention, daily
Feeding abalone enzymatic extract 2-3mL, continuous feeding 48 days have no that dead and undue toxicity reacts during test, off-test,
Dissection test mouse, observation liver,kidney,spleen, stomach, the heart, lung and intestines, is showed no abnormal change.According to acute toxicity criteria, embodiment 1-
3 product belongs to nontoxic grade.
Chronic toxicity test: it is fed using the feed of the product section substitution mouse of 1-3 of the embodiment of the present invention, daily
Abalone enzymatic extract 4-6mL is fed, continuously feeding 220 days, as the result is shown: feeding the small of the product of 1-3 of the embodiment of the present invention
Mouse fur is smooth, and behavioral activity is normal, and Normal-weight increases, and ingesting, it is normal to drink water, and hematology and blood parameters are just
Fluctuation in normal range, marrow and routine urinalysis index fluctuate in the normal range, organize internal organs no abnormality seen, generally speaking have no
Toxicity reaction.
By being tested above it is found that product of the invention is safe and nontoxic.
Embodiment 6: sensory evaluation
Abalone extract A-C made from abalone enzymatic extract and comparative example A-C to the embodiment of the present invention 1,2,3 into
Row sense organ flavor evaluation, full marks are 10 points, and 6 points are qualification, 50 for asking 10 professional subjective appreciation teachers respectively and inviting at random
Ordinary consumer carries out flavor sensory evaluation, and concrete outcome (average mark) see the table below 1:
1 flavor sensory score result of table
| Group | Embodiment 1 | Embodiment 2 | Embodiment 3 | Compare A | Compare B | Compare C |
| Profession evaluation teacher's scoring | 9.51 | 9.33 | 9.42 | 8.31 | 7.89 | 6.47 |
| Ordinary consumer scoring | 9.36 | 9.22 | 9.25 | 6.89 | 6.56 | 5.28 |
Wherein the product of embodiment 1-3 all has good flavor, and delicate flavour is more sufficient；And its flavor of the product of comparative example A omits
It is worse than the product of embodiment 1-3, comparative example B and C have certain bitter taste, especially comparison C, the bitter taste of aftertaste denseer.
Embodiment 7: animal experiment
Kunming mouse 140 are taken, half male and half female is randomly divided into 7 groups, and every group 10 hero 10 is female.Wherein group 1 is blank control group,
It is fed using conventional mouse feed；Group 2-4 be experimental group, using 1-3 of the embodiment of the present invention abalone enzymatic hydrolyzed extract with
Conventional mouse feed is in mouse is fed after the ratio mixing of 1:4, and group 5-7 is comparative test group, using below to having a competition
It tests after an A-C abalone extract A-C is mixed with conventional mouse feed in the ratio of 1:4 and mouse is fed, seven groups are removed feed
Composition is different outer, other feeding conditions are identical, persistently feeds 15 days.Then carry out every test:
(1) antioxidase activity horizontal checkout:
At lasting feeding the 10th day, a small amount of whole blood is taken by caudal vein respectively, measures whole blood GSH-PX activity peroxide
The vigor of enzyme (GSH-Px), through detection find, organize 1(blank control group) mouse GSH-Px vigor range be 25 ± 4U/mL
Whole blood；Group 2-4(, that is, embodiment 1-3) abalone enzymatic extract feeding mouse GSH-Px vigor range be 38 ± 5U/mL
Whole blood；Control group 5-7(, that is, comparative example A-C) abalone extract feeding mouse GSH-Px vigor range be 28 ± 3U/mL
Whole blood (comparative example A), 30 ± 4U/mL whole blood (comparative example B), 29 ± 2U/mL whole blood (comparative example C).It can be seen that the present invention is real
The abalone enzymatic hydrolyzed extract for applying a 1-3 can promote the antioxidant levels of mouse, compare blank control group and comparative experimental example A-C
Unexpected technical effect is achieved, antioxidant levels improve 30% or more.
(2) swimming with a load attached to the body experiment, last feeding function of physical fatigue alleviation zoopery: are carried out after lasting feeding 15 days
After 120min, mouse is set in swimming trunk, depth of water 40cm, water tank length and width are respectively 0.75m*0.5m, and water tank wall is smooth, water temperature
(25 ± 0.5) DEG C, the sheet lead of 5% weight of rat-tail root load.
Mouse is mouse swimming time since the time swimming to death, as a result such as the following table 2:
2 function of physical fatigue alleviation zoopery test result of table
| Group | Swimming time/min |
| Group 1(blank control) | 55.8±10.1 |
| Group 2(embodiment 1) | 232.5±15.7 |
| Group 3(embodiment 2) | 220.6±12.2 |
| Group 4(embodiment 3) | 210.5±11.3 |
| Group 5(comparative example A) | 150.3±11.2 |
| Group 6(comparative example B) | 138.9±9.7 |
| Group 7(comparative example C) | 147.5±10.5 |
By the above test result it is found that compared to the blank control group that normal diet is fed, the product energy pole of 1-3 of the embodiment of the present invention
The earth improves the time of mice burden swimming, feeds the close feeding normal diet of walking weight load of the mouse of product of the present invention
3-4 times of mouse illustrates that product of the invention has significant function of relieving physical fatigue；Although comparative example A-C can also be improved
The time of mice burden swimming, however product of its antifatigue effect not as good as 1-3 of the embodiment of the present invention, just because of the present invention
Mutual collaboration between each processing step so that product of the invention achieve it is unexpected anti-for relative contrast's example A-C
Tired effect.
Claims (6)
1. a kind of preparation method of abalone enzymatic extract, which is characterized in that the preparation method includes the following steps:
Pretreatment: step 1 takes fresh abalone, decladding takes abalone meat after removing internal organ, and abalone meat is immersed in ozone water solution
Carry out disinfection processing；
Step 2, the preparation of slurries: the abalone meat that step 1 has been pre-processed is mixed with mixture of ice and water, be placed in colloid mill into
Abalone pretreatment slurries are prepared in row defibrination process；
Step 3, self-dissolving hydrolysis: the abalone pretreatment slurries that step 2 is prepared are irradiated using ultraviolet light, in the process
And it is stirred continuously, self-dissolving hydrolysis；
Step 4, fermentation process: being added oligoisomaltose and glucose in the self-dissolving hydrolysate that step 3 is prepared, and
It is inoculated with saccharomyces cerevisiae and Lactobacillus casei thereto afterwards, carries out fermentation process, obtain tunning；
Impulse ultrasound pretreatment: step 5 carries out impulse ultrasound pretreatment to the tunning that step 4 obtains, it is super to obtain pulse
Sound preconditioning product；
Enzymatic hydrolysis: the pH of impulse ultrasound pretreatment product made from step 5 is adjusted to 7.5-8.2, is added by alkalinity by step 6
Protease, papain, neutral proteinase and compound fertilizer production composition complex enzyme, 40-42 DEG C is heated to, at enzymatic hydrolysis
Reason 3.5-4 hours, obtains enzymolysis liquid, complex enzyme neutral and alkali protease, papain, neutral proteinase and composite flavor albumen
The weight of enzyme is 3:(1-1.5): (1-2): (2-3), the additive amount of complex enzyme are 0.5-0.7%；
Step 7, enzyme deactivation, concentration, as abalone enzymatic extract.
2. the preparation method of abalone enzymatic extract according to claim 1, which is characterized in that this method includes following step
It is rapid:
Pretreatment: step 1 takes fresh abalone, decladding takes abalone meat after removing internal organ, and abalone meat is immersed in ozone water solution
Carry out disinfection processing, and the processing time is 20min, and treatment temperature is 5 DEG C, and ozone concentration is 2-3ppm in ozone water solution；
The preparation of slurries: step 2 abalone meat and mixture of ice and water that step 1 has pre-processed is mixed according to mass ratio 1:2-4
It closes, is placed in colloid mill and carries out defibrination process, cross 30-50 mesh, stir evenly, abalone pretreatment slurries are prepared；
Step 3, self-dissolving hydrolysis: at a temperature of the abalone pretreatment slurries that step 2 is prepared are placed in 2-4 DEG C, using wave
The ultraviolet light of a length of 200-280nm irradiates, and in the process and is stirred continuously, and self-dissolving hydrolyzes 3-4 hours, passes through abalone itself institute
The enzyme contained carries out self-dissolving hydrolysis to macromoleculars such as the protein of its own, obtains self-dissolving hydrolysate；
Step 4, fermentation process: being added oligoisomaltose and glucose in the self-dissolving hydrolysate that step 3 is prepared, from
The quality proportioning of molten hydrolysate, oligoisomaltose and glucose is 100:(0.5-1): (2-3), then inoculation wine brewing thereto
Yeast and Lactobacillus casei, are placed at a temperature of 28-32 DEG C, fermentation process 20-24 hours, obtain tunning；
Impulse ultrasound pretreatment: step 5 carries out impulse ultrasound pretreatment, ultrasonic frequency to the tunning that step 4 obtains
For 20kHz, power 200W, ultrasonic time 10-12 minutes, time impulse ultrasound working clearance ratio was 2s/2s, and it is super to obtain pulse
Sound preconditioning product；
Enzymatic hydrolysis: the pH of impulse ultrasound pretreatment product made from step 5 is adjusted to 7.5-8.2, is added by alkalinity by step 6
Protease, papain, neutral proteinase and compound fertilizer production composition complex enzyme, 40-42 DEG C is heated to, at enzymatic hydrolysis
Reason 3.5-4 hours, obtains enzymolysis liquid, complex enzyme neutral and alkali protease, papain, neutral proteinase and composite flavor albumen
The weight of enzyme is 3:(1-1.5): (1-2): (2-3), the additive amount of complex enzyme are 0.5-0.7%；
Step 7, enzyme deactivation, concentration: by enzymolysis liquid heating water bath made from step 6 to 80 DEG C, destroy the enzyme treatment 10 minutes, so that multiple
Synthase inactivation, is concentrated into the 1/3 of enzymolysis liquid volume, as abalone enzymatic extract after filtering.
3. a kind of abalone enzymatic extract, which is characterized in that be prepared by the following method to obtain:
Pretreatment: step 1 takes fresh abalone, decladding takes abalone meat after removing internal organ, and abalone meat is immersed in ozone water solution
Carry out disinfection processing；
Step 2, the preparation of slurries: the abalone meat that step 1 has been pre-processed is mixed with mixture of ice and water, be placed in colloid mill into
Abalone pretreatment slurries are prepared in row defibrination process；
Step 3, self-dissolving hydrolysis: the abalone pretreatment slurries that step 2 is prepared are irradiated using ultraviolet light, in the process
And it is stirred continuously, self-dissolving hydrolysis；
Step 4, fermentation process: being added oligoisomaltose and glucose in the self-dissolving hydrolysate that step 3 is prepared, and
It is inoculated with saccharomyces cerevisiae and Lactobacillus casei thereto afterwards, carries out fermentation process, obtain tunning；
Impulse ultrasound pretreatment: step 5 carries out impulse ultrasound pretreatment to the tunning that step 4 obtains, it is super to obtain pulse
Sound preconditioning product；
Enzymatic hydrolysis: the pH of impulse ultrasound pretreatment product made from step 5 is adjusted to 7.5-8.2, is added by alkalinity by step 6
Protease, papain, neutral proteinase and compound fertilizer production composition complex enzyme, 40-42 DEG C is heated to, at enzymatic hydrolysis
Reason 3.5-4 hours, obtains enzymolysis liquid, complex enzyme neutral and alkali protease, papain, neutral proteinase and composite flavor albumen
The weight of enzyme is 3:(1-1.5): (1-2): (2-3), the additive amount of complex enzyme are 0.5-0.7%；
Step 7, enzyme deactivation, concentration, as abalone enzymatic extract.
4. a kind of abalone enzymatic extract, which is characterized in that be prepared by the following method to obtain:
Pretreatment: step 1 takes fresh abalone, decladding takes abalone meat after removing internal organ, and abalone meat is immersed in ozone water solution
Carry out disinfection processing, and the processing time is 20min, and treatment temperature is 5 DEG C, and ozone concentration is 2-3ppm in ozone water solution；
The preparation of slurries: step 2 abalone meat and mixture of ice and water that step 1 has pre-processed is mixed according to mass ratio 1:2-4
It closes, is placed in colloid mill and carries out defibrination process, cross 30-50 mesh, stir evenly, abalone pretreatment slurries are prepared；
Step 3, self-dissolving hydrolysis: at a temperature of the abalone pretreatment slurries that step 2 is prepared are placed in 2-4 DEG C, using wave
The ultraviolet light of a length of 200-280nm irradiates, and in the process and is stirred continuously, and self-dissolving hydrolyzes 3-4 hours, passes through abalone itself institute
The enzyme contained carries out self-dissolving hydrolysis to macromoleculars such as the protein of its own, obtains self-dissolving hydrolysate；
Step 4, fermentation process: being added oligoisomaltose and glucose in the self-dissolving hydrolysate that step 3 is prepared, from
The quality proportioning of molten hydrolysate, oligoisomaltose and glucose is 100:(0.5-1): (2-3), then inoculation wine brewing thereto
Yeast and Lactobacillus casei, are placed at a temperature of 28-32 DEG C, fermentation process 20-24 hours, obtain tunning；
Impulse ultrasound pretreatment: step 5 carries out impulse ultrasound pretreatment, ultrasonic frequency to the tunning that step 4 obtains
For 20kHz, power 200W, ultrasonic time 10-12 minutes, time impulse ultrasound working clearance ratio was 2s/2s, and it is super to obtain pulse
Sound preconditioning product；
Enzymatic hydrolysis: the pH of impulse ultrasound pretreatment product made from step 5 is adjusted to 7.5-8.2, is added by alkalinity by step 6
Protease, papain, neutral proteinase and compound fertilizer production composition complex enzyme, 40-42 DEG C is heated to, at enzymatic hydrolysis
Reason 3.5-4 hours, obtains enzymolysis liquid, complex enzyme neutral and alkali protease, papain, neutral proteinase and composite flavor albumen
The weight of enzyme is 3:(1-1.5): (1-2): (2-3), the additive amount of complex enzyme are 0.5-0.7%；
Step 7, enzyme deactivation, concentration: by enzymolysis liquid heating water bath made from step 6 to 80 DEG C, destroy the enzyme treatment 10 minutes, so that multiple
Synthase inactivation, is concentrated into the 1/3 of enzymolysis liquid volume, as abalone enzymatic extract after filtering.
5. the preparation method of abalone enzymatic extract according to claim 3 or 4, which is characterized in that preferably, step 4
The quality proportioning of middle self-dissolving hydrolysate, oligoisomaltose and glucose is 100:1:2 or 100:0.5:3.
6. the preparation method of abalone enzymatic extract according to claim 3 or 4, which is characterized in that preferably, step 6
Middle complex enzyme neutral and alkali protease, papain, neutral proteinase and compound fertilizer production weight be 3:
1.5:2:3 or 3:1:2:2.
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