CN109730191A - Comprehensive multi-index weighted analysis method optimizes solid state fermentation dregs of beans method - Google Patents
Comprehensive multi-index weighted analysis method optimizes solid state fermentation dregs of beans method Download PDFInfo
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- CN109730191A CN109730191A CN201910083208.7A CN201910083208A CN109730191A CN 109730191 A CN109730191 A CN 109730191A CN 201910083208 A CN201910083208 A CN 201910083208A CN 109730191 A CN109730191 A CN 109730191A
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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Abstract
The invention discloses comprehensive multi-index weighted analysis methods to optimize solid state fermentation dregs of beans method, and steps are as follows for specific method: a. carries out material preparation first, including prepares fermenting microbe, fermentation substrate；B. orthogonal test is carried out to it；C. its index is measured；D. mathematical method, including mathematical statistics method is utilized to be analyzed and processed multiple indication informations；E carries out verification test according to optimal conditions of fermentation in orthogonal test；F handles data, and analyzes result.The present invention optimizes fermented bean dregs processing parameter from fermenting microbe, inoculum concentration, fermentation temperature, material-water ratio etc., and systematic analysis technique is carried out to multiple production targets such as the basic nutrition of tunning composition and fatty acid, amino acid, to instruct actual production to provide more scientific theoretical foundation.
Description
Technical field
The present invention relates to technical field of agricultural product process, especially comprehensive multi-index weighted analysis methods to optimize solid state fermentation beans
Dregs of rice method.
Background technique
Culture fishery development increases fish meal demand year by year, and the excessive amount of fishing of bait fishery resources, leads to fish meal in addition
Price constantly increases, and therefore, replaces the animal proteins such as fish meal very popular with vegetable protein.Dregs of beans is obtained after soybean processing
A kind of byproduct, protein rich in are the widest plant protein sources of aquatic feeds industrial application, but due in dregs of beans
Anti-nutritional factors, amino acid imbalance, the difference of palatability caused by compact structure and digestibility is relatively low etc. limits its application valence
Value.Many studies have shown that beneficial microbe fermented bean dregs can reduce anti-nutritional factors content, animal growth rate is improved；Greatly
Molecule protein can also be degraded to polypeptide, small peptide and the free amine group for being easy to be absorbed by cultivated animals under microbial action
Acid[8], improve efficiency of feed utilization；Microbial fermentation can also degrade portion carbohydrate, and dregs of beans compact texture is made to become thin
Pine, reduces the generation of bitter taste, to increase dregs of beans palatability, improves the efficiency of feed utilization of aquatic livestock.
According to the difference of production purpose, the fermentation pattern of dregs of beans can be divided into liquid fermentation and solid fermentation.Using liquid
Fermentation pattern prepares Nattokinase, and research show that optimal combination is when connecing bacterium amount 1%, soybean meal content 2%, pH 7.0, fermentation
Between be 48h, Nattokinase enzyme activity reaches 4429.6U/mL under this condition.Wu Shenghua etc.[14]Dregs of beans is carried out using saccharomycete
Solid state fermentation the study found that anti-nutritional factors content degradation rate reaches 56.2%, small peptide content improves 4.3 times.Solid-state hair
Ferment has many advantages, such as that small investment, low energy consumption, easy to operate, pollution is few relative to liquid state fermentation, therefore each manufacturer is more in the market
Using solid fermentation mode of manufacture fermented bean dregs.Since each manufacturer's fermented bean dregs product physical and chemical index is not quite identical, hair
Ferment Soybean Meal lacks comprehensive evaluation index.Previous research is simple using polypeptide conversion ratio, viable count and conventional nutrient content
Some parameter index, which is generally only chosen, when being added and be used as evaluation index, and studying fermented bean dregs technique optimizes evaluation.
Single index can evaluate fermentation process of bean dregs superiority and inferiority to a certain extent, but more nutritional ingredient be required to evaluate as feedstuff
It is comprehensive, therefore comprehensive multi-index weighted analysis method being capable of more objective comprehensive evaluation fermentation process of bean dregs.However, at present
Research using comprehensive multi-index weighted analysis method evaluation fermentation process of bean dregs is less.For above, herein it is proposed that referring to more
It marks integrated weighting analysis method and optimizes solid state fermentation dregs of beans method.
Summary of the invention
The present invention provides comprehensive multi-index weighted analysis method for technical deficiencies such as existing solid state fermentation dregs of beans
Optimize solid state fermentation dregs of beans method.
The present invention is to solve above-mentioned phenomenon, using the technical solution of following modification, the optimization of comprehensive multi-index weighted analysis method
Solid state fermentation dregs of beans method, steps are as follows for specific method:
A. material preparation is carried out first, including prepares fermenting microbe, fermentation substrate；
B. orthogonal test is carried out to it；
C. its index is measured；
D. mathematical method, including mathematical statistics method is utilized to be analyzed and processed multiple indication informations, it is excellent in the hope of its
Of inferior quality grade carries out integrated weighting analysis according to each index experimental data, is finally scored, and according to scoring height, determines orthogonal
Optimal fermentation test group in test；
E carries out verification test according to optimal conditions of fermentation in orthogonal test, and measures 8 synthesis of dregs of beans after fermentation
Then evaluation index carries out Data Analysis Services；
F handles data, and analyzes result.
As present invention further optimization mode, the fermenting microbe includes bacillus subtilis, 1.5 ×
109cfu·mL-1；Lactic acid bacteria, 2 × 109cfu·mL-1；Ocean rhodotorula bacterium, 8 × 108cfu·mL-1。
As present invention further optimization mode, the orthogonal test method includes, with inoculum concentration, fermentation temperature, material
Water ratio, fermenting microbe are investigation factor, wherein the horizontally selected bacillus subtilis of fermenting microbe, lactic acid bacteria, ocean rhodotorula bacterium
Three's different proportion, with the crude fat of dregs of beans, reduced sugar, lactic acid, neutral proteinase, total amino acid, dry matter recycling after fermentation
Rate, unsaturated fatty acid, crude protein are overall target, and each processing group sets 3 repetitions, and fermentation time 48h optimizes best hair
Ferment technique condition.
As present invention further optimization mode, index determining method described in step c includes content of reducing sugar measurement
Method and lactic acid content measuring method；The content of reducing sugar measuring method includes using 3,5- dinitrosalicylic acid system (DNS)
The reduced sugar in fermented sample is measured, the preparation of sample reduced sugar: accurately weighing 1.0g fermented sample in 100mL triangular flask,
About 50mL distilled water is added, mixes, keeps the temperature 20min in 50 DEG C of waters bath with thermostatic control, stir frequently, leach reduced sugar, filters, it will
Filtrate is all collected in the volumetric flask of 100mL, with distilled water constant volume, as reduced sugar extracting solution.
As present invention further optimization mode, the lactic acid content measuring method includes, first to 0.1 mol/L's
NaOH solution is demarcated, and each 10.0g of dregs of beans after taking fermentation adds distilled water 90ml, sets 120r/min shaking table 30min, is centrifuged,
Take 10ml supernatant, if supernatant color is deeper, be added certain multiple without CO2Distilled water is diluted, 2 drop 1% phenolphthalein instruction of drop
Agent is titrated with standard NaOH solution, be titrated to it is rose pink, to terminal record quota of expenditure NaOH volume, repeat 3
It is secondary, it is averaged, calculation formula:
In formula: NNaOH--- the concentration (mol/L) of standard NaOH after calibration；
VNaOH--- the volume (ml) of quota of expenditure NaOH liquid when titration；
The measurement of the dry matter rate of recovery (DMR)
Fermented feed samples immediately after preparing, and calculates by weight before and after feed fermentation and dry matter (DM) content；
In formula: W1--- dry matter weight after fermentation；W0--- dry matter weight before fermenting.
As present invention further optimization mode, the data processing includes first to multi objective in fermentation process of bean dregs
Component content carries out data processing using Multi-index test equation, obtains the comprehensive score of orthogonal test group；Then, it utilizes
Orthogonal design is analyzed in the field mouthful design of Minitab 17, and is scored using ANOVA comprehensive multi-index and carried out variance point
Analysis, P < 0.05 are significant difference.
Compared with prior art: the present invention optimizes from fermenting microbe, inoculum concentration, fermentation temperature, material-water ratio etc. ferments
Dregs of beans processing parameter, and multiple production targets such as the basic nutrition of tunning composition and fatty acid, amino acid are carried out
Systematic analysis technique, to instruct actual production to provide more scientific theoretical foundation.
Detailed description of the invention
Fig. 1 is each factor level mean value main effect figure of influence fermentation process of bean dregs of the invention.
Specific embodiment
Below in conjunction with the embodiment of the present invention, technical scheme in the embodiment of the invention is clearly and completely described,
Obviously, described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Based in the present invention
Embodiment, every other embodiment obtained by those of ordinary skill in the art without making creative efforts, all
Belong to the scope of protection of the invention.
Referring to Fig. 1, the present invention provides a kind of technical solution: comprehensive multi-index weighted analysis method optimizes solid state fermentation dregs of beans
Method, steps are as follows for specific method:
A. material preparation is carried out first, including prepares fermenting microbe, fermentation substrate；
B. orthogonal test is carried out to it；
C. its index is measured；
D. mathematical method, including mathematical statistics method is utilized to be analyzed and processed multiple indication informations, it is excellent in the hope of its
Of inferior quality grade carries out integrated weighting analysis according to each index experimental data, is finally scored, and according to scoring height, determines orthogonal
Optimal fermentation test group in test；
E carries out verification test according to optimal conditions of fermentation in orthogonal test, and measures 8 synthesis of dregs of beans after fermentation
Then evaluation index carries out Data Analysis Services；
F handles data, and analyzes result.
The fermenting microbe includes bacillus subtilis, and 1.5 × 109cfu·mL-1；Lactic acid bacteria, 2 × 109cfu·mL-1；
Ocean rhodotorula bacterium, 8 × 108cfu·mL-1。
The orthogonal test method includes, using inoculum concentration, fermentation temperature, material-water ratio, fermenting microbe as investigation factor, wherein
The horizontally selected bacillus subtilis of fermenting microbe, lactic acid bacteria, ocean rhodotorula bacterium three's different proportion, with ferment after dregs of beans it is thick
Fat, reduced sugar, lactic acid, neutral proteinase, total amino acid, the dry matter rate of recovery, unsaturated fatty acid, crude protein are that synthesis refers to
Mark, each processing group set 3 repetitions, and fermentation time 48h optimizes best technological condition for fermentation.
Index determining method described in step c includes content of reducing sugar measuring method and lactic acid content measuring method；It is described
Content of reducing sugar measuring method includes using the reduced sugar in 3,5- dinitrosalicylic acid system (DNS) measurement fermented sample, sample
The preparation of reduced sugar: accurately weighing 1.0g fermented sample in 100mL triangular flask, and about 50mL distilled water is added, and mixes, in 50 DEG C
20min is kept the temperature in water bath with thermostatic control, is stirred frequently, reduced sugar is leached, is filtered, and filtrate is all collected to the volumetric flask in 100mL
In, with distilled water constant volume, as reduced sugar extracting solution.
The lactic acid content measuring method includes demarcating first to the NaOH solution of 0.1mol/L, after taking fermentation
Each 10.0g of dregs of beans, adds distilled water 90ml, sets 120r/min shaking table 30min, and centrifugation takes 10ml supernatant, if supernatant color is deeper,
Be added certain multiple without CO2Distilled water is diluted, and 2 1% phenolphthalein indicator of drop of drop is titrated with standard NaOH solution,
Be titrated to it is rose pink, to terminal record quota of expenditure NaOH volume, be repeated 3 times, be averaged, calculation formula:
In formula: NNaOH--- the concentration (mol/L) of standard NaOH after calibration；
VNaOH--- the volume (ml) of quota of expenditure NaOH liquid when titration；
The measurement of the dry matter rate of recovery (DMR)；
Fermented feed samples immediately after preparing, and calculates by weight before and after feed fermentation and dry matter (DM) content；
In formula: W1--- dry matter weight after fermentation；W0--- dry matter weight before fermenting.
The data processing includes using Multi-index test formula to multi-target ingredient content in fermentation process of bean dregs first
Method carries out data processing, obtains the comprehensive score of orthogonal test group；Then, using the field of Minitab 17 mouthful design to orthogonal
Design is analyzed, and is scored using ANOVA comprehensive multi-index and carried out variance analysis, and P < 0.05 is significant difference.
Embodiment
The method design of the orthogonal test of the present embodiment is as shown in Table 1 and Table 2.The orthogonal test method includes, to connect
Kind amount, fermentation temperature, material-water ratio, fermenting microbe are investigation factor, wherein the horizontally selected bacillus subtilis of fermenting microbe, lactic acid
Bacterium, ocean rhodotorula bacterium three's different proportion.
1 orthogonal test factor level table of table
2 orthogonal test designs table of table
With XijIndicate that i-th tests measured value (the i.e. X of j-th of indexijIndicate the jth of the i-th test of orthogonal test
The measurement content of a index), first using the maximum value of each index as reference, place is standardized to each data of same index
Reason, DijIndicate the standardized data of i-th of measured value under j-th of index. Dij=Xij/(Xj)max, wherein i=1,2,
...9；J=1,2 ..., 8.Weight coefficient, F are determined by the significance level of each index and the relative standard deviation of each group of data againj
=Ej×RSD/RSDj(being shown in Table 3), wherein RSD is the relative standard deviation of crude fat, RSDjFor the relative standard of j-th of index
Difference, since the effect of each nutritive index is all important, therefore significance level EjIt is set to 1, Pi=∑ (Fj×Dij)/∑Fj, calculate institute
What is obtained takes into account indices comprehensive score formula PiIt is the bigger the better, data processed result is shown in Table (3~table of table 7).
3 orthogonal test comprehensive score P of tableiMean value responds table
4 orthogonal test comprehensive score P of tableiAnalysis of variance table
5 orthogonal test comprehensive score P of tableiMean value responds table
6 orthogonal test comprehensive score P of tableiAnalysis of variance table
Each index determining result of 7 optimised process confirmatory experiment of table
To sum up, the present invention produces work from fermenting microbe, inoculum concentration, fermentation temperature, material-water ratio etc. optimization fermented bean dregs
Skill parameter, and comprehensive analysis is carried out to multiple production targets such as the basic nutrition of tunning composition and fatty acid, amino acid and is commented
Valence, to instruct actual production to provide more scientific theoretical foundation.
The above shows and describes the basic principles and main features of the present invention and the advantages of the present invention, for this field skill
For art personnel, it is clear that invention is not limited to the details of the above exemplary embodiments, and without departing substantially from spirit of the invention or
In the case where essential characteristic, the present invention can be realized in other specific forms.Therefore, in all respects, should all incite somebody to action
Embodiment regards exemplary as, and is non-limiting, the scope of the present invention by appended claims rather than on state
Bright restriction, it is intended that including all changes that fall within the meaning and scope of the equivalent elements of the claims in the present invention
It is interior.
In addition, it should be understood that although this specification is described in terms of embodiments, but not each embodiment is only wrapped
Containing an independent technical solution, this description of the specification is merely for the sake of clarity, and those skilled in the art should
It considers the specification as a whole, the technical solutions in the various embodiments may also be suitably combined, forms those skilled in the art
The other embodiments being understood that.
Claims (6)
1. comprehensive multi-index weighted analysis method optimizes solid state fermentation dregs of beans method, which comprises the steps of:
A. material preparation is carried out first, including prepares fermenting microbe, fermentation substrate；
B. orthogonal test is carried out to it；
C. its index is measured；
D. mathematical method, including mathematical statistics method is utilized to be analyzed and processed multiple indication informations, in the hope of its superiority and inferiority etc.
Grade carries out integrated weighting analysis according to each index experimental data, is finally scored, and according to scoring height, determines orthogonal test
In optimal fermentation test group；
E carries out verification test according to optimal conditions of fermentation in orthogonal test, and 8 overall merits for measuring dregs of beans after fermentation refer to
Mark, then carries out Data Analysis Services；
F handles data, and analyzes result.
2. comprehensive multi-index weighted analysis method according to claim 1 optimizes solid state fermentation dregs of beans method, which is characterized in that
The fermenting microbe includes bacillus subtilis NHS1 (Bacillus subtilis NHS1), and 1.5 × 109cfu·mL-1；Cream
Sour bacterium (Lactic acid bacteria), 2 × 109cfu·mL-1；Ocean rhodotorula bacterium (Rhodotorula
Mucilaginosa), 8 × 108cfu·mL-1。
3. comprehensive multi-index weighted analysis method according to claim 1 optimizes solid state fermentation dregs of beans method, which is characterized in that
The method of orthogonal test described in step b includes, using inoculum concentration, fermentation temperature, material-water ratio, fermenting microbe as investigation factor,
The horizontally selected bacillus subtilis of middle fermenting microbe, lactic acid bacteria, ocean rhodotorula bacterium three's different proportion, with dregs of beans after fermenting
Crude fat, reduced sugar, lactic acid, neutral proteinase, total amino acid, the dry matter rate of recovery, unsaturated fatty acid, crude protein are synthesis
Index, each processing group set 3 repetitions, and fermentation time 48h optimizes best technological condition for fermentation.
4. comprehensive multi-index weighted analysis method according to claim 1 optimizes solid state fermentation dregs of beans method, which is characterized in that
Index determining method described in step c includes content of reducing sugar measuring method and lactic acid content measuring method；The reduced sugar contains
Quantity measuring method includes using the reduced sugar in 3,5- dinitrosalicylic acid system measurement fermented sample, the preparation of sample reduced sugar:
1.0g fermented sample is accurately weighed in 100mL triangular flask, about 50mL distilled water is added, mixes, is protected in 50 DEG C of waters bath with thermostatic control
Warm 20min, is stirred frequently, leaches reduced sugar, filtering, and filtrate is all collected in the volumetric flask of 100mL, fixed with distilled water
Hold, as reduced sugar extracting solution.
5. comprehensive multi-index weighted analysis method according to claim 4 optimizes solid state fermentation dregs of beans method, which is characterized in that
The lactic acid content measuring method includes demarcating first to the NaOH solution of 0.1mol/L, and the dregs of beans after taking fermentation is each
10.0g adds distilled water 90ml, sets 120r/min shaking table 30min, and centrifugation takes 10ml supernatant, if supernatant color is deeper, is added one
Determine multiple without CO2Distilled water is diluted, and 2 1% phenolphthalein indicator of drop of drop is titrated with standard NaOH solution, is titrated to
It is rose pink, to the volume of terminal record quota of expenditure NaOH, it is repeated 3 times, is averaged, calculation formula:
In formula: NNaOH--- the concentration (mol/L) of standard NaOH after calibration；
VNaOH--- the volume (ml) of quota of expenditure NaOH liquid when titration；
The measurement of the dry matter rate of recovery (DMR):
Fermented feed samples immediately after preparing, and calculates by weight before and after feed fermentation and dry matter (DM) content；
In formula: W1--- dry matter weight after fermentation；W0--- dry matter weight before fermenting.
6. comprehensive multi-index weighted analysis method according to claim 1 optimizes solid state fermentation dregs of beans method, which is characterized in that
The data processing includes being counted first to multi-target ingredient content in fermentation process of bean dregs using Multi-index test equation
According to processing, the comprehensive score of orthogonal test group is obtained；Then, orthogonal design is divided using the field of Minitab 17 mouthful design
Analysis, and scored using ANOVA comprehensive multi-index and carry out variance analysis, P < 0.05 is significant difference.
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