CN102342567B - Pepper peeling method - Google Patents
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Abstract
The invention discloses a pepper peeling method which comprises the following steps: mixing the pepper fruit to be peeled with solid culture of pectinase producing bacteria or bacteria suspension or spore suspension of the pectinase producing bacteria, and performing solid state fermentation; and after the fermentation is over, peeling to obtain the peeled pepper. The method disclosed by the invention is characterized that: the peeling time is short, the peculiar smell of the pepper product and the ambient environment caused by an immersion method and a liquid state fermentation method is avoided, the loss of the pepper components is reduced, and the sensuous quality of the pepper product is improved; and meanwhile, the environmental pollution is reduced, and the unavoidable shortcomings of peculiar smell, long production period and the like of the traditional immersion method are eliminated.
Description
Technical field
The present invention relates to a kind of method that makes pepper decortication.
Background technology
At present, the method for pepper decortication mainly contains: infusion process (being bubble method), Mechanical Method, enzyme process and bioanalysis.Generally adopt infusion process both at home and abroad, this method production cycle reaches 7-20d (becoming according to temperature), and water pollution is large, and product is often with obvious stink, and microbial count is higher; Mechanical Method is difficult to pericarp to remove thoroughly, and easily makes pepper corn profile impaired; Although there are some researches show that enzyme process decortication effect is better, as short in the time, efficiency is high, pollutes less, cost is too high, also just rests at present the laboratory research stage, is difficult to apply; Bioanalysis decortication is also in the laboratory Primary Study stage.
In recent years, though there is the research of biological peeling method, most constraints that are subject to traditional infusion process thinking, are confined to, in liquid state fermentation method, not obtain good effect, and liquid fermentation method equipment investment and power consumption higher, be difficult for promoting.
For this reason, urgently develop a kind of fast, efficient, low energy consumption, low pollution and the new method of pepper decortication cheaply.
Summary of the invention
An object of the present invention is to provide a kind of method that makes pepper decortication.
The method that makes pepper decortication provided by the present invention, comprise the steps: pepper fruit to be peeled with the solid-state culture of Pectinase Producing Strain or mix with bacteria suspension or the spore suspension of Pectinase Producing Strain, carrying out solid state fermentation, fermenting complete, peeling, obtains the pepper of peeling.
In said method, before described mixing, comprise the step that following blanching is processed: pepper fruit to be peeled is placed in to 60 ℃ of above or more than 100 ℃ environment and keeps 10s-3min, be specially 10-30s.
In above-mentioned arbitrary described method, described in the proportioning of solid-state culture of pepper fruit to be peeled and described Pectinase Producing Strain be 100g: (0.1g-10g), be specially 100g: (0.1g-5g) or 100g: (0.3g-5g);
In above-mentioned arbitrary described method, described in the proportioning of spore suspension of pepper fruit to be peeled and described Pectinase Producing Strain be 20g: (1-5) * 10
7individual.
In above-mentioned arbitrary described method, the temperature of described solid state fermentation is 20 ℃-40 ℃ or 25 ℃-40 ℃, is specially 28 ℃-37 ℃, then is specially 28-30 ℃ (der Pilz) or 30-37 ℃ (bacterium class).
In above-mentioned arbitrary described method, the time of described solid state fermentation is 24h-96h, is specially 36h-84h, then is specially 36h-60h or 60h-84h.
In above-mentioned arbitrary described method, the method for described peeling is for washing by rubbing with the hands.
In above-mentioned arbitrary described method, the solid-state culture of described Pectinase Producing Strain is prepared as follows and obtains: for the preparation of the solid medium of cultivating described Pectinase Producing Strain, described Pectinase Producing Strain is inoculated in to described solid medium, cultivate, extremely described bacterium is bred, and all substances in the culture vessel obtaining are the solid-state culture of described Pectinase Producing Strain;
In above-mentioned arbitrary described method, described solid medium is following 1) or 2) shown in:
1) by wheat bran and water with 10g: (8mL-20mL) or 10g: ratio (10mL-20mL) is mixed, and the mixture obtaining is described solid medium;
2) by wheat bran, water with at least one in following material with 5g: (8mL-20mL): the ratio of 5g is mixed: rice husk, bean cake powder, maize cob meal, pomelo peel powder, orange peel powder, bagasse, manioc waste, the mixture obtaining is described solid medium;
In above-mentioned arbitrary described method, the pH value of described solid medium is 3.5-8, is specially 5.5-7.0, then is specially nature pH value (6.6).
In above-mentioned arbitrary described method, described wheat bran is wheat bran;
In above-mentioned arbitrary described method, described Pectinase Producing Strain is aspergillus niger (Aspergillus niger) or aspergillus awamori (Aspergillus awamori) or head mold (Rizupus sp.) or mould (Pennicilium sp.) or Aureobasidium pullulans (Aureobasidium pullulans) or bacillus subtilis (Bacillus subtilis).
In above-mentioned arbitrary described method, described aspergillus niger (Aspergillus niger) is specially at least one in following aspergillus niger: aspergillus niger (Aspergillus niger) CICC 40273, aspergillus niger (Aspergillus niger) CICC 2317, aspergillus niger (Aspergillus niger) CICC 2214, aspergillus niger (Aspergillus niger) CICC 40616, aspergillus niger (Aspergillus niger) CICC 40097, aspergillus niger (Aspergillus niger) CICC 40094, aspergillus niger (Aspergillus niger) CICC40493, aspergillus niger (Aspergillus niger) CICC 40091, carbon black aspergillus (Aspergillus carbonarius) CICC 41254, carbon black aspergillus (Aspergillus carbonarius) CICC 40093, aspergillus niger (Aspergillus niger) CGMCC 3.316, aspergillus niger (Aspergillus niger) CGMCC 3.3289, aspergillus niger (Aspergillus niger) CGMCC 3.3287, aspergillus niger (Aspergillus niger) CGMCC 3.3284, aspergillus niger (Aspergillus niger) D-04,
Described aspergillus awamori (Aspergillus awamori) is specially at least one in following aspergillus awamori: aspergillus awamori (Aspergillus awamori) CICC 41467, aspergillus awamori (Aspergillus awamori) CICC 41466, aspergillus awamori (Aspergillus awamori) CICC 40499;
Described Aureobasidium pullulans (Aureobasidium pullulans) is Aureobasidium pullulans (Aureobasidium pullulans) CICC 40331;
Described bacillus subtilis (Bacillus subtilis) is as lower at least one: bacillus subtilis (Bacillus subtilis) CICC10076, bacillus subtilis (Bacillus subtilis) CICC21704, bacillus subtilis (Bacillus subtilis) CICC20220, bacillus subtilis (Bacillus subtilis) CICC20020, bacillus subtilis (Bacillus subtilis) ACCC10629, bacillus subtilis (Bacillus subtilis) ACCC03427,
Described mould (Pennicilium sp.) is as lower at least one: penicillium purpurogenum (Penicillium purpurogenum) 03005, mould (Penicillium) Y-03, mould (Penicillium) C-01, mould (Penicillium) A-012131, mould (Penicillium) A-08;
Described head mold (Rizupus sp.) is as lower at least one: Rhizopus stolonifer (Rhizopus stolonifer) CICC40326, Rhizopus stolonifer (Rhizopus Stolonifer) CICC40322, Rhizopus stolonifer (Rhizopus stolonifer) CICC40343, middle rhizopus chinensis (Rhizopus chinensis) ACCC30301.
In above-mentioned arbitrary described method, during described blanching is processed, described 60 ℃ of above or more than 100 ℃ environment are hot water, boiling water or steam ambient or microwave treatment.
In above-mentioned arbitrary described method, in the preparation of the solid-state culture of described Pectinase Producing Strain, the temperature of described cultivation is 20 ℃-40 ℃, is specially 28-32 ℃ (der Pilz) or 30-37 ℃ (bacterium class), the time of described cultivation is 48h-96h, is specially 72h-96h;
In above-mentioned arbitrary described method, described pepper is new fresh pepper or black pepper;
In above-mentioned arbitrary described method, in the spore suspension of described Pectinase Producing Strain, solute is spore, and solvent is water.
The present invention adopts solid state fermentation decortication, has significantly reduced the time that pepper contacts with water, and then has reduced the loss of the various compositions of pepper.
The outstanding especially feature of the present invention is to have avoided the white pepper product that causes because of infusion process and liquid fermentation method and the foreign odor taste of surrounding environment, reduced the loss of the various compositions of pepper, aesthetic quality and the quality of pepper products have been improved, reduce environmental pollution simultaneously, eliminated the shortcomings such as foreign odor taste that traditional infusion process is difficult to avoid and production cycle be long.
Accompanying drawing explanation
Fig. 1 is solid state fermentation decortication.A: aspergillus niger D-04 solid state fermentation makes pepper bean shelling; B: bacterium 11b045 solid state fermentation makes pepper bean shelling.
Fig. 2 is the comparison of different fermentations decortication mode.A: just washed the white pepper after decortication by rubbing with the hands; B: the white pepper of drying.
Fig. 3 is the impact that different material-water ratios reduce substrate dry weight.
Fig. 4 is the impact that different pH values reduce substrate dry weight.
Fig. 5 is different pretreatments decortication result.A: acid treatment; B: contrast; C: blanching is processed.
Fig. 6 connects the impact of bent amount on pepper bean shelling rate.
Fig. 7 is the impact of different fermentations time on pepper peeling rate.
Fig. 8 is solid state fermentation decortication amplification test.A: enlarged drawing; B: Local map.
Fig. 9 is the contrast of main chemical compositions content.
Figure 10 is white pepper sample GC figure.A: infusion process gained white pepper; B: solid state fermentation gained white pepper.
The specific embodiment
The experimental technique using in following embodiment if no special instructions, is conventional method.
In following embodiment, material used, reagent etc., if no special instructions, all can obtain from commercial channels.
Peeling rate computing formula in following embodiment is as follows: the total grain of peeling rate %=decortication grain number ÷ number * 100%.
The exploration of embodiment 1, solid state fermentation conditions
One, bacterial classification is selected
1, the growing state of bacterial strain on bran mass
Consider that in solid state fermentation, wheat bran is the base-material that conventional bacterial classification is cultivated, the 113 strain inoculation that 24 bacterial strains buying and laboratory are screened are in earlier stage to bran mass, cultivate after 72h, observe the growth and breeding situation of each bacterial strain and the situation of change of solid-state bran mass.The result of observable part bacterial strain is as shown in table 1 below, and all the other bacterial strains are not observed any variation on bran mass.
Bran mass preparation: 10g wheat bran, add 10mL distilled water, fully infiltrate, pack in the triangular flask of 250mL, jump a queue, wrap up, 121 ℃ of moist heat sterilization 20min.
The growing state of bacterial strain on table 1 bran mass
As can be seen from Table 1, aspergillus niger, mould, head mold can be observed the variation of the bran mass color causing because of mycelia or spore during the fermentation, bacterium can judge whether this bacterial strain can be at the good growth and breeding of solid state fermentation basal medium from viscosity and the smell of bran mass, other the bacterial strain that does not observe any variation is thought and can not be grown thereon, does not consider the bacterial classification as solid state fermentation.
2, the bacterial strain screening based on decortication effect
Method: be inoculated into after bacterial strain is activated in solid medium basal culture medium, 28 ℃ of cultivation 72h of der Pilz (37 ℃ of cultivation 72h of bacterium class), are wheat bran seed bent.Then by the pepper fresh fruit 20g of blanching 20s in boiling water (draining after scalding), be placed in 250mL triangular flask, according to the inoculum concentration of 0.5% (mass percent), add wheat bran bent, mix, breathable sealing film sealing, 28 ℃ of fermentations of der Pilz, 37 ℃ of fermentations of bacterium class, with the bran mass do not inoculated in contrast, according to above-mentioned peeling technology test, calculate peeling rate, observe color and luster, smell smelling.
The composition of solid medium basal culture medium: the mixture that wheat bran and water are mixed to get with the mass ratio of 1: 1.
Result: directly the bacterial strain in step 1 is carried out to pepper decortication experiment, peeling rate and pepper organoleptic parameters are in Table 2 and Fig. 1.Wherein, best with the decortication effect of the 15 strain aspergillus nigers such as CICC40273, CICC2317, D-04, after fermentation 60h, pepper peeling rate reaches 100%, and the color and luster of gained white pepper, smell, taste sensory evaluation are all higher, smells basic free from extraneous odour while hearing.Pepper decortication after aspergillus niger is processed blanching is very effective, and the decortication time is short, and peeling rate is high, and gained white pepper color is better, and smell is pure, substantially can eliminate traditional infusion process and in sense of smell, be difficult to the foreign odor taste of avoiding.
Bacillus subtilis (Bacillus subtilis) CICC10076, bacillus subtilis (Bacillus subtilis) CICC21704, bacillus subtilis (Bacillus subtilis) CICC20220, bacillus subtilis (Bacillussubtilis) CICC20020 are all purchased from Chinese industrial microorganism fungus kind preservation administrative center (CICC); Bacillus subtilis (Bacillus subtilis) ACCC10629 and bacillus subtilis (Bacillus subtilis) ACCC03427 are all purchased from Chinese agriculture microorganism fungus kind preservation administrative center (ACCC).
Penicillium purpurogenum (Penicillium purpurogenum) 03005, mould (Penicillium) Y-03, mould (Penicillium) C-01, mould (Penicillium) A-012131, mould (Penicillium) A-08 are University Of Hainan's Foodstuffs Academy laboratory from row filter preservation strain.
Rhizopus stolonifer (Rhizopus stolonifer) CICC40326, Rhizopus stolonifer (Rhizopus Stolonifer) CICC40322, Rhizopus stolonifer (Rhizopus stolonifer) CICC40343 are all purchased from Chinese industrial microorganism fungus kind preservation administrative center (CICC); Middle rhizopus chinensis (Rhizopus chinensis) ACCC30301 is purchased from Chinese agriculture microorganism fungus kind preservation administrative center (ACCC).
The decortication effect of table 2 different strains
From table 1, table 2 and Fig. 1 comprehensively, aspergillus niger growth and breeding in wheat bran solid-state fermentation culture medium is fast, and there is no peculiar smell; And bacterium class, head mold class and the decortication of mould class solid state fermentation, after fermentation 60h, not only peeling rate is low, and gained pepper products color and luster is poor.We select aspergillus niger D-04 to carry out subsequent experimental as pepper bean shelling bacterial classification.
Two, the contrast of fermentation mode
Liquid state fermentation: get 20g pepper fresh fruit fringe, blanching 40s, the zymotic fluid of inoculation 1mL penicillium purpurogenum 03005, adds 100mL distilled water, mixes, and after fermentation 4.5d, washes decortication by rubbing with the hands, dry.The preparation method of the zymotic fluid of penicillium purpurogenum 03005: be inoculated in liquid state fermentation seed culture medium after penicillium purpurogenum 03005 is activated, 28 ℃ of fermentation 24h, are seed liquor.Seed liquor is inoculated in liquid state fermentation culture medium, 28 ℃ of fermentation 72h, are zymotic fluid again.The composition of seed culture medium: (NH
4)
2sO
410.0g, K
2hPO
43H
2o1.0g, MgSO
47H
2o1.0g, FeSO
47H
2o0.1g, KCl0.5g, yeast extract 0.1g, pectin 2.0g, distilled water is settled to 1000mL, and pH nature, jumps a queue, wraps up, 115 ℃ of sterilizing 20min.The composition of culture medium: beef extract 10.0g, pectin 2.0g, NaCl 5.0g, KCl 1.0g, MgSO47H
2o 0.5g, FeSO
47H
2o 0.05g, distilled water is settled to 1000mL, and pH6.0 jumps a queue, wraps up, 115 ℃ of sterilizing 20min.
Solid state fermentation: get 20g pepper fresh fruit fringe, blanching 20s, adds 0.1g (0.5%) wheat bran bent, mixes, and washes decortication by rubbing with the hands after fermentation 60h, dry.The preparation of wheat bran song: be inoculated in bran mass after aspergillus niger D-04 is activated, 28 ℃ of fermentation 72h, are wheat bran song.The composition of bran mass: the mixture that wheat bran and water are mixed to get with the mass ratio of 1: 1.
Liquid state fermentation and solid state fermentation decortication result are as following table 3.
The comparison of the different decortication modes of table 3
As shown in Table 3, liquid state fermentation makes pepper bean shelling, needs strict aseptic condition early stage, and process is complicated, and investment, energy consumption are high; Pollute around water body and air; And the pepper bean shelling time is long, peel thorough not, cannot avoid the generation of peculiar smell etc.And solid state fermentation is the method generally adopting in China's traditional food fermentation.Solid state fermentation is by the bacterial classification advantage of inoculation, and without strict aseptic condition, sweat is extensive, and microorganism particularly mould easily grows, and enzyme activity is high, and enzyme system is abundant; Simple in equipment, small investment, energy consumption are low, easy and simple to handle, basic non-wastewater discharge, be particularly suitable for applying in rural area, this is the advantage of solid state fermentation uniqueness, and the pepper bean shelling time is short, thoroughly, pepper smells basic free from extraneous odour while hearing (solid state fermentation decortication is produced white pepper picture and seen Fig. 2) in decortication.
Two, the preparation of wheat bran song
1, the impact of material-water ratio on strain growth breeding
Experimental strain: aspergillus niger D-04.
According to the distilled water of every 10g wheat bran siccative interpolation 8,10,12,15,20mL, cultivate after 72h for 28 ℃, by measuring substrate dry weight, reduce to judge the situation of Aspergillus Niger Growth breeding, substrate dry weight reduction is as shown in Figure 3.
As shown in Figure 3, culture medium material-water ratio is between 1: 1 to 1: 2, and substrate dry weight reduces all larger, and now, aspergillus niger D-04 sprouts very fast, and the gas porosity of culture medium is better.When culture medium material-water ratio is 1: 0.8, water content is very few, cannot meet aspergillus niger spore and sprout mycelial growth desired moisture level, and strain growth metabolism is not slower; When culture medium material-water ratio is 1: 2, water content is excessive, and the humidity of growth of microorganism is just excessive, and the ventilation of culture medium, heat dispersion are had to impact, and as culture medium, easily caking is agglomerating, and then suppresses its growth and breeding.
2, the impact of pH value on strain growth breeding
Use certain density H
2sO
4prepare respectively the wheat bran solid medium of different initial pH value with NaOH solution, carry out solid-state culture experiment.Cultivate 72h for 28 ℃, measure substrate dry weight and reduce.Result as shown in Figure 4.
The composition of wheat bran solid medium: the distilled water that adds 10mL according to every 10g wheat bran siccative.
As shown in Figure 4, aspergillus niger D-04 can be at natural pH and growth and breeding well in the environment of slant acidity slightly, and alkali condition is unfavorable for the growth and breeding of this aspergillus niger.Culture medium nature pH value is 6.6 left and right, therefore, in experiment afterwards, no longer carries out the adjusting of pH value.
Two, the research of pepper fresh fruit form of feed pretreatment
Solid state fermentation conditions in this experiment is as follows:
Wheat bran is bent: 10g wheat bran siccative adds the distilled water of 10mL, pH value nature, and 121 ℃ of moist heat sterilization 20min, are bran mass.After aspergillus niger D-04 is activated, be inoculated in bran mass, 28 ℃ of fermentation 72h, are wheat bran song.
Fermentation decortication temperature: 28 ℃.Pepper connects bent amount 0.5%.
1, the impact of pepper fruit ear threshing on pepper bean shelling rate
Pepper corn after threshing is directly mixed to after fermentation with wheat bran song, observe the softening situation of pepper fruit, and from 36h, wash decortication by rubbing with the hands, calculating peeling rate.Result is as table 4.Bacterial strain uses therefor is aspergillus niger D-04.The method that threshing is processed: directly pepper fresh fruit fringe crumpled to threshing, remove stalk.
The impact on pepper bean shelling is processed in table 4 threshing
Table 4 result shows, experimental group and control group indifference, even if later observation is found fermentation 6 days, also only have several can peel, and we think that threshing processing can not promote pepper bean shelling.
2, the impact of pretreatment mode on pepper bean shelling rate
Pepper fresh fruit is at room temperature carried out respectively to acid treatment (3%HCl soaks 10min), alkali treatment (3%NaOH soaks 10min) and water blancing 20s to be processed, and consider that threshing processing may produce interactive impact to three kinds of processing modes, has designed following pepper peeling test.After strain fermentation 60h, wash decortication by rubbing with the hands and calculate its peeling rate, result is as table 5.
The impact of table 5 different pretreatments on pepper bean shelling rate
From table 5 and Fig. 5, can find out, in above-mentioned three kinds of processing, to only have blanching to process and make pepper bean shelling effective to aspergillus niger, other processing pepper fresh fruit after fermentation 60h does not almost change, and does not occur softening sign.And threshing is processed these three kinds of pretreatment modes be there is no to too large impact.
3, the impact of blanching processing time on pepper bean shelling rate
Pepper fresh fruit is carried out respectively to blanching 10s, 20s, 30s, 40s, 50s, 1min, 1.5min, 2min, 3min processing, in the different fermentations time, wash decortication by rubbing with the hands, calculate peeling rate, result is as following table 6.Bacterial strain uses therefor is aspergillus niger D-04.
The impact of the different blanching treatment time of table 6 on pepper bean shelling rate
As can be found from Table 6, usining aspergillus niger D-04 during as pepper bean shelling bacterial classification, as long as process through blanching, no matter blanching treatment time length, after fermentation 60h, the peeling rate of pepper fresh fruit can reach 100%.And, along with the prolongation in blanching processing time, fermentation decortication time shorten, peeling rate improves gradually.
Three, the impact of vaccination ways
Solid state fermentation conditions in this experiment is as follows:
Wheat bran is bent: 10g wheat bran siccative adds the distilled water of 10mL, pH value nature, and 121 ℃ of moist heat sterilization 20min, are bran mass.After aspergillus niger D-04, CICC40273, CICC2317 is activated, be inoculated in bran mass, 28 ℃ of fermentation 72h, are wheat bran song.
Fermentation decortication temperature: 28 ℃.Test organisms: aspergillus niger D-04, CICC 40273, CICC 2317.
Adopt two kinds of different modes to inoculate, the one, directly the spore suspension (10 of making is washed in the PDA inclined-plane from fresh activation
7individual/mL) be sprayed directly on blanching and process on the pepper after 20s, and mix thoroughly, inoculum concentration is about 10
6individual/g pepper.The 2nd, according to 0.5%, (be that 0.5g wheat bran is bent: the bent pepper of processing after 20s with blanching of wheat bran 100g pepper) mixes.Be placed in 28 ℃ of bottom fermentation decortications simultaneously.Every kind of mode do 4 parallel.48h starts to wash by rubbing with the hands decortication, and result is as following table 7.
The impact of table 7 different vaccination ways on pepper bean shelling rate
Table 7 result shows, this directly sprinkling by spore liquid and seed that need not solid-state wheat bran song, and the time of peeling completely, the decortication time was 3.5d completely approximately than the many 1d of wheat bran sort of quyi left and right.
Four, the impact of inoculum concentration
Solid state fermentation conditions in this experiment is as follows:
Wheat bran is bent: 10g wheat bran siccative adds the distilled water of 10mL, pH value nature, and 121 ℃ of moist heat sterilization 20min, are bran mass.After aspergillus niger D-04, CICC40273, CICC2317 is activated, be inoculated in bran mass, 28 ℃ of fermentation 72h, are wheat bran song.
Fermentation decortication temperature: 28 ℃.Test organisms: aspergillus niger D-04, CICC 40273, CICC 2317.
According to every 100 grams of cooled peppers of blanching 20s, connect the bent amount of wheat bran and be respectively 0.1g, 0.3g, 0.5g, 0.8g, 1g, 5g wheat bran song, to determine minimum inoculum concentration.The peeling rate of different fermentations after the time as shown in Figure 6.
As can be seen from Figure 6, before fermentation 60h, peeling rate increases along with connecing the increase of bent amount; But after fermentation 60h, there is not this positive correlation.When connecing bent amount 0.5% when above, the 60h rear peeling rate of fermenting can reach 100%.
Five, the impact of fermentation decortication temperature
All the other solid state fermentation conditionses in this experiment are as follows:
Wheat bran is bent: 10g wheat bran siccative adds the distilled water of 10mL, pH value nature, and 121 ℃ of moist heat sterilization 20min, are bran mass.After aspergillus niger D-04 is activated, be inoculated in bran mass, 28 ℃ of fermentation 72h, are wheat bran song.
Inoculum concentration: it is 0.5g that every 100 grams of cooled peppers of blanching 20s connect the bent amount of wheat bran.Test organisms: aspergillus niger D-04, CICC 40273, CICC 2317.
Consider the weather conditions that pepper harvest season and Hainan are special, general room temperature is 28-35 ℃, therefore choose the decortication experiment that slightly high temperature is carried out pepper fresh fruit.Experimental result is as shown in the table.
The impact of table 8 different fermentations temperature on pepper bean shelling
As can be seen from Table 8, at fermentation proecdysis, excess Temperature (more than 37 ℃) may have certain inhibitory action to microbial growth breeding, but after fermentation 60h, pepper fresh fruit can be peeled completely.Consider the climatic environment of Hainan uniqueness, room temperature just can meet microorganism fermentation well makes the softening decortication of pepper fresh fruit.
Six, the impact of fermentation decortication time
All the other solid state fermentation conditionses in this experiment are as follows:
Wheat bran is bent: 10g wheat bran siccative adds the distilled water of 10mL, pH value nature, and 121 ℃ of moist heat sterilization 20min, are bran mass.After aspergillus niger D-04 is activated, be inoculated in bran mass, 28 ℃ of fermentation 72h, are wheat bran song.
Inoculum concentration: it is 0.5g that every 100 grams of cooled peppers of blanching 20s connect the bent amount of wheat bran.
Fermentation decortication temperature: 28 ℃.
Pepper in fermentation decortication process is washed by rubbing with the hands to decortication at 24h, 36h, 48h, 60h respectively, calculate its peeling rate, result is as Fig. 7.As can be seen from Figure 7, along with the prolongation of fermentation time, pepper peeling rate increases gradually; In fermentation 48h rear peeling rate, almost can reach more than 95%.And after fermentation 60h, peeling rate just can reach 100%.
One, experiment material:
Wheat bran: wheat bran;
Pepper fresh fruit: pepper fresh fruit is in picking up from pepper orchard, Yangjiang town, Qionghai City, Hainan Province 2009/2010 year 5-8 month, fresh fruit maturity 70%～80% (hand is pinched pulp can be separated with pericarp, and kernel has certain degree of hardness).
Aspergillus niger (Aspergillus niger) CICC 40273, aspergillus niger (Aspergillus niger) CICC 2317, aspergillus niger (Aspergillus niger) CICC 2214, aspergillus niger (Aspergillus niger) CICC 40616, aspergillus niger (Aspergillus niger) CICC 40097, aspergillus niger (Aspergillus niger) CICC 40094, aspergillus niger (Aspergillus niger) CICC40493, aspergillus niger (Aspergillus niger) CICC 40091, carbon black aspergillus (Aspergillus carbonarius) CICC 41254, carbon black aspergillus (Aspergillus carbonarius) CICC 40093, aspergillus awamori (Aspergillus awamori) CICC 41467, aspergillus awamori (Aspergillus awamori) CICC41466, aspergillus awamori (Aspergillus awamori) CICC 40499, Aureobasidium pullulans (Aureobasidium pullulans) CICC 40331 is all purchased from Chinese industrial microorganism fungus kind preservation administrative center (CICC),
Aspergillus niger (Aspergillus niger) CGMCC 3.316, aspergillus niger (Aspergillus niger) CGMCC 3.3289, aspergillus niger (Aspergillus niger) CGMCC 3.3287, aspergillus niger (Aspergillus niger) CGMCC 3.3284 are all purchased from Chinese common micro-organisms DSMZ (CGMCC).
Solid state fermentation container: the little pail of plastics.
Two, experimental technique
The basic step of the inventive method is as follows:
1, prepare wheat bran song or spore suspension
1-1, prepare bacterial classification solid medium: be following 1) or 2) shown in:
1) wheat bran is mixed with certain proportion with water, the mixture obtaining is described solid medium; Solid medium is divided and installed in 250mL triangular flask, each 20g, breathable sealing film sealing, puts autoclaving 20min at 121 ℃, or atmospheric cooking 40-60min.
2) wheat bran, water are mixed with certain proportion with at least one in following material: rice husk, bean cake powder, maize cob meal, pomelo peel powder, orange peel powder, bagasse, manioc waste etc., the mixture obtaining is described solid medium; Solid medium is divided and installed in 250mL triangular flask, each 20g, breathable sealing film sealing, puts autoclaving 20min at 121 ℃, or atmospheric cooking 40-60min.
The preparation of spore suspension: the PDA inclined-plane of getting fresh activation, add 0.05% (v/v) Tween-80 aqueous solution of sterilized water or sterilizing, wash lower spore to being equipped with in the triangular flask of bead, vibration 10min, with blood counting chamber counting, with sterilized water, adjust spore concentration 1 * 10
7individual/mL.
1-2, bacterial classification are cultivated
In above-mentioned solid medium, access bacterial classification, cultivate, extremely described bacterium obtains growth and breeding, and all substances in the triangular flask obtaining are wheat bran song.
2, pepper fresh fruit pretreatment (blanching processing)
The environment that pepper fresh fruit wait peeling is placed in more than 60 ℃ or more than 100 ℃ kept after a period of time, and nature or flowing water are cooled to room temperature.Specifically can adopt the methods such as hot water blanching, water blancing, steam blanching, microwave blanching.
Specifically can be as follows: pepper fresh fruit is wrapped with bundle, be placed in the boiling water of ebuillition of heated in advance, start timing, and keep after a period of time in boiling water environment, take out pepper, flowing water is cooling or naturally cooling.
3, solid state fermentation
Vaccination ways 1 (wheat bran is bent): the pepper after blanching is processed mixes according to a certain percentage with above-mentioned wheat bran song, is placed in 250mL triangular flask, mixes thoroughly, and breathable sealing film sealing, carries out solid state fermentation.
Vaccination ways 2 (spore suspension): the pepper after blanching is processed mixes (as spore suspension is sprayed onto on pepper fruit) according to a certain percentage with spore suspension, be placed in 250mL triangular flask, mixes thoroughly, and breathable sealing film sealing, carries out solid state fermentation.
4, peeling
Pepper fruit after fermentation is washed by rubbing with the hands, obtained the white pepper of decortication.
5, dry
Pepper seed after decortication is cleaned, be placed in and on straw mat or concrete floor, dry or dry, can obtain the just white pepper of system.
Specific experiment is as follows
(1), experimental group 1:
1, prepare wheat bran song
Wheat bran: water=10g: 8mL; The pH value nature (6.6) of mixture.
Bacterium: aspergillus niger (Aspergillus niger) CICC 40273.
Cultivation temperature: 28 ℃.
Incubation time: 72h.
2, pepper pretreatment
Water blancing, 10s.
3, solid state fermentation
Vaccination ways 1 (wheat bran is bent): the pepper after blanching is processed: wheat bran song=100g: 0.3g; Pepper after blanching is processed is 20g.Fermentation temperature: 25 ℃.
Vaccination ways 2 (spore suspension): the pepper after blanching is processed: spore suspension=20g: 1mL concentration is 1 * 10
7the spore suspension of individual/mL.Fermentation temperature: 25 ℃.
Control group 1-1, undressed pepper fresh fruit (being left intact);
In pepper after control group 1-2, blanching, do not inoculate wheat bran song or spore liquid, all the other are identical with experimental group;
Control group 1-3, the pepper after blanching is mixed with the solid medium that does not add bacterium, all the other are identical with experimental group.
Result is as follows:
The experimental group of inoculation wheat bran song, fermentation 36h, its peeling rate 80%; Fermentation 60h, its peeling rate reaches 100%.
The experimental group of inoculating spores suspension, fermentation 48h, its peeling rate 50%; Fermentation 84h, its peeling rate reaches 100%.
Control group 1-1,1-2,1-3 place 60h, only have only a few pepper bean shelling.
(2), experimental group 2:
1, prepare wheat bran song
Wheat bran: water=10g: 10mL; The pH value nature of mixture.
Bacterium: aspergillus niger (Aspergillus niger) CICC 2317.
Cultivation temperature: 28 ℃.Incubation time: 84h.
2, pepper pretreatment
Water blancing, 20s.
3, solid state fermentation
Vaccination ways 1 (wheat bran is bent): the pepper after blanching is processed: wheat bran song=100g: 0.5g; Pepper after blanching is processed is 20g.Fermentation temperature: 28 ℃.
Vaccination ways 2 (spore suspension): the pepper after blanching is processed: spore suspension=20g: 2mL concentration is 1 * 10
7the spore suspension of individual/mL.Fermentation temperature: 28 ℃.
Control group 2-1, undressed pepper fresh fruit (being left intact);
In pepper after control group 2-2, blanching, do not inoculate wheat bran song or spore liquid, all the other are identical with experimental group;
Control group 2-3, the pepper after blanching is mixed with the solid medium that does not add bacterium, all the other are identical with experimental group.
Result is as follows:
The experimental group of inoculation wheat bran song, fermentation 36h, its peeling rate 85%; Fermentation 60h, its peeling rate reaches 100%.
The experimental group of inoculating spores suspension, fermentation 48h, its peeling rate 60%; Fermentation 84h, its peeling rate reaches 100%.
Control group 2-1,2-2,2-3 place 60h, only have only a few pepper bean shelling.
(3), experimental group 3:
1, prepare wheat bran song
Wheat bran: water=10g: 20mL; The pH value nature of mixture.
Bacterium: aspergillus niger (Aspergillus niger) D-04.
Cultivation temperature: 28 ℃.Incubation time: 96h.
2, pepper pretreatment
Water blancing, 30s.
3, solid state fermentation
Vaccination ways 1 (wheat bran is bent): the pepper after blanching is processed: wheat bran song=100g: 5g; Pepper after blanching is processed is 20g.Fermentation temperature: 40 ℃.
Vaccination ways 2 (spore suspension): the pepper after blanching is processed: spore suspension=20g: 5mL concentration is 1 * 10
7the spore suspension of individual/mL.Fermentation temperature: 40 ℃.
Control group 3-1, undressed pepper fresh fruit (being left intact);
In pepper after control group 3-2, blanching, do not inoculate wheat bran song or spore liquid, all the other are identical with experimental group;
Control group 3-3, the pepper after blanching is mixed with the solid medium that does not add bacterium, all the other are identical with experimental group.
Result is as follows:
The experimental group of inoculation wheat bran song, fermentation 36h, its peeling rate 85%; Fermentation 60h, its peeling rate can reach 100%.
The experimental group of inoculating spores suspension, fermentation 48h, its peeling rate 60%; Fermentation 84h, its peeling rate can reach 100%.
Control group 3-1,3-2,3-3 place 60h, only have only a few pepper bean shelling.
Three, effect assessment
(1) after decortication, pepper main chemical compositions is analyzed
The content of pipering, volatile oil and fixedness ethereal extract in pepper after test experience group 1, experimental group 2 and experimental group 3 decortications; And compare with commercially available pepper of the same race.Result is as shown in the table.Three kinds of component contents in the pepper that three experimental group obtain are all high or approaching than corresponding component content in commercially available pepper.Show, the inventive method does not reduce the quality of pepper.
The mensuration of pipering is with reference to GB GB/T17528-2009;
The mensuration of volatile oil (pepper essential oil) is with reference to GB GB/T17527-2009;
The mensuration of fixedness ethereal extract is with reference to GB GB/T 12729.12-2008.
Table 9
(2) detect the content of n-butyric acid, p-Cresol, m-cresol, three-methyl indol in pepper
Content with n-butyric acid, p-Cresol, m-cresol, three-methyl indol in the white pepper obtaining after the 60h that ferments in gas-chromatography (GC) method test experience group 1,2,3.Compare with natural infusion process simultaneously.
The standard items that use are:
N-butyric acid (Cas:107-92-6) is pure for analyzing, and is purchased from Aladdin reagent (China) Co., Ltd (Aladdin Chemistry Co.Ltd.), and production number is 1111946.
P-Cresol (Cas:106-44-5) is pure for analyzing, and is purchased from Aladdin reagent (China) Co., Ltd (Aladdin Chemistry Co.Ltd.), and production number is 1134007.
M-cresol (Cas:108-39-4) is pure for analyzing, and is purchased from Aladdin reagent (China) Co., Ltd (Aladdin Chemistry Co.Ltd.), and production number is 1133329.
3-methyl indol (Cas:83-34-1) is pure for analyzing, and is purchased from A Faaisha Chemical Co., Ltd. (Alfar Aesar, A Johnson Matthey Company), and production number is L03890.
Gas-chromatography (GC) method is as follows:
Instrument is the U.S. 7890A of Agilent Technologies company gas chromatograph, hand sampling-headspace solid-phase microextraction (SPME, U.S. Supelco company), sample injection time 15min.Before experiment every day, by the aging 2h of the extracting head of SPME, aging temperature is 250 ℃.
GC condition: DB-WAX 122-7032 capillary column (30m * 0.25mm * 0.25 μ tm); 60 ℃ of initial column temperatures, 20 ℃/min of temperature programming, 200 ℃ of lasting 10min, rear operation 2min, flow 1.5mL/min, does not shunt, and carrier gas is High Purity Nitrogen, 250 ℃ of injector temperatures.
The step of nature infusion process: pack woven bag into after ripe pepper fruit ear is plucked and be directly placed in pond, fish pond, or be directly placed in mobile little river and soak, 7～15d is processed in spontaneous fermentation, and black pepper peel can directly be washed decortication by rubbing with the hands after fully rotting.Then with clear water, rinse, remove the foreign material such as carpopodium wherein, after being fully dried, obtain white pepper.
(A is nature infusion process result to result as shown in figure 10; B is the inventive method result).Result shows, 1, the pepper that natural infusion process is produced detects three kinds of stink compositions, n-butyric acid 1.19mg/g (retention time is 5.829min), p-Cresol 0.18 μ tg/g (retention time is 8.152min), m-cresol 0.047 μ g/g (retention time is 8.561min), and n-butyric acid content is higher.2, the pepper that the inventive method is produced only detects a kind of stink composition of n-butyric acid (retention time is 5.83min), and content is extremely low, only has 18.9% of nature infusion process, experimental group 1:0.231mg/g, experimental group 2:0.225mg/g, experimental group 3:0.227mg/g.Illustrate that the inventive method does not produce or produce the stink composition of minute quantity, smell news odorless, free from environmental pollution.
Nature infusion process and the inventive method group all do not detect 3-methyl indol.
(3) the blanching processing time is determined
According to the experiment of peeling of the method for experimental group 2, the blanching processing time is 10s, 30s, 50s, 90s, 120s, 180s, and all the other are all identical with the method for experimental group 2.
The pepper, the commercially available pepper of the same race that detect after decortication carry out main component analysis (pipering, volatile oil, fixedness ethereal extract).
Result as shown in Figure 9.
Result shows:
1, blanching treatment time has larger impact to fixedness ethereal extract, should shorten blanching treatment time as far as possible, is controlled at 30s and is advisable with interior.
2, the almost not impact of blanching treatment time on pipering and volatile oil.
3, the pepper products that the inventive method is produced has superiority than commercially available white pepper on pipering, volatile oil, fixedness ethereal extract content.
The inventive method also can be used for the pepper decortication of other form, as the decortication of black pepper.Because black pepper is to adopt the lower fresh pepper of maturity to obtain after drying, the fresh pepper that its pericarp composition and maturity are higher there is no too large difference.
In above-mentioned experimental group 1,2 or 3, when solid medium being replaced with when following, decortication effect and experimental group 1,2 or 3 be without significant difference, and the quality of the pepper obtaining and experimental group 1,2 or 3 be without significant difference, and the stink substance content of pepper and experimental group 1,2 or 3 are without significant difference.
Solid medium: by wheat bran, water with at least one in following material with 5g: 10mL: 5g ratio is mixed: rice husk, bean cake powder, maize cob meal, pomelo peel powder, orange peel powder, bagasse, manioc waste etc., the mixture obtaining is described solid medium; Solid medium is divided and installed in 250mL triangular flask, each 20g, breathable sealing film sealing, puts autoclaving 20min at 121 ℃, or atmospheric cooking 40-60min.
In above-mentioned experimental group 1,2 or 3, when bacterium being replaced with to any in following bacterium, decortication effect and experimental group 1,2 or 3 be without significant difference, and the quality of the pepper obtaining and experimental group 1,2 or 3 be without significant difference, and the stink substance content of pepper and experimental group 1,2 or 3 are without significant difference.
Aspergillus niger (Aspergillus niger) CICC 2214, aspergillus niger (Aspergillus niger) CICC 40616, aspergillus niger (Aspergillus niger) CICC 40097, aspergillus niger (Aspergillus niger) CICC 40094, aspergillus niger (Aspergillus niger) CICC40493, aspergillus niger (Aspergillus niger) CICC 40091, carbon black aspergillus (Aspergillus carbonarius) CICC 41254, carbon black aspergillus (Aspergillus carbonarius) CICC40093, aspergillus awamori (Aspergillus awamori) CICC 41467, aspergillus awamori (Aspergillus awamori) CICC 41466, aspergillus awamori (Aspergillus awamori) CICC 40499, Aureobasidium pullulans (Aureobasidium pullulans) CICC 40331 is all purchased from Chinese industrial microorganism fungus kind preservation administrative center (CICC), aspergillus niger (Aspergillus niger) D-04.
Aspergillus niger (Aspergillus niger) CGMCC 3.316, aspergillus niger (Aspergillus niger) CGMCC3.3289, aspergillus niger (Aspergillus niger) CGMCC 3.3287, aspergillus niger Aspergillus niger) CGMCC3.3284 is all purchased from Chinese common micro-organisms DSMZ (CGMCC).
Claims (16)
1. a method that makes pepper decortication, comprise the steps: pepper fruit to be peeled with the solid-state culture of Pectinase Producing Strain or mix with bacteria suspension or the spore suspension of Pectinase Producing Strain, carrying out solid state fermentation, fermenting complete, peeling, obtains the pepper of peeling; Before described mixing, comprise the step that following blanching is processed: pepper fruit to be peeled is placed in to 60 ℃ of above environment and keeps 10s-3min; Described Pectinase Producing Strain is at least one in following aspergillus niger: at least one in aspergillus niger (Aspergillus niger) CICC40273, aspergillus niger (Aspergillus niger) CICC2317 and aspergillus niger (Aspergillus niger) D-04;
The solid-state culture of described Pectinase Producing Strain is prepared as follows and obtains: for the preparation of the solid medium of cultivating described Pectinase Producing Strain, described Pectinase Producing Strain is inoculated in to described solid medium, cultivate, extremely described bacterium is bred, and all substances in the culture vessel obtaining are the solid-state culture of described Pectinase Producing Strain;
Described solid medium is following 1) or 2) shown in:
1) by wheat bran with water with 10g:(8mL-20mL) ratio mix, the mixture obtaining is described solid medium;
2) by wheat bran, water with at least one in following material with 5g:(8mL-20mL): the ratio of 5g is mixed: rice husk, bean cake powder, maize cob meal, pomelo peel powder, orange peel powder, bagasse, manioc waste, the mixture obtaining is described solid medium;
The pH value of described solid medium is 3.5-8.
2. method according to claim 1, is characterized in that: in described method, before described mixing, comprise the step that following blanching is processed: pepper fruit to be peeled is placed in to 60 ℃ of above environment and keeps 10-30s.
3. method according to claim 1 and 2, is characterized in that: described in the proportioning of solid-state culture of pepper fruit to be peeled and described Pectinase Producing Strain be 100g:(0.1g-10g);
The proportioning of pepper fruit described to be peeled and the spore suspension of described Pectinase Producing Strain is 20g:(1-5) * 10
7individual.
4. method according to claim 1 and 2, is characterized in that: the temperature of described solid state fermentation is 20 ℃-40 ℃.
5. method according to claim 4, is characterized in that: the temperature of described solid state fermentation is 28-30 ℃.
6. method according to claim 4, is characterized in that: the temperature of described solid state fermentation is 30-37 ℃.
7. method according to claim 1 and 2, is characterized in that: the time of described solid state fermentation is 24h-96h.
8. method according to claim 1 and 2, is characterized in that: the method for described peeling is for washing by rubbing with the hands.
9. method according to claim 1, is characterized in that: 1) described solid medium for by wheat bran with water with 10g:(10mL-20mL) ratio mix, the mixture obtaining is described solid medium.
10. method according to claim 9, is characterized in that: the pH value of described solid medium is 5.5-7.0.
11. methods according to claim 9, is characterized in that: the pH value of described solid medium is nature pH value.
12. methods according to claim 9, is characterized in that: described wheat bran is wheat bran.
13. methods according to claim 1 and 2, is characterized in that: during described blanching is processed, described 60 ℃ of above environment are hot water, boiling water or steam ambient.
14. methods according to claim 9, is characterized in that: in the preparation of the solid-state culture of described Pectinase Producing Strain, the temperature of described cultivation is 20 ℃-40 ℃, and the time of described cultivation is 48h-96h;
In the spore suspension of described Pectinase Producing Strain, solute is spore, and solvent is water.
15. methods according to claim 14, is characterized in that: in the preparation of the solid-state culture of described Pectinase Producing Strain, the temperature of described cultivation is 28-30 ℃, and the time of described cultivation is 72h-96h.
16. methods according to claim 14, is characterized in that: in the preparation of the solid-state culture of described Pectinase Producing Strain, the temperature of described cultivation is 30-37 ℃, and the time of described cultivation is 72h-96h.
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| CN104256862B (en) * | 2014-08-08 | 2016-11-02 | 中国热带农业科学院农业机械研究所 | A kind of Fructus Piperis fresh fruit associating peeling technology |
| CN104432423B (en) * | 2014-12-23 | 2017-02-01 | 海南省农业科学院农产品加工设计研究所 | Color protecting method applied to quick pepper decrustation conducted with biological enzyme method |
| CN105768029B (en) * | 2016-03-17 | 2020-09-15 | 中华全国供销合作总社南京野生植物综合利用研究所 | New method for processing white pepper |
| CN108634170B (en) * | 2018-02-28 | 2021-05-28 | 李赵生 | Method for decoloring white pepper by adopting microbial fermentation |
| CN111054307B (en) * | 2019-11-28 | 2022-07-05 | 海南大学 | Modified pepper stem adsorbing material and preparation method and application thereof |
| CN113151055B (en) * | 2021-03-17 | 2022-07-22 | 武汉轻工大学 | Pectinase-producing bacillus, fermentation method thereof and pepper peeling method |
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