CN103190585A - Novel bean sprout production process - Google Patents

Novel bean sprout production process Download PDF

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CN103190585A
CN103190585A CN2013101304758A CN201310130475A CN103190585A CN 103190585 A CN103190585 A CN 103190585A CN 2013101304758 A CN2013101304758 A CN 2013101304758A CN 201310130475 A CN201310130475 A CN 201310130475A CN 103190585 A CN103190585 A CN 103190585A
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bud
bud dish
dish
bacterium
fermentation
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CN103190585B (en
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左勇
鞠帅
边名鸿
叶阳
刘利平
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YIBIN SHUANGYI FUKANG FOOD CO., LTD.
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Sichuan University of Science and Engineering
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Abstract

The invention belongs to the technical field of bean sprout preparation methods, and in particular relates to a novel bean sprout production process. The process adopts an artificially inoculated and selected composite strain, which is prepared from lactobacilli and orange red yellow yeast in the ratio of (1-2):(1-2) in weight percent, wherein the inoculum size of the strain is 0.9-1.1% of the mass content of input bean sprout. By adopting the process, the fermentation time can be reduced by about 2 months, and the produced ester content is highest, and the variety and total content of volatile fragrant substances are largest, so that the Yibin bean sprout is richer in mouthfeel, and heavier in fragrance.

Description

A kind of new bud vegetable production technique
Technical field
The invention belongs to the preparation method technical field of bud dish, be specifically related to shorten fermentation time about 2 months, and the ester class content that produces is maximum, the kind of volatility aroma-producing substance and total content are also maximum, can make the mouthfeel of Yibin bud dish fuller, a kind of new bud vegetable production technique that fragrance is stronger.
Background technology
Yibin bud dish and " Fuling hot pickled mustard tube ", " Nanchong preserved vegetable ", " inland river root-mustard " also are called Sichuan four big catsup and pickled vegetables, and long history is arranged.Yibin bud dish is because having unique style and excellent extremely favors of consumer of quality such as " fragrant, sweet, crisp, tender, aquatic foods ".
At present, Yibin bud dish mainly adopts traditional aging process production, and the bud dish is left in fermentation vat or the ceramic jar, ferments by work bacterial classifications such as lactic acid bacteria, saccharomycete.At the bud dish fermentation initial stage, saccharomycete amount in the bud dish is few, mainly be lactobacillus-fermented, but along with fermentation time prolongs, lactic acid bacteria cross reasons such as low owing to being subjected to pH value and carbon source concentration, can not the recycling carbon source, and saccharomycete still can utilize well, until carbon source being depleted to least concentration, the organic acid generation esterification in the metabolite alcohols of its generation and bud dish after-ripening stage, produce abundant flavor substance, also the storage after bud dish local flavor and quality and the fermentation is had material impact.Fermentation period generally needs 4-6 month time, and traditional Yibin bud dish fermenting and producing cycle is long and quality is unstable, is unfavorable for large-scale industrial production, different saccharomycete, and the local flavor that its metabolism produces is also different with nutriment.
Summary of the invention
The present invention provides and can shorten fermentation period nearly 2 months, and improve the quality of products just at above technical problem, makes detectable flavor substance composition from 14 kinds of a kind of new bud vegetable production techniques that are increased to 22 kinds.
Concrete technical scheme of the present invention is as follows:
A kind of new bud vegetable production technique, this process using artificial infection and the composite bacteria that filters out, this composite bacteria is lactobacillus and orange red yellow yeast by mass ratio is that the proportionate relationship proportioning of 1-2:1-2 forms, and the inoculum concentration of its bacterial classification is the 0.9-1.1% into pond bud dish mass content.
A kind of new bud vegetable production technique, this technology specifically may further comprise the steps:
(1), will be ripe bud dish harvesting, its head is cut away, branches and leaves are removed, by hand its bar is cut into uniform strip, build the canopy airing in the sun, the bud dish bar of well cutting is put transpiring moisture thereon, evaporation capacity of moisture content is the 50%-60% of quality percentage composition in the bud dish;
(2) stirring is salted
Bud dish after reclaiming from the canopy that dries in the air stirs salted, and the suitable time of airing is strong and weak and weather decision by wind-force.Target is just passable when feeling that softness does not have hard core when holding green vegetables stem silk.Then that airing is good bud dish bar is cleaned, and it is dropped in the pond of preparing for fermentation, stirs salted method to be determined by the consumption of one deck bud dish one deck salt salinity and the situation of bud dish, and the consumption of salt is generally the 5-6% of bud dish weight percentage this moment;
(3) fermentation
In going into the bud dish in pond, add composite bacteria in proportion, ferment.Fermentation condition is that salinity is 5-6%, and the pH value is 6.5-7.0 in the fermentation vat environment, and temperature is controlled at 29-31 ℃, and fermentation time is that 90-100 days composite bacterias are by the inoculation of 0.9-1.1% amount;
(4) clean, dewater
Reject yellow leaf, go mouldy and other rotten dish, behind the foreign material that weed a garden, after the running water soaking and washing, guarantee no-sundries, no silt stone is removed excessive moisture in the bud dish by squeezing then, the 74-78% of squeezing back bud dish moisture control mass percent of bud dish after squeezing;
(5) cut the dish, screening and spice:
By cutting the bud dish is cut fineness degree to needs. separate defective bud dish by screening.Then in this moment bud dish mass percent the ratio of 0.1-0.2% add Chinese prickly ash, how anise and three waits spices, uniform mixing, how Chinese prickly ash, anise and three wait the addition of spices in right amount, only for seasoning;
(6) can, sterilization, test package
To can the packaging bag of bud dish vacuumize, utilize pasteurization to boil bag sterilization, sterilising temp 85-95 ℃.Sterilization time, single bag 500g weight following 25-30 minute, single bag 500g weight above 30-40 minute; Bottom pour ladle, mashed bag defective work, vanning are rejected in check.
The isolation and identification method that saccharomycete, lactic acid bacteria separate is:
One, the culture medium of seed selection
(1) 1%CaCO 3The MRS solid medium
Adopt peptone 10g, beef extract 10g, yeast extract 5g, dibasic ammonium citrate 2g, sodium acetate 5g, K 2HPO 42g, MnSO 44H 2O0.25g, MgSO 47H 2O0.58g, glucose 20g, 10g CaCO 3, Tween 80 1mL, agar 25g, water 1000mL.Transfer pH6.2～6.4,121 ℃ sterilization 15min.
(2) PY basal medium
Peptone 0.5g, pancreatin solution casein 0.5g, yeast extract 10g, salting liquid 4mL.Distilled water 100mL. adjust pH about 6.0,115 ℃ of sterilization 20min.
Salting liquid composition: anhydrous CaCl 20.2g, MgSO 47H 2O0.48g, K 2HPO 41.0g, KH 2PO 41.0g, NaHCO 310.0g NaCl2.0g is with CaCl 2And MgSO 4Be dissolved in together in the 300mL distilled water, add 500mL distilled water again, slowly add other salts while stir. continue to stir up to whole dissolvings, adding distil water is settled to 1000mL, mixes the back in 4 ℃ of storages, and is standby.
(3), glucose peptone water culture medium
Peptone 5g, glucose 5g, K 2HPO 42g, distilled water 1000mL.Adjust pH 7.0-7.2 filters, 112 ℃ of sterilization 30min.
(4), gelatin-based basal culture medium
Peptone 1.0g, yeast extract 1.0g, glucose 0.1g, gelatin 12.0g, salting liquid 4.0mL, distilled water 100mL.7.0,113 ℃-115 ℃ sterilizations of pH value 15min-20min.
(5), fermentation medium
Beef extract 10g/L, peptone 10g/L, yeast extract 10g/L, glucose 20g/L, Tween-80 0.5g/L, pH7.0,120 ℃ of sterilization 15min.
(6), litmus milk enrichment
With the upper strata cream of centrifuge removal milk, sub-cloud skim milk, it is the reindeer moss of 25g/L that the skim milk of every 100mL adds 4mL concentration, packing test tube, milk height 4-5cm.113 ℃ of high pressure steam sterilization 15-20min.
Two, strain separating, purifying
1, separates
Under aseptic condition, with the 1%CaCO of sterilization 3Be added in the MRS solid medium, pour into immediately in the culture dish after mixing shakes up, cool off standby.
Sterilized water is joined in the bud dish of fermentation Yibin, wash Yibin bud dish function bacterial classification, carry out gradient dilution then.Choose suitable dilution factor, coating is fallen flat board in 30 ℃ of cultivation 48h, and picking has bacterium colony and the colony counting of molten calcium circle.On the MRS flat board, rule repeatedly up to single bacterium colony; Repeat the dull and stereotyped line separation of going up again, purifying repeatedly, (the single size of bacterium colony is consistent to purifying through Gram, gram stain microscopy, thalline size solid colour), isolates 16 kinds of different strains altogether, be numbered AA → AP respectively, it is transferred on the MRS slant medium preservation under 4 ℃ of conditions respectively.
From I, II, 3 samples of III, get each 1mL of bacteria suspension respectively, these 3 kinds of bacteria suspensions are diluted to 10 respectively -8, coating, result such as following table 1.
Table 1 plate count result
Figure BDA00003054321500041
Can find out that according to table table 1 along with the bacteria suspension dilution factor increases, the bacterium colony on the flat board is fewer and feweri, and dilution factor is 10 -5Flat board afterwards can both be observed the morphological feature of single bacterium colony clearly.Morphological feature and microscopy result according to bacterium colony filter out 16 kinds of bacterium altogether, to its numbering, see Table 2.
Table 2 morphological feature
Figure BDA00003054321500052
2, saccharomycetic evaluation
2.1 saccharomycete morphology and physics and chemistry are identified
With AB, AE, the AF3 strain bacterium that obtains in separation in the table 2, choose suitable dilution factor, coating, fall dull and stereotyped in 28 ℃ of cultivation 48h, observation colony characteristics and microscopy, picking list bacterium colony, rule repeatedly on the PDA culture medium flat plate up to single bacterium colony, pure bacterial strain moves into slant medium.Carry out following serial qualification test:
2.1.1 morphology is identified
The bacterial classification of separation, purifying is coated onto respectively in the PDA culture medium, cultivates 48h for 28 ℃, observe colonial morphology, and do methylene blue dye liquor water logging sheet, water one iodine liquid water logging sheet, under high power lens, observe the individual morphology of thalline.Its form result such as table 3 and Fig. 1,2,3.
Table 3 colony morphology characteristic
Figure BDA00003054321500061
Learn by table 3, by colony morphology characteristic, can judge tentatively that this three strains bacterium may be saccharomycete.Microscopic morphology result
(1) methylene blue dye liquor water logging sheet
Get 1 of 0.05% methylene blue dyeing liquor, put slide central authorities, and get 3 kinds of bacterium and dyeing liquor mixing with oese respectively, dyeing 2～3min adds cover glass, observes thalli morphology under high power lens, the results are shown in Figure 4,5,6.
Find out that from Fig. 4,5,6 AB, AE bacterial cell are rounded, AF bacterial cell ovalize all has cell to be in the gemmation state.Known that by figure some cell is transparence, some is blue because methylene blue has oxidisability, and just carrying out in the living cell body metabolism have reducing property methylene blue is reduced into colourless, so, be the expression living cells of transparence, blueness is dead cell.
(2) water-iodine liquid water logging sheet
1 iodine liquid is placed slide central authorities, get a bacteria suspension again, mixing, covered is observed under the oily mirror, the results are shown in Figure 7,8,9.
Learnt that by Fig. 7,8,9 each cell edges is the transparent interior color and deepens to be yellowish-brown gradually.Cell has sprout in the bacterium, and cell is dyed yellowish-brown and sprout is transparence.From the AB bacterium, can be observed two cells, one of them half be yellowish-brown half be transparent, another cell is yellowish-brown up and down, it is transparent that the centre is, and it is investigated and read related data as can be known.Two cells may be in the latter stage in the cell division state, are about to be split into two cells by a cell.
2.1.2 physics and chemistry is identified
(1) produces pure ability comparative experiments
Under sterile working, in TTC lower floor culture medium, place 38 ℃ to cultivate 48h each inoculation of separation and purification, pour TTC upper strata culture medium into, insulation (shady place) 2～3h after growing bacterium colony.By change in color, compare the product alcohol ability between each bacterial strain, the more dark explanation producing and ethanol of redness ability is more strong.
(2) kind of starch hydrolysis hydrolysis
Under sterile working, in each the inoculation PYD fluid nutrient medium with separation and purification, place 200r/min, 28 ℃ of constant temperature culture 48h get a nutrient solution and drop in the white color board, add a Ge Shi iodine liquid mixing, observe change in color.
(4) produce the uncut jade test
Under the sterile working, the bacterial classification that activated is inoculated into aseptic brewer's wort solid medium examination respectively by 2% inoculum concentration, behind 28 ℃ of constant temperature culture 48h, observes, record its surface and whether produce uncut jade.
(5) physics and chemistry qualification result
By the different physical and chemical experiment of bacterial classification is drawn, the results are shown in Table 4.
Table 4 physics and chemistry qualification result
Figure BDA00003054321500071
Figure BDA00003054321500081
Annotate :+expression can be produced alcohol/products uncut jade, ++ number is represented to produce what ,-represent not produce.
Draw by table 4, the product alcohol ability of AE bacterium is the strongest, and the AB bacterium takes second place, and the product alcohol ability of AF bacterium is the poorest; The nutrient solution color all is black-and-blue, illustrates that these three kinds of bacterial strains have all produced the kind of starch compound in growth course; AB bacterium, AE bacterium, AF bacterium all produce denseer wine flavour; The bacterium uncut jade that AF produces is maximum, and AB takes second place, and the AE bacterium does not produce the bacterium uncut jade.
In actual production, flower is given birth on the surface of curing food, long tunica albuginea should be avoided aborning as far as possible.Because living flower, long tunica albuginea not only can influence the normal fermentation of Yibin bud dish, produce bad smell, and the appearance of product is also bad, can test by producing uncut jade, eliminate those and produce the strong bacterial strain of uncut jade abilities.
2.1.3Biolog Automatic Analyzer for Microbes is identified saccharomycete
(1) BUY culture medium flat plate line
Select the single bacterium colony of cultivating on the 28h inclined-plane, carry out " cross " sectional streak at the BUY culture medium, cultivate 48h at 28 ℃ of following constant temperature and humidities.
(2) adjust turbidity
Single bacterium colony with in aseptic bamboo let picking " cross " line is inoculated under aseptic condition in the turbidity pipe.Blank transfers to 100%, and saccharomycete suspension standard pipe transfers to 47%.Measure with postvaccinal turbidity pipe, can constantly adjust turbidity with special-purpose sterilized water, to reach saccharomycete suspension standard turbidity value (47%).
(3) preparation bacteria suspension
Get aseptic bamboo let in inoculation liquid, dip in wet, with the bamboo let sticking thalline of getting that slowly slides on the bacterium colony surface.Inclination inoculation liquid pipe also rotates bamboo let along inwall, and thalline is attached on the inwall, simultaneously thalline is evenly broken up.Adjust turbidity value by adding thalline or blank inoculation liquid, make its respective standard turbidity value ± 2% scope in.
(4) inoculation microplate
To adjust the bacteria suspension of turbidity, bacteria suspension is added in respectively in each hole of different identification plates with 8 electronic pipettors, every hole application of sample 100 μ L, totally 96 holes are placed in the pallet, and keep certain humidity, cultivate 24h, 48h and 72h down for 26 ℃.
(5) read data
Take out and identify that microplate is placed on the readout instrument, open identification systems, computer reads data, provides the title of 10 ID according to the possibility size.
Biolog microorganism automatic identifying system qualification result
Biolog software will read 96 hole microplate reaction results according to listing 10 results with the matching degree of database, each result all shows 3 kinds of important parameters, be probable value Probability(PROB), similitude Similarity(SIM) and distance of positions Distance(DIS).Wherein SIM and DIS are 2 most important parameters, the similarity degree of SIM value representation test result and database corresponding data bar, the distance of positions of DIS value representation test result and database corresponding data bar.Biolog system regulation: saccharomycete was cultivated 24 o'clock, and the SIM value answers 〉=0.75, when cultivating 48h or 72h, and SIM value 〉=0.50, the qualification result that system provides automatically is kind of a name, and the SIM value is more near 1.00, and the reliability of qualification result is more high; When SIM value＜0.5, but the result's that generic name is identical in the qualification result SIM value sum is greater than 0.5 o'clock, and the qualification result that provides automatically is generic name.3 strain bacterium see Table 5,6,7 at the biolog of Best Times system qualification result.
Table 5Biolog system is to AB dientification of bacteria result (24h)
Figure BDA00003054321500091
Table 6Biolog system is to AE dientification of bacteria result (48h)
Figure BDA00003054321500092
Figure BDA00003054321500101
Table 7Biolog system is to AF dientification of bacteria result (48h)
Figure BDA00003054321500102
By table 5,6,7 as can be known, the AB bacterium is that the possibility of pale yellow Cryptococcus bacterium is 99%, and similarity is 0.828; The AE bacterium is that the possibility of orange rhodotorula bacterium B is 96%, and similarity is 0.748; Because in the AF bacterium incubation, its YT plate database in the biolog system does not have the database of the AF bacterium of coupling, and after using the GP plate instead, the AF bacterium is that the possibility of staphylococcus xylosus is 100%, similarity is 0.502.Can find out, two kinds of bacterium of AB, AE be saccharomycetic possibility all greater than 96%, similarity is all more than 0.5, so tentatively judge that preceding 2 kinds of bacterium all are saccharomycete, and AF bacterium possibility is 100%, similarity all is higher than 0.5, is staphylococcus xylosus.
Identify that according to biolog AB, AE, AF are saccharomycete, in conjunction with the physico-chemical analysis result of three strain bacterium, interim AE bacterium does not produce the bacterium uncut jade, and the AE bacterium produces denseer wine flavour, therefore the best saccharomycete that ferments for the bud dish.
3, the screening of lactic acid bacteria and evaluation
Other 13 kinds of bacterium of remainder are cultivated, analyzed its metabolite.With anti-high effective liquid chromatography for measuring Lactic Acid from Fermentation Broth content, by measuring work strain fermentation product lactic acid, improve the accuracy of differentiating lactic acid bacteria, the method can be qualitative again can quantitative analysis, simultaneously fast, sensitive, detection range is wide.By determination of lactic acid, determine which lactic acid producing of strain separated with this.
3.1 the quantitative and qualitative analysis of bacterial classification metabolite
3.1.1 lactic acid titer regression equation and collection of illustrative plates
Standard items are diluted suitable multiple, with the 10 μ L of sample introduction behind the filtering with microporous membrane of 0.45 μ m, carry out chromatography by the chromatographic condition after optimizing, with mass concentration X (mg/mL) to peak area Y(mv) to try to achieve equation of linear regression be Y=6772X-5.62, R2=0.9973, the range of linearity is 0.02-0.1mg/mL, and the method detection line of detection limit (S/N=3) is 0.05 μ g/ml, shows to have good linear relationship thus.The lactic acid titer the results are shown in shown in Figure 10 at the chromatogram of liquid chromatogram.
As shown in Figure 10, the retention time of standard lactic acid is 4.025min, can whether identical or approaching with the retention time of lactic acid titer according to the retention time of each zymotic fluid, can judge whether this metabolite is lactic acid.
3.1.2 the analysis result of sample
The metabolite of 13 kinds of bacterial strains is gathered post processing, determining the analysis of chromatographic condition sample introduction, under the same conditions, the parallel sample introduction twice of each sample, each sample introduction 10 μ L, record result.By liquid chromatogram, the zymotic fluid of 13 kinds of bacterial classifications of mensuration, retention time and lactic acid titer retention time (4.025min) contrast according to each group sample can qualitatively judge zymotic fluid and whether contain lactic acid.The liquid chromatogram that obtains behind 13 kinds of zymotic fluid sample introductions, contrast with lactic acid titer liquid chromatogram, have to contain lactic acid to 4 kinds of zymotic fluids, be respectively AH bacterium (4.029min), AJ bacterium (4.048min), AK bacterium (4.008min), AN bacterium (3.982min), the results are shown in Figure 11, Figure 12, Figure 13, Figure 14.
The retention time that draws the retention time of AH, AJ, AK, AN bacterium and lactic acid titer from Figure 11,12,13,14 approaches, and these four kinds of bacterium may be lactic acid bacteria.According to lactic acid titer regression equation, determine Lactic Acid from Fermentation Broth content with external standard method, the results are shown in Table 8.
Table 8 sample analysis result
Figure BDA00003054321500111
In conjunction with liquid chromatogram and sample analysis result, according to the time of staying of every kind of zymotic fluid, can qualitatively judge this bacterium lactic acid producing, can quantitatively draw this bacterium lactic acid producing amount according to its peak area.The time of staying of AH bacterium and the standard lactic acid time of staying are the most approaching, and AN bacterium lactic acid producing amount is maximum, and AK bacterium lactic acid producing amount is taken second place, and AJ bacterium lactic acid producing amount is minimum.
3.2 Physiology and biochemistry experiment
To AH, AJ, four kinds of bacterium of AK, AN, carry out the Physiology and biochemistry experimental result and see Table 9.
Table 9 Physiology and biochemistry experimental result
Figure BDA00003054321500122
Annotate: "+" expression positive reaction, " one " represents negative reaction.
Draw by table, these four kinds of bacterium show positive in the gelatin liquefaction experiment, show negative in the hydrogen peroxide experiment, in the litmus milk experiment pink, solidification phenomenon is arranged, show positive in the glucan experiment, these four kinds of bacterium are satisfied the biochemical characteristics of lactic acid bacteria, in conjunction with liquid phase measurement result and morphological feature, can judge tentatively that AJ, AK, AN bacterium are satisfied the morphological feature of lactic acid bacteria substantially, the AH bacterium might be the Bacillus acidi lactici that produces pigment.
Maximum at AN bacterium lactic acid producing amount, it is carried out 16S rDNA identify
3.3 the AN bacterium is carried out 16S rDNA identifies
3.3.1PCR product electrophoresis result
Be that 1.0% Ago-Gel is made electrophoresis detection to the pcr amplification product of the 16S rDNA of AN bacterial classification with mass fraction, after nucleic acid staining agent ethidium bromide (EB) dyeing, be placed on gel imaging system and observe and take a picture.The agarose gel electrophoresis figure of 16S rDNA pcr amplification the results are shown in Figure 15.
As can be seen from Figure 15, the fluorescence band occurs at about 1600bp place, and do not have obvious conditions of streaking, the pcr amplification success is described, satisfy the requirement of follow-up 16S rDNA sequencing.
2.3.2.216S rDNA testing result
The AN sequencing result is as follows.
Figure BDA00003054321500131
With sequencing result at GeneBank(http: // BLAST on www.ncbi.nlm.nih.gov/) mates, the result shows with lactic acid bacteria 16SrDNA sequence and presents higher homology that comparison result is as shown in figure 16.
Figure BDA00003054321500141
BLAST sequence on Figure 16 AN and the GeneBank compares
As shown in Figure 16, the row that check order by BLAST comparison, with the 16S rDNA homology of lactobacillus (Lactobacilus) be 99%, and according to Kuttzman﹠amp; The of the same race interior different strains differences of Robnet defined generally is no more than 1% standard, therefore assert that AN is lactobacillus.
Three, the fermented bacterium inoculative proportion determines
With the bacterial classification mixed culture fermentation of screening previously, lactic acid bacteria culturers AH and lactobacillus AN, with saccharomycete AE, AF mixed culture fermentation, four kinds of bacterium are pressed the different proportion collocation, after the 24h Liquid Culture, get each 5mL of zymotic fluid of 9 kinds of samples respectively, dilute 10 times after, the method that adopts GB/T10345-2007 to measure total ester is carried out titration, consumes the H of 0.1mol/L 2SO 4Liquor capacity the results are shown in Table 10.
Table 10 mixed culture fermentation consumes H 2SO 4Volume (unit: mL)
Figure BDA00003054321500142
Figure BDA00003054321500151
Concentration is 40% ethyl lactate solution, measures the method for total ester according to GB/T10345-2007 and carries out titration, and the volume that consumes 0.1mol/L sulfuric acid is 12.21mL, and again according to nine groups of zymotic fluids, the sulfuric acid volume of consumption can be obtained the content of total ester in the zymotic fluid.The results are shown in Table 11.
Total ester content (the unit: g/L) of table 11 zymotic fluid
Figure BDA00003054321500152
The total ester content of bacterial classification mixed culture fermentation liquid can be found out, except 1 group of data exception, other eight groups of data illustrate bacterial classification mixed culture fermentation, and product ester ability is stronger.More lactic acid and ethanol synthesis are namely arranged, generate Ester.Data are carried out range analysis of orthogonal experiment, obtain a result, the results are shown in Table 12.
Table 12 orthogonal test analysis table
Figure BDA00003054321500153
As seen from the above table, in the mixed culture fermentation experiment of these four factors, three levels, AE, the AN bacterium is principal element, optimum mixed culture fermentation ratio is: AH:AN:AE:AF=3:1:1:3.
Because AE, AN is principal element, and the saccharomycete AE that lactobacillus AN is strong with producing pure ability is inoculated in the bud dish, and preferred compositions bacterial strain 1:1 ferments.
Good effect of the present invention is embodied in:
(1), in the sweat of Yibin bud dish, add composite bacteria AN:AE, can shorten fermentation time about 2 months;
(2), the ester class content that produces is maximum, the kind of volatility aroma-producing substance and total content are also maximum, have constituted the local flavor of high-quality Yibin bud dish uniqueness, make the mouthfeel of Yibin bud dish fuller, fragrance is stronger.
Description of drawings
Fig. 1 is B colonial morphology figure;
Fig. 2 is E colonial morphology figure;
Fig. 3 is F colonial morphology figure;
Fig. 4 is AB bacterium microscopic morphology figure;
Fig. 5 is AE bacterium microscopic morphology figure;
Fig. 6 is AF bacterium microscopic morphology figure;
Fig. 7 is strains A B at water-iodine liquid microscopy figure as a result;
Fig. 8 is strains A E at water-iodine liquid microscopy figure as a result;
Fig. 9 is strains A F at water-iodine liquid microscopy figure as a result;
Figure 10 is lactic acid titer HPLC figure
Figure 11 is AH fermented liquid HPLC figure
Figure 12 is AJ fermented liquid HPLC figure;
Figure 13 is AK fermented liquid PHLC figure;
Figure 14 is AN fermented liquid HPCL figure;
Figure 15 pcr amplified fragment electrophoresis is M:DNA molecular mass standard wherein; 1,2 represents the amplified production of AN
Figure 16 is the schematic flow sheet of bud vegetable production technique among the present invention.
The specific embodiment
In order to make purpose of the present invention, technical scheme and advantage clearer, the present invention is described in further detail below in conjunction with the specific embodiment, but this should be interpreted as that the scope of the above-mentioned theme of the present invention only limits to following embodiment.
Embodiment 1:
Adopt traditional zymotic technology respectively, and AN:AE press bud dish product sample 1 that 1:1 compatibility work bacterial classification forced fermentation technology obtains, reaches sample 2.Two kinds of product sample 1, sample 2 subjective appreciations and physics and chemistry comparison, organic acid content, volatile matter contents are compared as follows:
Table 13 subjective appreciation result
Figure BDA00003054321500171
Table 14 physical and chemical index
Figure BDA00003054321500172
Organic acid content (unit: mg/g) in table 15 sample
Figure BDA00003054321500173
The flavor substance component list of the different Yibin of table 16 bud dish
Figure BDA00003054321500181
Figure BDA00003054321500191
From above-mentioned each index more as can be seen, in the sample 2, not only the maximum mouthfeels of ester class content are good, and volatility becomes kind and the total content of flavor material also maximum, constituted the local flavor of high-quality Yibin bud dish uniqueness, it becomes the flavor material to mainly contain 3,7-dimethyl-1,6-octadiene-3-alcohol, ethyl palmitate, ethyl linoleate, ethyl linolenate, anethole, phenylethanol, 4-terpenol, methyl linolenate, isoamyl formate, 2-butanols, alpha-terpineol, allyl isothiocyanate, ethyl stearte etc.
Yibin bud dish to different samples and different fermentations period, adopt while distillation extraction (SDE) and gas chromatography mass spectrometry (GC-MS) to detect the flavor substance of Yibin bud dish, Yibin bud dish cycle of AN:AE bacterial classification compatibility fermentation is the shortest, shorten one month, and kind and the total content of generation volatility aroma-producing substance are maximum.
In the sweat of Yibin bud dish, add composite bacteria AN:AE, it not only can shorten fermentation time, and the ester class content of its generation is maximum, the kind of volatility aroma-producing substance and total content are also maximum, constituted the local flavor of high-quality Yibin bud dish uniqueness, make the mouthfeel of Yibin bud dish fuller, fragrance is stronger.
Embodiment 2:
Adopt traditional zymotic technology respectively, and AN:AE presses bud dish product sample 1, sample 2, sample 3 samples 4 of 1:1,1:2, the acquisition of 2:1 compatibility work bacterial classification forced fermentation technology.Four kinds of product volatile matter contents are compared as follows:
The flavor substance component list of the different Yibin of table 17 bud dish
Figure BDA00003054321500201
Figure BDA00003054321500211
From above-mentioned each index more as can be seen, in the sample 2, not only the maximum mouthfeels of ester class content are good, and volatility becomes kind and the total content of flavor material also maximum, constituted the local flavor of high-quality Yibin bud dish uniqueness, it becomes the flavor material to mainly contain 3,7-dimethyl-1,6-octadiene-3-alcohol, ethyl palmitate, ethyl linoleate, ethyl linolenate, anethole, phenylethanol, the 4-terpenol, methyl linolenate, isoamyl formate, the 2-butanols, alpha-terpineol, allyl isothiocyanate, the sample of ethyl stearte etc. and other scheme all is not so good as sample 2 aspect the kind and content of flavor substance.
Yibin bud dish to different samples and different fermentations period, adopt while distillation extraction (SDE) and gas chromatography mass spectrometry (GC-MS) to detect the flavor substance of Yibin bud dish, it is the shortest that AN:AE presses Yibin bud dish cycle of 1:1 bacterial classification compatibility fermentation, shorten one month, and kind and the total content of generation volatility aroma-producing substance are maximum.
In the sweat of Yibin bud dish, add composite bacteria AN:AE, it not only can shorten fermentation time, and the ester class content of its generation is maximum, the kind of volatility aroma-producing substance and total content are also maximum, constituted the local flavor of high-quality Yibin bud dish uniqueness, make the mouthfeel of Yibin bud dish fuller, fragrance is stronger.

Claims (7)

1. new bud vegetable production technique, it is characterized in that: this process using artificial infection and the composite bacteria that filters out, this composite bacteria is lactobacillus and orange red yellow yeast by mass ratio is that the proportionate relationship proportioning of 1-2:1-2 forms, and the inoculum concentration of its bacterial classification is into 0.9-1.1% of pond bud dish mass content.
2. bud vegetable production technique according to claim 1 is characterized in that this technology specifically may further comprise the steps:
(1) its head is cut away in will be ripe bud dish harvesting, and branches and leaves are removed, and manual its bar is cut into uniform strip, builds the canopy airing in the sun, and the bud dish bar of well cutting is put transpiring moisture thereon;
(2) stirring is salted
Bud dish after reclaiming from the canopy that dries in the air stirs salted, and then that airing is good bud dish bar is cleaned, and it is dropped into in the pond of preparing of fermenting, and stirring salted method is one deck bud dish one deck salt;
(3) fermentation
Add composite bacteria in proportion in going into the bud dish in pond, ferment, fermentation condition is that salinity is 5-6%, and the pH value is 6.5-7.0 in the fermentation vat environment, and temperature is controlled at 29-31 ℃, and fermentation time is 90-100 days, and composite bacteria is pressed the amount inoculation of 0.9-1.1%;
(4) clean, dewater
The rotten dish that reject yellow leaf, goes mouldy behind the foreign material that weed a garden, after the running water soaking and washing, guarantees no-sundries, and no silt stone is removed excessive moisture in the bud dish by squeezing then;
(5) cut the dish, screening and spice:
By cutting the bud dish is cut fineness degree to needs. separating defective bud dish by screening, is how 0.1% ratio adds Chinese prickly ash, anise and three, uniform mixing in bud dish mass percent then;
(6) can, sterilization, test package
To can the packaging bag of bud dish vacuumize, utilize pasteurization to boil the bag sterilization, bottom pour ladle, mashed bag defective work, vanning are rejected in the check of sterilization back.
3. require 2 described bud vegetable production techniques according to claim, it is characterized in that: the bud dish bar of well cutting described in the step (1) is put transpiring moisture thereon, and evaporation capacity of moisture content is the 50%-60% of bud dish quality percentage composition.
4. require 2 described bud vegetable production techniques according to claim, it is characterized in that: the stirring described in the step (2) is salted, and the consumption of its salt is the 5-6% of bud dish weight.
5. require 2 described bud vegetable production techniques according to claim, it is characterized in that: excessive moisture in the bud dish is removed in the squeezing of passing through described in the step (4), the 74-78% of its squeezing back moisture control mass percent of bud dish after squeezing.
6. require 2 described bud vegetable production techniques according to claim, it is characterized in that: the pasteurization described in the step (6) boils bag sterilization, 85-95 ℃ of its sterilising temp.
7. require 2 described bud vegetable production techniques according to claim, it is characterized in that: the pasteurization described in the step (6) boils the bag sterilization, and its sterilization time is single bag 500g weight following 25-30 minute, single bag 500g weight above 30-40 minute.
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CN104605300A (en) * 2015-02-03 2015-05-13 四川理工学院 Cellar type bean sprout fermentation method
CN107927662A (en) * 2017-12-12 2018-04-20 建始县好硒奇农产品开发有限公司 A kind of eddo lotus stem curing food and its processing method
CN109349494A (en) * 2018-12-10 2019-02-19 四川师范大学 Instant spicy bean sprout packet and preparation method thereof
CN110279086A (en) * 2019-07-30 2019-09-27 山东省农业科学院农产品研究所 A kind of coarse cereals bud pickled vegetables and its processing method

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104605300A (en) * 2015-02-03 2015-05-13 四川理工学院 Cellar type bean sprout fermentation method
CN104605300B (en) * 2015-02-03 2018-03-27 四川理工学院 Cellar type bean sprout fermentation process
CN107927662A (en) * 2017-12-12 2018-04-20 建始县好硒奇农产品开发有限公司 A kind of eddo lotus stem curing food and its processing method
CN109349494A (en) * 2018-12-10 2019-02-19 四川师范大学 Instant spicy bean sprout packet and preparation method thereof
CN110279086A (en) * 2019-07-30 2019-09-27 山东省农业科学院农产品研究所 A kind of coarse cereals bud pickled vegetables and its processing method

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