CN103190585B - Novel bean sprout production process - Google Patents

Novel bean sprout production process Download PDF

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CN103190585B
CN103190585B CN201310130475.8A CN201310130475A CN103190585B CN 103190585 B CN103190585 B CN 103190585B CN 201310130475 A CN201310130475 A CN 201310130475A CN 103190585 B CN103190585 B CN 103190585B
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bud
bud dish
dish
bacterium
fermentation
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CN103190585A (en
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左勇
鞠帅
边名鸿
叶阳
刘利平
刘川秀
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YIBIN SHUANGYI FUKANG FOOD CO., LTD.
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YIBIN SHUANGYI FUKANG FOOD Co Ltd
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 approximately 2 months, and the ester class content producing 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 " are also called the large catsup and pickled vegetables in Sichuan four, have long history.Yibin bud dish is because having the style of uniquenesses such as " fragrant, sweet, crisp, tender, fresh " and excellent extremely consumer's favor of quality.
At present, Yibin bud dish mainly adopts traditional aging process to produce, and bud dish is left in fermentation vat or ceramic jar, ferments by the work such as lactic acid bacteria, saccharomycete bacterial classification.At the bud dish fermentation initial stage, saccharomycete amount in bud dish is few, be mainly lactobacillus-fermented, but along with fermentation time extends, lactic acid bacteria is owing to being subject to the reasons such as pH value and carbon source concentration be too low, can not recycle carbon source, and saccharomycete still can utilize well, until carbon source is 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 to the storage important after bud dish local flavor and quality and fermentation.Fermentation period generally needs the time of 4-6 month, 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 and nutriment are also different.
Summary of the invention
The present invention, just for above technical problem, provides and can shorten fermentation period nearly 2 months, and improve the quality of products, and makes detectable flavor substance composition be increased to a kind of new bud vegetable production technique of 22 kinds from 14 kinds.
Concrete technical scheme of the present invention is as follows:
A kind of new bud vegetable production technique, this process using artificial infection the composite bacteria that filters out, this composite bacteria be lactobacillus and orange rhodotorula in mass ratio for the proportionate relationship proportioning of 1-2:1-2 forms, the inoculum concentration of its bacterial classification is the 0.9-1.1% into pond bud dish mass content.
A new bud vegetable production technique, this technique specifically comprises the following steps:
(1), by ripe bud dish harvesting, its head is cut away, branches and leaves are removed, by hand its bar is cut into uniform strip, build in the sun canopy airing, the bud dish bar of well cutting is put to transpiring moisture thereon, the evaporation capacity of moisture is the 50%-60% of quality percentage composition in bud dish;
(2) stir salted
Bud dish from the canopy that dries in the air reclaims stirs salted, and the suitable time of airing is determined by wind-force power and weather.Target is just passable while feeling that softness does not have hard core when holding green vegetables stem silk.Then bud dish bar good airing is cleaned, is dropped into as fermenting in the pond of preparing, stir salted method and determined by the consumption of one deck bud dish one deck salt salinity and the situation of bud dish, the consumption of salt is generally the now 5-6% of bud dish weight percentage;
(3) fermentation
In the bud dish that enters pond, add in proportion composite bacteria, ferment.Fermentation condition is that salinity is 5-6%, and in fermentation vat environment, pH value is 6.5-7.0, and temperature is controlled at 29-31 DEG C, 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, weed a garden after foreign material, after running water soaking and washing, ensure no-sundries, without silt stone, then remove excessive moisture in bud dish by squeezing, after squeezing, bud dish moisture is controlled at the 74-78% of the mass percent of the rear bud dish of squeezing;
(5) cut the dish, screening and spice:
By cutting, bud dish is switched to the fineness degree needing. separate defective bud dish by screening.Then add Chinese prickly ash, anise and three spices such as how in the ratio of the 0.1-0.2% of bud dish mass percent now, uniform mixing, how Chinese prickly ash, anise and three addition of spices such as are appropriate, only for seasoning;
(6) filling, sterilizing, test package
Packaging bag to filling bud dish vacuumizes, and utilizes pasteurization to boil bag sterilizing, sterilising temp 85-95 DEG C.Sterilization time, the following 25-30 minute of single bag 500g weight, the above 30-40 minute of single bag 500g weight; Bottom pour ladle, rotten bag defective work are rejected in inspection, vanning.
The isolation and identification method that saccharomycete, lactic acid bacteria separate is:
One, the culture medium of seed selection
(1) 1%CaCO 3mRS 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.Adjust pH6.2～6.4,121 DEG C of sterilizing 15min.
(2) PY basal medium
Peptone 0.5g, pancreatin solution casein 0.5g, yeast extract 10g, salting liquid 4mL.Distilled water 100mL. adjust pH 6.0 left and right, 115 DEG C of sterilizing 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, by CaCl 2and MgSO 4be dissolved in together in 300mL distilled water, then add 500mL distilled water, stir while slowly add other salts. continue to stir until all dissolve, adding distil water is settled to 1000mL, in 4 DEG C of storages, for subsequent use after mixing.
(3), glucose peptone water culture medium
Peptone 5g, glucose 5g, K 2hPO 42g, distilled water 1000mL.Adjust pH 7.0-7.2, filters 112 DEG C of sterilizing 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 DEG C of-115 DEG C of sterilizing 15min-20min of pH value.
(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 DEG C of sterilizing 15min.
(6), litmus milk enrichment
Remove the upper strata cream of milk with centrifuge, 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 DEG C of high pressure steam sterilization 15-20min.
Two, strain separating, purifying
1, separate
Under aseptic condition, by the 1%CaCO of sterilizing 3be added in MRS solid medium, after mixing shakes up, pour into immediately in culture dish, cooling for subsequent use.
Sterilized water is joined in the bud dish of fermentation Yibin, wash lower Yibin bud dish function bacterial classification, then carry out gradient dilution.Choose suitable dilution factor, coating, is down flat plate in 30 DEG C of cultivation 48h, and picking has bacterium colony the colony counting of molten calcium circle.On MRS flat board, repeatedly rule until single bacterium colony; Repeating flat lining out separates again, purifying repeatedly, through Gram's staining, to purifying, (bacterium colony is single in the same size, gram stain microscopy, thalline size solid colour), isolate altogether 16 kinds of different strains, be numbered respectively AA → AP, it is transferred to respectively on MRS slant medium to preservation under 4 DEG C of conditions.
From I, II, 3 samples of III, get respectively the each 1mL of bacteria suspension, these 3 kinds of bacteria suspensions are diluted to respectively to 10 -8, coating, result is as following table 1.
Table 1 plate count result
Figure GDA0000407966880000041
Figure GDA0000407966880000051
Can find out according to table table 1, along with bacteria suspension dilution factor increases, the bacterium colony on flat board is fewer and feweri, and dilution factor is 10 -5flat board afterwards, can both observe the morphological feature of single bacterium colony clearly.According to the morphological feature of bacterium colony and microscopy result, filter out altogether 16 kinds of bacterium, to its numbering, in table 2.
Table 2 morphological feature
2, saccharomycetic qualification
2.1 saccharomycete morphology and physics and chemistry qualification
By the AB obtaining in separation in table 2, AE, AF3 strain bacterium, choose suitable dilution factor, coating, be down flat plate and cultivate 48h, observation colony characteristics microscopy, picking list bacterium colony in 28 DEG C, on PDA culture medium flat plate, line is until single bacterium colony repeatedly, and pure bacterial strain moves into slant medium.Carry out following serial qualification test:
2.1.1 Morphological Identification
The bacterial classification of separation, purifying is coated onto respectively in PDA culture medium, cultivates 48h for 28 DEG C, observe colonial morphology, and do methylene blue dye liquor water logging sheet, water one iodine liquid water logging sheet, in the individual morphology of high power Microscopic observation thalline.Its form result is as table 3 and Fig. 1,2,3.
Table 3 colony morphology characteristic
Figure GDA0000407966880000061
Learn by table 3, by colony morphology characteristic, can tentatively judge 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 liquid, put slide central authorities, and get 3 kinds of bacterium with oese respectively and dyeing liquor mixes, dyeing 2～3min, adds cover glass, at high power Microscopic observation thalli morphology, the results are shown in Figure 4,5,6.
From Fig. 4,5,6, find out, AB, AE bacterial cell are rounded, and AF bacterial cell ovalize, all has cell to be in gemmation state.Known by figure, some cell is transparence, and some is blue because methylene blue has oxidisability, and in living cell body, just carrying out 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 in to slide central authorities, then gets a bacteria suspension, mix, covered, oily Microscopic observation, the results are shown in Figure 7,8,9.
Learnt by Fig. 7,8,9, each cell edges is transparent interior color and deepens to be gradually yellowish-brown.In bacterium, cell has sprout, and cell is dyed to yellowish-brown and sprout is transparence.From AB bacterium, can be observed two cells, one of them half is yellowish-brown half and is transparent, and another cell is yellowish-brown up and down, and it is transparent that centre is, and it is investigated that to read related data known.Two cells may be in the latter stage in cell division state, are about to be split into two cells by a cell.
2.1.2 physics and chemistry qualification
(1) produce the comparative experiments of alcohol ability
Under sterile working, each inoculation of separation and purification, in TTC lower floor culture medium, is placed in to 38 DEG C and cultivates 48h, pour TTC upper strata culture medium into after growing bacterium colony, insulation (shady place) 2～3h.By the variation of color, the product alcohol ability between more each bacterial strain, redness illustrates that producing and ethanol ability is stronger more deeply.
(2) kind of starch hydrolysis hydrolysis
Under sterile working, by each inoculation PYD fluid nutrient medium of separation and purification, be placed in 200r/min, 28 DEG C of constant temperature culture 48h, get a nutrient solution and drop in white color board, add a Ge Shi iodine liquid and mix, and observe the variation of color.
(4) produce uncut jade test
Under sterile working, the bacterial classification having activated is inoculated into respectively to aseptic brewer's wort solid medium examination by 2% inoculum concentration, after 28 DEG C 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 GDA0000407966880000081
Note :+represent can produce alcohol/products uncut jade, ++ number represent produce number ,-expression do not produce.
Draw by table 4, the product alcohol ability of AE bacterium is the strongest, and AB bacterium takes second place, and the product alcohol ability of AF bacterium is the poorest; Nutrient solution color is all black-and-blue, illustrates that these three kinds of bacterial strains have all produced 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 AE bacterium does not produce bacterium uncut jade.
In actual production, the raw flower in the surface of curing food, long tunica albuginea should be avoided aborning as far as possible.The normal fermentation that spend because making a living, long tunica albuginea not only can affect Yibin bud dish, produce bad smell, and the appearance of product is also bad, can be by producing uncut jade test, and superseded those produce the strong bacterial strain of uncut jade abilities.
2.1.3Biolog Automatic Analyzer for Microbes qualification saccharomycete
(1) BUY culture medium flat plate line
Select the single bacterium colony of cultivating on 28h inclined-plane, carry out " cross " sectional streak on BUY culture medium, at 28 DEG C, constant temperature and humidity is cultivated 48h.
(2) adjust turbidity
With the single bacterium colony in aseptic bamboo let picking " cross " line, under aseptic condition, be inoculated in turbidity pipe.Blank is adjusted to 100%, and saccharomycete suspension standard pipe is adjusted to 47%.Measure with postvaccinal turbidity pipe, can constantly adjust turbidity with special sterile water, to reach saccharomycete suspension standard turbidity value (47%).
(3) prepare bacteria suspension
Get aseptic bamboo let and in inoculation liquid, dip in wetly, the sticky thalline of getting slowly slides bamboo let on bacterium colony surface.Inclination inoculation liquid pipe also rotates bamboo let along inwall, and thalline is attached on inwall, thalline is evenly broken up simultaneously.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, with 8 electric pipettors, bacteria suspension is added in respectively in each hole of different identification plates, every hole application of sample 100 μ L, totally 96 holes, are placed in pallet, and keep certain humidity, cultivate 24h, 48h and 72h at 26 DEG C.
(5) reading out data
Take out qualification microplate and be placed on readout instrument, open identification systems, computer reading out data, provide the title of 10 ID according to 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 is cultivated 24 o'clock, SIM value answers >=0.75, while cultivating 48h or 72h, SIM value >=0.50, the qualification result that system provides is automatically kind of a name, and SIM value more approaches 1.00, and the reliability of qualification result is higher; As SIM value < 0.5, but the SIM value sum of the result that in qualification result, generic name is identical is greater than at 0.5 o'clock, and the qualification result automatically providing is generic name.3 strain bacterium the results are shown in Table 5,6,7 at the biolog of Best Times system identification.
Table 5Biolog system is to AB dientification of bacteria result (24h)
Figure GDA0000407966880000091
Table 7Biolog system is to AF dientification of bacteria result (48h)
Figure GDA0000407966880000102
From table 5,6,7, AB bacterium is that the possibility of pale yellow Cryptococcus bacterium is 99%, and similarity is 0.828; AE bacterium is that the possibility of orange Rhodotorula sp B is 96%, and similarity is 0.748; In AF bacterium incubation, the database of its AF bacterium that YT plate database does not mate in biolog system, and use instead after GP plate, AF bacterium is that the possibility of staphylococcus xylosus is 100%, similarity is 0.502.Can find out, AB, two kinds of bacterium of AE are that saccharomycetic possibility is all greater than 96%, and similarity is all more than 0.5, so tentatively judgement, front 2 kinds of bacterium are all saccharomycete, and AF bacterium possibility is 100%, and similarity, all higher than 0.5, is staphylococcus xylosus.
According to biolog qualification, AB, AE, AF are saccharomycete, and in conjunction with the physico-chemical analysis result of three strain bacterium, interim AE bacterium does not produce bacterium uncut jade, and AE bacterium produces denseer wine flavour, the best saccharomycete therefore fermenting for bud dish.
3, the screening of lactic acid bacteria and qualification
Other 13 kinds of bacterium of remainder are cultivated, analyzed its metabolite.With lactic acid content in anti-Fermentation Liquor by High Performance Liquid Chromatography, 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 quick, sensitive, detection range is wide.By the mensuration of lactic acid, determine with this which lactic acid producing of bacterial strain separating.
The quantitative and qualitative analysis of 3.1 bacterial classification metabolites
3.1.1 lactic acid titer regression equation and collection of illustrative plates
Standard items are diluted to suitable multiple, with the 10 μ L of sample introduction after 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 thus to have good linear relationship.Lactic acid titer, at the chromatogram of liquid chromatogram, the results are shown in shown in Figure 10.
As shown in Figure 10, the retention time of standard lactic acid is 4.025min, can be 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 to 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, whether the zymotic fluid of 13 kinds of bacterial classifications of mensuration, according to the retention time of each group sample and lactic acid titer retention time (4.025min) contrast, can qualitatively judge zymotic fluid containing lactic acid.The liquid chromatogram obtaining after 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.
Show that from Figure 11,12,13,14 retention time of AH, AJ, AK, AN bacterium and the retention time of lactic acid titer approach, these four kinds of bacterium, may be lactic acid bacteria.According to lactic acid titer regression equation, determine Lactic Acid from Fermentation Broth content by external standard method, the results are shown in Table 8.
Table 8 sample analysis result
Figure GDA0000407966880000111
Figure GDA0000407966880000121
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 experiments
To AH, AJ, AK, tetra-kinds of bacterium of AN, carry out Physiology and biochemistry experimental result in table 9.
Table 9 Physiology and biochemistry experimental result
Figure GDA0000407966880000122
Note: "+" represents positive reaction, and " one " represents negative reaction.
Draw by table, these four kinds of bacterium are aobvious positive in gelatin liquefaction experiment, aobvious negative in hydrogen peroxide experiment, in litmus milk experiment, have pink, solidification phenomenon, aobvious positive in glucan experiment, these four kinds of bacterium meet the Biochemical Characteristics of lactic acid bacteria, in conjunction with liquid phase measurement result and morphological feature, can tentatively judge, AJ, AK, AN bacterium meet the morphological feature of lactic acid bacteria substantially, and AH bacterium is likely the Bacillus acidi lactici that produces pigment.
Maximum for AN bacterium lactic acid producing amount, it is carried out to 16S rDNA qualification
3.3 pairs of AN bacterium carry out 16S rDNA qualification
3.3.1PCR product electrophoresis result
The pcr amplification product of the 16S rDNA of the Ago-Gel that is 1.0% with mass fraction to AN bacterial classification is made electrophoresis detection, after nucleic acid staining agent ethidium bromide (EB) dyeing, is placed on gel imaging system and observes 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, there is fluorescence band at about 1600bp place, and without obvious conditions of streaking, pcr amplification success is described, meet the requirement of follow-up 16S rDNA sequencing.
2.3.2.216S rDNA testing result
AN sequencing result is as follows.
Figure GDA0000407966880000131
By sequencing result at GeneBank(http: the BLAST //www.ncbi.nlm.nih.gov/) mates, and result shows with lactic acid bacteria 16SrDNA sequence and present higher homology, and comparison result is as shown in figure 16.
Figure GDA0000407966880000141
BLAST sequence on Figure 16 AN and GeneBank contrasts
As shown in Figure 16, the row that check order are compared by BLAST, with the 16SrDNA homology of lactobacillus (Lactobacilus) be 99%, and be generally no more than 1% standard according to difference between the of the same race interior different strains of Kuttzman & Robnet defined, therefore assert that AN is lactobacillus.
Three, determining of fermented bacterium inoculative proportion
By the bacterial classification mixed culture fermentation of screening above, lactic acid bacteria culturers AH and lactobacillus AN, with saccharomycete AE, AF mixed culture fermentation, four kinds of bacterium are pressed different proportion collocation, after 24h Liquid Culture, get respectively the each 5mL of zymotic fluid of 9 kinds of samples, after diluting 10 times, 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 GDA0000407966880000142
Figure GDA0000407966880000151
Concentration is 40% ethyl lactate solution, measures the method for total ester carry out titration according to GB/T10345-2007, and the volume that consumes 0.1mol/L sulfuric acid is 12.21mL, then according to nine groups of zymotic fluids, and the sulfuric acid volume of consumption can be obtained the content of total ester in zymotic fluid.The results are shown in Table 11.
The total ester content (unit: g/L) of table 11 zymotic fluid
Figure GDA0000407966880000152
The total ester content of bacterial classification mixed culture fermentation liquid, can find out, except 1 group of data exception, other eight groups of data, illustrate bacterial classification mixed culture fermentation, and product ester ability is stronger.There are more lactic acid and ethanol synthesis, generate Ester.Data are carried out to range analysis of orthogonal experiment, obtain a result, the results are shown in Table 12.
Table 12 orthogonal test analysis table
Figure GDA0000407966880000153
As seen from the above table, in the mixed culture fermentation experiment of these four factors, three levels, AE, AN bacterium is principal element, optimum mixed culture fermentation ratio is: AH:AN:AE:AF=3:1:1:3.
Due to AE, AN is principal element, by the lactobacillus AN saccharomycete AE strong with producing alcohol ability, is inoculated in 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 approximately 2 months;
(2), produce ester class content maximum, the kind of volatility aroma-producing substance and total content are also maximum, have formed the local flavor of high-quality Yibin bud dish uniqueness, make the mouthfeel of Yibin bud dish fuller, fragrance is stronger.
Brief description of the 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 that strains A B is at water-iodine liquid microscopy result figure;
Fig. 8 is that strains A E is at water-iodine liquid microscopy result figure;
Fig. 9 is that strains A F is at water-iodine liquid microscopy result figure;
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;
Wherein M:DNA molecular mass standard of Figure 15 pcr amplified fragment electrophoresis; 1,2 represents the amplified production of AN
Figure 16 is the schematic flow sheet of bud vegetable production technique in the present invention.
Detailed description of the invention
In order to make object of the present invention, technical scheme and advantage clearer, below in conjunction with detailed description of the invention, the present invention is described in further detail, but this should be interpreted as to the scope of the above-mentioned theme of the present invention only limits to following embodiment.
Embodiment 1:
Adopt respectively traditional zymotic technique, and AN:AE presses bud dish product sample 1 and sample 2 that 1:1 compatibility work bacterial classification forced fermentation technique obtains.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 results of sensory evaluation
Figure GDA0000407966880000171
Table 14 physical and chemical index
Figure GDA0000407966880000172
Organic acid content in table 15 sample (unit: mg/g)
The flavor substance component list of the different Yibin of table 16 bud dish
Figure GDA0000407966880000181
Relatively can find out from above-mentioned each index, in sample 2, not only the maximum mouthfeels of ester class content are good, and volatility become the kind of taste material and total content also maximum, form the local flavor of high-quality Yibin bud dish uniqueness, it becomes taste 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.
To Yibin bud dish in 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, form the local flavor of high-quality Yibin bud dish uniqueness, make the mouthfeel of Yibin bud dish fuller, fragrance is stronger.
Embodiment 2:
Adopt respectively traditional zymotic technique, 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 technique.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 GDA0000407966880000201
Figure GDA0000407966880000211
Relatively can find out from above-mentioned each index, in sample 2, not only the maximum mouthfeels of ester class content are good, and volatility become the kind of taste material and total content also maximum, form the local flavor of high-quality Yibin bud dish uniqueness, it becomes taste 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, the sample of ethyl stearte etc. and other scheme is aspect the kind and content of flavor substance, all be not so good as sample 2.
To Yibin bud dish in 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, form the local flavor of high-quality Yibin bud dish uniqueness, make the mouthfeel of Yibin bud dish fuller, fragrance is stronger.

Claims (6)

1. a bud vegetable production technique, is characterized in that this technique specifically comprises the following steps:
(1) by ripe bud dish harvesting, its head is cut away, branches and leaves are removed, and manual its bar are cut into uniform strip, build in the sun canopy airing, and the bud dish bar of well cutting is put to transpiring moisture thereon;
(2) stir salted
Bud dish from the canopy that dries in the air reclaims stirs salted, then bud dish bar good airing is cleaned, and is dropped in the pond of preparing for fermentation, and stirring salted method is one deck bud dish one deck salt;
(3) fermentation
In the bud dish that enters pond, add in proportion composite bacteria, ferment, fermentation condition is that salinity is 5-6%, and in fermentation vat environment, pH value is 6.5-7.0, and temperature is controlled at 29-31 DEG C, 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, weeds a garden after foreign material, after running water soaking and washing, ensures no-sundries, without silt stone, then removes excessive moisture in bud dish by squeezing;
(5) cut the dish, screening and spice:
By cutting, bud dish is switched to the fineness degree needing. separate defective bud dish by screening, how the ratio that is then 0.1% in bud dish mass percent adds Chinese prickly ash, anise and three, uniform mixing;
(6) filling, sterilizing, test package
Packaging bag to filling bud dish vacuumizes, and utilizes pasteurization to boil bag sterilizing, and after sterilizing, bottom pour ladle, rotten bag defective work are rejected in inspection, vanning;
This process using artificial infection the composite bacteria that filters out, this composite bacteria is that lactobacillus and orange rhodotorula are in mass ratio for the proportionate relationship proportioning of 1-2:1-2 forms.
2. require the bud vegetable production technique described in 1 according to claim, it is characterized in that: described in step (1), the bud dish bar of well cutting is put to transpiring moisture thereon, the evaporation capacity of moisture is the 50%-60% of bud dish quality percentage composition.
3. require the bud vegetable production technique described in 1 according to claim, it is characterized in that: the stirring described in step (2) is salted, the consumption of its salt is the 5-6% of bud dish weight.
4. require the bud vegetable production technique described in 1 according to claim, it is characterized in that: described in step (4), removes excessive moisture in bud dish by squeezing, the rear moisture of its squeezing is controlled at the 74-78% of the mass percent of squeezing rear bud dish.
5. require the bud vegetable production technique described in 1 according to claim, it is characterized in that: the pasteurization described in step (6) boils bag sterilizing its sterilising temp 85-95 DEG C.
6. require the bud vegetable production technique described in 1 according to claim, it is characterized in that: the pasteurization described in step (6) boils bag sterilizing, its sterilization time is the following 25-30 minute of single bag 500g weight, the above 30-40 minute of single bag 500g weight.
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