CN1122452C - Method for suppressing generation of 'white spot' on fermented bean curd - Google Patents
Method for suppressing generation of 'white spot' on fermented bean curd Download PDFInfo
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- CN1122452C CN1122452C CN99114527A CN99114527A CN1122452C CN 1122452 C CN1122452 C CN 1122452C CN 99114527 A CN99114527 A CN 99114527A CN 99114527 A CN99114527 A CN 99114527A CN 1122452 C CN1122452 C CN 1122452C
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Abstract
The present invention discloses a method for inhibiting the generation of white spots on fermented bean curd. Optimized fermented bean curd mucor bacterial strains are made by taking bean curd mucor bacterial strains for use, preparing spore suspension, carrying out the heat treatment of spores, separating of singular bacterial strains, prescreening a casein flat plate, testing proteinase vitality of the singular bacterial strains and optimizing the bacterial strains by fermented bean curd mucor. An improved secondary fermentation technology comprises: taking fermented bean curd mucor test tube bacterial strains for use, generally cultivating bran koji in a triangular flask, preparing the spore suspension, inoculating the bacterial strains onto a bean curd blank for primary fermentation, pickling the bean curd blank, bottling the bean curd blank (with feed liquid) for secondary fermentation, ripening the bean curd in fermentation, exciting the activity of inactive enzyme by the method of water bath or electric heating, etc., and storing the bean curd in a bin or selling the bean curd. The white spot generation resistance period of the normally-bottled fermented bean curd is prolonged by the present invention from three months to one year or even more than one year.
Description
The present invention relates to a kind of method that suppresses generation of ' white spot ' on fermented bean curd, exactly, it is the store method of edible fermented bean curd better quality.
Fermented bean curd is one of traditional fermented food of China, it be the bean curd base through mucor by primary fermentation, pickle with after fermentation and form.At present, China production of preserved beancurd enterprise exist more following problem 1. the fermented bean curd base bottle and produce more " hickie " in back 3 months, had a strong impact on the aesthetic quality of 1 year shelf selling period of fermented bean curd, some enterprise even only produce " red " fermented bean curd, the way of adding with red colouring agent for food, also used as a Chinese medicine is covered the bad sense organ phenomenon of generation of ' white spot ' on fermented bean curd; 2. bacterial strain poor heat resistance, summer can not ordinary production; 3. the strain protein enzymatic activity a little less than, some factory's fermented bean curd after fermentation was for up to 2-3 month, fermented bean curd is organized fine and smooth inadequately.Therefore, optimize fermented bean curd Mucor bacterial strain and carry out sufu fermentation, and the ferment process suppresses and the control generation of ' white spot ' on fermented bean curd after fermented bean curd, will be to the production of preserved beancurd of enterprise, promotion, increasing economic efficiency has positive meaning.
" hickie " of fermented bean curd is the formed tyrosine crystal of fermented bean curd mucor protease decomposing soya-bean protein process.Tyrosine is the extremely low amino acid of a kind of solubility (25 degree, 100 gram water down can only dissolve 0.045 gram), and in the ferment process, along with the continuation of time lengthening enzymolysis, tyrosine increases gradually after fermented bean curd, tends to the supersaturation crystallization at last and separates out.
Mr. Lei Shifen of Guangxi light industry research institute thinks: fermented bean curd long " hickie " is " old, big, the difficulty " problem that exists in the production of preserved beancurd, and it not only influences outward appearance, also makes the people suspect whether mildew of fermented bean curd.
By retrieval as can be known: be published in " China brewages " 1993 (4) 18-24 pages or leaves, name is called in " seed selection of fermented bean curd mucor strain " literary composition to be introduced: because the restriction of technical conditions, separation to existing bacterial classification, rejuvenation, screening and seed selection work acquire a certain degree of difficulty, and cause spawn degeneration, resistance poor, enzyme system is few, it is low that enzyme is lived, and quality is undesirable after the fermented bean curd maturation, and particularly white side's fermented bean curd exists " small particles " and color and luster " brown stain " phenomenon more obvious.
Though the enterprise that some technical force is stronger has adopted strain breeding method, the screening prolease activity is strong, but the bacterial strain of tyrosine growing amount restriction, but from using, bacterial classification has degradation phenomena again after using one section, and soybean material also has destabilizing factor, thereby is difficult to eradicate.And concerning the middle-size and small-size manufacturing enterprise of major part, fermented bean curd is long, and " hickie " is the No.1 difficult problem of production of preserved beancurd.
The object of the invention is optimized screening on bacterial classification, improve the heat resistance of bacterial classification prolease activity and bacterial classification, thereby shortens after the fermented bean curd ferment time and enlarge production of preserved beancurd season.And on technology, improve simultaneously, after the sufu fermentation maturation, use the proteinase activity of thermal inactivation fermented bean curd Mucor, thereby fundamentally solved length " hickie " problem of fermented bean curd.
The present invention is realized that by following technical proposals technological process of the present invention is carried out the after fermentation process modification then for optimizing fermented bean curd Mucor bacterial strain earlier.It optimizes fermented bean curd Mucor bacterial strain process: the dull and stereotyped primary dcreening operation of fermented bean curd Mucor bacterial strain → preparation spore suspension → spore heat treatment → single strain separation → casein → single strain prolease activity mensuration → fermented bean curd Mucor is optimized bacterial strain.
After fermentation technology: fermented bean curd Mucor test tube slant → wheat bran triangular flask spreads cultivation → prepare on spore suspension → inoculation bean curd base, and primary fermentation → method inactive enzyme activity → warehouse entry storehouse or sale such as base → bottling (adding feed liquid) after fermentation → fermenting-ripening → water-bath or electric heating salt down.
Fermented bean curd Mucor slant medium adopts the potato agar culture medium, the single strain isolation medium: be to add 0.4% casein and 0.1% deoxysodium cholate in the potato culture medium.Dull and stereotyped primary dcreening operation culture medium: casein 0.4%, agar 1.5-2% is with the preparation of PH7.4 borate buffer.The wheat bran culture medium; 25 gram wheat brans add water 130-140%, mix the packing triangular flask thoroughly.Medium sterilization removes the wheat bran culture medium and adopts 1kg/cm
2Outside sterilizing 30 minutes, other three kinds of culture mediums all adopt 1kg/cm
2Sterilized 20 minutes.
Its basic process is:
(1) preparation spore suspension and spore heat treatment: fermented bean curd Mucor bacterial strain moved be connected on the potato slant medium, 30 degree were cultivated 3-4 days, add the 10ml SPSS, scrape spore, pour in the triangular flask that 20ml SPSS and bead are housed, vibration makes spore disperse the back filtered through gauze, adopt haemocytometer counting spore concentration, and the control spore concentration is 10
6Individual/more than the ml.Draw spore suspension 5ml and place test tube, processing cooling after 10 minutes in 45 degree water-baths.This processing can be screened the stronger spore of acquisition vigor, and this is very helpful to seed selection fermented bean curd Mucor high-temperature resistant strain, if need, and all right repeatable operation 2-3 time.
(2) bacterial strain separates and primary dcreening operation: heat treated spore liquid is diluted to 10
-3-10
-4, to get 0.1ml and coat on the isolation medium agar plate, 30 degree were cultivated 1-2 days, the bacterial strain spore inclined-plane that picking transparent circle diameter is big is cultivated, obtain high-temperature resistant strain and can improve temperature step by step and cultivate domestication, cultivate the back switching as 30 degree and cultivate, transfer after 32 degree are cultivated and cultivate in 34 degree etc. at 32 degree.Can select resistant to elevated temperatures fermented bean curd Mucor bacterial strain by this operation.It is fast to grow, the fermented bean curd Mucor separated strain that the temperature adaptive faculty is strong is inoculated in the triangular flask bran mass, 30 degree were cultivated 4 days, extracted enzyme liquid, with diameter 10mm sterilization filter paper, putting into Mucor wheat bran leaching liquor (enzyme liquid) soaked 1 hour, be attached on the casein agar flat board after taking out airing, each bacterial strain liquid is handled 4-6 filter paper, is attached on the flat board, place 40 degree, 6 hours mensuration of insulation transparent circle diameter down, choose the big bacterial strain of transparent circle diameter and carry out multiple sieve.Separating plate and primary dcreening operation flat board all add 0.1% deoxysodium cholate, too spread growth because of it can suppress mycelia, make single strain be separated into possibility, and the bacterial strain inclined-plane that the picking transparent circle is big on flat board is cultivated.
(3) multiple sieve: multiple sieve is to measure prolease activity, adopts folin's methods.At PH7.0, under temperature 40 degree, producing 1 microgram tyrosine with per minute enzymatic reaction decomposition casein is that a protease activity unit of force (μ g/min) selects the high bacterial strain of prolease activity to produce.But because required reagent in the prolease activity assay method, equipment and mensuration are comparatively complicated, and general enterprise may not all satisfy the requirements.Thereby production unit also can directly be heat-treated fermented bean curd Mucor spore, separation and purification improves temperature acclimatisation and cultivates, and uses the dull and stereotyped primary dcreening operation of casein repeatedly, select more high temperature resistant, fast growth, mycelia is white and fine and close, and the bacterial strain that transparent circle is bigger is produced, this operation is simple and easy, be easy to grasp, production unit separates final election as long as regularly the fermented bean curd Mucor is produced bacterium, and one improves the quality of production of preserved beancurd surely and shortens fermentation period.
The after fermentation process modification is that fermented bean curd Mucor test tube bacterial strain → wheat bran triangular flask spreads cultivation → prepare on spore suspension → inoculation bean curd base that primary fermentation → inactive enzyme activity → warehouse entry storehouse or sale such as base → bottling (adding feed liquid) after fermentation → fermenting-ripening → water-bath or electric heating salt down.
Seed selection of the present invention is high temperature resistant and the strong fermented bean curd Mucor bacterial strain of prolease activity carries out sufu fermentation, the bacterial classification preparation spore suspension that spreads cultivation routinely, inoculation bean curd base primary fermentation, salted, the feed liquid after fermentation is added in bottling, after fermented bean curd after fermentation maturation (fermented bean curd is organized the soft local flavor deliciousness of piece), adopt steam heat, methods such as electric heating or water-bath heat are come passivation fermented bean curd mucorin enzymatic activity, fermented bean curd is bottled by fermented bean curd base under the routine to be produced " hickie " in back 3 months and is deferred to 1 year even the longer time is not grown " hickie ", and look, fragrant, sense organs such as flavor are good, thereby fundamentally solving " old; big; the difficulty " problem of fermented bean curd long " hickie ".
As for heat fermented bean curd mucor protease effect of vigor is measured, can help to select the condition of heat treated, the present invention adopts and optimizes fermented bean curd Mucor bacterial strain, behind wheat bran cultivation lixiviate enzyme liquid, respectively with 50 degree, 55 spend to enzyme liquid, 60 degree, handle different time under the 70 degree temperature, measure its neutral protease vigor, the result is as shown in table 1.
| Temperature ℃ | Time (h) | Neutral proteinase inactivation rate (%) | Temperature ℃ | Time (h) | Neutral proteinase inactivation rate (%) |
| 50 | 6 12 24 48 | 28 39 57 81 | 60 | 6 12 | 42 65 |
| 55 | 6 12 24 | 38 53 72 | 70 | 1 2 | 57 81 |
Find out that from last table temperature has significant effects to prolease activity.These experimental datas have been arranged, can be according to actual conditions, select suitable temperature and heat time heating time that the product of sufu fermentation maturation is heat-treated, vigor with a passivation part or quite a few enzyme, make fermented bean curd in long sale and storage, reduce the growing amount of tyrosine, thereby effectively controlled the tyrosine crystal problem of preserved beancurd.Simultaneously, can find that the short-term after preliminary maturation of sufu fermentation or the full maturity does not have " hickie " in the time, have only after placing 2-3 month just to produce hickie.Therefore, select to heat-treat behind the fermenting-ripening, and control enzyme deactivation rate makes surplus enzyme continue to keep the follow-up fermentation of fermented bean curd in the 60%-80% scope.This consideration is to be of value to the mouthfeel of preserved beancurd and quality.The packing cartonning fermentation of fermented bean curd bottling back, after maturation, be extended down to some times again, can electrothermal tube intensification heating be installed in indoor (sealing), this way is implemented comparatively easy, concerning every bottle of fermented bean curd finished product, it is very little that cost improves, and (also can with additive methods such as steam heat) be though heating process also brings many procedures to production, but from product quality and factory's benefit, it is necessary increasing this operation.
Below the present invention is described in further detail by specific embodiment.
Suspension miospore content 10 during spore heat treatment
6Individual/more than the ml, place 45 degree water-baths to handle 10 minutes the pityrosporion ovale suspension, can screen the stronger spore of vigor, this is very helpful to seed selection fermented bean curd Mucor high-temperature resistant strain, if need, the all right repeatable operation of this operation 2-3 time, specific practice is: one time the heat treatment rear plate is cultivated, preparation spore suspension secondary heat treatment.The screening high-temperature resistant strain can also progressively improve temperature acclimatisation in incubation and cultivate.
The bacterial strain of the energetic and fast growth of screening fermented bean curd mucor protease, when separating single bacterium colony of fermented bean curd Mucor and primary dcreening operation, all interpolation 0.4% casein and 0.1% deoxysodium cholate are made flat board in culture medium, and form at bacterium colony, when mycelia too spreads growth, single bacterium colony inclined-plane that the picking transparent circle is big is cultivated, separating plate interpolation deoxysodium cholate has inhibitory action to the growth that too spreads of Mucor mycelia, and in flat board, add casein, prolease activity that can its different strains of Preliminary detection.
Use and optimize bacterial strain production fermented bean curd, the base that generally salts down bottling after fermentation in 20～30 days is just ripe.But for the delicious local flavor that makes fermented bean curd better, the after fermentation phase can be extended down to one month, then bottled fermented bean curd was placed 70～80 degree water bath processing 1 hour, or handle half a day to one day with 60 methods such as degree above hot-air steam heat etc., this processing can make fermented bean curd mucorin enzyme activity inactivation, its level of deactivation is subject to processing temperature, factor affecting such as time and medium heat transfer, be as the criterion with the work of passivation 60-80% enzyme after the general fermented bean curd maturation, after the fermented bean curd after fermentation maturation, with by the use of thermal means passivation fermented bean curd mucorin enzyme activity, the temperature that the temperature that is adopted will make fermented bean curd be heated reaches more than 50 degree, if temperature can be quickened proteinase activity when 35-45 spends, thereby generates more fermented bean curd " hickie ".
Embodiment:
(1) preparation spore suspension
Fermented bean curd Mucor bacterial strain moved be connected to the potato slant medium, 30 degree were cultivated 3 days, in an inclined-plane, add the 10ml SPSS, scrape spore, pour in the triangular flask that SPSS and bead are housed and vibrate, the back is with the filtrations of four layers of sterile gauze, collect filtrate and to control spore be 5.6 * 10
6Individual/ml.
(2) spore heat treatment and more high temperature resistant fermented bean curd Mucor bacterial strain separates
Draw the sterile test tube that spore suspension 5ml places 15 * 15mm, place 45 degree water-baths to handle 10 minutes, instant cooling is diluted to 10
-3, get 10
-2, 10
-3Dilution tube 0.1ml is coated on the separating plate, 30 degree were cultivated after 1 day, very fast and the intensive single bacterium colony inclined-plane of picking 11 strain mycelial growths is cultivated, the inclined-plane is shifted in the growth back, progressively improve temperature acclimatisation and cultivate screening acquisition also well-grown bacterial classification 5 strains under 34 degree, improved about 10% than the highest well-grown temperature of former bacterial strain.
(3) the dull and stereotyped primary dcreening operation of casein
5 strain single strains are connected to respectively in the triangular flask wheat bran skin culture medium together with former bacterial strain, 30 degree were cultivated 4 days, each triangular flask adds the 100ml sterilized water, smash culture to pieces, lixiviate is 1 hour in 40 degree water-baths, the back is filtered with filter paper and is collected filtrate, soaked the wheat bran leaching liquor 1 hour with 10mm sterilization filter paper, take out filter disc paper airing, be attached to respectively on the casein flat board, each sample liquid repeats 4-6 time, each dull and stereotyped 4-6 piece filter paper that pastes, put 40 and spend insulation down 6 hours, the transparent circle diameter is measured in the back, chooses the big bacterial strain of 3 strain transparent circles and carries out prolease activity mensuration.
(4) bacterial classification sieves again: in the single strain prolease activity is measured, the bacterial strain that 3 strain transparent circles are big carries out the triangular flask wheat bran together with former bacterial strain and cultivates, the same method in back is extracted enzyme liquid, adopt folin's methods to measure neutral proteinase, acid protease and basic protein enzyme activity, screen 1 strain neutral proteinase, acid protease and basic protein enzyme activity are higher than the fermented bean curd hair enzyme of former bacterial strain 19.2%, 30.8% and 15% respectively and optimize bacterial strain.
(5) sufu fermentation;
The fermented bean curd mucor strain of optimizing is inoculated in the triangular flask bran mass 30 degree cultivated 2 days, every bottle adds cold boiling water 500ml and soaks half an hour, and vibration evenly back is filtered with sterile gauze and obtained spore inoculating liquid.Soybean legal system routinely is equipped with the bean curd base, and the control water content is 68-70%, after the cutting spore liquid is sparged on the beans piece, (26-30 degree) cultivated 30 hours under the room temperature, the back is with salt down base 24 hours or directly use salted 2-7 days of 18 Baume degrees salt solution, take out back bottling and reinforced liquid sealing, after fermentation to a month maturation under the room temperature.
(6) passivation fermented bean curd mucorin enzyme activity:
1. the fermented bean curd 20 bottles (every bottle 400 grams) with fermenting-ripening places 80 degree water-bath heating 1 hour, and store back cooling naturally, and the result is: fermented bean curd has been stored and do not grown " hickie " in nearly 2 years, from sense organ, organizes piece complete, and faint yellow, mouthfeel is good, local flavor U.S..
2. 40 bottles in the fermented bean curd of fermenting-ripening (every bottle 400 gram) is placed incubator, with 55-60 degree hot air treatment 1 day, back cooling storage naturally, the also same immersion method of result, long preservation, the quality of fermented bean curd is good.
3. in placing the bottled fermented bean curd of fermenting-ripening between airtight storage, handled 1 day 1 year all not long hickie of fermented bean curd preservation to the 55--60 degree with the electrothermal tube heat temperature raising.
From as can be known above-mentioned: with the method for heat after the sufu fermentation maturation with the passivation of fermented bean curd proteinase activity or make its considerable part enzyme deactivation, be to guarantee that fermented bean curd not grow " hickie ", the effective measures of assurance fermented bean curd better quality for a long time.Say that from actual result this method has contained that fermented bean curd " hickie " forms, and does not influence the tissue of fermented bean curd again, color and local flavor, and ripe fermented bean curd does not take heat treated sample to generate " hickie " in 3-4 month storage period, has looked close and numerous to half a year " hickie ", and sense organ is extremely bad.This method will be considered the heat transfer of medium from production application, and factors such as heat distribution uniformity are formulated heating condition.Adopt the fermented bean curd mucor protease of thermal inactivation fermenting-ripening to live, the temperature of fermented bean curd is reached more than 50 degree,, can not play inactive enzyme and live, suppress the effect that " hickie " generates if temperature is too low.
Claims (1)
1. the method that suppresses generation of ' white spot ' on fermented bean curd comprises and optimizes fermented bean curd Mucor bacterial strain, after fermentation technology two steps, it is characterized in that:
(1) method of optimization fermented bean curd Mucor bacterial strain: fermented bean curd Mucor bacterial strain, prepare spore suspension and spore heat treatment then, fermented bean curd Mucor bacterial strain moved be connected on the potato slant medium, 30 degree were cultivated 3-4 days, added the 10ml SPSS, scraped spore, pour in the triangular flask that 20ml SPSS and bead are housed, vibration makes spore disperse the back filtered through gauze, adopt haemocytometer counting spore concentration, and the control spore concentration is 10
6Individual/as more than the ml, to draw spore suspension 5ml and place test tube, processing cooling after 10 minutes is carried out bacterial strain again and is separated and primary dcreening operation in 45 degree water-baths, and heat treated spore liquid is diluted to 10
-3-10
-4Getting 0.1ml coats on the isolation medium agar plate, 30 degree were cultivated 1-2 days, the bacterial strain spore inclined-plane that picking transparent circle diameter is big is cultivated, it is fast to grow, and the fermented bean curd Mucor separated strain that the temperature adaptive faculty is strong is inoculated in the triangular flask bran mass, and 30 degree were cultivated 4 days, extract enzyme liquid, with diameter 10mm sterilization filter paper, put into Mucor wheat bran leaching liquor and soaked 1 hour, be attached on the casein agar flat board behind the taking-up airing, each bacterial strain liquid is handled 4-6 filter paper, be attached on the flat board, place 40 degree, 6 hours mensuration of insulation transparent circle diameter down, choosing the big bacterial strain of transparent circle diameter carries out multiple sieve, carry out the strain protein enzyme activity determination at last, the fermented bean curd Mucor is optimized bacterial strain;
(2) zymotechnique: the fermented bean curd mucor strain of optimizing is inoculated in the triangular flask bran mass 30 degree cultivated 2 days, every bottle adds cold boiling water 500ml and soaks half an hour, vibration evenly back obtains spore inoculating liquid with the sterile gauze filtration, soybean legal system routinely is equipped with the bean curd base, and the control water content is 68-70%, after the cutting spore liquid is sparged on the beans piece, cultivated 30 hours under the room temperature, the back is with salt down base 24 hours or directly use salted 2-7 days of 18 Baume degrees salt solution, take out back bottling and reinforced liquid sealing, after fermentation to one a month maturation under the room temperature, the fermented bean curd of fermenting-ripening is placed 80 degree water-bath heating 1 hour, store back cooling naturally, or the fermented bean curd of fermenting-ripening placed incubator, and with 55-60 degree hot air treatment 1 day, back cooling storage naturally, in also can placing the bottled fermented bean curd of fermenting-ripening between airtight storage, handle 1 day with electrothermal tube heat temperature raising to 55--60 degree.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN99114527A CN1122452C (en) | 1999-10-30 | 1999-10-30 | Method for suppressing generation of 'white spot' on fermented bean curd |
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|---|---|---|---|
| CN99114527A CN1122452C (en) | 1999-10-30 | 1999-10-30 | Method for suppressing generation of 'white spot' on fermented bean curd |
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| CN1122452C true CN1122452C (en) | 2003-10-01 |
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Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101396099B (en) * | 2008-10-24 | 2013-01-23 | 北京二商王致和食品有限公司 | Production method of preserved beancurd |
| CN104798905B (en) * | 2015-04-12 | 2018-02-13 | 浙江大学 | The method for eliminating fermented bean curd hickie |
| CN106106806B (en) * | 2016-07-22 | 2019-06-11 | 福建省潘氏食品有限公司 | A method of inhibit white dot in fermented bean curd storage to generate |
| CN112375690B (en) * | 2020-11-18 | 2023-10-03 | 成都国酿食品股份有限公司 | Mucor preserved beancurd with low white point rate and application thereof |
| CN113974079A (en) * | 2021-11-04 | 2022-01-28 | 广东广中皇食品有限公司 | Method for improving quality of red-rot milk and application thereof |
| CN114058670A (en) * | 2021-11-15 | 2022-02-18 | 鹤山市东古调味食品有限公司 | Microbial source research method for weeping juice of swollen pot with fermented bean curd |
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