CN106591280A - Mutation screening method of Mucor sufu strain, fermented bean curd, and production method of fermented bean curd - Google Patents
Mutation screening method of Mucor sufu strain, fermented bean curd, and production method of fermented bean curd Download PDFInfo
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Abstract
The invention relates to a mutation screening method of Mucor sufu strain, fermented bean curd and a production method of the fermented bean curd. The screening method comprises the following steps: preparing a strain suspension from mucor strains to be selected; carrying out ultraviolet mutation operation on the strain suspension; preliminarily culturing the ultraviolet mutated strain suspension with a medium to obtain a mucor strain colony; respectively simultaneously carrying out high temperature screening culture operation and low temperature screening culture operation on the mucor strain colony; judging whether the mucor strain colony subjected to the high temperature screening culture operation accords with a high-temperature preset spore production condition or not; judging whether the mucor strain colony subjected to the low temperature screening culture operation accords with a low-temperature preset spore production condition or not; and selecting the mucor strain colony as the Mucor sufu strain if the mucor strain colonies accord with the conditions. The screening method allows the mutated strain to generate few spores in the fermented bean curd production process, reduce glass crystals in the fermented bean curd, and improves the quality of the fermented bean curd.
Description
Technical field
The present invention relates to production of preserved beancurd technical field, more particularly to a kind of mutagenesis screening method of Mucor sufu strain,
Fermented bean curd and its production method.
Background technology
Fermented bean curd, also known as fermented bean curd, is the Chinese characteristic Traditional Folk cuisines for spreading thousands of years, because which is in good taste, nutrition is high,
Smelling has strand smelly, and special fragrance of tasting is liked by Chinese common people and the south east asia people deeply, is prolonged one
The delicious food not waned.
At present there is the production of fermented bean curd China various places, although they due to not of uniform size, dispensing is different, and variety name is numerous
It is many, but producing principle is mostly identical.Semen sojae atricolor is made into bean curd first, then pressed compact is divided into fritter, put and can connect in tub
The very strong rhizopus of prolease activity or mucoraceous strain, carry out fermenting, the peculiar flavour of fermented bean curd is exactly in fermentation storage
Formed in process.
However, existing production of preserved beancurd easily produces more glass crystal, the quality of fermented bean curd can be had a strong impact on so that rotten
The taste bad of breast.
And the generation of glass crystal, and the spore that mucor strain is produced has an important relation, mucor strain produces spore quantity
The more, then in fermented bean curd, glass crystal is the more.
The content of the invention
Based on this, it is necessary to provide the mutagenesis screening method and glass of a kind of Mucor sufu strain for producing spore negligible amounts
Crystal is less and quality is higher fermented bean curd and its production method.
A kind of mutagenesis screening method of Mucor sufu strain, comprises the steps：Mucor strain to be selected is made into bacteria suspension；
Ultraviolet mutagenesis operation is carried out to the bacteria suspension, wherein, the persistent period of the ultraviolet mutagenesis operation is 2 minutes～24 minutes；
Under 15 DEG C～20 DEG C temperature conditionss, using culture medium to the bacteria suspension operated through the ultraviolet mutagenesis is carried out tentatively
Culture operation, obtains mucor strain bacterium colony；The mucor strain bacterium colony is simultaneously carried out high temperature screening and culturing operation and low respectively
Warm screening and culturing operation, wherein, the high temperature screening and culturing operation is carried out under 24 DEG C～28 DEG C temperature conditionss, the low temperature sieve
Choosing culture operation is carried out under 15 DEG C～20 DEG C temperature conditionss；Judge the Mucor after the high temperature screening and culturing is operated
Whether strain bacterium colony meets the default product spore situation of high temperature, obtains the first judged result, wherein, the default spore situation of producing of the high temperature is
Every gram of thallospore number is less than 104Individual/gram；Judge that the mucor strain bacterium colony after the low temperature screening and culturing is operated is
It is no to meet the default product spore situation of low temperature, the second judged result is obtained, wherein, the default spore situation of producing of the low temperature is every gram of mycelia spore
Subnumber is more than 108Individual/gram；First judged result is with second judged result when being, then choose the mucormycosiss
It is the Mucor sufu strain to plant bacterium colony.
Wherein in one embodiment, the culture medium is Rhizoma Solani tuber osi culture medium.
Wherein in one embodiment, the spore concentration of mucor strain to be selected described in the bacteria suspension for (0.1～
9.99)×106Individual/milliliter.
Wherein in one embodiment, using ultraviolet mutagenesis case, the ultraviolet mutagenesis operation is carried out.
Wherein in one embodiment, the ultraviolet case is provided with uviol lamp, and the power of the uviol lamp is 15 watts, described
The wavelength of the ultraviolet that uviol lamp sends is 253.7 nanometers.
Wherein in one embodiment, the mucor strain to be selected is made using 0.01% sodium dodecyl sulfate solution
Bacteria suspension.
Wherein in one embodiment, when the ultraviolet mutagenesis operation is carried out, behaviour is stirred to the bacteria suspension also
Make.
Wherein in one embodiment, the stirring operation is operated for magnetic agitation.
The present invention also provides a kind of production method of fermented bean curd, sieves including the mutation by as above arbitrary Mucor sufu strain
Mucor sufu strain prepared by choosing method is seeded on fermented bean curd embryo, prepares fermented bean curd.
Wherein in one embodiment, it is described the operation that Mucor sufu strain is seeded on fermented bean curd embryo is specifically included it is as follows
Step：
The Mucor sufu strain is mixed with yeast, mixed bacteria liquid is prepared；
The mixed bacteria liquid is seeded on the fermented bean curd embryo.
The present invention also provides a kind of fermented bean curd, adopts and obtains prepared by the as above production method of arbitrary fermented bean curd.
The mutagenesis screening method of above-mentioned Mucor sufu strain, can mutation go out at 15 DEG C～20 DEG C well-grown, produce spore
It is sub normally spore not to be produced at 24 DEG C～28 DEG C or sporogenic Mucor sufu strain is produced less, can produce in the production of fermented bean curd
The less spore of life such that it is able to reduce the glass crystal in fermented bean curd, improves the quality of fermented bean curd.
Description of the drawings
The step of Fig. 1 is the mutagenesis screening method of the Mucor sufu strain of embodiment flow chart；
The step of Fig. 2 is the mutagenesis screening method of the Mucor sufu strain of another embodiment flow chart.
Specific embodiment
It is understandable to enable the above objects, features and advantages of the present invention to become apparent from, below in conjunction with the accompanying drawings to the present invention
Specific embodiment be described in detail.Elaborate many details in order to fully understand this in the following description
It is bright.But the present invention can be implemented with being much different from alternate manner described here, and those skilled in the art can be not
Similar improvement is done in the case of running counter to intension of the present invention, therefore the present invention is not embodied as being limited by following public.
For example, a kind of mutagenesis screening method of Mucor sufu strain, comprises the steps：Mucor strain to be selected is made into bacterium
Suspension；Ultraviolet mutagenesis operation is carried out to the bacteria suspension, wherein, the persistent period of the ultraviolet mutagenesis operation is 2 minutes～24
Minute；Under 15 DEG C～20 DEG C temperature conditionss, using culture medium to the bacteria suspension operated through the ultraviolet mutagenesis is entered
The preliminary culture operation of row, obtains mucor strain bacterium colony；The mucor strain bacterium colony is carried out into high temperature screening and culturing behaviour respectively simultaneously
Make and the operation of low temperature screening and culturing, wherein, the high temperature screening and culturing operation is carried out under 24 DEG C～28 DEG C temperature conditionss, described
The operation of low temperature screening and culturing is carried out under 15 DEG C～20 DEG C temperature conditionss；Judge the institute after the high temperature screening and culturing is operated
State whether mucor strain bacterium colony meets the default product spore situation of high temperature, obtain the first judged result, wherein, the high temperature is default to produce spore
Situation is that every gram of thallospore number is less than 104/gram；Judge the mucor strain after the low temperature screening and culturing is operated
Whether bacterium colony meets the default product spore situation of low temperature, obtains the second judged result, wherein, the default spore situation of producing of the low temperature is per gram
Thallospore number is more than 108/gram；First judged result is with second judged result when being, then choose described
Mucor strain bacterium colony is the Mucor sufu strain.The mutagenesis screening method of above-mentioned Mucor sufu strain, can mutation go out 15
DEG C～20 DEG C at well-grown, to produce spore normal and spore is not produced at 24 DEG C～28 DEG C or sporogenic Mucor sufu bacterium is produced less
Kind, less spore can be produced in the production of fermented bean curd such that it is able to reduce the glass crystal in fermented bean curd, improve the product of fermented bean curd
Matter.
Refer to Fig. 1, the mutagenesis screening method and the producer of fermented bean curd of the Mucor sufu strain of an embodiment of the present invention
Method, comprises the steps：
S110：The strain that sets out is mucor strain to be selected, also referred to as mucor strain.
Mucor is commonly used for the fermentation technologys such as fermented bean curd, Semen Sojae Preparatum, and by setting out, strain is chosen for mucor strain, can pass
On the basis of system mucor strain, mutation goes out spore can not be produced in production of preserved beancurd or produces less sporogenic Mucor sufu strain,
The problem of the content of glass crystal in fermented bean curd is reduced with this.As, in traditional processing technology, mucor strain is more commonly used, strain
Source is easier to obtain, and is chosen for mucor strain by the strain that will directly set out, is easy in the production of fermented bean curd on the spot to Mucor
Strain carries out mutation, can effectively reduce the strain purchase cost in production.
In one embodiment, the Mucor is Mucor racemosus Fres, Mucor mucedo, at least one in Zygosaccharomyces rouxii.Preferably,
The Mucor is Mucor mucedo strain, in such manner, it is possible to further improve the stability of strain after mutation, that is to say, that, it is not easy to
Generation back mutation.
In order that mucor strain reaches preferable growth conditions, to improve the success rate of mutation, for example, to the mucormycosiss
Planting carries out preculture using culture medium, and the preculture operation persistent period is 24 hours～72 hours, treats the plentiful standby of spore
With and for example, the preculture operation persistent period is 42 hours～54 hours, and and for example, the preculture operation persistent period is
48 hours, and for example, under 25 DEG C of temperature conditionss, the preculture is carried out, in such manner, it is possible to make the mucor strain reach preferably
Growth conditions, to improve the success rate of mutation.And for example, the spore is plentiful refers to that every gram of thallospore number is more than 108Individual/gram.
In one embodiment, in the preculture operating process, purification process is carried out to the mucor strain also, that is,
Say, mucor strain is carried out into purification, select single bacterium colony strain to carry out mutation, in such manner, it is possible to reduce strain after follow-up mutation
Screening operation amount.And for example, the purification process is specially：The mucor strain is rule or coating is incubated at culture medium
On, after the culture operation terminates, the mucor strain for selecting single bacterium colony is standby, that is to say, that carried out S120 steps
It is rapid to operate.
In one embodiment, the culture medium is solid medium.And for example, the culture medium is Rhizoma Solani tuber osi culture medium.And for example,
The Rhizoma Solani tuber osi culture medium contains each component of following mass parts：180 parts～220 parts of Rhizoma Solani tuber osi, 18 parts～22 parts of glucose, agar 15
Part～20 parts and 950 parts～1050 parts of water.And for example, the collocation method of the Rhizoma Solani tuber osi culture medium is as follows：Potato decortication is shredded, is boiled
Acquirement murphy juice after rotten, adds glucose, adds agar, adds water, and after heating for dissolving, bottles and in 121 DEG C of sterilizings is within 20 minutes
Can.
In one embodiment, the Rhizoma Solani tuber osi culture medium contains each component of following mass parts：200 parts of Rhizoma Solani tuber osi, glucose 20
1000 parts of part, 15 parts～20 parts of agar and water.
It is understood that mutation has randomness, the probability that usual strain produces site directed mutagenesiss is relatively low, in order to further improve
The successful of mucor strain mutation, for example, the Rhizoma Solani tuber osi culture medium contains each component of following mass parts：Rhizoma Solani tuber osi 180 parts～220
It is part, 18 parts～22 parts of glucose, 15 parts～20 parts of agar, 0.2 part～0.6 part of leucine, 0.1 part～0.22 part of arginine, different
950 parts～1050 parts of 0.15 part～0.27 part of leucine, 0.12 part～0.38 part of L-Valine and water.And for example, the Rhizoma Solani tuber osi culture
The collocation method of base is as follows：Potato decortication is shredded, well-done rear acquirement murphy juice adds glucose, adds agar, add water, plus
After heat of solution, bottling and 121 DEG C sterilizing 20 minutes after, after which is cooled to 60 DEG C, add L-Valine, isoleucine, arginine,
Leucine aqueous solution is simultaneously mixed, wherein, the L-Valine, isoleucine, arginine, leucine aqueous solution pass through filtration sterilization.
Above-mentioned Rhizoma Solani tuber osi culture medium can make the growth of Mucor more stable using each component of as above mass parts, by adding
Enter 0.2 part～0.6 part of leucine, 0.15 part～0.27 part of isoleucine, 0.12 part～0.38 part of L-Valine, Mucor can be made to exist
Injury of the ultraviolet to Mucor cell mitochondrial is reduced in follow-up mutagenic processes, the radical chain caused because of ultraviolet is reduced
Injury of the reaction to Mucor cell, enables the fast quick-recovery of the Mucor after mutation to enter trophophase, so as to reduce hair after mutation
Mould adaptive time.By add 0.1 part～0.22 part of arginine, and with 0.2 part～0.6 part of leucine, isoleucine 0.15
When part～0.27 part, 0.12 part～0.38 part of L-Valine are used together, it is possible to increase the success rate of Mucor mutation, reduce Mucor and exist
Mortality rate in ultra-vioket radiation mutation.Above-mentioned Rhizoma Solani tuber osi culture medium can make the growth of Mucor using each component of as above mass parts
It is more stable, moreover it is possible to reduce the adaptive time of Mucor after mutation, it is possible to increase the success rate of Mucor mutation, Mucor is reduced ultraviolet
Mortality rate in irradiation mutation.
In one embodiment, the Rhizoma Solani tuber osi culture medium contains each component of following mass parts：200 parts of Rhizoma Solani tuber osi, glucose 20
Part, 15 parts～20 parts of agar, 0.35 part～0.5 part of leucine, 0.15 part～0.2 part of arginine, 0.18 part of isoleucine～
1000 parts of 0.22 part, 0.22 part～0.28 part of L-Valine and water, each component of the above-mentioned Rhizoma Solani tuber osi culture medium using as above mass parts, energy
It is enough further to make the growth of Mucor more stable, moreover it is possible to further to reduce the adaptive time of Mucor after mutation, it is possible to increase Mucor
The success rate of mutation, reduces mortality rate of the Mucor in ultra-vioket radiation mutation.
In one embodiment, the Rhizoma Solani tuber osi culture medium contains each component of following mass parts：200 parts of Rhizoma Solani tuber osi, glucose 20
Part, 15 parts～20 parts of agar, 0.45 part of leucine, 0.18 part of arginine, 0.19 part of isoleucine, 0.26 part of L-Valine and water
1000 parts, above-mentioned Rhizoma Solani tuber osi culture medium can further make the growth of Mucor more stable using each component of as above mass parts, also
The adaptive time of Mucor after mutation can further be reduced, it is possible to increase the success rate of Mucor mutation, Mucor is reduced in ultra-vioket radiation
Mortality rate in mutation.
The industry characteristics of mucor strain in one embodiment, are determined, follow-up generation time and growth time is adjusted, or
Person says that the estimation mucor strain grows to the spore plentiful required time.And for example, the growth characteristics are：In different temperatures
Under spore growth situation, prolease activity, the characteristic of the opportunistic pathogen such as the speed of growth.
S120：The mucor strain to be selected is made into bacteria suspension.
By the mucor strain is made bacteria suspension, it is easy to carry out mutation to mucor strain.For example, in the bacteria suspension
The spore concentration of the mucor strain is (0.1～9.99) × 106Individual/milliliter, in such manner, it is possible to operate in follow-up ultraviolet mutagenesis
During, select few sporogenic Mucor sufu strain during production of preserved beancurd.Specifically, and for example, using 0.01% ten
The mucor strain is made bacteria suspension by sodium dialkyl sulfate solution, and for example, will using 0.01% sodium dodecyl sulfate solution
The mucor strain makes spore concentration for (0.1～9.99) × 106The bacteria suspension of individual/milliliter, so, the dodecane of low concentration
Base sodium sulfate can affect spore, and punching effect can be played to the cell wall of Mucor such that it is able to improve mucormycosiss
Plant the success rate of mutation.
In one embodiment, adopt 0.01% sodium dodecyl sulfate solution by the mucor strain make spore concentration for
(1.1～7.5) × 106The bacteria suspension of individual/milliliter, and for example, using 0.01% sodium dodecyl sulfate solution by the mucormycosiss
Plant spore concentration is made for (2.2～6.1) × 106The bacteria suspension of individual/milliliter, and for example, using 0.01% sodium lauryl sulphate
The mucor strain is made spore concentration for 3.2 × 10 by solution6The bacteria suspension of individual/milliliter is in such manner, it is possible to further improve hair
The success rate of mould species mutation.
In order that the spore in the bacteria suspension can be broken up, for example, the mucor strain is being made into bacteria suspension
During, bead is added in bacteria suspension, and carries out concussion operation, so that the spore of Mucor can solely be dispersed in bacterium and hang
In liquid.In order to remove the spore ball in the bacteria suspension, for example, the bacteria suspension after the concussion operation is carried out into filtration behaviour
Make, to remove agglomerating larger spore ball, and for example, the bacteria suspension after the concussion operation is carried out into filtration behaviour using with absorbent cotton
Make, so, remove the effect of spore ball preferably, and operation is also more convenient.
In one embodiment, the bacteria suspension is carried out into filter operation, to remove undispersed spore ball, and for example, by institute
State bacteria suspension to be filtered using absorbent cotton, that is to say, that in the case of without concussion operation, directly enter the bacteria suspension
Row filter operation.
It should be noted that when carrying out shaking operation and/or filter operation, above-mentioned spore concentration is all concussion operation
And/or the spore concentration of the bacteria suspension after filter operation.
S130：Ultraviolet mutagenesis operation is carried out to the bacteria suspension, wherein, the parameter of the ultraviolet mutagenesis operation is as follows：Institute
The wavelength of the ultraviolet that ultraviolet mutagenesis operation is adopted is stated for 253.7 nanometers, the persistent period for stating ultra-vioket radiation operation is 2 points
Clock～24 minute.
By carrying out ultra-vioket radiation operation to the bacteria suspension, to carry out ultraviolet mutagenesis to mucor strain.Need explanation
It is that the mutation process of microorganism has randomness so that the orthomutation repeatability of mucor strain is poor.By will be described ultraviolet
The parameter of mutation operation is as follows：The wavelength of the ultraviolet that the ultraviolet mutagenesis operation is adopted is for 253.7 nanometers, described to state ultraviolet photograph
Penetrate operation persistent period be 2 minutes～24 minutes, can make mucor strain orthomutation it is reproducible.And for example, adopt
Ultraviolet mutagenesis case, carries out the ultraviolet mutagenesis operation, and specifically, and for example, the ultraviolet case is provided with uviol lamp, the uviol lamp
Power be 15 watts, the wavelength of the ultraviolet that the uviol lamp sends is 253.7 nanometers, in such manner, it is possible to further make Mucor
The orthomutation of strain it is reproducible.
In one embodiment, the ultra-vioket radiation operation persistent period is 2 minutes, 2.5 minutes, 3 minutes, 3.5 minutes, 4
Minute, 4.5 minutes, 5 minutes, 5.5 minutes, 6 minutes, 6.5 minutes, 7 minutes, 7.5 minutes, 8 minutes, 8.5 minutes, 9 minutes,
9.5 minutes, 10 minutes, 10.5 minutes, 11 minutes, 11.5 minutes, 12 minutes, 12.5 minutes, 13 minutes, 13.5 minutes, 14 points
Clock, 14.5 minutes, 15 minutes, 15.5 minutes, 16 minutes, 16.5 minutes, 17 minutes, 17.5 minutes, 18 minutes, 18.5 minutes,
19 minutes, 19.5 minutes, 20 minutes, 20.5 minutes, 21 minutes, 21.5 minutes, 22 minutes, 22.5 minutes, 23 minutes, 23.5 points
Clock or 24 minutes.And for example, the ultra-vioket radiation operation execution time is 2 minutes～24 minutes, and for example, the ultra-vioket radiation operation
The execution time is 4 minutes～16 minutes, and and for example, the ultra-vioket radiation operation execution time is 8 minutes～12 minutes, and for example, institute
It is 9 minutes～11 minutes to state ultra-vioket radiation and operate the execution time, and and for example, the ultra-vioket radiation operation execution time is 10 minutes,
In such manner, it is possible to the Mucor sufu strain that spore negligible amounts are produced in production of preserved beancurd is obtained, and it is reproducible.
The fermented bean curd hair that spore negligible amounts are produced in production of preserved beancurd obtain after ultraviolet mutagenesis operation in order to further be improved
The quantity of mould species, for example, when the ultraviolet mutagenesis operation is carried out, is also stirred operation to the bacteria suspension, and for example, institute
State stirring operation and operate for magnetic agitation, and for example, the agitation revolution of magnetic agitation operation is 100 revs/min～150 turns/
Minute, and for example, the agitation revolution of the magnetic agitation operation is 120 revs/min～140 revs/min, and and for example, the magnetic force is stirred
The agitation revolution for mixing operation is 130 revs/min, in such manner, it is possible to the reproducible of the orthomutation of mucor strain is further made,
Can also further improve and the Mucor sufu strain that spore negligible amounts are produced in production of preserved beancurd is obtained after ultraviolet mutagenesis operation
Quantity.
In one embodiment, in ultra-vioket radiation operation, the bacteria suspension is carried out in being positioned over the culture dish of opening
The ultra-vioket radiation operation, and for example, a diameter of 80mm of the culture dish, and for example, the bacteria suspension is positioned over the culture dish
In volume be 10 milliliters, and for example, the culture dish be glass culture dish, in such manner, it is possible to further make determining for mucor strain
To the reproducible of mutation, the mutagenesis screening method of the Mucor sufu strain can be made relatively stable, that is to say, that can be compared with
Stably to obtain the Mucor sufu strain that spore negligible amounts are produced in production of preserved beancurd.
S140：Using culture medium to the bacteria suspension operated through the ultraviolet mutagenesis is carried out preliminary culture operation,
Mucor strain bacterium colony is obtained, wherein, under 15 DEG C～20 DEG C temperature conditionss, carry out the preliminary culture operation.
, obtain to the bacteria suspension operated through the ultraviolet mutagenesis is carried out preliminary culture operation by using culture medium
To mucor strain bacterium colony, wherein, under 15 DEG C～20 DEG C temperature conditionss, the preliminary culture operation is carried out, so that through described
Mucor in the bacteria suspension of ultraviolet mutagenesis operation recovers from unfavorable ultraviolet environments.
In one embodiment, by after the bacteria suspension that the ultraviolet mutagenesis are operated is coated in culture medium, then enter
The row preliminary culture operation, to filter out the single bacterium colony after ultraviolet mutagenesis, is easy to follow-up screening and identification.By by purple
The bacteria suspension after external exposure carries out the preliminary culture operation in coating culture medium, so as to grasp through the ultraviolet mutagenesis
Mucor in the bacteria suspension made recovers from unfavorable ultraviolet environments, meanwhile, by grasping through the ultraviolet mutagenesis
The bacteria suspension made is coated, and can filter out the single bacterium colony after ultraviolet mutagenesis, be easy to follow-up screening and identification.
In one embodiment, the culture medium is Rhizoma Solani tuber osi culture medium.It should be noted that Rhizoma Solani tuber osi culture specified herein
Base is consistent with Rhizoma Solani tuber osi culture medium above, that is, consistent with the Rhizoma Solani tuber osi culture medium in step S110, is not repeating secondary.
In one embodiment, the Rhizoma Solani tuber osi culture medium contains each component of following mass parts：180 parts～220 parts of Rhizoma Solani tuber osi, Portugal
18 parts～22 parts of grape sugar, 15 parts～20 parts of agar, 0.2 part～0.6 part of leucine, 0.1 part～0.22 part of arginine, isoleucine
950 parts～1050 parts of 0.15 part～0.27 part, 0.12 part～0.38 part of L-Valine and water.Above-mentioned Rhizoma Solani tuber osi culture medium is using as above matter
The each component of amount part, can make the growth of Mucor more stable, by adding 0.2 part～0.6 part of leucine, isoleucine 0.15
Part～0.27 part, 0.12 part～0.38 part of L-Valine, the Mucor after mutation can be made to reduce the free radical chain caused because of ultraviolet
Formula reacts the injury to Mucor cell, enables the fast quick-recovery of the Mucor after mutation to enter trophophase, so as to reduce after mutation
The adaptive time of Mucor.By add 0.1 part～0.22 part of arginine, and with 0.2 part～0.6 part of leucine, isoleucine
0.15 part～0.27 part, 0.12 part～0.38 part of L-Valine is when being used together, it is possible to increase the success rate of Mucor mutation, reduce hair
The mould mortality rate after ultra-vioket radiation mutation.Above-mentioned Rhizoma Solani tuber osi culture medium can make Mucor using each component of as above mass parts
Growth is more stable, moreover it is possible to reduce the adaptive time of Mucor after mutation, it is possible to increase the success rate of Mucor mutation, reduces Mucor and exists
Mortality rate in ultra-vioket radiation mutation.
In one embodiment, the persistent period of the preliminary culture operation is 1 day～4 days, and for example, the preliminary culture behaviour
The persistent period of work is 2 days～4 days.
S150：The mucor strain bacterium colony is carried out the operation of high temperature screening and culturing and low temperature screening and culturing behaviour respectively simultaneously
Make, wherein, under 24 DEG C～28 DEG C temperature conditionss, the high temperature screening and culturing operation is carried out, in 15 DEG C～20 DEG C temperature conditionss
Under, carry out the low temperature screening and culturing operation.For example, by the mucor strain bacterium colony carry out respectively the operation of high temperature screening and culturing and
Low temperature screening and culturing is operated, wherein, under 24 DEG C～28 DEG C temperature conditionss, the high temperature screening and culturing operation is carried out, at 15 DEG C
Under～20 DEG C of temperature conditionss, the low temperature screening and culturing operation is carried out.And for example, the mucor strain bacterium colony is carried out into high temperature simultaneously
Screening and culturing is operated and the operation of low temperature screening and culturing, wherein, under 24 DEG C～28 DEG C temperature conditionss, carry out the high temperature screening training
Operation is supported, under 15 DEG C～20 DEG C temperature conditionss, the low temperature screening and culturing operation is carried out.
By the mucor strain bacterium colony to be carried out the operation of high temperature screening and culturing and the operation of low temperature screening and culturing respectively simultaneously,
Wherein, under 24 DEG C～28 DEG C temperature conditionss, the high temperature screening and culturing operation is carried out, under 15 DEG C～20 DEG C temperature conditionss,
Carry out low temperature screening and culturing operation, can filter out at 15 DEG C～20 DEG C well-grown, produce spore it is normal and at 24 DEG C
Spore is not produced at～28 DEG C or sporogenic Mucor sufu strain is produced less such that it is able to the glass crystal in fermented bean curd is reduced, to carry
The quality of high fermented bean curd.
It should be noted that normal temperature fermentation in the production of fermented bean curd, is typically carried out, and southern weather, such as Guangdong, year
Temperature is usually above 20 degrees Celsius, more with 24 DEG C～more than 28 DEG C of temperature.And pass through to filter out at 15 DEG C～20 DEG C
Well-grown, product spore normally do not produce spore at 24 DEG C～28 DEG C or produce sporogenic Mucor sufu strain less, Neng Gou
The glass crystal in fermented bean curd is reduced in the production process of the room temperature of fermented bean curd, to improve the quality of fermented bean curd.Certainly, northern temperature is relatively low
When, the strain for filtering out can also pass through the fermentation temperature for adjusting fermented bean curd at 24 DEG C～28 DEG C, reduce in north production fermented bean curd
Glass crystal, to improve the quality of fermented bean curd.
In one embodiment, the mucor strain bacterium colony is put respectively and is connected to two inventionculture tools equipped with the culture medium
In after, carry out high temperature screening and culturing operation and low temperature screening and culturing operation at the same time, that is to say, that first culture
Equipment is cultivated at 15 DEG C～20 DEG C, and second inventionculture tools is cultivated at 24 DEG C～28 DEG C.It should be noted that the point connects operation
It is that each mucor strain bacterium colony is inoculated in two inventionculture tools equipped with the culture medium respectively, that is to say, that Mei Yimao
Mould species bacterium colony two culture medium equipment of correspondence, identify with this whether mucor strain bacterium colony can be during production of preserved beancurd
Spore is produced less or does not produce spore.And for example, the inventionculture tools are culture dishs.Certainly, the inventionculture tools can also be other trainings
Foster utensil, as long as Mucor single bacterium colony growth wherein can be made.
In one embodiment, the persistent period of the high temperature screening and culturing operation is 1 day～4 days, the low temperature screening training
The persistent period for supporting operation is 1 day～4 days, and and for example, the persistent period of the high temperature screening and culturing operation is 2 days～4 days, described
The persistent period of low temperature screening and culturing operation is 2 days～4 days, and for example, under 26 DEG C～28 DEG C temperature conditionss, carries out the high temperature
Screening and culturing is operated, and under 15 DEG C～17 DEG C temperature conditionss, carries out the low temperature screening and culturing operation, and for example, warm at 27.5 DEG C
Under the conditions of degree, the high temperature screening and culturing operation is carried out, under 15.5 DEG C of temperature conditionss, carry out the low temperature screening and culturing behaviour
Make, in such manner, it is possible to filter out well-grown, product spore at 15 DEG C～20 DEG C normally not produce at 24 DEG C～28 DEG C
Spore produces sporogenic Mucor sufu strain less, and strain stability is preferable such that it is able to reduce the glass crystal in fermented bean curd, with
Improve the quality of fermented bean curd.
It should be noted that in high temperature screening and culturing operation and low temperature screening and culturing operation, the training of employing
Foster base is consistent with the culture medium mentioned in S110.
S160：Judge whether the mucor strain bacterium colony after the high temperature screening and culturing is operated meets high temperature and preset
Spore situation is produced, is then execution step S170, wherein, the default spore situation of producing of the high temperature is that every gram of thallospore number is less than 104Individual/
Gram.
Preset by judging whether the mucor strain bacterium colony after the high temperature screening and culturing is operated meets high temperature
Spore situation is produced, is then execution step S170, wherein, the default spore situation of producing of the high temperature is that every gram of thallospore number is less than 104Individual/
Gram, do not produce in being chosen at the high temperature screening and culturing operation or produce sporogenic mucor strain bacterium colony alternately bacterium less
Kind such that it is able to the glass crystal in fermented bean curd is reduced, to improve the quality of fermented bean curd.Otherwise, when through the high temperature screening and culturing
When the mucor strain bacterium colony after operation does not meet that high temperature is default to produce spore situation, then given up.
S170, judge through the low temperature screening and culturing operate after the mucor strain bacterium colony whether meet low temperature preset
Spore situation is produced, and is the mucor strain bacterium colony to be chosen for the Mucor sufu strain, wherein, the low temperature is default to produce spore situation
It is that every gram of thallospore number is more than 108Individual/gram.
Preset by judging whether the mucor strain bacterium colony after the low temperature screening and culturing is operated meets low temperature
Spore situation is produced, and is the mucor strain bacterium colony to be chosen for the Mucor sufu strain, wherein, the low temperature is default to produce spore situation
It is that every gram of thallospore number is more than 108Individual/gram, do not produce in being chosen at the high temperature screening and culturing operation or produce spore less
The mucor strain bacterium colony of son is used as strain such that it is able to reduce the glass crystal in fermented bean curd, to improve the quality of fermented bean curd.
And for example, exchange the execution sequence of step S160 and step S170.
Certainly, in S160 can also be：Judge that the mucor strain bacterium colony after the high temperature screening and culturing is operated is
It is no to meet the default product spore situation of high temperature, the first judged result is obtained, wherein, the default spore situation of producing of the high temperature is every gram of mycelia spore
Subnumber is less than 104Individual/gram；In S170 can also be：Judge the mucor strain bacterium after the low temperature screening and culturing is operated
Fall to whether meeting the default product spore situation of low temperature, obtain the second judged result, wherein, the default spore situation of producing of the low temperature is every gram of bacterium
Silk spore count is more than 108Individual/gram；First judged result is with second judged result when being, then choose the hair
Mould species bacterium colony is the Mucor sufu strain.
In one embodiment, described selection mucor strain bacterium colony, as Mucor sufu strain, can both be to choose through institute
The bacterium colony after the operation of low temperature screening and culturing is stated as Mucor sufu strain, is alternatively chosen and is grasped through the high temperature screening and culturing
Bacterium colony after work can also obtain mucor strain bacterium colony as Mucor sufu to choose after preliminary culture operation as strain
Strain.In view of first two bacterium colony is all obtained by obtaining mucor strain bacterium colony culture after preliminary culture operation, it is therefore contemplated that it
Be same bacterial strain.
Preferably, choose mucor strain bacterium colony is obtained as Mucor sufu strain, so, energy after preliminary culture operation
The stability of Mucor sufu strain is improved enough.In other words, step S150, step S160 and step S170 are training after step S140
The authentication step of foster mucor strain bacterium colony.
Wherein in one embodiment, Fig. 2 is referred to, also included after step S170：
S180：The Mucor sufu strain is carried out into 5 Secondary Cultures, each Secondary Culture repeat step S150,
Step S160, step S170.
The strain of mutation after unfavorable environment is eliminated is susceptible to back mutation, is unfavorable for the production of fermented bean curd.In order to
The preferable Mucor sufu strain of stability is obtained, by the Mucor sufu strain is carried out 5 Secondary Cultures, training is passed on every time
Support and repeat step S150, S160, step S170, in such manner, it is possible to obtain the preferable Mucor sufu strain of stability.And for example,
Once the Secondary Culture time is 1 day～5 days, and for example, the incubation time phase one of the Secondary Culture time and step S150
Cause, in such manner, it is possible to obtain to obtain the preferable Mucor sufu strain of stability, that is to say, that filter out and pass on training at least five times
The strain of back mutation will not occur in supporting, i.e., well-grown, product spore are normally and at 24 DEG C～28 DEG C at 15 DEG C～20 DEG C
Under do not produce spore or produce sporogenic Mucor sufu strain less.
Further, and for example, also include after S180 steps：The Mucor sufu bacterium cultivated at choosing 15 DEG C～20 DEG C
Kind, with normal spawn culture program culture spore bacterium solution, bacterium, fermentation are sprayed, and makes fermented bean curd, the Jing multiple batches of Different climate times
Section test, picking up product do not produce the excellent species of glass crystallization body, in such manner, it is possible to obtain stably more preferably, being more suitable for production should
Excellent species.
It should be noted that normal spawn culture program culture spore bacterium solution, sprays bacterium, fermentation, and makes the tool of fermented bean curd
Body flow process is referring to prior art.
The mutagenesis screening method of above-mentioned Mucor sufu strain can mutation go out at 15 DEG C～20 DEG C well-grown, produce spore
Normally spore is not produced at 24 DEG C～28 DEG C or sporogenic Mucor sufu strain is produced less, can be produced in the production of fermented bean curd
Less spore such that it is able to reduce the glass crystal in fermented bean curd, improves the quality of fermented bean curd.It is more particularly suitable for southern room temperature fermented bean curd
Production.
The present invention also provides a kind of production method of fermented bean curd, sieves including the mutation by as above arbitrary Mucor sufu strain
Mucor sufu strain prepared by choosing method is seeded on fermented bean curd embryo, prepares fermented bean curd.
It should be noted that existing production of preserved beancurd process is as follows：Soybean soaking, mill bean, bean dreg separation, cooked beans slurry
(or boiled), selects slurry solidification, squeezing and water (bean curd embryo), stripping and slicing (fermented bean curd embryo), row's block are removed to fermenting
Frame, spray bacterium, primary fermentation, rub that hair, the salt that salts down, bubble breast desalt, dewatering and flavouring agent or coloring juice mix (can save) with the hands, bottle (or its
Its container), plus decoction or wine etc., after fermentation storage, wash bottle body, change lid, labeling, vanning.Spray on the good endosperm embryo of stripping and slicing, row's block
Fermenting house is just sent to after bacterium and removes primary fermentation.After the completion of primary fermentation, thick Mucor on fermented bean curd embryo, is grown, used for rear operation.
The more detailed production technology of fermented bean curd refers to prior art, and the present invention will not be described here.
In the production method of the fermented bean curd of the present embodiment, selection-breeding well-grown at 15 DEG C～20 DEG C produces spore, and
26-30 DEG C does not produce or produces less sporogenic bacterial strain when being cultivated, the formation number of Mucor spore when greatly reducing fermentation
Amount so as to from the 10 of opportunistic pathogen spore count in production fermentation8It is individual/gram more than, drop to 104It is individual/gram below, meanwhile, substantially product
In there is no the appearance of glass crystallization body.
It is understood that in the production process of fermented bean curd, strain configuration be generally all by manually carrying out, will cultured hair
Mould species are washed with water, and are diluted to finite concentration, form mucormycosiss spore suspension, are used in production.Due to existing corruption
Newborn mucor strain spore suspension standing time during spray bacterium is long, and easy microbiological contamination is totally unfavorable to the production technology of fermented bean curd.
In order to solve the above problems, for example, it is described that the operation that Mucor sufu strain is seeded on fermented bean curd embryo is specifically included into following steps：
The Mucor sufu strain is mixed with yeast, mixed bacteria liquid is prepared；The mixed bacteria liquid is seeded to into the fermented bean curd
On embryo, so, by adding yeast liquid, yeast can be made to form good symbiosiss with Mucor sufu strain, effectively
Ground suppresses the growth of other miscellaneous bacterias, moreover it is possible to improves nutrition and the quality of fermented bean curd, can make mixed bacteria liquid in 7 hours～8 for using
During hour, it is greatly lowered total number of bacteria, reduces the gradient of infection of antibacterial.
And for example, the mixed liquor includes Mucor sufu strain suspension and yeast bacterium suspension, in the yeast bacteria suspension
Saccharomycetic concentration is 107It is more than individual/milliliter, the mass ratio of the Mucor sufu strain suspension and the yeast bacterium suspension
(95～105)：1, so, by the yeast bacteria suspension for adding aforementioned proportion, yeast can be made to be formed with Mucor sufu strain
Good symbiosiss, effectively suppress the growth of other miscellaneous bacterias, moreover it is possible to improve nutrition and the quality of fermented bean curd, can make mixed vaccine
Liquid is in 7 hours for using～during 8 hours, make total number of bacteria by original 105More than individual/milliliter, it is reduced to 103Individual/milli
Rise following, fall below significantly the gradient of infection of antibacterial.And for example, the yeast is Lu Shi yeast.And for example, the Mucor sufu bacterium
Plant the mass ratio 99 of bacteria suspension and yeast bacteria suspension：1, in such manner, it is possible to further suppress the growth of other miscellaneous bacterias in mixed bacteria liquid.
And for example, the Mucor sufu bacteria suspension is obtained by the following method：By the Mucor sufu strain sterilized water
Make Mucor sufu strain bacteria suspension.
In order to improve the saccharomycetic activity, for example, the yeast bacteria suspension is obtained by the following method：Use beerwort
The yeast is cultivated by culture medium, makes yeast cell number reach 107More than individual/milliliter, yeast bacteria suspension is obtained final product, this
Sample, it is possible to increase saccharomycetic activity, can further suppress the growth of other miscellaneous bacterias in mixed bacteria liquid.
The production method of above-mentioned fermented bean curd, can reduce the content of the glass crystal in fermented bean curd, improve the quality of fermented bean curd so that
The mouthfeel of fermented bean curd is all good.
The present invention also provides a kind of fermented bean curd, adopts and obtains prepared by the as above production method of arbitrary fermented bean curd.
The fermented bean curd that the production method of above-mentioned fermented bean curd is prepared, can reduce the content of the glass crystal in fermented bean curd, improve
The quality of fermented bean curd so that the mouthfeel of fermented bean curd is all good.
Provide a specific embodiment below again, the present invention will be described for continuation.
Embodiment 1：
A kind of mutagenesis screening method of Mucor sufu strain, comprises the steps：
(1) strain that, sets out is Mucor mucedo strain.Selectively, the characteristic of its opportunistic pathogen is determined, as at different temperatures
The characteristic of the opportunistic pathogens such as spore growth situation, prolease activity, the speed of growth.
(2), using Rhizoma Solani tuber osi culture medium, purification is carried out to Mucor mucedo strain, and is cultivated 48 hours, treat the plentiful standby of spore
With.
(3), adopt 0.01% sodium dodecyl sulfate solution by Mucor mucedo strain make spore concentration for (0.1～
9.99)×106The spore bacteria suspension of individual/milliliter；Specially：With 0.01% sodium dodecyl sulfate solution spore bacteria suspension 15
Milliliter, is put in triangular flask, and adds bead, and concussion breaks up spore.Filtered to equipped with 30 milliliter 0.01% with absorbent cotton
In sodium dodecyl sulfate solution, spore concentration is made to be (0.1～9.99) × 106Individual/milliliter, standby, as spore bacteria suspension.
(4) 10 milliliters of spore bacteria suspensions, are taken in a diameter of 90 millimeters of culture dish, is made and multiple is loaded with spore suspension
Culture dish.
(5), each culture dish places bar magnet respectively, the culture dish placed after bar magnet is uncapped and is positioned over mutation case
On magnetic stirring apparatuss, the use of power is 15W, wavelength is irradiated for the uviol lamp of 253.7nm, adjustment stir speed (S.S.) is 100～
150 revs/min, ultra-vioket radiation obtained each treatment fluid after 4,6,8,10,12,16 minutes to each culture dish respectively；1 milliliter of process is taken respectively
Liquid, Jing suitably dilute (make every flat board have 10～12 bacterium colonies to be advisable), take 0.1 milliliter of coating culture, cultivate at 15 DEG C～20 DEG C
2-4 days.
(6), primary election, by the single mucormycosiss of above grown on flat dishes, each bacterium selects respectively and is connected to two Rhizoma Solani tuber osi culture medium
On plate, one is cultivated at 15 DEG C～20 DEG C, and another is cultivated at 24 DEG C～28 DEG C, incubation time 2-4 days,
The well-grown at 15 DEG C～20 DEG C is selected, spore is produced normally, and is cultivated at 24 DEG C～28 DEG C, do not produce or produce less
The strain of spore is used as Mucor sufu strain.
(7), by the strain of above-mentioned selection, more than 5 times Secondary Cultures are carried out, and carries out (6) identical viewing test, screened
The strain of growth and stability of characteristics is used as stablizing Mucor sufu strain.
Select 12 plants of Low- temperature cultures normal, i.e., every gram thallospore number is more than 108Individual/gram, and 24 DEG C～28 DEG C are carried out
During culture, every gram of thallospore number is less than 104It is individual/gram below bacterial strain.
(8) pilot production, is carried out, with the strain cultivated at 15 DEG C～20 DEG C, with normal spawn culture program culture spore
Daughter bacteria liquid, sprays bacterium, fermentation, and makes fermented bean curd, and the multiple batches of Different climate time period tests of Jing, picking up product do not produce glass crystallization
The excellent species of body.
In the production and using method of the Mucor sufu strain of the present embodiment, selection-breeding grows good at 15 DEG C～20 DEG C
It is good, spore is produced, is cultivated at 26-30 DEG C, is not produced or produce less sporogenic bacterial strain, Mucor spore when greatly reducing fermentation
The quantity of formation of son so as to from the 10 of opportunistic pathogen spore count in production fermentation8It is individual/gram more than, drop to 104It is individual/gram below, meanwhile,
Substantially there is no the appearance of glass crystallization body in product.
In the present embodiment, the bacterial strain of the Mucor sufu strain obtained after 8 minutes～12 minutes time of ultra-vioket radiation is more,
Its strain stability is preferable.
It should be noted that " part " of the respective embodiments described above including kilogram, gram, milligram, rise and the measurement unit such as milliliter,
And in each embodiment, the implication that described " part " represents is identical or different.
Embodiment described above only expresses the several embodiments of the present invention, and its description is more concrete and detailed, but and
Therefore the restriction to the scope of the claims of the present invention can not be interpreted as.It should be pointed out that for one of ordinary skill in the art
For, without departing from the inventive concept of the premise, some deformations and improvement can also be made, these belong to the guarantor of the present invention
Shield scope.Therefore, the protection domain of patent of the present invention should be defined by claims.
Claims (10)
1. a kind of mutagenesis screening method of Mucor sufu strain, it is characterised in that comprise the steps：
Mucor strain to be selected is made into bacteria suspension；
Ultraviolet mutagenesis operation is carried out to the bacteria suspension, wherein, the persistent period of the ultraviolet mutagenesis operation is 2 minutes～24 points
Clock；
Under 15 DEG C～20 DEG C temperature conditionss, using culture medium to the bacteria suspension operated through the ultraviolet mutagenesis is carried out
Preliminary culture operation, obtains mucor strain bacterium colony；
The mucor strain bacterium colony is carried out the operation of high temperature screening and culturing and the operation of low temperature screening and culturing respectively simultaneously, wherein, institute
Stating the operation of high temperature screening and culturing is carried out under 24 DEG C～28 DEG C temperature conditionss, and the low temperature screening and culturing is operated at 15 DEG C～20 DEG C
Carry out under temperature conditionss；
Judge whether the mucor strain bacterium colony after the high temperature screening and culturing is operated meets the default product spore situation of high temperature,
The first judged result is obtained, wherein, the default spore situation of producing of the high temperature is that every gram of thallospore number is less than 104Individual/gram；
Judge whether the mucor strain bacterium colony after the low temperature screening and culturing is operated meets the default product spore situation of low temperature,
The second judged result is obtained, wherein, the default spore situation of producing of the low temperature is that every gram of thallospore number is more than 108Individual/gram；
First judged result is with second judged result when being, then choose the mucor strain bacterium colony for the corruption
Newborn mucor strain.
2. the mutagenesis screening method of Mucor sufu strain according to claim 1, it is characterised in that the culture medium is soil
Bean culture medium.
3. the mutagenesis screening method of Mucor sufu strain according to claim 1, it is characterised in that institute in the bacteria suspension
The spore concentration for stating mucor strain to be selected is (0.1～9.99) × 106Individual/milliliter.
4. the mutagenesis screening method of Mucor sufu strain according to claim 1, it is characterised in that adopt ultraviolet mutagenesis
Case, carries out the ultraviolet mutagenesis operation.
5. the mutagenesis screening method of Mucor sufu strain according to claim 4, it is characterised in that the ultraviolet case is arranged
There is uviol lamp, the power of the uviol lamp is 15 watts, the wavelength of the ultraviolet that the uviol lamp sends is 253.7 nanometers.
6. the mutagenesis screening method of Mucor sufu strain according to claim 1, it is characterised in that using 0.01% 12
The mucor strain to be selected is made bacteria suspension by sodium alkyl sulfate solution.
7. the mutagenesis screening method of Mucor sufu strain according to claim 1, it is characterised in that carry out it is described ultraviolet
When mutation is operated, operation is stirred to the bacteria suspension also.
8. a kind of production method of fermented bean curd, it is characterised in that include the Mucor sufu as any one of claim 1 to 7
Mucor sufu strain obtained by the mutagenesis screening method of strain is seeded on fermented bean curd embryo, prepares fermented bean curd.
9. production method according to claim 8, it is characterised in that described that Mucor sufu strain is seeded on fermented bean curd embryo
Operation specifically include following steps：
The Mucor sufu strain is mixed with yeast, mixed bacteria liquid is prepared；
The mixed bacteria liquid is seeded on the fermented bean curd embryo.
10. a kind of fermented bean curd, it is characterised in that using such as claim 8 and Mucor sufu strain any one of claim 9
Production method prepare.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107873857A (en) * | 2017-11-22 | 2018-04-06 | 桂林国农生态农业有限公司 | The manufacture craft of low-salt flavor fermented bean curd |
| CN107889894A (en) * | 2017-11-22 | 2018-04-10 | 桂林国农生态农业有限公司 | The manufacture craft of low-salt fermented bean curd |
| CN108740062A (en) * | 2018-08-20 | 2018-11-06 | 江南大学 | A kind of preparation method of the oil preserved bean curd of the high yield fragrance of a flower and fruity |
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2016
- 2016-12-13 CN CN201611147411.9A patent/CN106591280A/en active Pending
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107873857A (en) * | 2017-11-22 | 2018-04-06 | 桂林国农生态农业有限公司 | The manufacture craft of low-salt flavor fermented bean curd |
| CN107889894A (en) * | 2017-11-22 | 2018-04-10 | 桂林国农生态农业有限公司 | The manufacture craft of low-salt fermented bean curd |
| CN108740062A (en) * | 2018-08-20 | 2018-11-06 | 江南大学 | A kind of preparation method of the oil preserved bean curd of the high yield fragrance of a flower and fruity |
| CN108740062B (en) * | 2018-08-20 | 2021-10-22 | 江南大学 | Preparation method of oil fermented bean curd with high flowery flavor and fruit flavor yield |
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Application publication date: 20170426 |