CN109439583A - The microbial inoculum and fermentation process of composite fermentation papainase - Google Patents
The microbial inoculum and fermentation process of composite fermentation papainase Download PDFInfo
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Abstract
The present invention relates to the microbial inoculum of composite fermentation papainase and fermentation process, belong to ferment preparation technical field.The microbial inoculum of composite fermentation papainase, which is characterized in that the microbial inoculum includes lactobacillus plantarum BC299, Bayer Zygosaccharomyces Y4 and the wooden coltfoal shape bacillus C82.Bacteria fermentation acid producing ability provided by the invention is strong, can greatly shorten fermentation time, and is able to suppress harmful bacteria growth, ferments more stable, obtained papainase is with rich flavor.
Description
Technical field
The present invention relates to ferment preparation technical fields, and in particular to the microbial inoculum of composite fermentation papainase and fermentation side
Method.
Background technique
Ferment is with animal, plant, mushroom etc. for raw material, adds or does not add auxiliary material, through containing made from microbial fermentation
There is the product of particular organisms active constituent.According to fermenting microbe difference, pure-blood ferment ferment can be divided into, group plants fermentation ferment and multiple
Close fermentation ferment.Pure-blood ferment ferment refers to the product as made from the microbial fermentation for having clear specific name manually cultivated,
Group's kind fermentation ferment refers to that the product as made from the specific microbial fermentation in food materials surface, composite fermentation ferment refer to purebred hair
Ferment and group plant two kinds of techniques of fermentation ferment product obtained jointly.Composite fermentation mode, which had both been utilized under spontaneous fermentation state, to be enriched
Microbiologic population, further through access beneficial bacterium promote fermentation.Composite fermentation mode fermentation process relatively easily controls, and is better than it
His two kinds of zymotechniques.
Papaya contains the minerals such as mineral calcium, magnesium, phosphorus, potassium, and vitamin also rich in is especially enriched in β Hu trailing plants
Bu Su.Oleanolic acid ingredient contained by mature papaya pulp has protect liver decompression and other effects.Papaya is full of nutrition, taste
Road is fresh and sweet, and the converted products of papaya is less in existing market, is even more more rare in terms of ferment product development.
Summary of the invention
The purpose of the present invention is to provide the microbial inoculum of composite fermentation papainase and fermentation process.Bacterium provided by the invention
Agent fermentation and acid ability is strong, can greatly shorten fermentation time, and is able to suppress harmful bacteria growth, ferments more stable, obtains
Papainase is with rich flavor.
The present invention provides the microbial inoculum of composite fermentation papainase, the microbial inoculum includes lactobacillus plantarum BC299, Bayer
The Zygosaccharomyces Y4 and wooden coltfoal shape bacillus C82；
The deposit number of the lactobacillus plantarum BC299 is CGMCC6326；
The deposit number of the Bayer Zygosaccharomyces Y4 is CGMCC16183；
The deposit number of the wood coltfoal shape bacillus C82 is CGMCC1618.
Preferably, in the microbial inoculum lactobacillus plantarum BC299, Bayer Zygosaccharomyces Y4 and the wooden coltfoal shape bacillus C82 it is dense
Degree is than being (1~10): (1~10): (1~10).
The present invention also provides the methods that the bacteria fermentation prepares papainase based on the above-mentioned technical proposal, including with
Lower step:
1) papaya is successively tiled with white granulated sugar, obtains fermentation raw material；
2) microbial inoculum as claimed in claim 1 or 2 of activation is inoculated in the step 1) fermentation raw material, 30 DEG C of fermentations 30
It, obtains papainase.
Preferably, the step 1) papaya is cleaning, removes the papaya after son and slice.
Preferably, the mixing mass ratio of the step 1) papaya and white granulated sugar is (2~4): (1~3).
Preferably, the amount of the step 2) inoculation is 4~10%.
Preferably, the step 2) activation is to lactobacillus plantarum BC299, Bayer Zygosaccharomyces Y4 and the wooden coltfoal shape bacillus
Bacterium C82 is activated respectively.
Preferably, the step 2) activation method are as follows:
Lactobacillus plantarum BC299 is inoculated in MRS solid medium, 37 DEG C of 48~72h of fermentation, the inoculation of picking single colonie
It is expanded culture in MRS fluid nutrient medium, the lactobacillus plantarum BC299 after being activated；
Bayer Zygosaccharomyces Y4 is inoculated in YPD solid medium, 28 DEG C of 24~48h of fermentation, the inoculation of picking single colonie
It is expanded culture in YPD fluid nutrient medium, the Bayer Zygosaccharomyces Y4 after being activated.
The wooden coltfoal shape bacillus C82 is inoculated in bromocresol purple solid medium, 30 DEG C of 48~72h of fermentation, picking single colonie
It is inoculated in bromocresol purple fluid nutrient medium and expands culture, the wooden coltfoal shape bacillus C82 after being activated.
The present invention provides the microbial inoculums of composite fermentation papainase.Microbial inoculum provided by the invention combines three kinds of advantages
Bacterium keeps fermentation and acid ability stronger, shortens fermentation time, and acidic environment effectively inhibits the growth and breeding of harmful bacteria,
Keep papainase safer；The present invention also makes fermentation process and fermentation end products index more steady by the use of above-mentioned microbial inoculum
It is fixed；The product that bacteria fermentation obtains through the invention does not add any food additives, can obtain the health care food of green, health
Product.Test result shows that the papainase acidity obtained through microbial inoculum of the present invention can reach 15.2gkg-1, pH value 3.5.
Detailed description of the invention
Fig. 1 is the result of variations figure for the papainase fermentation process pH that the embodiment of the present invention 4 provides；
Fig. 2 is the result of variations figure for the papainase fermentation process total acid content that the embodiment of the present invention 4 provides.
Preservation explanation
Lactobacillus plantarum BC299, Latin literary fame were known as Lactobacillus plantarum, on 07 10th, 2012
It is deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, address is BeiChen West Road, Chaoyang District, BeiJing City 1
Number institute 3, Institute of Microorganism, Academia Sinica, biological deposits number is CGMCC No.6326；
Bayer Zygosaccharomyces Y4, Latin literary fame were known as Zygosaccharomyces bailii, on 07 30th, 2018
It is deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, address is BeiChen West Road, Chaoyang District, BeiJing City 1
Number institute 3, Institute of Microorganism, Academia Sinica, biological deposits number is CGMCC No.16183；
The wooden coltfoal shape bacillus C82, Latin literary fame are known as Komagataeibacter xylinus, protect on 07 30th, 2018
China Committee for Culture Collection of Microorganisms's common micro-organisms center is ensconced, address is BeiChen West Road, Chaoyang District, BeiJing City 1
Institute 3, Institute of Microorganism, Academia Sinica, biological deposits number is CGMCC No.16184.
Specific embodiment
The present invention provides the microbial inoculum of composite fermentation papainase, the microbial inoculum includes lactobacillus plantarum BC299, Bayer
The Zygosaccharomyces Y4 and wooden coltfoal shape bacillus C82；
The deposit number of the lactobacillus plantarum BC299 is CGMCC6326；
The deposit number of the Bayer Zygosaccharomyces Y4 is CGMCC16183；
The deposit number of the wood coltfoal shape bacillus C82 is CGMCC16184.
In the present invention, lactobacillus plantarum BC299, Bayer Zygosaccharomyces Y4 and the wooden coltfoal shape bacillus C82 in the microbial inoculum
Concentration is than being preferably (1~10): (1~10): (1~10).
The present invention also provides the methods that the bacteria fermentation prepares papainase based on the above-mentioned technical proposal, including with
Lower step:
1) papaya is successively tiled with white granulated sugar, obtains fermentation raw material；
2) microbial inoculum as claimed in claim 1 or 2 of activation is inoculated in the step 1) fermentation raw material, 30 DEG C of fermentations 30
It, obtains papainase.
The present invention successively tiles papaya with white granulated sugar, obtains fermentation raw material.In the present invention, the papaya is preferred
To clean, going the papaya after son and slice.The present invention is not particularly limited the thickness of the slice, using normal fermentation kind
The thickness being sliced when pawpaw.In the present invention, the mixing mass ratio of the papaya and white granulated sugar is preferably (2~4): (1
~3).The present invention does not have special restriction to the source of the papaya and white granulated sugar, and use is well known to those skilled in the art
Conventional commercial papaya and white granulated sugar.
After obtaining fermentation raw material, microbial inoculum described in above-mentioned technical proposal of the present invention by activation is inoculated in the fermentation raw material
In, 30 DEG C ferment 30~700 days, obtain papainase.In the present invention, the amount of the inoculation is preferably 4~10%, more excellent
It is selected as 6~8%.
In the present invention, the activation is preferably to plant lactobacillus plantarum BC299, Bayer Zygosaccharomyces Y4 and the wooden coltfoal shape bar
Bacterium C82 is activated respectively.
In the present invention, the activation method is preferred are as follows: lactobacillus plantarum BC299 is inoculated in MRS solid medium,
37 DEG C of 48~72h of fermentation, picking single colonie are inoculated in MRS fluid nutrient medium and expand culture, the plant after being activated
Lactobacillus 299；
Bayer Zygosaccharomyces Y4 is inoculated in YPD solid medium, 28 DEG C of 24~48h of fermentation, the inoculation of picking single colonie
It is expanded culture in YPD fluid nutrient medium, the Bayer Zygosaccharomyces Y4 after being activated；
The wooden coltfoal shape bacillus C82 is inoculated in bromocresol purple solid medium, 30 DEG C of 48~72h of fermentation, picking single colonie
It is inoculated in bromocresol purple fluid nutrient medium and expands culture, the wooden coltfoal shape bacillus C82 after being activated.
In the present invention, in the activation lactobacillus plantarum BC299, the composition of the MRS fluid nutrient medium is preferably as follows:
Peptone 1% (w/v), beef extract 1% (w/v), yeast extract 0.5% (w/v), Triammonium citrate 0.2% (w/v), glucose 2%
(w/v), Tween 80 0.1% (w/v), sodium acetate 0.5% (w/v), dipotassium hydrogen phosphate 0.2% (w/v), 0.005% (w/ of manganese sulfate
V), it is obtained after 6.2~6.4,121 DEG C of sterilizing 20min of magnesium sulfate 0.02% (w/v), pH；The MRS solid medium preferably exists
2~2.5% agar powders are added in MRS fluid nutrient medium, are obtained after 121 DEG C of sterilizing 20min.
In the present invention, in the activation Bayer Zygosaccharomyces Y4, the YPD fluid nutrient medium composition is preferably as follows: ferment
Mother's leaching powder 1% (w/v), peptone 2% (w/v), glucose 2% (w/v), pH after 115 DEG C of high pressure sterilization 20min naturally, obtain；
2~2.5% agar powders are added in the YPD solid medium preferably in YPD fluid nutrient medium, after 115 DEG C of sterilizing 20min
It arrives.
In the present invention, in the wooden coltfoal shape bacillus C82 of the activation, the bromocresol purple fluid nutrient medium composition is preferably such as
Under: glucose 1% (w/v), yeast extract 1% (w/v), 1.6% bromocresol purple 4ml/L adjust pH value to 7.2~7.4,115 DEG C
20min sterilizes to obtain；1.5~2% fine jades are added in the bromocresol purple solid medium preferably in bromocresol purple fluid nutrient medium
Rouge obtains after conventional sterilant.
The microbial inoculum and fermentation process of composite fermentation papainase of the present invention are done combined with specific embodiments below
Further details of introduction, technical solution of the present invention include but is not limited to following embodiment.
Embodiment 1
Papaya is selected, clean, is successively laid in sealing in the ratio of 4:1 (w/w) with white granulated sugar after deseeding slice holds
In device.8% (v/v) activated microbial inoculum is accessed, microbial inoculum is lactobacillus plantarum BC299 (CGMCC6326), Bayer Zygosaccharomyces Y4
(CGMCC16183) He Muju shape bacillus C82 (CGMCC16184) is with 1:1:1 ratio combined inoculation.It ferments 30 days, obtains at 30 DEG C
To the papainase.
Any food additives for being used to increase acid are not added into papainase for fermentation process of the present invention, that is, can reach excellent
The papainase of matter, gained papainase acidity is up to 15.2gkg-1, pH value 3.5.
Embodiment 2
Papaya is selected, clean, is successively laid in sealing in the ratio of 4:1 (w/w) with white granulated sugar after deseeding slice holds
In device.6% (v/v) activated microbial inoculum is accessed, microbial inoculum is lactobacillus plantarum BC299 (CGMCC6326), Bayer Zygosaccharomyces Y4
(CGMCC16183) He Muju shape bacillus C82 (CGMCC16184) is with 1:1:1 ratio combined inoculation.It ferments 30 days, obtains at 30 DEG C
To the papainase.
Any food additives for being used to increase acid are not added into papainase for the method for the invention, that is, can reach excellent
The papainase of matter.Gained papainase acidity is up to 14.6gkg-1, pH value 3.6.
Embodiment 3
Papaya is selected, clean, is successively laid in sealing in the ratio of 4:1 (w/w) with white granulated sugar after deseeding slice holds
In device.8% (v/v) activated microbial inoculum is accessed, microbial inoculum is lactobacillus plantarum BC299 (CGMCC6326), Bayer Zygosaccharomyces Y4
(CGMCC16183) He Muju shape bacillus C82 (CGMCC16184) is with 1:1:1 ratio combined inoculation.It ferments 30 days, obtains at 30 DEG C
To the papainase.
Any food additives for being used to increase acid are not added into papainase for fermentation process of the present invention, that is, can reach excellent
The papainase of matter.Gained papainase acidity is up to 10.67gkg-1, pH value 3.88.
The resulting composite fermentation papainase of the present invention has the integrated function of papaya and beneficial bacterial metabolic product,
It is with a wide range of applications and huge economic benefit in the fields such as food fermentation and health care of food.
Embodiment 4
Spontaneous fermentation papainase and present invention fermentation papainase are compared, to pH value in its fermentation process
And acidity is monitored, the result of variations of papainase fermentation process pH is as shown in Figure 1, papainase fermentation process total acid
The result of variations of content is as shown in Figure 2.The result shows that with existing spontaneous fermentation technology as a comparison, utilizing bacteria fermentation of the present invention
Acid producing ability is stronger, and pH is lower.This explanation, the addition of microbial inoculum of the present invention can reinforce fermentation, and the initial stage energy that ferments
PH is down to 4 hereinafter, this just provides good condition for subsequent fermentation, harmful microbe can be inhibited to breed in the early stage, be
Fermentation provides the environmental condition of safety and stability, and beneficial microorganism can be promoted to grow, and accelerates course of fermentation.
Embodiment 5
It using microbial inoculum of the present invention, ferments at different temperatures to papaya respectively, measures different fermentations temperature respectively
The pH value and acidity of lower papainase liquid, the results are shown in Table 1.
Influence of 1 fermentation temperature of table to pH and total acid
As seen from the results in Table 1, check experiment has been made at a temperature of different fermentations, fermentation temperature is too high or too low all to be inhibited
The growth of bacterial strain, and then influence acid producing ability.When fermentation temperature is 30 DEG C, the pH value of papainase liquid reaches 3.48 and total acid
Reach 12.83gkg-1, optimal with other sample comparative effectiveness.Comprehensive check experiment is as a comparison, determine most suitable fermentation temperature
It is 25~40 DEG C.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (8)
1. the microbial inoculum of composite fermentation papainase, which is characterized in that the microbial inoculum includes that lactobacillus plantarum BC299, Bayer connect
Close yeast Y4 and the wooden coltfoal shape bacillus C82；
The deposit number of the lactobacillus plantarum BC299 is CGMCC6326；
The deposit number of the Bayer Zygosaccharomyces Y4 is CGMCC16183；
The deposit number of the wood coltfoal shape bacillus C82 is CGMCC16184.
2. microbial inoculum according to claim 1, which is characterized in that lactobacillus plantarum BC299, Bayer engage ferment in the microbial inoculum
The concentration ratio of female Y4 and the wooden coltfoal shape bacillus C82 are (1~10): (1~10): (1~10).
3. the method for preparing papainase based on bacteria fermentation as claimed in claim 1 or 2, comprising the following steps:
1) papaya is successively tiled with white granulated sugar, obtains fermentation raw material；
2) microbial inoculum as claimed in claim 1 or 2 of activation is inoculated in the step 1) fermentation raw material, 30 DEG C of fermentations 30~700
It, obtains papainase.
4. fermentation process according to claim 3, which is characterized in that the step 1) papaya is cleaning, removes son and cut
Papaya after piece.
5. fermentation process according to claim 3, which is characterized in that the mixing matter of the step 1) papaya and white granulated sugar
Amount is than being (2~4): (1~3).
6. the fermentation process according to claim 3 or 5, which is characterized in that the amount of the step 2) inoculation is 4~10%.
7. fermentation process according to claim 3, which is characterized in that the step 2) activation is to lactobacillus plantarum
BC299, Bayer Zygosaccharomyces Y4 and wood coltfoal shape bacillus bacterium C82 are activated respectively.
8. fermentation process according to claim 3 or 7, which is characterized in that the step 2) activation method are as follows:
Lactobacillus plantarum BC299 is inoculated in MRS solid medium, 37 DEG C of 48~72h of fermentation, picking single colonie is inoculated in
It is expanded culture in MRS fluid nutrient medium, the lactobacillus plantarum BC299 after being activated；
Bayer Zygosaccharomyces Y4 is inoculated in YPD solid medium, 28 DEG C of 24~48h of fermentation, picking single colonie is inoculated in YPD
It is expanded culture in fluid nutrient medium, the Bayer Zygosaccharomyces Y4 after being activated；
The wooden coltfoal shape bacillus C82 is inoculated in bromocresol purple solid medium, 30 DEG C of 48~72h of fermentation, the inoculation of picking single colonie
It is expanded culture in bromocresol purple fluid nutrient medium, the wooden coltfoal shape bacillus C82 after being activated.
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| CN110025014A (en) * | 2019-04-18 | 2019-07-19 | 宝康士(中国)保健食品销售有限公司 | The method of substrate liquid of the hybrid bacterial strain fermentation preparation rich in probiotics and NK cell |
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