CN115074294A - Pickle starter culture and preparation method and application method thereof - Google Patents
Pickle starter culture and preparation method and application method thereof Download PDFInfo
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- C12N1/20—Bacteria; Culture media therefor
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract
The invention discloses a pickle starter culture and a preparation method and an application method thereof. The pickle starter culture is characterized by containing Weissella cibaria AP6 and arabinoxylo-oligosaccharide. The invention takes the grain bran enzymatic hydrolysate as the strain protective agent, is beneficial to the comprehensive utilization of byproducts, and can strengthen the economic development and promote the environmental protection; the grain bran enzymolysis liquid is used as a natural protective agent and an excipient of Weissella sinus AP6, and can keep the survival rate and stability of Weissella sinus AP6 in the processing and storage processes on the premise of not adding other drying protective agents, thereby reducing the production cost; the starter is added in the preparation process of the pickle, and the arabinoxylo-oligosaccharide can be used as a carbon source, so that the activity of heterotypic lactic acid bacteria in the fermentation process of the pickle can be protected, the heterotypic fermentation time can be prolonged, and the pickle is endowed with better flavor.
Description
Technical Field
The invention belongs to the technical field of leavening agents, and particularly relates to a pickle leavening agent as well as a preparation method and an application method thereof.
Background
The cereal bran not only contains abundant vitamins, minerals, proteins and essential amino acids for human body, but also has polysaccharide with high content of araboxylan. But because of the high fiber content, the difficult milling, the rough and bitter taste and other reasons in the cereal bran, the palatability of the bran product is poor, and the consumer acceptance is low. Bran has been mainly used in feed processing industry for a long time, and the utilization rate of food is less than 20%.
At present, the pickle in China is mostly produced by adopting a natural fermentation mode under a high-salt condition, although the direct vat set starter is used for fermentation in a small range to realize artificial inoculation, the production cost is high from the currently developed direct vat set starter, and although the fermentation period of the pickle is shortened, the accumulation of flavor substances of the pickle is not facilitated.
Therefore, the cereal bran is utilized to develop the pickle starter culture with low cost and good flavor, and the method has important economic and social significance.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a pickle starter culture and a preparation method and application thereof, aiming at solving the technical problems of low utilization rate of cereal bran food, high cost of the pickle direct vat set starter culture and less accumulation of flavor substances in the prior art.
In order to achieve the purpose, the invention adopts the following technical scheme:
in a first aspect, the pickle starter culture comprises Weissella sinus AP6 and arabinoxylo-oligosaccharide, wherein the Weissella sinus AP6 is preserved in Guangdong collection of microorganisms with the preservation number GDMCC No62461 and the preservation date 2022, 5 months and 11 days.
Further, the concentration of live bacteria of Weissella sinus AP6 in the pickle starter culture is more than or equal to 10 9 CFU/g, the content of the arabinoxylo-oligosaccharide is 51.22-65.03%.
In a second aspect, the invention provides a preparation method of a pickle starter culture, which comprises the following steps:
(1) preparing Weissella civora AP6 thallus: activating Weissella sinus AP6, inoculating the activated Weissella sinus AP6 into a culture medium for culture, centrifuging the culture medium to obtain Weissella sinus AP6 thallus, wherein the Weissella sinus AP6 is preserved in Guangdong microbiological collection center with the preservation number of GDMCC No. 62461 and the preservation date of 2022, 5 months and 11 days;
(2) preparing a grain bran enzymolysis liquid: crushing delignified cereal bran, performing enzymolysis, centrifuging and sterilizing to obtain cereal bran enzymatic hydrolysate;
(3) and uniformly mixing the Weissella cibaria AP6 thallus with the grain bran enzymatic hydrolysate, and drying to obtain the pickle starter.
Further, the preparation of Weissella cibaria AP6 thallus in the step (1) comprises the following steps:
s1, inoculating activated Weissella sinus AP6 thalli into an MRS culture medium according to the inoculation amount (v/v) of 1%, culturing for 24h at 37 ℃, and continuously carrying out passage for three times to obtain a culture solution containing Weissella sinus AP6 thalli;
s2, centrifuging the culture solution at 4 ℃ at 8000-10000 r/min for 10min, pouring out the supernatant, then resuspending the thallus by using sterile normal saline, and repeating the steps for three times to obtain the Weissella sinus AP6 thallus.
Further, the preparation of the grain bran enzymolysis liquid in the step (2) comprises the following steps:
A. crushing delignified grain bran to 30-80 meshes;
B. mixing grain bran powder and water with the mass 10-20 times of that of the grain bran powder to form a suspension, adjusting the temperature of the suspension to 45-55 ℃, adjusting the pH value to 4.5-6.0, adding xylanase into the suspension according to 80-120U/g of grain bran, and stirring for enzymolysis for 3-5 hours;
C. after the enzymolysis is finished, centrifuging the suspension, taking the supernatant, and sterilizing at high temperature to obtain a cereal bran enzymolysis liquid, wherein the content of the arabinoxylo-oligosaccharide in the cereal bran enzymolysis liquid is 1.05-1.86%;
further, the delignified cereal bran in the step A is crushed into 50 meshes; in the step B, the mass ratio of the delignified cereal bran powder to the water is 1:15, the temperature is 50 ℃, the pH value is adjusted to 4.8, the addition amount of the xylanase is 100U/g cereal bran, and the enzymolysis time is 4 hours.
Further, the mass ratio of the Weissella sinus AP6 thallus to the grain bran enzymolysis liquid in the step (3) is 1: 3.
Further, the drying mode of the step (3) is spray drying or vacuum freeze drying.
In a third aspect, the present invention provides kimchi fermented with the kimchi starter culture according to the first aspect or the kimchi starter culture prepared by the method for preparing the kimchi starter culture according to the second aspect.
In a fourth aspect, the invention provides an application method of the pickle starter culture, wherein the pickle starter culture is prepared by the preparation method of the second aspect, and the pickle starter culture is used for preparing pickle, and the addition amount of the pickle starter culture is 0.5-2% of the total mass of the pickle water.
Further, the addition amount is 1.5% of the total mass of the vegetable water.
Compared with the prior art, the invention has the beneficial effects that:
1. the invention takes the grain bran enzymatic hydrolysate as the strain protective agent, which is beneficial to the comprehensive utilization of byproducts and improves the economic benefit;
2. the Weissella sinus AP6 has strong acid resistance and substrate specificity, has strong activity in an MRS culture medium which takes xylo-oligosaccharide as a single carbon source, takes grain bran enzymolysis liquid as a Weissella natural protective agent and an excipient, can keep the survival rate and stability of the Weissella in the drying and storage processes on the premise of not adding a drying protective agent, is convenient to use, and reduces the production cost;
3. the direct vat set starter is added in the preparation process of the pickle, the arabinoxylo-oligosaccharide in the grain bran enzymatic hydrolysate can be used as a carbon source, the activity of Weissella sinus AP6 can be improved, the special fermentation time can be prolonged, and the pickle is endowed with better flavor.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the principles of the invention and not to limit the invention.
FIG. 1 is a graph showing a comparison of viable bacteria in the fermenters of example 2 of the present invention and comparative example 1;
FIG. 2 is a graph showing the growth of Weissella antrioea in the starter cultures of example 2 of the present invention and comparative example 2;
fig. 3 is a graph showing pH changes during fermentation of kimchi according to example 2, comparative example 1 and comparative example 2 of the present invention.
The different letters in FIGS. 2 and 3 indicate that the difference in time is significant (p ≦ 0.05).
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be described in detail below. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. All other embodiments, which can be derived by a person skilled in the art from the examples given herein without any inventive step, are within the scope of the present invention.
In the present document,% represents wt% unless otherwise specified, and the proportional relationship is mass ratio; the raw materials used are all commercial products unless otherwise specified.
Example 1
Firstly, preparing a pickle starter culture, which comprises the following steps:
1. preparation of Weissella cibaria AP6 thallus
1) Activating Weissella fossa AP6 with the preservation number of GDMCC.No62461, inoculating the Weissella fossa AP6 into an MRS culture medium according to the inoculation amount (v/v) of 1%, culturing for 24h at 37 ℃, and continuously carrying out passage for three times to obtain a culture solution containing Weissella fossa AP 6;
2) centrifuging the culture solution at 4 deg.C and 9000r/min for 10min, pouring out supernatant, re-suspending thallus with sterile normal saline, and repeating the above steps for three times to obtain Weissella sinus AP6 thallus.
2. Preparation of enzymatic hydrolysate of cereal bran
1) Crushing the delignified cereal bran and sieving the crushed delignified cereal bran with a 30-mesh sieve;
2) taking 10g of cereal bran powder, adding 100g of water, adjusting the temperature of the suspension to 45 ℃, adjusting the pH value to 4.5, adding 800U of xylanase, stirring and performing enzymolysis for 5 hours;
3) and after the enzymolysis is finished, centrifuging the suspension, taking supernatant, and sterilizing at 100 ℃ for 5min to obtain the grain bran enzymolysis liquid, wherein the content of the arabinoxylo-oligosaccharide in the grain bran enzymolysis liquid is 1.05%.
3. Taking 10g of Weissella AP6 thallus prepared in the step 1 and 30g of cereal bran enzymolysis liquid prepared in the step 2, uniformly stirring, and then performing spray drying on the mixed liquid to obtain the pickle starter culture, wherein the air inlet temperature of a spray drying device is 118 ℃; the obtained pickle starter culture has live Weissella bacteria count of 9.45 × 10 9 CFU/g, arabinoxylo-oligosaccharide content of 51.22%.
Secondly, preparing pickled vegetables
Putting the pickle into a fermentation tank, adding a pickle starter with the total mass of 2% of the pickle water, and uniformly mixing; the kimchi may be prepared according to a conventional method.
Example 2
Firstly, preparing a pickle starter culture, which comprises the following steps:
1. preparation of Weissella cibaria AP6 thallus
1) Activating Weissella sinus AP6 with the preservation number of GDMCC.No62461, inoculating the Weissella sinus AP6 into an MRS culture medium according to the inoculation amount (v/v) of 1%, culturing for 24h at 37 ℃, and continuously carrying out passage for three times to obtain a culture solution of Weissella sinus AP 6;
2) centrifuging the culture solution at 4 deg.C and 8000r/min for 10min, pouring out the supernatant, re-suspending the thallus with sterile normal saline, and repeating the above steps for three times to obtain Weissella sinus AP6 thallus.
2. Preparation of enzymatic hydrolysate of cereal bran
1) Crushing the delignified cereal bran and sieving the crushed delignified cereal bran with a 50-mesh sieve;
2) taking 10g of cereal bran powder, adding 150g of water, adjusting the temperature of the suspension to 50 ℃, adjusting the pH value to 4.8, adding 1000U of xylanase, stirring and performing enzymolysis for 4 hours;
3) and after the enzymolysis is finished, centrifuging the suspension, taking supernatant, and sterilizing at 100 ℃ for 5min to obtain the grain bran enzymolysis liquid, wherein the content of the arabinoxylo-oligosaccharide in the grain bran enzymolysis liquid is 1.5%.
3. Taking 10g of Weissella AP6 thallus prepared in the step 1 and 30g of cereal bran enzymolysis liquid prepared in the step 2, uniformly stirring, and then performing spray drying on the mixed liquid to obtain the pickle starter culture, wherein the air inlet temperature of a spray drying device is 130 ℃; the obtained pickle starter culture has live Weissella bacteria count of 1.07 × 10 10 CFU/g, the content of the arabinoxylo-oligosaccharide is 60.10 percent.
Secondly, preparing pickled vegetables
Putting the pickle into a fermentation tank, adding a pickle starter with the total mass of 1.5 percent of the pickle, and uniformly mixing; the kimchi may be prepared according to a conventional method.
Example 3
Firstly, preparing a pickle starter culture, which comprises the following steps:
1. preparation of Weissella cibaria AP6 thallus
1) Activating Weissella civorans AP6 with the preservation number of GDMCC.No62461, inoculating the Weissella civorans AP6 into an MRS culture medium according to the inoculation amount (v/v) of 1%, culturing for 24 hours at 37 ℃, and continuously carrying out passage for three times to obtain a culture solution containing Weissella civorans AP 6;
2) centrifuging the culture solution at 4 deg.C and 10000r/min for 10min, pouring out the supernatant, re-suspending the thallus with sterile normal saline, and repeating the above steps for three times to obtain Weissella sinus AP6 thallus.
2. Preparation of enzymatic hydrolysate of cereal bran
1) Crushing the delignified cereal bran and sieving the crushed delignified cereal bran with a 80-mesh sieve;
2) taking 10g of cereal bran powder, adding 200g of water, adjusting the temperature of the suspension to 50 ℃, adjusting the pH value to 6.0, adding 1200U of xylanase, stirring and performing enzymolysis for 3 hours;
3) and after the enzymolysis is finished, centrifuging the suspension, taking supernatant, and sterilizing at 100 ℃ for 5min to obtain the grain bran enzymolysis liquid, wherein the content of the arabinoxylo-oligosaccharide in the grain bran enzymolysis liquid is 1.86%.
3. Taking 10g of Weissella AP6 thallus prepared in the step 1 and 30g of the grain bran enzymolysis liquid prepared in the step 2, uniformly stirring, placing in a refrigerator with the temperature of 30 ℃ below zero for pre-freezing for 3h, then placing in a pre-cooled vacuum freeze dryer, starting a vacuum pump, and drying for 24h to obtain the pickle starter; the obtained pickle starter culture has live Weissella bacteria count of 7.8 × 10 10 CFU/g, arabinoxylo-oligosaccharide content 65.03%.
Secondly, preparing pickled vegetables
Putting the pickle into a fermentation tank, adding a pickle starter with the total mass of 0.5 percent of the pickle water, and uniformly mixing; the kimchi may be prepared according to a conventional method.
Comparative example 1
Preparation of pickle starter culture
The preparation method of the invention in example 2 is adopted, wherein the difference between the comparative example 1 and the example 2 is as follows: replacing the bran hydrolysate with skim milk powder; the other conditions are the same as example 2, and the viable count of Weissella in the prepared starter is 9.22 × 10 9 CFU/g。
Secondly, preparing pickled vegetables
Putting the pickle into a fermentation tank, adding a pickle starter with the total mass of 1.5 percent of the pickle, and uniformly mixing; the kimchi may be prepared according to a conventional method.
Comparative example 2
Preparation of pickle starter culture
The preparation method of the invention in the example 2 is adopted, wherein the comparative example 2 is different from the example 2 in that: weissella sinus TG3 was used to replace Weissella sinus AP 6; the other conditions are the same as example 2, and the viable count of Weissella in the prepared starter is 9.41 × 10 9 CFU/g。
Secondly, preparing pickled vegetables
Putting the pickle into a fermentation tank, adding a pickle starter with the total mass of 1.5 percent of the pickle, and uniformly mixing; the kimchi may be prepared according to a conventional method.
Examples of the experiments
1. The results of comparing the starter of example 2 and the starter of comparative example 1 for the percentage of viable bacteria of Weissella cibaria AP6 are shown in FIG. 1.
As can be seen from FIG. 1, the percentage of viable bacteria of Weissella antrioea AP6 in the leavening agent of example 2 is slightly higher than that in comparative example 1, which indicates that the protection effect of the enzymatic hydrolysate of cereal bran is slightly higher than that of the skim milk powder, and the enzymatic hydrolysate of cereal bran can be used to replace the skim milk powder.
2. Growth curves of weissella antrioides AP6 in the starter of comparative example 2 and weissella antrioides TG3 in the starter of comparative example 2 in MRS medium with xylo-oligosaccharide as single carbon source were obtained, and the results are shown in fig. 2.
As shown in FIG. 2, Weissella sinus AP6 is highly active, and xylooligosaccharide has selective specificity to promote growth of Weissella sinus AP 6.
3. The kimchi of example 2, comparative example 1 and comparative example 2 was examined for pH change during fermentation, and the result is shown in fig. 3.
As can be seen from fig. 3, the pH of the kimchi of comparative example 1 decreased significantly faster than that of the kimchi of example 2, indicating that homolactic acid bacteria rapidly increased to produce acid, and the addition of the grain bran enzymatic hydrolysate is beneficial to the growth of heterolactic acid bacteria and the reduction of acid production rate, thereby being beneficial to the accumulation of flavor substances; in the pickle fermentation process, the pH dropping rate of the example 2 is different from that of the comparative example 2, and the growth rate of the Weissella sinus AP6 is better than that of the Weissella sinus TG3, so that the growth of homolactic bacteria is delayed, and the acid production rate is reduced.
4. After the fermentation was completed, the kimchi of example 2, comparative example 1 and comparative example 2 was subjected to sensory evaluation, and the results are as follows:
| average score/score | Example 2 | Comparative example 1 | Comparative example 2 |
| Form of the composition | 8.45±0.38 a | 7.64±1.03 b | 8.24±051 a |
| Color | 7.61±0.84 a | 6.45±1.19 b | 7.51±0.20 a |
| Fragrance | 8.94±0.56 a | 7.15±0.61 c | 8.16±0.86 b |
| Taste of the product | 9.03±0.22 a | 8.35±1.51 b | 8.99±0.57 a |
| Degree of brittleness | 8.93±0.43 a | 7.56±0.61 c | 8.47±0.51 b |
| Overall mouthfeel | 9.53±0.61 a | 7.59±1.21 b | 9.22±1.09 a |
Note: different letters indicate significant differences between different treatments (p ≦ 0.05)
As can be seen from the table above, the shapes, colors, aromas, tastes, crispness and overall mouthfeel of the pickles of the example 2 and the comparative example 2 are all obviously superior to those of the comparative example 1, and the aromas and crispness of the pickles of the example 2 are obviously superior to those of the comparative example 2, which shows that the addition of the leavening agent containing the combination of the arabinoxylo-oligosaccharide and the Weissella sinus AP6 in the vegetables is beneficial to the growth of Weissella during the fermentation process, thereby being beneficial to the accumulation of flavor substances and the sensory quality.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.
Claims (10)
1. A pickle starter culture is characterized in that: contains Weissella sinus AP6 and arabinoxylo-oligosaccharide, wherein the Weissella sinus AP6 is preserved in Guangdong collection of microorganisms with the preservation number of GDMCC No. 62461 and the preservation date of 2022, 5 months and 11 days.
2. The kimchi starter culture of claim 1, wherein: the concentration of live bacteria of Weissella sinus-feeding AP6 in the pickle starter culture is more than or equal to 10 9 CFU/g, the content of the arabinoxylo-oligosaccharide is 51.22-65.03%.
3. A preparation method of a pickle starter culture is characterized by comprising the following steps:
(1) preparing Weissella civora AP6 thallus: activating Weissella sinus AP6, inoculating the activated Weissella sinus AP6 into a culture medium for culture, centrifuging the culture medium to obtain Weissella sinus AP6 thallus, wherein the Weissella sinus AP6 is preserved in Guangdong microbiological collection center with the preservation number of GDMCC No. 62461 and the preservation date of 2022, 5 months and 11 days;
(2) preparing a grain bran enzymolysis liquid: crushing delignified cereal bran, performing enzymolysis, centrifuging and sterilizing to obtain cereal bran enzymatic hydrolysate;
(3) and uniformly mixing the Weissella cibaria AP6 thallus with the grain bran enzymatic hydrolysate, and drying to obtain the pickle starter.
4. The preparation method of kimchi starter culture according to claim 3, wherein the preparation of Weissella assidus AP6 biomass in step (1) comprises the steps of:
s1, inoculating activated Weissella sinus AP6 thalli into an MRS culture medium according to the inoculation amount (v/v) of 1%, culturing for 24h at 37 ℃, and continuously carrying out passage for three times to obtain a culture solution containing Weissella sinus AP6 thalli;
s2, centrifuging the culture solution at 4 ℃ at 8000-10000 r/min for 10min, pouring out the supernatant, then resuspending the thallus by using sterile normal saline, and repeating the steps for three times to obtain the Weissella sinus AP6 thallus.
5. The preparation method of kimchi starter culture according to claim 3, wherein the preparation of the grain bran enzymatic hydrolysate in the step (2) comprises the steps of:
A. crushing the delignified cereal bran to 30-80 meshes;
B. mixing grain bran powder and water with the mass 10-20 times of that of the grain bran powder to form a suspension, adjusting the temperature of the suspension to 45-55 ℃, adjusting the pH value to 4.5-6.0, adding xylanase into the suspension according to 80-120U/g of grain bran, and stirring for enzymolysis for 3-5 hours;
C. and after the enzymolysis is finished, centrifuging the suspension, taking the supernatant, and sterilizing at high temperature to obtain the grain bran enzymolysis liquid, wherein the content of the arabinoxylo-oligosaccharide in the grain bran enzymolysis liquid is 1.05-1.86%.
6. The preparation method of kimchi starter culture according to claim 5, wherein the delignified cereal bran in step a is crushed to 50 mesh; in the step B, the mass ratio of the delignified cereal bran powder to the water is 1:15, the temperature is 50 ℃, the pH value is adjusted to 4.8, the addition amount of the xylanase is 100U/g cereal bran, and the enzymolysis time is 4 hours.
7. The preparation method of the kimchi starter culture according to claim 3, wherein a mass ratio of the Weissella sinus AP6 biomass to the grain bran enzymatic hydrolysate in the step (3) is 1: 3.
8. A kind of pickled vegetable, characterized by that: the kimchi is fermented by using the kimchi starter culture according to claim 1 or 2 or the kimchi starter culture prepared by the method for preparing the kimchi starter culture according to any one of claims 3 to 7.
9. An application method of a pickle starter culture is characterized in that: the pickle starter culture is prepared by the preparation method of any one of claims 2 to 7, is used for fermenting pickle, and is added in an amount of 0.5 to 2 percent of the total mass of the pickle water.
10. The method of applying kimchi starter culture in accordance with claim 9, comprising: the addition amount of the pickle starter is 1.5 percent of the total mass of the pickle.
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