CN113604393A - Pediococcus pentosaceus ZF604 and application thereof - Google Patents

Pediococcus pentosaceus ZF604 and application thereof Download PDF

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CN113604393A
CN113604393A CN202110945633.XA CN202110945633A CN113604393A CN 113604393 A CN113604393 A CN 113604393A CN 202110945633 A CN202110945633 A CN 202110945633A CN 113604393 A CN113604393 A CN 113604393A
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soy sauce
fermentation
pediococcus pentosaceus
lactic acid
crude oil
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CN113604393B (en
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黄艳
栗连会
周其洋
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Foshan Haitian Flavoring and Food Co Ltd
Foshan Haitian Gaoming Flavoring and Food Co Ltd
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Foshan Haitian Gaoming Flavoring and Food Co Ltd
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Abstract

The invention discloses pediococcus pentosaceus ZF604, which is preserved in 26.7.2021 in Guangdong province of microbial strain collection center with the preservation number GDMCC No: 61837. The pediococcus pentosaceus ZF604 has the characteristics of high yield of lactic acid and salt tolerance, can normally grow in high-salt fermented soy sauce mash, obviously improves the lactic acid level of fermented crude oil, and provides a sufficient substrate for crude oil fermentation. The pediococcus pentosaceus ZF604 is used for soy sauce fermentation by compounding aspergillus oryzae and yeast in the prior art, so that the composite aroma-enhancing effect can be achieved, lactic acid and flavor substances such as alcohols in crude oil are subjected to composite reaction, the content of flavor substances in the soy sauce is integrally increased, the types of the flavor substances in the soy sauce are enriched, and the flavor components such as 3-methylthiopropanol, 2-methyl-6- [ [ (E) -1-propenyl ] pyrazine and 1,3,5, 7-cyclooctanoate are newly added into the soy sauce.

Description

Pediococcus pentosaceus ZF604 and application thereof
Technical Field
The invention belongs to the technical field of microbial fermentation, and particularly relates to a pediococcus pentosaceus ZF604 with high lactic acid yield and salt tolerance and application thereof.
Background
Soy sauce is mainly divided into two main categories, namely high-salt liquid state fermented soy sauce and low-salt solid state fermented soy sauce according to different fermentation processes. Since the two processes have great differences in fermentation temperature, brine concentration, fermentation time and the like, the obtained soy sauce has obvious differences in sensory quality and flavor. Research shows that the types and relative contents of aroma substances in the high-salt dilute fermented soy sauce are obviously higher than those of the low-salt solid soy sauce. Therefore, the high-salt dilute fermented soy sauce can better meet the requirements of consumers and is a main development direction for producing soy sauce in the future.
The aroma characteristics are important indexes for evaluating the quality of the high-salt dilute soy sauce, but the prior research on the aroma of the high-salt dilute soy sauce in China is not systematic and deep. In modern large-scale brewing, single aspergillus oryzae is mostly adopted for pure culture starter propagation, and although the process of protein decomposition and lactic acid bacteria acid production to quickly reduce pH after mash preparation is carried out, the diversity of microorganisms is insufficient, so that the soy sauce has single flavor and insufficient flavor richness. In addition, in recent years, the emphasis on the flavor of soy sauce has been on the components of 4-ethylguaiacol and furanone. However, 3-methylthiopropanol, known by the English name 1-Propanol,3- (methylthio) -, known by the Chinese name 3- (methylthio) -1-Propanol, has the formula C4H10OS, molecular weight is 106.18, the substance has strong onion and meat odor, and soy sauce odor appears after dilution, and the substance is mainly used for sauce seasoning. The inventor of the invention finds that the 3-methylthio propanol can enhance the fragrance of the soy sauce and improve the richness of the fragrance of the soy sauce, but no report that the content of the 3-methylthio propanol in the fermented crude oil is enhanced and improved by lactic acid bacteria exists at present is found.
Disclosure of Invention
In order to solve the defects and shortcomings in the prior art, the invention aims to provide the pediococcus pentosaceus ZF604 with high lactic acid yield and salt tolerance, the lactic acid level of the fermented crude oil is improved by utilizing the pediococcus pentosaceus ZF604, a sufficient substrate is provided for crude oil fermentation, and the soy sauce has more types of aroma components, higher total content of the aroma components and stronger soy sauce aroma.
In order to realize the purpose, the technical scheme adopted by the invention is as follows:
a Pediococcus pentosaceus ZF604(Pediococcus pentosaceus) is preserved in 26 months 7 in 2021 in Guangdong province of microbial strain collection with the preservation number GDMCC No. 61837. The preservation address is No. 59 building 5 of No. 100 Dazhong Jie of Jifurao, Guangzhou, China.
The pediococcus pentosaceus ZF604 has the characteristics of high yield of lactic acid and salt tolerance, and the bacterial colony characteristics are as follows: the colony is round, the color is milk white at the initial culture stage, the color is turned to milk yellow at the later stage, the colony is not transparent, but the periphery is a transparent ring.
The Pediococcus pentosaceus ZF604 is obtained by conditionally screening fermented soy sauce mash by the inventor, and is identified to belong to Pediococcus pentosaceus (Pediococcus pentosaceus), and the lactic acid content of the fermented crude oil can be obviously improved by adding the Pediococcus pentosaceus ZF into the soy sauce mash fermentation.
The invention also provides application of the pediococcus pentosaceus ZF604 in food fermentation. In some embodiments, the food product comprises soy sauce or fermented paste.
The invention also provides a fermentation method of food, which comprises the step of adding the pediococcus pentosaceus ZF604 for fermentation. In some embodiments, the food product comprises soy sauce or fermented paste.
The invention also provides a method for improving the content of flavor substances in soy sauce, which comprises the step of adding the pediococcus pentosaceus ZF604 for fermentation.
Preferably, the method further comprises the step of adding aspergillus oryzae and yeast in a compound manner for fermentation.
Preferably, the flavour substances comprise at least one of 3-methylthiopropanol, 2-methyl-6- [ [ (E) -1-propenyl ] pyrazine, 1,3,5, 7-cyclooctanoate, ethyl octanoate, ethyl nonanoate, ethyl laurate.
The inventor finds that the pediococcus pentosaceus ZF604, aspergillus oryzae and saccharomycetes are combined and applied to soy sauce fermentation to achieve the effect of compound aroma enhancement, and lactic acid and flavor substances such as alcohols in crude oil are subjected to compound reaction, so that not only can new aroma components such as 3-methylthiopropanol, 2-methyl-6- [ [ (E) -1-propenyl ] pyrazine and 1,3,5, 7-cyclooctanoate be added to the soy sauce, but also the content of the flavor substances in the soy sauce can be integrally increased.
The invention also provides a fermentation inoculant which comprises the pediococcus pentosaceus ZF 604.
Preferably, the zymophyte agent also comprises aspergillus oryzae and yeast.
Compared with the prior art, the invention has the beneficial effects that: the pediococcus pentosaceus ZF604 provided by the invention has the characteristics of high yield of lactic acid and salt tolerance, can normally grow in high-salt fermented soy sauce mash, obviously improves the lactic acid level of fermented crude oil, and provides a sufficient substrate for crude oil fermentation. The pediococcus pentosaceus ZF604 is used for soy sauce fermentation by compounding aspergillus oryzae and yeast in the prior art, so that the composite aroma-enhancing effect can be achieved, lactic acid and flavor substances such as alcohols in crude oil are subjected to composite reaction, the content of flavor substances in the soy sauce is integrally increased, the types of the flavor substances in the soy sauce are enriched, and the flavor components such as 3-methylthiopropanol, 2-methyl-6- [ [ (E) -1-propenyl ] pyrazine and 1,3,5, 7-cyclooctanoate are newly added into the soy sauce.
Drawings
FIG. 1 is a morphological microscopic image of the colony of Pediococcus pentosaceus ZF 604;
FIG. 2 is a graph showing the difference analysis of aroma components between groups 1 and 3 in example 3.
Detailed Description
To better illustrate the objects, aspects and advantages of the present invention, the present invention is further illustrated by the following examples. It is apparent that the following examples are only a part of the embodiments of the present invention, and not all of them. It should be understood that the embodiments of the present invention are only for illustrating the technical effects of the present invention, and are not intended to limit the scope of the present invention. Unless otherwise indicated, the methods and test conditions employed in the examples of the present invention are those conventionally used in the art, and the reagents, equipment and culture media used therein are those conventionally used in the art, and can be prepared by existing methods or commercially available. In the examples, the percentages are by mass unless otherwise specified. In the examples, Aspergillus oryzae used was Hu Niang 3.042, and the yeast used was ZB118 (accession number GDMCC No. 60280, Guangdong Collection of microorganisms).
Example 1 acquisition of Pediococcus pentosaceus ZF604 Strain
Performing conditional breeding (salt resistance and acid production) on sauce mash production, screening a plate by using an MRS culture medium added with 2% calcium carbonate, screening bacterial colonies with a lysis ring, and obtaining 7 strains, wherein the salt resistance of 5 strains is improved relative to a reference strain, 4 strains have obvious lactic acid production improvement (acid production activity is cultured in a salt-containing bean juice liquid) relative to the reference strain (Lactobacillus plantarum Shanghai 1.08), and the total 3 strains with improved comprehensive salt resistance and lactic acid production are respectively RS1703, RS1710 and ZF 604; the fermentation index analysis is shown in Table 1.
TABLE 1 results of strain screening and fermented lactic acid mass analysis
Figure BDA0003213948550000041
Remarking: the control strain value is 1, and other strains are converted into corresponding ratios.
Example 2 Effect of Pediococcus pentosaceus ZF604 Strain on increasing lactic acid content of crude oil
The bred 3 strains with high lactic acid yield and high salt tolerance are compounded with soy sauce main fermentation strain aspergillus oryzae for use, the effect comparison in a fermentation crude oil culture medium is carried out, and the obtained indexes are shown in the following table 2:
as can be seen from the table, the total acid content and the lactic acid content of the crude oil are improved to a certain extent by the 3 strains, wherein the effect of the strain ZF604 on improving the lactic acid content of the crude oil is optimal. However, lactic acid is milder in sourness than other organic acids (acetic acid, citric acid, etc.) and is not irritating, but excessive production of total acid also makes crude oil sour in taste and is not favorable for flavor enhancement. Therefore, the invention performs composite aroma enhancement on the strain ZF604, yeast and aspergillus oryzae, and performs composite reaction on lactic acid and flavor substances such as alcohols in crude oil.
TABLE 2 lactic acid bacteria strain screening and fermented lactic acid quality analysis results
Figure BDA0003213948550000042
Note: and calculating the relative lifting content by taking each index as a reference.
Example 3 enhancing the composite flavor enhancing effect of Pediococcus pentosaceus ZF604 and yeast and Aspergillus oryzae
(1) The pediococcus pentosaceus ZF604 is used in combination with aspergillus oryzae and/or yeast for comparison of effects in a fermentation crude oil culture medium, and the obtained indexes are shown in the following table 3:
as can be seen from the table, the total acid content of the crude oil was higher when Aspergillus oryzae was used alone; after yeast is added in the aspergillus oryzae fermentation, the fermentation liquor has a certain inhibition effect on the generation of total acid; after pediococcus pentosaceus ZF604 is added in aspergillus oryzae fermentation, the contents of total acid and lactic acid are increased; after yeast and pediococcus pentosaceus ZF604 are added in the aspergillus oryzae fermentation, the pediococcus pentosaceus ZF604 and the yeast are compounded for use, on one hand, irritant acid generated by rapid propagation of other acid-producing bacteria can be inhibited by the yeast, on the other hand, the generation of lactic acid is enhanced at the later stage, and the lactic acid is beneficial to lactic acid generation, aroma generation and flavor generation.
TABLE 3 analysis of ZF604 and Yeast composite fermentation quality
Figure BDA0003213948550000051
Note: and calculating the relative lifting content by taking each index as a reference.
Sensory evaluation of the aroma was performed on the crude oils obtained in groups 1 to 3, and the evaluation of each sample was described by a highly experienced sensory evaluator, and the results are shown in table 4.
TABLE 4 sensory evaluation of crude oil aroma
Figure BDA0003213948550000052
(2) And analyzing the aroma components of the groups 1 and 3 by using a gas chromatography-mass spectrometer for the crude oil obtained by fermentation. The specific GC-MS analysis method is as follows:
equipment name: agilent 8890-5977B gas phase mass spectrometer
Using DB-INNOWAX 60 mm × 0.25 μm gas chromatography column;
the temperature of a sample inlet is 250 ℃;
the temperature rise conditions are as follows: the initial temperature is 40 deg.C, keeping for 5min, heating to 240 deg.C at 5 deg.C per minute, keeping for 15min, and the carrier gas flow rate is 1.2mL/min, and no flow-dividing sampling is performed.
The electron impact ionization (EI) ion source has the ion source temperature of 250 ℃, the transmission line temperature of 280 ℃ and the mass scanning range of 30-500 m/z.
The results are shown in FIG. 2 (in FIG. 2, Aspergillus oryzae + yeast represents group 1, and Aspergillus oryzae + yeast + ZF604 represents group 3). As can be seen from fig. 2, strengthening pediococcus pentosaceus ZF604 during soy sauce fermentation can make the main flavor substances in soy sauce: the content of esters and phenols is increased, and the content of alcohols, acids and aldehydes is reduced.
(3) By comparing the difference of the different aroma components in the control group 1 and the group 3, the increase, decrease and addition of the components on the different aroma substances of the control group and the experimental group are found, and the specific results are shown in table 5. As can be seen from the data in the table, the addition of Pediococcus pentosaceus ZF604 increases the content of ester substances in the soy sauce to a certain extent, reduces the content of acetic acid, furfural, furfuryl alcohol, phenethyl alcohol, maltol and ethyl palmitate to a certain extent, and enhances the content of 3-methylthiopropanol, 2-methyl-6- [ [ (E) -1-propenyl in the soy sauce]Pyrazine and 1,3,5, 7-cyclooctanoate are novel aroma components. Wherein the 3-methylthiopropanol has a high content, the 3-methylthiopropanol has an English name of 1-Propanol,3- (methylthio) -, a Chinese name of 3- (methylthio) -1-Propanol and a molecular formula of C4H10OS, molecular weight 106.18, which has a strong onion and meat odor, and which develops a soy sauce odor upon dilution, is currently used mainly for sauce seasoning, and its presence enhances the soy sauce aroma.
TABLE 5 control and test samples aroma part key material composition change
Figure BDA0003213948550000061
Figure BDA0003213948550000071
Note: in the table, "-" indicates that the component is not contained. The other components are not all listed.
Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention and not for limiting the protection scope of the present invention, and although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.

Claims (10)

1. A Pediococcus pentosaceus ZF604 is preserved in 26.7.2021 in Guangdong province microbial strain preservation center with the preservation number of GDMCC No. 61837 and the preservation address of No. 59 building 5 of Mirabilitum 100 of Guangzhou City of China.
2. Use of pediococcus pentosaceus ZF604 according to claim 1 in food fermentation.
3. The use of claim 2, wherein the food product comprises soy sauce or fermented paste.
4. A method for fermenting a food, comprising adding the Pediococcus pentosaceus ZF604 of claim 1 to the fermentation.
5. The fermentation process of claim 4, wherein the food product comprises soy sauce or fermented paste.
6. A fermentation inoculant comprising the pediococcus pentosaceus ZF604 of claim 1.
7. The fermentation inoculant of claim 6 further comprising Aspergillus oryzae and yeast.
8. A method for increasing the content of flavor substances in soy sauce, which comprises adding the pediococcus pentosaceus ZF604 of claim 1 to the soy sauce for fermentation.
9. The method of claim 8, further comprising adding aspergillus oryzae and yeast for fermentation.
10. The method of claim 8, wherein said flavor comprises at least one of 3-methylthiopropanol, 2-methyl-6- [ [ (E) -1-propenyl ] pyrazine, 1,3,5, 7-cyclooctanoate, ethyl octanoate, ethyl nonanoate, ethyl laurate.
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115886233A (en) * 2022-11-15 2023-04-04 佛山市海天(高明)调味食品有限公司 Soy sauce aroma-enhancing base material, soy sauce and preparation method thereof

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WO2013073249A1 (en) * 2011-11-18 2013-05-23 キッコーマン株式会社 Soy sauce-like seasoning
CN103614325A (en) * 2013-11-26 2014-03-05 江南大学 Pediococcus pentosaceus and application thereof
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