CN106834181B - A kind of Pediococcus acidilactici and its application - Google Patents
A kind of Pediococcus acidilactici and its application Download PDFInfo
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- CN106834181B CN106834181B CN201710106981.1A CN201710106981A CN106834181B CN 106834181 B CN106834181 B CN 106834181B CN 201710106981 A CN201710106981 A CN 201710106981A CN 106834181 B CN106834181 B CN 106834181B
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- pediococcus acidilactici
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- 241000191998 Pediococcus acidilactici Species 0.000 title claims abstract description 100
- 238000000034 method Methods 0.000 claims abstract description 35
- 241000894006 Bacteria Species 0.000 claims description 39
- 240000006439 Aspergillus oryzae Species 0.000 claims description 36
- 235000002247 Aspergillus oryzae Nutrition 0.000 claims description 36
- 238000000855 fermentation Methods 0.000 claims description 31
- 230000004151 fermentation Effects 0.000 claims description 29
- 150000001875 compounds Chemical class 0.000 claims description 28
- 235000013555 soy sauce Nutrition 0.000 claims description 26
- 241000131386 Aspergillus sojae Species 0.000 claims description 23
- 239000000463 material Substances 0.000 claims description 22
- 239000002054 inoculum Substances 0.000 claims description 20
- 235000010469 Glycine max Nutrition 0.000 claims description 17
- 244000068988 Glycine max Species 0.000 claims description 16
- 235000015067 sauces Nutrition 0.000 claims description 16
- 150000003839 salts Chemical class 0.000 claims description 14
- 239000000203 mixture Substances 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- 238000004321 preservation Methods 0.000 claims description 9
- 241000228212 Aspergillus Species 0.000 claims description 8
- 241000192001 Pediococcus Species 0.000 claims description 4
- 235000013527 bean curd Nutrition 0.000 claims description 4
- 235000021419 vinegar Nutrition 0.000 claims description 4
- 239000000052 vinegar Substances 0.000 claims description 4
- 238000011534 incubation Methods 0.000 claims description 3
- 238000010563 solid-state fermentation Methods 0.000 claims description 2
- 241000193830 Bacillus <bacterium> Species 0.000 abstract description 26
- 230000001580 bacterial effect Effects 0.000 abstract description 8
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 20
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 18
- 239000010779 crude oil Substances 0.000 description 16
- 235000002639 sodium chloride Nutrition 0.000 description 16
- 244000046052 Phaseolus vulgaris Species 0.000 description 15
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 15
- 239000000796 flavoring agent Substances 0.000 description 15
- 235000019634 flavors Nutrition 0.000 description 15
- 241000209140 Triticum Species 0.000 description 13
- 235000021307 Triticum Nutrition 0.000 description 13
- 235000001014 amino acid Nutrition 0.000 description 11
- 150000001413 amino acids Chemical class 0.000 description 11
- 230000000694 effects Effects 0.000 description 11
- 239000002253 acid Substances 0.000 description 10
- 235000013312 flour Nutrition 0.000 description 10
- 229910052757 nitrogen Inorganic materials 0.000 description 10
- 238000001514 detection method Methods 0.000 description 9
- 235000014655 lactic acid Nutrition 0.000 description 9
- 239000004310 lactic acid Substances 0.000 description 9
- 235000015096 spirit Nutrition 0.000 description 9
- 240000006024 Lactobacillus plantarum Species 0.000 description 8
- 235000013965 Lactobacillus plantarum Nutrition 0.000 description 8
- 229940072205 lactobacillus plantarum Drugs 0.000 description 8
- 244000005700 microbiome Species 0.000 description 8
- 210000000697 sensory organ Anatomy 0.000 description 8
- 108090000145 Bacillolysin Proteins 0.000 description 7
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 7
- 108091005507 Neutral proteases Proteins 0.000 description 7
- 102000035092 Neutral proteases Human genes 0.000 description 7
- 235000013922 glutamic acid Nutrition 0.000 description 7
- 239000004220 glutamic acid Substances 0.000 description 7
- 238000002360 preparation method Methods 0.000 description 7
- 230000007423 decrease Effects 0.000 description 6
- 238000004519 manufacturing process Methods 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 102000004190 Enzymes Human genes 0.000 description 5
- 108090000790 Enzymes Proteins 0.000 description 5
- 238000002156 mixing Methods 0.000 description 5
- 239000002994 raw material Substances 0.000 description 5
- 235000019640 taste Nutrition 0.000 description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- 241001478240 Coccus Species 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 3
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 3
- 230000001413 cellular effect Effects 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 239000003921 oil Substances 0.000 description 3
- 239000013641 positive control Substances 0.000 description 3
- 230000035755 proliferation Effects 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 235000012976 tarts Nutrition 0.000 description 3
- 241000233866 Fungi Species 0.000 description 2
- 241000191938 Micrococcus luteus Species 0.000 description 2
- 235000019658 bitter taste Nutrition 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- 238000005138 cryopreservation Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 235000013399 edible fruits Nutrition 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000002386 leaching Methods 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 235000013599 spices Nutrition 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 235000019605 sweet taste sensations Nutrition 0.000 description 2
- 230000003462 zymogenic effect Effects 0.000 description 2
- 108020004465 16S ribosomal RNA Proteins 0.000 description 1
- MNKRTDOUBUSQHX-UHFFFAOYSA-N 2,4-dihydroxy-2-methyl-3-oxopentanoic acid Chemical compound CC(O)C(=O)C(C)(O)C(O)=O MNKRTDOUBUSQHX-UHFFFAOYSA-N 0.000 description 1
- 108091005508 Acid proteases Proteins 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 241001269238 Data Species 0.000 description 1
- 101100311938 Dictyostelium discoideum phesA gene Proteins 0.000 description 1
- 101100423325 Dictyostelium discoideum phesB gene Proteins 0.000 description 1
- 239000004278 EU approved seasoning Substances 0.000 description 1
- 241000726221 Gemma Species 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000000843 anti-fungal effect Effects 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 239000013065 commercial product Substances 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 239000012531 culture fluid Substances 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 235000011194 food seasoning agent Nutrition 0.000 description 1
- 230000003760 hair shine Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 101150080777 pheS gene Proteins 0.000 description 1
- SBNFWQZLDJGRLK-UHFFFAOYSA-N phenothrin Chemical compound CC1(C)C(C=C(C)C)C1C(=O)OCC1=CC=CC(OC=2C=CC=CC=2)=C1 SBNFWQZLDJGRLK-UHFFFAOYSA-N 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000012797 qualification Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 235000014347 soups Nutrition 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 235000015099 wheat brans Nutrition 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12J—VINEGAR; PREPARATION OR PURIFICATION THEREOF
- C12J1/00—Vinegar; Preparation or purification thereof
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Medicinal Chemistry (AREA)
- Mycology (AREA)
- Botany (AREA)
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Soy Sauces And Products Related Thereto (AREA)
Abstract
The present invention relates to a kind of Pediococcus acidilactici and its purposes in flavouring is being prepared, the method for further relating to prepare flavouring using the Pediococcus acidilactici.The bacterial strain is Pediococcus acidilactici (Pediococcus acidilactici) ZF559, and deposit number is CCTCC M 2016690.The bacterial strain is applied when preparing flavouring, enough significantly inhibits the growth of bacillus, significantly reduces the bacillus number in product.
Description
Technical field
The invention belongs to microorganisms technical fields, and in particular to a kind of Pediococcus acidilactici and its prepare the use in flavouring
On the way, the method for further relating to prepare flavouring using the Pediococcus acidilactici.
Background technique
In the production process of the traditional zymotics type flavouring such as sauce or soy sauce, a large amount of bacillus can be bred.Yeast making process
In, the growth of bacillus can consume carbohydrate, organic acid and amino acid in raw material, generate bad smell, can also produce
It gives birth to antifungal activity substance and consumes oxygen rapidly, so that aspergillus be inhibited to grow, reduce the protease activity of aspergillus, seriously affect subsequent
Fermented quality.Researcher also found, the major microorganisms that bacillus still causes sack expanding of sauce rotten.Bacillus generates
Gemma, heat resistance is very strong, needed before product sterilization packaging using high temperature for a long time inactivate, this will cause product special flavour
The increase of loss and production energy consumption with nutrition, it is also possible to the risk for bringing product total plate count exceeded.Therefore, in koji-making
The proliferation of stage control bacillus seems especially important for promoting koji-making and fermented quality.
In starter-making stage, micrococcus luteus and streptococcic proliferation produce acid, pH value can be made to decline, thus the life to bacillus
Length plays inhibiting effect, and still, it is too fast and unfavorable to karusen generation that a large amount of increments of micrococcus luteus may result in pH value decline
It influences (referring to Bao Qian, soy sauce science and brewing technique, China Light Industry Press, 2011).
CN105524905A discloses a kind of soy sauce complex enzyme containing acid protease, contains in the complex enzyme and is able to suppress
The spice extract of bacillus subtilis, spice extract can prevent sauce fermentation process living contaminants, guarantee fermentation just
Often carry out.
In the production process of the traditional zymotics type flavouring such as sauce or soy sauce, the proliferation of bacillus how is controlled, is promoted
The growth of Aspergillus strain promotes the prolease activity of aspergillus, is always that this field endeavours to solve the problems, such as.
Summary of the invention
Inventor outer obtains one plant of lactic acid sheet from Foshan Haitian's fermentation moromi is favorite
Coccus.The bacterial strain is added in koji-making, the growth of bacillus can be significantly inhibited, not will cause the decline of pH value, it will not be right
Subsequent fermentation is brought a negative impact, it can be ensured that the harmonic growth of multi-cultur es promotes the growth of Aspergillus strain, promotes the egg of aspergillus
White enzyme activity improves fermentation index and flavor taste quality, significantly reduces the bacillus number in product.
The first aspect of the present invention is related to a kind of Pediococcus acidilactici (Pediococcus acidilactici) ZF559,
Deposit number is CCTCC M 2016690.
The bacterial strain has been preserved in China typical culture collection center on November 28th, 2016, and (abbreviation CCTCC, address are
Wuhan University, Wuhan City, postcode: 430072), deposit number is CCTCC M 2016690, and classification naming is Pediococcus acidilactici
(Pediococcus acidilactici), strain name ZF559.
Inventor's separation from Foshan Haitian's fermentation moromi obtains one plant of bacterial strain, will
It is named as ZF559, and screening process is referring to Fig. 1.According to cellular morphology, physiological and biochemical property, 16S rRNA gene sequencing, pheS
The experimental datas comprehensive analysis such as gene sequencing, the bacterial strain are accredited as Pediococcus acidilactici (Pediococcus
acidilactici).Qualification result is shown in that " Institute of Microorganism, Academia Sinica's test sensitivity reports (2015) micro- searching the 127th
Number ".The results are shown in Table 1 for the cellular morphology and physical and chemical experiment of the bacterial strain.
1 cellular morphology of table and physical and chemical experiment result
Pediococcus acidilactici ZF559 of the present invention derives from traditional zymotic system, belongs to and is conventionally used to food production
The strain of processing.According to European Food Safety Authority (EFSA) for the logical of food microorganism fungus kind (MFC) version publication in 2013
Cross security rights certification (QPS) microorganism list, Pediococcus acidilactici (Pediococcus acidilactici) without use limit
Condition processed belongs to highly-safe microorganism fungus kind.
Utilize the conventional method culture Pediococcus acidilactici (Pediococcus of the present invention of culture lactic acid bacteria
Acidilactici) ZF559, can obtain vigor height, and bacteria concentration reaches 108The bacteria culture fluid of CFU/mL or more.Preferably
Cultivation temperature is 30 DEG C~35 DEG C, and preferred incubation time is 16 hours~48 hours,
The second aspect of the present invention is related to a kind of compound bacteria, wherein including Pediococcus acidilactici described in first aspect present invention
(Pediococcus acidilactici) ZF559, and other optional strains.
In one embodiment, compound bacteria of the present invention, wherein other described strains are that can be used for preparing
The strain of fermented type flavouring, such as aspergillus oryzae (Aspergillus oryzae) or Aspergillus sojae (Aspergillus
sojae).In compound bacteria of the present invention, the mass ratio of the Pediococcus acidilactici ZF559 and other strains is preferably 1:
10~1:2 (such as 1:3,1:4,1:5,1:6,1:7,1:8 or 1:9).
In another embodiment, compound bacteria of the present invention is by by Pediococcus acidilactici of the present invention
(Pediococcus acidilactici) ZF559 and aspergillus oryzae (Aspergillus oryzae) composition, Pediococcus acidilactici
The mass ratio of ZF559 and aspergillus oryzae is preferably 1:10~1:2 (such as 1:3,1:4,1:5,1:6,1:7,1:8 or 1:9).
In another embodiment, compound bacteria of the present invention is by by Pediococcus acidilactici of the present invention
(Pediococcus acidilactici) ZF559 and Aspergillus sojae (Aspergillus sojae) composition, Pediococcus acidilactici
The mass ratio of ZF559 and Aspergillus sojae is preferably 1:10~1:2 (such as 1:3,1:4,1:5,1:6,1:7,1:8 or 1:9).
In another embodiment, compound bacteria of the present invention is by by Pediococcus acidilactici of the present invention
(Pediococcus acidilactici) ZF559, aspergillus oryzae (Aspergillus oryzae) and Aspergillus sojae
(Aspergillus sojae) composition, wherein the mass ratio of aspergillus oryzae and Aspergillus sojae is preferably 1:9~9:1, Pediococcus acidilactici
ZF559 and the mass ratio of mixing aspergillus (aspergillus oryzae and Aspergillus sojae mixture) be preferably 1:10~1:2 (such as 1:3,1:4,1:
5,1:6,1:7,1:8 or 1:9).
The third aspect of the present invention is related to Pediococcus acidilactici (Pediococcus described in first aspect present invention
Acidilactici) ZF559 or the compound bacteria are preparing the purposes in flavouring.
The fourth aspect of the present invention is related to a kind of method for preparing flavouring, including uses Pediococcus acidilactici of the present invention
The step of (Pediococcus acidilactici) ZF559 or the compound bacteria prepare flavouring.
In one embodiment, preparation method described in fourth aspect present invention, wherein the Pediococcus acidilactici
(Pediococcus acidilactici) ZF559 or compound bacteria use in koji-making step.
In aforementioned fourth aspect present invention any embodiment, the preparation method the following steps are included:
1) by bacterium needed for Pediococcus acidilactici (Pediococcus acidilactici) ZF559 and other koji-makings
Kind is seeded in starter-making materials, or the compound bacteria is seeded in starter-making materials, and culture obtains into song；
2) at addition salt water post-fermentation in song.
In aforementioned fourth aspect present invention any embodiment, lactic acid sheet described in the step 1) of the preparation method
The inoculum concentration of coccus (Pediococcus acidilactici) ZF559 is 1 × 103~1 × 107CFU/g starter-making materials, preferably
It is 1 × 103~9.9 × 106CFU/g starter-making materials, further preferably 1 × 103~9.9 × 105CFU/g starter-making materials.
Preparation method described in aforementioned fourth aspect present invention any embodiment, the culture in step 1) can be at this
It is cultivated under the common koji-making condition in field field.Cultivation temperature is preferred are as follows: 28 DEG C~38 DEG C.Incubation time is preferably 24 small
When~60 hours, such as 48 hours.
Preparation method described in aforementioned fourth aspect present invention any embodiment can use this field in step 2)
Common fermentation process ferments, such as high-salt fermentation, high-salt dilute naturally shine and make fermentation, low-salt solid-state fermentation, low
Salt heat-preservation fermentation.The technological parameter of fermentation process can be adjusted according to practical condition.
Preparation method described in aforementioned fourth aspect present invention any embodiment, wherein the starter-making materials are selected from
Soybean, defatted soybean, wheat and wheat bran.The wheat is preferably wheat flour.
Flavouring of the present invention is primarily referred to as being fermented and being generated the fermented type of flavor using microorganism and its enzyme
Flavouring, such as soy sauce, sauce, vinegar, fermented soya bean or fermented bean curd etc..
In a specific embodiment, the present invention utilizes the Pediococcus acidilactici (Pediococcus
Acidilactici) ZF559 prepares soy sauce or sauce.It is in koji-making, soybean/defatted soybean and wheat/wheat flour is routinely square
After method carries out boiling processing, it is inoculated with Pediococcus acidilactici (Pediococcus acidilactici) ZF559 of the present invention and other systems
Qu Suoxu strain (such as aspergillus oryzae and/or Aspergillus sojae), the inoculum density of Pediococcus acidilactici ZF559 are 103CFU/g material~
107The inoculum concentration of CFU/g material, aspergillus oryzae or Aspergillus sojae is the 0.01%~5% of raw material weight.The koji-making time is 24~60
Hour.Obtained Cheng Qu, bacillus is few, pH value does not decline, prolease activity is high, and bacillus number reduces a quantity
Grade or more, neutral protease vigor improves 30% or more, at bent pH value between 6.0~6.4, in the reasonable scope.
Above-mentioned Cheng Qu is fermented using solarization pond or fermentor, salinity management, temperature management and fermentation period management are pressed
More solito carries out, and moromi bacillus number when fermentation terminates reduces 1~2 order of magnitude or more, and amino-acid nitrogen mentions
High by 5% or more, moromi flavor is improved.
Advantageous effects of the invention
Pediococcus acidilactici (Pediococcus acidilactici) ZF559 provided by the invention is easy to cultivate, safety
It is high.Flavouring is prepared using the strain, has the advantages that following one or more:
1) the bacillus quantity of Cheng Quzhong can be significantly reduced；
2) it is obviously improved the neutral protease vigor of Cheng Quzhong；
3) pH value of Cheng Qu does not reduce；
4) bacillus quantity significantly reduces in gained flavouring；
5) amino acid nitrogen content is obviously improved in gained flavouring；
6) gained flavouring flavor be improved significantly；
7) product quality of gained flavouring gets a promotion.
Detailed description of the invention
The screening process figure of Fig. 1 Pediococcus acidilactici (Pediococcus acidilactici) ZF559；
The sense organ appraise result of high-salt fermentation crude oil prepared by Fig. 2 embodiment 2；
The sense organ appraise result of lower salt, heat preservation and fermented crude oil prepared by Fig. 3 embodiment 3；
The sense organ appraise result of fermentation beans unstrained spirits prepared by Fig. 4 embodiment 4.
Specific embodiment
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and should not be taken as limiting the scope of the invention.Unless stated otherwise, implement
Reagent, the method and apparatus used in example is the art conventional reagent, method and apparatus.It is not specified in embodiment specific
Condition person carries out according to conventional conditions or manufacturer's recommended conditions.Production firm is not specified in material therefor, reagent or instrument
Person, being can be with conventional products that are commercially available.
Unless stated otherwise, used medium of the embodiment of the present invention and experimental condition are this field conventional medium and test
Condition.
In following example:
The detection method of total plate count is referring to GB 4789.2-2010.
In the detection method of bacillus number, for routine operation referring to GB 4789.2-2010, special operation is after dilution
Select 2~3 suitable dilutions, 92 DEG C of ± 1 DEG C of maintenances 15min of water-bath, then press GB 4789.2-2010 in method into
Row inoculated and cultured and counting.
In the detection method of lactic acid bacteria number, for routine operation referring to GB 4789.2-2010, special operation is culture medium selection
MRS culture medium is cultivated after inoculation using anaerobic culture box.
The detection method of neutral protease vigor referring to " fermented seasonings production technology ", (compile by Shanghai brewing science research institute
Write revised edition) Section six of chapter 1.
The detection method of total acid is referring to GB/T 12456-2008.
The detection method of amino-acid nitrogen is referring to the first method and GB/T 5009.40- in GB/T 5009.39-2003
2003。
The detection method of reduced sugar is referring to GB 5009.7-2008.
The detection method of glutamic acid is referring to GB/T 5009.124-2003
The sense organ appraise method of soy sauce crude oil and beans unstrained spirits: convening 10 appraise personnel with abundant sense organ appraise experience,
2~5 samples of design carry out appraise, and appraise content includes delicate flavour, sweet taste, tart flavour, bitter taste, saline taste and dense sense totally 5 sides
Face scores to each dimension, and with 5 points for full marks, the higher preference degree of score is better.Points for attention in appraise are referring to GB/T
12315-2008。
The aspergillus oryzae used in following example has microorganism resource library by oneself from company.
Embodiment one
MRS meat soup commercial product culture medium is weighed, distilled water dissolution, cooling after 121 DEG C of sterilizing 15min is added by product description
It is spare.Pediococcus acidilactici (ZF559) and the lactobacillus plantarum cryopreservation tube strain (making 1.08 in Shanghai) for taking out preservation, respectively by both
Bacterial strain thaws rapidly 1min in 40 DEG C of water-baths, then aseptically by 1v/v% inoculum concentration by Pediococcus acidilactici (ZF559)
It is inoculated into MRS broth bouillon respectively, 30 DEG C~35 DEG C stationary cultures with lactobacillus plantarum cryopreservation tube strain (making 1.08 in Shanghai)
24 hours, i.e., obtaining concentration respectively is 108The Pediococcus acidilactici culture solution and lactobacillus plantarum culture solution of CFU/mL or more, is put in
4 DEG C of refrigerators are spare.
Embodiment lactyl-lactic acid piece coccus ZF559 is preparing the application in high-salt diluted state fermentation soy
1. koji-making
Use defatted soybean and wheat flour for raw material koji-making, wherein the mass ratio of defatted soybean and wheat flour is 6.5:3.5.
Blank control example and experimental example is respectively set, blank control example is only inoculated with aspergillus oryzae, and experimental example is inoculated with aspergillus oryzae and Ben Fa simultaneously
The bright Pediococcus acidilactici ZF559.Blank control example is identical with the inoculum concentration of aspergillus oryzae in experimental example, is 0.5wt%.It is real
The inoculum concentration of Pediococcus acidilactici ZF559 in example is tested referring to table 2.
The method of koji-making are as follows: defatted soybean is first added to the water of 1 times of weight, is moistened water 1 hour after mixing thoroughly, high pressure under 0.1Mpa
When being cooled to 40 DEG C, wheat flour, aspergillus oryzae and Pediococcus acidilactici ZF559 (blank control example is not added) is added in boiling 30min, into
Row aerated koji making, control zymogenic temperature obtains into song when cultivating 48 hours between 28 DEG C~38 DEG C in entire yeast making process.
Total plate count, bacillus number, lactic acid bacteria number and the neutral protease vigor for detecting Cheng Quzhong, the results are shown in Table 2.Knot
Fruit shows, compared with blank control example, does not cause into bent lactic acid bacteria sum after being added to Pediococcus acidilactici ZF559 of the invention
It increases and the significant decrease of pH value, total plate count is in the same order of magnitude, bacillus reduces by 1~2 order of magnitude, neutral protein
Enzyme activity is promoted significant.Meanwhile the higher bacillus number of Pediococcus acidilactici ZF559 inoculum concentration is lower in a certain range, but connects
Kind amount is excessively high to be brought a negative impact at bent pH value and prolease activity.Wherein, 105The inoculum concentration of CFU/g material is corresponding
Bacillus reduce by two orders of magnitude, neutral protease vigor promotes 32% or more, and pH value is between 6.0~6.4, in just
Normal range.
2 soy sauce of table is at bent Quality Comparison result
2. fermentation
It is separately added into the salt water that 2 times of weight concentrations are 19wt% at song, high-salt dilute is carried out after mixing and naturally shines system hair
Ferment stirs evenly in fermentation process on the the 2nd, 5 day, behind it is primary every stirring in 1 week, fermentation terminates for 60 days, filters soy sauce crude oil.
Total acid, amino-acid nitrogen, reduced sugar and the glutamic acid of detection gained soy sauce crude oil, the results are shown in Table 3.And to soy sauce original
Oil carries out sense organ appraise, as a result sees Fig. 2.
The results show that being added to after Pediococcus acidilactici ZF559 of the invention, soy sauce crude oil compared with blank control example
Total acid, ammonia nitrogen have the promotion of certain amplitude, reduced sugar and the variation of glutamic acid index are little.Wherein, 105CFU/g material
The amino-acid nitrogen index highest of soy sauce crude oil fermented of inoculum concentration, promote 7% or more.
Compared with blank control example, different Pediococcus acidilactici ZF559 inoculum concentrations are in delicate flavour, sweet taste, saline taste, bitter taste and dense
Thickness sense aspect has different degrees of improvement to be promoted, in addition to 106Other than the inoculum concentration of CFU/g material, other tart flavour no significant differences.
Wherein, 105The soy sauce crude oil comprehensive flavour that the inoculum concentration of CFU/g material is fermented is optimal, the Improving flavor effect of soy sauce crude oil
Fruit is significant.
The soy sauce crude oil physical and chemical index testing result that 3 high-salt dilute of table obtains
Three Pediococcus acidilactici ZF559 of embodiment is preparing the application in lower salt, heat preservation and fermented soy sauce
1. koji-making
Use defatted soybean and wheat flour for raw material koji-making, wherein the mass ratio of defatted soybean and wheat flour is 6.5:3.5.
Blank control example and experimental example is respectively set, blank control example is only inoculated with aspergillus oryzae, and experimental example is inoculated with aspergillus oryzae and Ben Fa simultaneously
The bright Pediococcus acidilactici ZF559.Blank control example is identical with the inoculum concentration of aspergillus oryzae in experimental example, is 0.5wt%.It is real
The inoculum concentration for testing Pediococcus acidilactici ZF559 in example is 105CFU/g material.
The method of koji-making is the same as described in embodiment two.
2. fermentation
The present embodiment is fermented using lower salt, heat preservation and fermented technique at Qu Jinhang.1.6 times of weight are separately added into Cheng Quzhong
Concentration is the salt water of 15wt%, and 50 DEG C of heat preservation less salt fermentations are carried out after mixing, and fermentation carries out leaching oil on the the 2nd, 5,12,19 day, every time
Leaching oil mass is to add the 4wt% of salt water, and filtering obtains soy sauce crude oil after fermentation 30 days.
Total acid, amino-acid nitrogen, reduced sugar and the glutamic acid in soy sauce crude oil are detected, the results are shown in Table 4.Meanwhile to soy sauce
Crude oil carries out sense organ appraise, as a result sees Fig. 3.
The results show that being added to after Pediococcus acidilactici ZF559, use is lower salt, heat preservation and fermented compared with blank control example
Technique, the total acid of obtained soy sauce crude oil, reduced sugar, glutamic acid variation are little, and amino-acid nitrogen promotes 5% or more.
As shown in figure 3, being vaccinated with the soy sauce that the material of Pediococcus acidilactici ZF559 ferments compared with blank control example
Crude oil improves obviously in terms of delicate flavour and dense sense, other aspect differences are unobvious.
The lower salt, heat preservation and fermented obtained soy sauce crude oil physical and chemical index testing result of table 4
Application of the example IV Pediococcus acidilactici ZF559 in preparation beans sauce
1. koji-making
Use soybean and wheat flour for raw material koji-making, wherein the mass ratio of soybean and wheat flour is 7:3.Blank is respectively set
Reference examples, positive Control example and experimental example, blank control example are only inoculated with aspergillus oryzae, positive Control example while inoculated plant lactobacillus
And aspergillus oryzae, experimental example are inoculated with aspergillus oryzae and Pediococcus acidilactici ZF559 of the present invention simultaneously.Blank control example, the positive are right
It is identical with the inoculum concentration of aspergillus oryzae in experimental example as usual, it is 0.4wt%.The inoculum concentration of lactobacillus plantarum is in positive Control example
105CFU/g material.The inoculum concentration of Pediococcus acidilactici ZF559 is 10 in experimental example5CFU/g material.
The method of koji-making are as follows: first add 40 DEG C of water of 3 times of weight to impregnate 2 hours soybean, autoclaving under 0.1Mpa
10min when being cooled to 40 DEG C, is added wheat flour, aspergillus oryzae, lactobacillus plantarum or Pediococcus acidilactici ZF559, is aerated system
Song, for control zymogenic temperature between 28 DEG C~38 DEG C, culture obtained beans sauce Cheng Qu after 40 hours in entire yeast making process.
Total plate count, bacillus number, lactic acid bacteria number and the neutral protease vigor for detecting beans sauce Cheng Quzhong, the results are shown in Table
5。
5 beans sauce of table is at bent Quality Comparison result
The results show that compared with blank control example, after adding lactobacillus plantarum, although bacillus number has obtained centainly
The inhibition of degree, but significantly reducing occurs in the pH value of beans sauce Cheng Qu, and there is significant raising in total plate count and lactic acid bacteria number, in
Property prolease activity appearance decline by a small margin.And it is added to after Pediococcus acidilactici ZF559 of the invention, beans sauce Cheng Quzhong bacterium colony
Sum and lactic acid bacteria number difference are little, are in the same order of magnitude, and the pH value of Cheng Qu does not significantly reduce, maintain essentially in same
One is horizontal, and bacillus reduces by 1 order of magnitude or more, and neutral protease vigor promotes 41%, and pH value is between 6.0~6.4, place
In in normal range (NR).
2. fermentation
It is separately added into the salt water that 1.8 times of weight concentrations are 19wt% to Cheng Quzhong, natural shine is carried out after mixing and makes fermentation, hair
It stirs evenly within ferment the 2nd, 5,15,25,36 day, fermentation obtained beans unstrained spirits after 45 days.
Total acid, amino-acid nitrogen, reduced sugar and the glutamic acid for detecting beans unstrained spirits, the results are shown in Table 6.Meanwhile to gained beans unstrained spirits into
Row sense organ appraise, is as a result shown in Fig. 4.
The fermentation beans unstrained spirits physical and chemical index testing result of table 6
The results show that compared with blank control example, after adding lactobacillus plantarum, although bacillus is effectively suppressed,
But the total acid for the beans unstrained spirits that ferments increases, and downward trend is presented in amino-acid nitrogen；And be added to after Pediococcus acidilactici ZF559, it ferments
Total acid, reduced sugar, glutamic acid difference in beans unstrained spirits is little, and amino acid nitrogen content promotes 10% or more, and bacillus is had
Effect control.
As shown in figure 4, saline taste is weaker after adding lactobacillus plantarum, but mouthfeel meta-acid, and adds compared with blank control example
After Pediococcus acidilactici ZF559, saline taste and dense sense are significantly improved, and tart flavour and other flavor have no significant effect, and show
To the beneficial effect of fermentation beans unstrained spirits Improving flavor.
Claims (24)
- Pediococcus acidilactici 1. (Pediococcus acidilactici) ZF559, deposit number is CCTCC NO:M 2016690。
- 2. a kind of compound bacteria, wherein including Pediococcus acidilactici described in claim 1 (Pediococcus acidilactici) ZF559, and other optional strains.
- 3. compound bacteria as claimed in claim 2, wherein other described strains are the strain for being used to prepare fermented type flavouring.
- 4. compound bacteria as claimed in claim 3, wherein the fermented type flavouring is soy sauce, sauce, vinegar, fermented soya bean or fermented bean curd.
- 5. compound bacteria as claimed in claim 3, wherein other described strains be aspergillus oryzae (Aspergillus oryzae) or Aspergillus sojae (Aspergillus sojae).
- 6. compound bacteria as claimed in claim 2, wherein the compound bacteria is by Pediococcus acidilactici described in claim 1 (Pediococcus acidilactici) ZF559 and aspergillus oryzae (Aspergillus oryzae) composition, orThe compound bacteria is by Pediococcus acidilactici described in claim 1 (Pediococcus acidilactici) ZF559 and sauce Oily aspergillus (Aspergillus sojae) composition, orThe compound bacteria is by Pediococcus acidilactici described in claim 1 (Pediococcus acidilactici) ZF559, meter Qu Mould (Aspergillus oryzae) and Aspergillus sojae (Aspergillus sojae) composition.
- 7. compound bacteria as claimed in claim 6, wherein the compound bacteria is by Pediococcus acidilactici described in claim 1 (Pediococcus acidilactici) ZF559 and aspergillus oryzae (Aspergillus oryzae) composition, Pediococcus acidilactici The mass ratio of ZF559 and aspergillus oryzae is 1:10~1:2.
- 8. compound bacteria as claimed in claim 6, wherein the compound bacteria is by Pediococcus acidilactici described in claim 1 (Pediococcus acidilactici) ZF559 and Aspergillus sojae (Aspergillus sojae) composition, Pediococcus acidilactici The mass ratio of ZF559 and Aspergillus sojae is 1:10~1:2.
- 9. compound bacteria as claimed in claim 6, wherein the compound bacteria is by Pediococcus acidilactici described in claim 1 (Pediococcus acidilactici) ZF559, aspergillus oryzae (Aspergillus oryzae) and Aspergillus sojae (Aspergillus sojae) composition, the mass ratio of aspergillus oryzae and Aspergillus sojae are 1:9~9:1, Pediococcus acidilactici ZF559 with The mass ratio of the mixture of aspergillus oryzae and Aspergillus sojae is 1:10~1:2.
- 10. (Pediococcus acidilactici) ZF559 of Pediococcus acidilactici described in claim 1 or claim 2 to 9 times Compound bacteria described in one is preparing the purposes in flavouring.
- 11. purposes described in any one of claim 10, wherein the flavouring is fermented type flavouring.
- 12. purposes described in claim 11, wherein the fermented type flavouring is soy sauce, sauce, vinegar, fermented soya bean or fermented bean curd.
- 13. a kind of method for preparing flavouring, including use Pediococcus acidilactici (Pediococcus described in claim 1 Acidilactici the step of) ZF559 or the described in any item compound bacterias of claim 2 to 9 prepare flavouring.
- 14. the method for preparing flavouring described in claim 13, wherein the flavouring is fermented type flavouring.
- 15. the method for preparing flavouring described in claim 14, wherein the fermented type flavouring is soy sauce, sauce, vinegar, fermented soya bean Or fermented bean curd.
- 16. the method for preparing flavouring described in claim 13 the, wherein Pediococcus acidilactici (Pediococcus Acidilactici) ZF559 or compound bacteria use in koji-making step.
- 17. the described in any item methods for preparing flavouring of claim 13 to 16, comprising the following steps:1) strain needed for Pediococcus acidilactici (Pediococcus acidilactici) ZF559 and other koji-makings is connect Kind is seeded in starter-making materials into starter-making materials, or by the compound bacteria, and culture obtains into song；2) at addition salt water post-fermentation in song.
- 18. the method for preparing flavouring described in claim 17, Pediococcus acidilactici (Pediococcus described in step 1) Acidilactici) inoculum concentration of ZF559 is 1 × 103~1 × 107CFU/g starter-making materials.
- 19. the method for preparing flavouring described in claim 18, wherein Pediococcus acidilactici described in step 1) The inoculum concentration of (Pediococcus acidilactici) ZF559 is 1 × 103~9.9 × 106CFU/g starter-making materials.
- 20. the method for preparing flavouring described in claim 19, wherein Pediococcus acidilactici described in step 1) The inoculum concentration of (Pediococcus acidilactici) ZF559 is 1 × 103~9.9 × 105CFU/g starter-making materials.
- 21. the method for preparing flavouring described in claim 17, wherein cultivation temperature is 28 DEG C~38 DEG C.
- 22. the method for preparing flavouring described in claim 17, wherein incubation time is 24 hours~60 hours.
- 23. the method for preparing flavouring described in claim 17, the method wherein fermented in step 2) is selected from: high-salt dilute hair Ferment, low-salt solid-state fermentation and lower salt, heat preservation and fermented.
- 24. the method for preparing flavouring described in claim 17, the method wherein fermented in step 2) is that high-salt dilute is natural Shine system fermentation.
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