CN106987546A - Tetramethylpyrazine bacillus amyloliquefaciens and the special wheat bran purposes of distilled spirit with sesame flavour - Google Patents
Tetramethylpyrazine bacillus amyloliquefaciens and the special wheat bran purposes of distilled spirit with sesame flavour Download PDFInfo
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- CN106987546A CN106987546A CN201710374672.2A CN201710374672A CN106987546A CN 106987546 A CN106987546 A CN 106987546A CN 201710374672 A CN201710374672 A CN 201710374672A CN 106987546 A CN106987546 A CN 106987546A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
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Abstract
The invention discloses the purposes of a kind of Tetramethylpyrazine bacillus amyloliquefaciens and its special wheat bran of distilled spirit with sesame flavour, it is characterized in utilize the bacterial strains of Tetramethylpyrazine bacillus amyloliquefaciens Bacillus amyloliquefaciens XJB 104, pass through the preparation of primary seed solution, the preparation of secondary seed solution, carry out wheat bran solid substrate fermentation, finally obtained ripe zymogenic aeration-drying, the special wheat bran of Tetramethylpyrazine distilled spirit with sesame flavour is obtained.The special wheat bran of Tetramethylpyrazine distilled spirit with sesame flavour prepared using the inventive method has bright, in dark brown, the feature of smell typical case, slightly ammonia taste, Tetramethylpyrazine content ＞ 380mg/kg are beneficial to the quality-improving of distilled spirit with sesame flavour.
Description
Technical field
The invention belongs to field of engineering technology of making wine, and in particular to Tetramethylpyrazine bacillus amyloliquefaciens and use it for
Prepare the purposes of the special wheat bran of distilled spirit with sesame flavour.
Background technology
According to《Pharmacopeia》Introduce, Tetramethylpyrazine, also known as rhizome of chuanxiong piperazine are heterocycle, nitrogenous colorless crystalline compound,
It is separated and extracts first in the rhizome of Chinese herbal medicine Ligusticum wallichii, is a kind of conventional medicine for treating a variety of diseases.According to China《China makes
Make》(the tenth phase in 2007,60-61 pages) with《Brewing science and technology》(third phase in 2012,17-22 pages) is introduced, distilled spirit with sesame flavour
Middle Tetramethylpyrazine content is higher, and it not only has important contribution to the local flavor of distilled spirit with sesame flavour, while also giving sesame
The wholesome function of flavor white spirit, the content for improving Tetramethylpyrazine in distilled spirit with sesame flavour is conducive to sesame flavor style liquor wine product
The lifting of quality.Flavor substance and its manufacture commercial guild provide that Tetramethylpyrazine (coding 3237) is a kind of safe food wind
Taste additive.But GB/T 20824-2007《Distilled spirit with sesame flavour》Regulation, distilled spirit with sesame flavour do not allow addition edible alcohol with
The flavor and fragrance substance that non-liquor fermentation is produced.So, Tetramethylpyrazine in white wine can only be improved by production technology improvement
Content.Wheat bran is that sesame perfume (or spice) brewages the saccharifying ferment used in production process, improves the content of Tetramethylpyrazine in wheat bran,
It is the important channel for improving Tetramethylpyrazine content in sesame-flavor white spirit.It is special on Tetramethylpyrazine distilled spirit with sesame flavour at present
There is not been reported for the research of wheat bran.The entitled bacillus amyloliquefaciens of Chinese patent Authorization Notice No. CN102766592B and its use
One plant of culture presevation number is disclosed in the patent on way for CGMCC No.5740 bacillus amyloliquefaciens Bacillus
Amyloliquefaciens JNC-002, the ability with production Tetramethylpyrazine, but so far there are no that it is used for the fragrant wheat bran of sesame
Production report.
The content of the invention
The present invention is intended to provide a kind of Tetramethylpyrazine bacillus amyloliquefaciens and its special wheat bran of distilled spirit with sesame flavour
Purposes, Tetramethylpyrazine distilled spirit with sesame flavour is prepared using Tetramethylpyrazine bacillus amyloliquefaciens fermentation wheat bran solid medium
Special wheat bran.
The Tetramethylpyrazine bacillus amyloliquefaciens of the present invention, it is characterised in that be that the name obtained is separated from distiller's yeast is brewageed
For bacillus amyloliquefaciens Bacillus amyloliquefaciens XJB-104 bacterial strains, in preservation on March 20 in 2017
In China typical culture collection center, address：Wuhan City, Hubei Province Wuchang District Bayi Road Luo Jia Shan, culture presevation number is
CCTCC M 2017133；The strain morphology is shaft-like, long 2.6-3.5 μm, 1.0-1.3 μm of diameter, and colony colour is white；Should
Bacterial strain has the physiological and biochemical property of typical bacillus amyloliquefaciens, the gyrB gene orders of the bacterial strain and other solution starch buds
The homology of spore bacillus gyrB genes reaches more than 99%.
The Tetramethylpyrazine bacillus amyloliquefaciens of the present invention be used to preparing the purposes of the special wheat bran of distilled spirit with sesame flavour-
That is application method, it is characterised in that comprise the following steps：
1st, the preparation of Tetramethylpyrazine bacillus amyloliquefaciens primary seed solution
With oese in the Tetramethylpyrazine bacillus amyloliquefaciens that culture presevation number is CCTCC M 2017133
Picking colony is placed in first order seed culture in the pure culture inclined-plane of Bacillus amyloliquefaciens XJB-104 bacterial strains
In base, in 37 DEG C, 160rpm, 24h is cultivated, primary seed solution is obtained；
The formula of the primary-seed medium is：Peptone 10g/L, yeast extract 5g/L, NaCl 15g/L；
2nd, the preparation of Tetramethylpyrazine bacillus amyloliquefaciens secondary seed solution
Above-mentioned primary seed solution is inoculated in secondary seed medium by the inoculum concentration of 1-2% volume ratios, 37 DEG C,
160rpm, cultivates 20-25h, obtains secondary seed solution；
The formula of the secondary seed medium is：Glucose 70g/L, peptone 30g/L, yeast extract 10g/L, phosphoric acid hydrogen
Two ammonium 30g/L, pH to 7.4-7.6 is adjusted with 20-40g/L sodium hydrate aqueous solutions；
3rd, the preparation of the special wheat bran of Tetramethylpyrazine distilled spirit with sesame flavour
Above-mentioned secondary seed solution is accessed into wheat bran solid medium with every kg broth 0.06-0.10 inoculum concentrations risen
In, 37 DEG C of incubated 2-2.5d obtain ripe zymogenic, then the aeration-drying at 35-45 DEG C, to water content ＜ 13%, produce four
The special wheat bran of methylpyrazine distilled spirit with sesame flavour.
The culture presevation number is CCTCC M 2017133 bacillus amyloliquefaciens XJB-104 bacterial strains, and it separates and sieved
Choosing actual specific operation process be：Wine high temperature 80 DEG C of water-bath 30min of wrapped starter will be declared, spore solution, 30 DEG C of constant temperature is obtained
Cultivate and nutrient broth flat board is coated with after 24h, select feature single bacterium colony, it is anti-by skim milk Selective agar medium proteolysis circle
Answer, acetyl methyl carbinol (V-P) reacts and tunning Analyze ＆ separate, screen the obtained mesh with production Tetramethylpyrazine ability
Bacterial strain is marked, bacillus amyloliquefaciens are identified as.
The wheat bran solid medium can be using common bran mass, and preferred formula is in the present invention：Wheat bran
650-750g, bean cake powder 250-350g, NaOH 1-3g, water 1-1.4L.
Compared with the prior art, beneficial effects of the present invention are embodied in：
1st, purebred bacterial bran koji prepared by the inventive method makes controllability height, product compared with existing multi-cultur es wheat bran
It is stable, it is to avoid the complicated interspecies relation of multi-cultur es wheat bran incubation, definite functions, it is intended to increase Tetramethylpyrazine in wheat bran
Content.
2nd, bacterial bran koji prepared by the inventive method is added in distilled spirit with sesame flavour brewing process, grain is not only directly increased
Tetramethylpyrazine in unstrained spirits, the vigor bacterial strain of production Tetramethylpyrazine is also provided for brewing process, is further improved in wine body
The content of Tetramethylpyrazine, is one-time process technological innovation to brewageing for distilled spirit with sesame flavour, with very high theory and application
Value.
3rd, bean cake powder substitution part wheat bran, Ke Yiwei are added in the special wheat bran of Tetramethylpyrazine distilled spirit with sesame flavour of the present invention
The generation of Tetramethylpyrazine provides abundant protein precursor, is conducive to improving the content of Tetramethylpyrazine in wheat bran, while rich
Rich protein provides the precursor substance of abundance for Maillard reaction, is conducive to generation higher alcohol, senior ester and sesame-flavor
Distinctive sulfide -3 methylthiol propyl alcohol, increases the richness and stereovision of sesame cordiale body, to improving distilled spirit with sesame flavour
Yield and quality play positive role.
4th, sodium hydroxide is with the addition of in the special wheat bran of Tetramethylpyrazine distilled spirit with sesame flavour of the invention, is not only tetramethyl
The generation of pyrazine provides favourable weakly alkaline environment, and has neutralized part Tetramethylpyrazine bacillus amyloliquefaciens in fermentation
During the acids metabolite that produces, make that the fragrance of finished product wheat bran is purer, typical case, the raising to product quality is beneficial.
5th, the present invention has that technique is simple, be easy to operation, the wheat bran bright that is made, in dark brown, smell typical case, slightly
Band ammonia taste, Tetramethylpyrazine content ＞ 380mg/kg.
Embodiment
Embodiment 1：
The preparation method of the special wheat bran of Tetramethylpyrazine distilled spirit with sesame flavour is as follows in the present embodiment：
1st, the preparation of Tetramethylpyrazine bacillus amyloliquefaciens primary seed solution
With oese in the Tetramethylpyrazine bacillus amyloliquefaciens that culture presevation number is CCTCC M 2017133
Picking colony is placed in first order seed culture in the pure culture inclined-plane of Bacillus amyloliquefaciens XJB-104 bacterial strains
In base, 37 DEG C, 160rpm, 24h obtain primary seed solution；Wherein primary-seed medium：Peptone 10g/L, yeast extract 5g/
L, NaCl 15g/L；
2nd, the preparation of Tetramethylpyrazine bacillus amyloliquefaciens secondary seed solution
Above-mentioned primary seed solution is inoculated in secondary seed medium, 37 DEG C, 160rpm, training by 1% (V/V) inoculum concentration
20-25h is supported, secondary seed solution is obtained；Wherein secondary seed medium：Glucose 70g/L, peptone 30g/L, yeast extract 10g/L,
Diammonium hydrogen phosphate 30g/L, pH 7.5；
3rd, the preparation of the special wheat bran of Tetramethylpyrazine distilled spirit with sesame flavour
By above-mentioned secondary seed solution so that in 6% (L/kg) inoculum concentration access wheat bran solid medium, 37 DEG C incubated
2.5d produces ripe zymogenic, and ripe zymogenic aeration-drying at 40 DEG C, to water content ＜ 13%, obtains Tetramethylpyrazine sesame fragrant
The special wheat bran of type white wine；
The wheat bran solid medium can use the wheat bran solid-state employed in common bran mass, the present embodiment
The formula of culture medium is：Wheat bran 700g, bean cake powder 300g, NaOH 3g, water 1L.
Addition dregs of beans substitution part wheat bran, Ke Yiwei in the special wheat bran of Tetramethylpyrazine distilled spirit with sesame flavour of the present invention
The generation of Tetramethylpyrazine provides abundant protein precursor, is conducive to improving the content of Tetramethylpyrazine in wheat bran, while rich
Rich protein provides the precursor substance of abundance for Maillard reaction, is conducive to generation higher alcohol, senior ester and sesame-flavor
Distinctive sulfide -3 methylthiol propyl alcohol, increases the richness and stereovision of sesame cordiale body, to improving distilled spirit with sesame flavour
Yield and quality play positive role.
Gained Tetramethylpyrazine distilled spirit with sesame flavour special wheat bran bright in the present embodiment, in dark brown, smell allusion quotation
Type, slightly ammonia taste, Tetramethylpyrazine content are 391mg/kg.
Embodiment 2：
The preparation method of the special wheat bran of Tetramethylpyrazine distilled spirit with sesame flavour is as follows in the present embodiment：
1st, the preparation of Tetramethylpyrazine bacillus amyloliquefaciens primary seed solution
With oese in the Tetramethylpyrazine bacillus amyloliquefaciens that culture presevation number is CCTCC M 2017133
Picking colony is placed in first order seed culture in the pure culture inclined-plane of Bacillus amyloliquefaciens XJB-104 bacterial strains
In base, 37 DEG C, 160rpm, 24h obtain primary seed solution；Wherein primary-seed medium：Peptone 10g/L, yeast extract 5g/
L, NaCl 15g/L；
2nd, the preparation of Tetramethylpyrazine bacillus amyloliquefaciens secondary seed solution
Above-mentioned primary seed solution is inoculated in secondary seed medium, 37 DEG C, 160rpm, training by 1% (V/V) inoculum concentration
20-25h is supported, secondary seed solution is obtained；Wherein secondary seed medium：Glucose 70g/L, peptone 30g/L, yeast extract 10g/L,
Diammonium hydrogen phosphate 30g/L, pH 7.5；
3rd, the preparation of the special wheat bran of Tetramethylpyrazine distilled spirit with sesame flavour
By above-mentioned secondary seed solution so that in 10% (L/kg) inoculum concentration access wheat bran solid medium, 37 DEG C incubated
2.5d produces ripe zymogenic, and ripe zymogenic aeration-drying at 40 DEG C, to water content ＜ 13%, obtains Tetramethylpyrazine sesame fragrant
The special wheat bran of type white wine；
Described wheat bran solid medium is：Wheat bran 700g, bean cake powder 300g, NaOH 2g, water 1.2L；
Gained Tetramethylpyrazine distilled spirit with sesame flavour special wheat bran bright in the present embodiment, in dark brown, smell allusion quotation
Type, slightly ammonia taste, Tetramethylpyrazine content are 492mg/kg.
Embodiment 3：
The preparation method of the special wheat bran of Tetramethylpyrazine distilled spirit with sesame flavour is as follows in the present embodiment：
1st, the preparation of Tetramethylpyrazine bacillus amyloliquefaciens primary seed solution
With oese in the Tetramethylpyrazine bacillus amyloliquefaciens that culture presevation number is CCTCC M 2017133
Picking colony is placed in first order seed culture in the pure culture inclined-plane of Bacillus amyloliquefaciens XJB-104 bacterial strains
In base, 37 DEG C, 160rpm, 24h obtain primary seed solution；Wherein primary-seed medium：Peptone 10g/L, yeast extract 5g/
L, NaCl 15g/L；
2nd, the preparation of Tetramethylpyrazine bacillus amyloliquefaciens secondary seed solution
Above-mentioned primary seed solution is inoculated in secondary seed medium, 37 DEG C, 160rpm, training by 1% (V/V) inoculum concentration
20-25h is supported, secondary seed solution is obtained；Wherein secondary seed medium：Glucose 70g/L, peptone 30g/L, yeast extract 10g/L,
Diammonium hydrogen phosphate 30g/L, pH 7.5；
3rd, the preparation of the special wheat bran of Tetramethylpyrazine distilled spirit with sesame flavour
By above-mentioned secondary seed solution so that in 8% (L/kg) inoculum concentration access wheat bran solid medium, 37 DEG C incubated
2.5d produces ripe zymogenic, and ripe zymogenic aeration-drying at 40 DEG C, to water content ＜ 13%, obtains Tetramethylpyrazine sesame fragrant
The special wheat bran of type white wine；
Described wheat bran solid medium is：Wheat bran 700g, bean cake powder 300g, NaOH 2g, water 1.2L；
Gained Tetramethylpyrazine distilled spirit with sesame flavour special wheat bran bright in the present embodiment, in dark brown, smell allusion quotation
Type, slightly ammonia taste, Tetramethylpyrazine content are 505mg/kg.
Sequence table
<110>Anhui Xuanjiu Group Co., Ltd.；HeFei University of Technology
<120>Bacillus amyloliquefaciens gyrB genes
<130> 2017
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1085
<212> DNA
<213> Bacillus amyloliquefaciens
<400> 1
acgataaacg ccttgtcgac cactcttgac gttacggttc atcgtgacgg aaaaatccac 60
tatcaggcgt acgagcgcgg tgtacctgtg gccgatcttg aagtgatcgg tgatactgat 120
aagaccggaa cgattacgca cttcgttccg gatccggaaa ttttcaaaga aacaactgta 180
tatgactatg atctgctttc aaaccgtgtc cgggaattgg ccttcctgac aaaaggcgta 240
aacatcacga ttgaagacaa acgtgaagga caagaacgga aaaacgagta ccactacgaa 300
ggcggaatca aaagctatgt tgagtactta aaccgttcca aagaagtcgt tcatgaagag 360
ccgatttata tcgaaggcga gaaagacggc ataacggttg aagttgcatt gcaatacaac 420
gacagctata caagcaatat ttattctttc acaaataata tcaacacata cgaaggcggc 480
acgcacgagg ccggatttaa aaccggtctg acccgtgtca taaacgacta tgcaagaaga 540
aaagggattt tcaaagaaaa tgatccgaat ttaagcgggg atgatgtgag agaagggctg 600
actgccatta tttcaattaa gcaccctgat ccgcaattcg aagggcagac gaaaacgaag 660
ctcggcaact ccgaagcgag aacgatcact gatacgctgt tttcttctgc gctggaaaca 720
ttccttcttg aaaatccgga ctcagcccgc aaaatcgttg aaaaaggttt aatggccgca 780
agagcgcgga tggcggcgaa aaaagcgcgg gaattgaccc gccgcaaaag tgcgcttgag 840
atttccaatc tgccgggcaa actggcggac tgttcttcta aagatccgag catttccgag 900
ctgtatatcg tagagggtga ctctgcgggc ggatcagcga aacagggacg ggaccgtcat 960
ttccaagcca ttctgccgct gcgcggtaag attctgaacg ttgagaaagc cagacttgat 1020
aagattctct caaacaatga ggtcagatca atgatcacgg ccctcggaac aggaatcggc 1080
gaaga 1085
Claims (4)
1. one plant of Tetramethylpyrazine bacillus amyloliquefaciens, it is characterised in that entitled Bacillus amyloliquefaciens
XJB-104, form is shaft-like, long 2.6-3.5 μm, 1.0-1.3 μm of diameter, and colony colour is white, and the bacterial strain has typical
The physiological and biochemical property of bacillus amyloliquefaciens, the gyrB gene orders of the bacterial strain and other bacillus amyloliquefaciens gyrB genes
Homology reach more than 99%, the strain has been preserved in China typical culture collection center, and culture presevation number is：CCTCC
M 2017133。
2. the purposes of the special wheat bran of Tetramethylpyrazine distilled spirit with sesame flavour is prepared using Tetramethylpyrazine bacillus amyloliquefaciens, its
It is characterised by comprising the following steps：
(1) preparation of Tetramethylpyrazine bacillus amyloliquefaciens primary seed solution
With oese in flat board picking bacillus amyloliquefaciens XJB-104 bacterium colonies in primary-seed medium, 37 DEG C,
160rpm, cultivates 24h, obtains primary seed solution；
The formula of the primary-seed medium is：Peptone 10g/L, yeast extract 5g/L, NaCl 15g/L；
(2) preparation of Tetramethylpyrazine bacillus amyloliquefaciens secondary seed solution
Above-mentioned primary seed solution is inoculated in secondary seed medium by the inoculum concentration of 1-2% volume ratios, in 37 DEG C, 160rpm,
20-25h is cultivated, secondary seed solution is obtained；
The formula of the secondary seed medium is：Glucose 70g/L, peptone 30g/L, yeast extract 10g/L, diammonium hydrogen phosphate
30g/L, pH to 7.4-7.6 is adjusted with 20-40g/L sodium hydrate aqueous solution；
(3) preparation of the special wheat bran of Tetramethylpyrazine distilled spirit with sesame flavour
Above-mentioned secondary seed solution is accessed in wheat bran solid medium with every kg broth 0.06-0.10 inoculum concentrations risen, 37
DEG C incubated 2-2.5d, obtains ripe zymogenic, then the aeration-drying at 35-45 DEG C, to water content ＜ 13%, produces tetramethyl pyrrole
The special wheat bran of piperazine distilled spirit with sesame flavour.
3. prepare Tetramethylpyrazine distilled spirit with sesame flavour using Tetramethylpyrazine bacillus amyloliquefaciens according to claim 2
The purposes of special wheat bran, is characterised by described bacillus amyloliquefaciens XJB-104, is that will declare 80 DEG C of water of wine high temperature wrapped starter
30min is bathed, obtains being coated with nutrient broth flat board after spore solution, 30 DEG C of incubated 24h, selects feature single bacterium colony, by de-
The reaction of fat milk Selective agar medium proteolysis circle, acetyl methyl carbinol reaction and tunning Analyze ＆ separate, what screening was obtained
Aimed strain with production Tetramethylpyrazine ability, is identified as bacillus amyloliquefaciens.
4. prepare Tetramethylpyrazine distilled spirit with sesame flavour using Tetramethylpyrazine bacillus amyloliquefaciens according to claim 2
The purposes of special wheat bran, the formula for being characterised by the wheat bran solid medium is：Wheat bran 650-750g, bean cake powder 250-
350g, NaOH 1-3g, water 1-1.4L.
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Cited By (4)
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| CN109706097A (en) * | 2019-01-10 | 2019-05-03 | 江南大学 | Produce the strain of sesame fragrant bacterium and its application in liquor production |
| CN109880720A (en) * | 2017-12-06 | 2019-06-14 | 安徽宣酒集团股份有限公司 | The production method that a kind of Tetramethylpyrazine loses poor function wheat bran |
| CN110760404A (en) * | 2019-11-27 | 2020-02-07 | 四川轻化工大学 | Bacillus mixed bran koji and preparation process and application thereof |
| CN114854617A (en) * | 2022-03-01 | 2022-08-05 | 四川大学 | Bacillus amyloliquefaciens for high-yield tetramethylpyrazine and solid state fermentation simulation method thereof |
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