CN104388329B - A kind of application of Lactobacillus plantarum degradation biological amine in wine production - Google Patents
A kind of application of Lactobacillus plantarum degradation biological amine in wine production Download PDFInfo
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- CN104388329B CN104388329B CN201410467452.0A CN201410467452A CN104388329B CN 104388329 B CN104388329 B CN 104388329B CN 201410467452 A CN201410467452 A CN 201410467452A CN 104388329 B CN104388329 B CN 104388329B
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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
- 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
- C12R2001/225—Lactobacillus
- C12R2001/25—Lactobacillus plantarum
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- C—CHEMISTRY; METALLURGY
- 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
- C12G1/00—Preparation of wine or sparkling wine
- C12G1/02—Preparation of must from grapes; Must treatment and fermentation
- C12G1/0203—Preparation of must from grapes; Must treatment and fermentation by microbiological or enzymatic treatment
Abstract
The invention discloses the method for the biogenic amine during a kind of degraded wine production of utilization Lactobacillus plantarum, belong to biotechnology and fermentation arts.The bacterial strain is that screening is obtained from vinifera surface, the strain can not only effectively reduce the content of the biogenic amine in the MRS culture mediums that with the addition of putrescine, histamine and tyrasamine, during more directly applying to brewing grape wine, tolerance with preferable peracid, high alcohol content and SO2 high, the conversion ratio of malolactic acid is high, can effectively reduce the content of the biogenic amine during wine production.
Description
Technical field
The present invention relates to a kind of application of Lactobacillus plantarum degradation biological amine during wine production, belong to biological skill
Art and fermentation arts.
Background technology
Biogenic amine（Biogenic amines,BAs）It is nitrogenous low molecular weight organic compound that a class has bioactivity
General name.It is prevalent in numerous food and alcoholic fermented beverage, such as cheese, meat products, aquatic products, beer, grape
Wine and yellow rice wine are medium.Biogenic amine is the normal active component in biological organic body, and important physiological action is played in body.
It is the precursor of the material such as synthetic hormones, alkaloid, nucleic acid, protein in eukaryotic, is important neurotransmission, people
Take in a small amount of biogenic amine will not on human body produce influence.But when the content of biogenic amine reaches certain concentration, can cause
A series of adverse reaction, such as headache, disordered breathing, palpitaition ill symptomses.The toxicity of all kinds of amine is of different sizes, and histamine is to people
The influence of body health is maximum, although next to that tyrasamine, putrescine and cadaverine are without direct toxic action, can strengthen histamine
Toxicity.In addition, biogenic amine is to produce the indicator of sanitary condition and relevant with alcoholic beverages " drinking head more ".Therefore, part
The maximum Limited Doses of European Union member countries' setting section biogenic amine, related limitation of the China also to biogenic amine in Partial Food is done
Clearly specify.
Grape wine, as a kind of health drink, is to increase one of very fast, most active industry in the national economic development.Closely
The annual growth of grape wine is more than 20% within several years.2013, vintage was 117.83 ten thousand kilolitres, income from sales 408.17
Hundred million yuan, 43.81 hundred million yuan of profit.Although quickly growing in recent years, impacted by macroeconomy slowdown in growth and import grape wine etc.
The influence of factor, no matter domestic grape wine outlet is in quantity, or on the amount of money, compared with import grape wine, proportion is all
It is very low；And there is the big defect of product homogeneity, security-hidden trouble in China's wine industry, badly influenced industry
Develop in a healthy way.Therefore, the quality of the domestic grape wine of raising, reduction production cost, elimination security-hidden trouble are to improve grape wine
The main policies of international competitiveness, are also the key point of China's Grape Wine Industry sustainable and stable development.
Grape wine is used as a kind of fermented wine, shadow of its quality by factors such as raw material, leavening, fermentation condition and holding conditions
Ring.Used as the class in fermentation byproduct, biogenic amine is produced by decarboxylate of microorganisms enzymatic amino acid decarboxylase in brewing process
A class alkaline organic compound.The safety and sanitation quality of how many pairs of grape wine of its content has a significant impact, wherein histamine and junket
The toxicity of amine is maximum.External scientific research personnel has been detected by having more than 20 kinds of biogenic amine in grape wine；Part European Union member countries are
The maximum Limited Doses of histamine in grape wine are set, such as Germany：2mg/L；Belgium：6 mg/L；Holland：4 mg/L；France：8
mg/L；Switzerland and Ao Li ground：10 mg/L.China's health ministry is also promulgated and has formulated determination of biogenic amines standard in fermented food
（GB/T 5009.208-2008）, emphasis detection eight kinds of biogenic amines such as histamine and tyrasamine.It can be seen that, carry out biogenic amine in grape wine
Prevention and control it is critical that.
Because biogenic amine can produce certain toxic action in people's cylinder accumulation, then reduce the research of biogenic amine in food
It is critical that.The content for reducing biogenic amine in food mainly includes two aspects：One is the generation for controlling biogenic amine, but
The biogenic amine that degraded has been generated.
The content of the invention
It is an object of the invention to provide a kind of utilization Lactobacillus plantarum during wine production degradation biological amine
Method.
The present invention is achieved by the following technical solutions：
The Lactobacillus plantarum of degradation biological amine provided by the present invention, Latin is entitledlactobacillus plantarum,
Numbering L9, be it is isolated from vinifera surface, it is common in China Committee for Culture Collection of Microorganisms at present
The preservation of microorganism center, depositary institution address is：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, deposit number is CGMCC
No.9263, preservation date is：On 06 04th, 2014.The Physiology and biochemistry property of bacterial strain is as follows：Gram-positive bacteria, cell shape
State is rod long, single raw or opposite, word order, colonial morphology：Bacterium colony 1.0-1.2mm, projection, circle, smaller, smooth, side
Edge is neat, milky.
16S rDNA Sequence Identifications are carried out using bacterial universal primers 27F/1492R, is Lactobacillus plantarum.
The present invention has mainly carried out the application of Lactobacillus plantarum degradation biological amine by two ways：One is by plant breast
Bacillus is cultivated in being inoculated in the MRS culture mediums for the addition of monosomic amine, with blank, observes the change of Content of Biogenic Amines
Change situation.Two is that Lactobacillus plantarum is inoculated in into grape juice through carrying out fermentation test in the zymotic fluid after alcoholic fermentation, with sky
White control, observes the situation of change of Content of Biogenic Amines.
Three kinds of biogenic amines are with the addition of used by the present invention（Histamine, tyrasamine, putrescine）Modified MRS culture medium, its contained group
It is divided into：Peptone 10.0g, beef extract 10.0g, yeast extract 5.0g, glucose 20.0g,（NH4）2HC6H5O7 2.0 g、
CH3COONa·3H2O 5.0g、K2HPO4·3H2O 2.0g、MgSO4·7H2O 0.58g、MnSO4·H2The g of O 0.25, Tween 80
1.0 mL, putrescine 1mol/L, histamine 1 mol/L, tyrasamine 1mol/L, distilled water 1L, pH=5.5.
It is raw material, grape juice total sugar content that vinous fermentation liquid used by the present invention uses Changli area cabernet sauvignon grape juice
It is 208.45g/L.After being cultivated 6 days under the conditions of 28 DEG C, used after separating yeast.
Alcohol, pH, the SO of the present invention to the bacterial strain2Wine brewing compatibility test design：The MRS trainings of bacterial strain alcohol adaptability
Support alcoholic strength in base（Volume fraction）It is 10%, 12%, 14%；The pH of the MRS culture mediums of bacterial strain pH adaptability is 3.4,3.2,3.0；
Bacterial strain SO2SO in the MRS culture mediums of adaptability2Concentration is 20mg/L, 30mg/L, 50mg/L.The plant is carried out using above-mentioned culture medium
The fermenting experiment of thing lactobacillus, determines its OD600Value changes.
The method that the present invention determines the bacterial strain malic acid-lactic acid characteristic：Take activation bacteria suspension 1.0mL and access 100mL grapes
Juice through in the zymotic fluid after alcoholic fermentation, quiescent culture at 20 DEG C, using ion-chromatographic determination is initial, 5d, 10d, 15d, 20d
When malic acid, lactic acid content.
Specific embodiment：
Embodiment one：The biogenic amine that Lactobacillus plantarum is used for during degraded with the addition of the MRS culture mediums of monosomic amine
From picking Lactobacillus plantarum on MRS inclined-planes, activated 2-3 times in MRS culture mediums, picking 3-5 rings through overactivation plant
Thing lactobacillus is accessed in sterilized 10mL MRS culture mediums, and 18h is cultivated under the conditions of 30 DEG C, takes this bacteria suspension 1.0mL accesses
In the sterilized MRS culture mediums that with the addition of biological amine monomers of 100mL, after being cultivated 5 days under the conditions of 25 DEG C in detection nutrient solution
The change of histamine, tyrasamine and putrescine content.Testing result such as following table：
Content of Biogenic Amines change in the MRS culture mediums of table 1（mg/L）
| Strain | Putrescine | Histamine | Tyrasamine | Summation |
| L9 | -12.64 | -6.36 | -14.76 | -33.76 |
Note：Data are three kinds of differences of amine, negative value table in putrescine, histamine and tyrasamine in nutrient solution and blank cultures in table
Show Content of Biogenic Amines reduction.
Embodiment two：Lactobacillus plantarum is for the biogenic amine in vinous fermentation liquid of degrading
From picking Lactobacillus plantarum on MRS inclined-planes, activated 2-3 times in MRS culture mediums, picking 3-5 rings through overactivation plant
Thing lactobacillus is accessed in sterilized 10mL MRS culture mediums, and 18h is cultivated under the conditions of 30 DEG C, takes this bacteria suspension 1.0mL accesses
During 100mL grape juices are through the zymotic fluid after alcoholic fermentation, histamine, tyrasamine in nutrient solution are detected after being cultivated 20 days under the conditions of 20 DEG C
With the change of putrescine content.Testing result is as shown in the table：
Influence of the Lactobacillus plantarum of table 2 to Content of Biogenic Amines change during malic-lactic acid fermentation（Unit：mg/L）
| Strain | Putrescine | Histamine | Tyrasamine | Summation |
| Blank | 3.116 | 0.926 | 2.872 | 6.914 |
| L9 | 2.298 | 0.154 | 0.342 | 2.794 |
Embodiment three：The wine brewing adaptability and malic acid of Lactobacillus plantarum-lactic acid conversion are evaluated
The picking Lactobacillus plantarum from inclined-plane, activates 2-3 times in MRS culture mediums, and picking 3-5 rings are newborn through the plant of overactivation
Bacillus is accessed in MRS culture mediums, the quiescent culture 18h under the conditions of 30 DEG C；This bacteria suspension is taken according to the access sterilized spies of 100mL
Determine MRS culture mediums（Alcoholic strength, SO2It is as described above with pH culture mediums）In, the quiescent culture under the conditions of 25 DEG C determines initial and training
Support the OD of 10d600Value, OD600Value increases by 500 and then strain growth is in order above.Result shows that the Lactobacillus plantarum has
Good wine brewing adaptability, can tolerate alcoholic strength is 12 ° (V/V), and pH is 3.2, SO2Concentration is 30mg/L, i.e. the bacterial strain can be
Peracid, SO high2Grown with the grape wine environment of high alcohol content.
Further to analyze the ability of the bacterial strain malic acid-lactic acid conversion, malic-lactic acid fermentation process is simulated：Grape juice is at 28 DEG C
Under the conditions of cultivate 6 days, complete alcoholic fermentation after, separate bottom yeast, by activated Lactobacillus plantarum L9 according to 107cfu
The amount of/ml is inoculated into zymotic fluid, and 25 DEG C ferment 20 days, determine malic acid, lactic acid content during being somebody's turn to do, and as a result such as table 3, make breast
Between the 4.5~5.0g/L of increment of acid, malic acid reduction amount is between 2.0~2.5g/L.
The malic acid of the Lactobacillus plantarum of table 3-lactic acid conversion capability is determined（Unit：g/L）
| Strain | Malic acid | Lactic acid |
| 0d | 5.810 | 7.119 |
| 5d | 4.987 | 8.523 |
| 10d | 4.003 | 9.013 |
| 15d | 3.894 | 10.984 |
| 20d | 3.695 | 11.748 |
The wine brewing adaptability of the L9 Lactobacillus plantarums used by the present invention is good, and malic acid-rotational rate of lactic acid is high, malic acid
2.115g/L is reduced, lactic acid increases by 4.629 g/L, and wine body is softer, and fragrance is coordinated, and in the vinous fermentation liquid that can degrade
Histamine, putrescine and tyrasamine, degradation rate are up to 59.59%, therefore can do the promoter of malolactic fermentation.
Claims (5)
1. a kind of Lactobacillus plantarum（lactobacillus plantarum）L9, its deposit number is CGMCC No.9263, is protected
Ensconce China Committee for Culture Collection of Microorganisms's common micro-organisms center.
2. the application of the Lactobacillus plantarum L9 bacterial strains degradation biological amine in vinous fermentation described in claim 1, the biology
Amine is histamine, tyrasamine and putrescine.
3. application as claimed in claim 2, is from picking Lactobacillus plantarum L9 bacterial strains on MRS inclined-planes, in MRS fluid nutrient mediums
Activation 2-3 times, picking 3-5 rings are accessed in sterilized 10mL MRS culture mediums through the Lactobacillus plantarum of overactivation, in 30 DEG C of bars
18h is cultivated under part, in taking this bacteria suspension 1.0mL access 100mL grape juices through the zymotic fluid after alcoholic fermentation, under the conditions of 20 DEG C
Culture 20 days.
4. the Lactobacillus plantarum L9 bacterial strains described in claim 1 carry out the application of malic acid-lactic acid conversion in vinous fermentation.
5. application as claimed in claim 4, is cultivated 6 days under the conditions of 28 DEG C of grape juice, after completing alcoholic fermentation, separates bottom
Yeast, by activated Lactobacillus plantarum L9 according to 107Cfu/ml is inoculated into zymotic fluid, and 25 DEG C ferment 20 days, and analysis is every
It malic acid and lactic acid content, makes the g/L of growth by 4.629 of lactic acid, malic acid reduction 2.115g/L.
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Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104694332B (en) * | 2015-03-17 | 2017-10-31 | 江南大学 | A kind of yellow wine production technology of low yield biogenic amine |
| CN106244383B (en) * | 2016-10-13 | 2019-04-02 | 福建农林大学 | The integrated processing method of Content of Biogenic Amines in a kind of reduction yellow rice wine |
| CN109401998B (en) * | 2018-10-25 | 2020-09-04 | 江南大学 | Lactobacillus mindendori for degrading biogenic amine and application thereof |
| CN110846242B (en) * | 2019-05-17 | 2022-08-16 | 宁夏大学 | Lactobacillus hilgardii strain and application thereof |
| CN111363699B (en) * | 2020-03-18 | 2022-07-15 | 鲁东大学 | Lactobacillus plantarum with both biological amine degradation activity and biological acid reduction activity and application of lactobacillus plantarum in fruit wine |
| CN111793584B (en) * | 2020-08-05 | 2021-11-02 | 四川省农业科学院农产品加工研究所 | Lactobacillus rhamnosus strain D2, and preparation method and application thereof |
| CN112760257B (en) * | 2021-01-27 | 2022-09-16 | 宁夏农产品质量标准与检测技术研究所(宁夏农产品质量监测中心) | Lactobacillus plantarum strain YC21 and application thereof |
| CN112795448B (en) * | 2021-02-19 | 2023-09-26 | 宁夏农产品质量标准与检测技术研究所(宁夏农产品质量监测中心) | Mixed bacteria fermentation process based on lactobacillus plantarum and saccharomyces cerevisiae |
| CN114395512B (en) * | 2022-02-22 | 2024-01-16 | 宁夏大学 | Lactic acid bacteria capable of degrading biogenic amine and application thereof |
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| CN101724591A (en) * | 2009-11-27 | 2010-06-09 | 南京农业大学 | Lactobacillus plantarum strain and application thereof |
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| CN101724591A (en) * | 2009-11-27 | 2010-06-09 | 南京农业大学 | Lactobacillus plantarum strain and application thereof |
Non-Patent Citations (2)
| Title |
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| Occurrence of lactic acid bacteria and biogenic amines in biologically aged wines.;M.Victoria Moreno-Arribas et al.;《Food Microbiology》;20080523;第25卷(第7期);第875-881页 * |
| 葡萄酒中生物胺含量的影响因素研究;张敬等;《中国酿造》;20121031;第31卷(第10期);第18-21页 * |
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