CN102899262B - Lactobacillus plantarum and method for fast degrading nitrite produced in fermentation by the lactobacillus plantarum - Google Patents

Lactobacillus plantarum and method for fast degrading nitrite produced in fermentation by the lactobacillus plantarum Download PDF

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CN102899262B
CN102899262B CN201210180129.6A CN201210180129A CN102899262B CN 102899262 B CN102899262 B CN 102899262B CN 201210180129 A CN201210180129 A CN 201210180129A CN 102899262 B CN102899262 B CN 102899262B
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China
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fermentation
pepper
lactobacillus plantarum
huangdi
nitrite
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CN201210180129.6A
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Chinese (zh)
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CN102899262A (en
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诸葛斌
李汉文
方慧英
孙进
龚星慧
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浙江正味食品有限公司
江南大学
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    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02ATECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/90Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in food processing or handling, e.g. food conservation

Abstract

The invention discloses lactobacillus plantarum and a method for fast reducing content of nitrite produced in Huangdi pepper fermentation by inoculation of the lactobacillus plantarum. The method comprises that in Huangdi pepper fermentation, a lactic acid bacterium strain broth which is subjected to activation and enlarge cultivation is introduced and is used for fermentation. The method is characterized in that the Huangdi pepper is a high-pungency degree pepper produced only in Hainan; the lactic acid bacterium strain broth is a liquid of lactobacillus plantarum; the lactobacillus plantarum is screened from natural fermentation microbes of the high-pungency Huangdi pepper; the inoculation quantity of the lactobacillus plantarum is in a range of 3 to 5% (by volume); and after fermentation, 12% of a high-content salt is added into the fermentation products and is used for seasoning and killing residual microbes. Through the method, in Huangdi pepper fermentation, nitrite content is low and an original local flavor and nutrients of the Huangdi pepper are kept. The method has simple processes, greatly reduces a Huangdi pepper product production period, obviously improves product safety and has a wide application prospect.

Description

A kind of method of plant lactobacillus and fast degradation fermenting process nitrite thereof
Technical field
The present invention relates to a kind of plant lactobacillus and application thereof, particularly a kind of fermenting process nitrite plant lactobacillus capable of being fast degraded and application thereof.
Technical background
Fermented vegetables is one of traditional fermented food of China, has long edible history.Traditional vegetable fermentation technique is mainly spontaneous fermentation.Vegetables are easy to accumulate nitrogen element in soil and nitrate in process of growth, nitrate in vegetables is in the process of anaerobically fermenting, can be because nitrate reduction forms nitrite, microbe species in spontaneous fermentation process is mixed and disorderly, by it, be that earlier fermentation exists and much can reduce the microorganism of nitrate, can greatly increase product nitrite.Nitrite has very large injury to human body.The intake of 0.2-0.5g can make people poisoning, and 3g is lethal.In addition, nitrite or the precursor of strong carcinogen nitrosamine, it can with secondary amine substance in vivo or external synthetic nitrosamine; Nitrite in human body has strong oxidizing property, can be in conjunction with Fe 2+and Fe 3+, force oxyphorase to lose oxygen carrying capacity.
Lack amino acid decarboxylase and Terminal oxidase system in most lactic acid bacteria, can not reduce nitrate, do not produce ammonia.During the fermentation, milk-acid bacteria can produce rapidly acid and reduce environment pH value, suppresses the growth of miscellaneous bacteria, reduces the generation of nitrite, and low pH value can be also that self-degraded occurs nitrite.Milk-acid bacteria can also produce nitrite reductase, and degrading nitrite lowers product nitrite.
Traditional leavened food is generally by extending fermentation time, treats that superior microorganism is lowering the content of product nitrite after forming by superior microorganism effect, and the growing amount of this process nitrite exists peak value, and the cycle is also relatively long.At present the research of leavened food nitrite is mainly concentrated on to the dynamic change of nitrite content and nitrate reductase activity etc. upper, relatively less to the research of direct reduction nitrite content technological method.The today that becomes and become more and more important in food safety, the zymotechnique of inventing a kind of energy fast degradation product nitrite is significant.
Summary of the invention
The milk-acid bacteria that the object of this invention is to provide a kind of Yellow Emperor, a legendary ruler green pepper product nitrite capable of being fast degraded, to reduce the content of Yellow Emperor, a legendary ruler green pepper nitrite, improves the security of product.
The milk-acid bacteria of using in the present invention be screening to the natural fermented micro-raw bacterium in the Yellow Emperor, a legendary ruler green pepper of Hainan, bacterial strain is plant lactobacillus through Physiology and biochemistry experiment and 16S rDNA evaluation.
The method of fast reducing Yellow Emperor, a legendary ruler green pepper nitrite content of the present invention comprises the steps:
The first step, the preparation of substratum:
Bacterial strain activation medium: MRS substratum (g/L) (extractum carnis 10, peptone 10, yeast powder 5, K 2hPO 42, ammonium citrate 2, sodium acetate 5, glucose 20, tween 80 1ml, MgSO 47H 2o 0.58, MnSO 44H 2o 0.25, pH=6.2-6.4,20g agar.)
Liquid seed culture medium: MRS substratum, does not add agar and be liquid nutrient medium.
Second step, the activation of bacterial strain: by the plant lactobacillus of the preserving activation of ruling on solid MRS substratum.
The 3rd step, the preparation of seed liquor: in liquid MRS substratum, in 37 ℃ of standing cultivation 24-30h, centrifugal collection thalline, washs thalline with aseptic physiological saline, is diluted to concentration 10 by the inoculation having activated in second step 9-10 10cFU/mL.
The 4th step, by fresh Yellow Emperor, a legendary ruler green pepper clean, chopping, the cold water of quality such as add, and by inoculum size 3%-5%(mass volume ratio) access the seed liquor preparing.
The 5th step, sealing and fermenting: by the material mixing up 30 ℃ of fermentation 20-24h in the container of sealing, stop fermentation.
The 6th step, high salt is antibacterial: after fermentation ends, add 12%(mass ratio) high salt suppress remaining microorganism.
The main body of fermenting in the inventive method bacterial strain is clear and definite, and fermenting process and quality product are all to control, and with short production cycle, technique is simple, and nitrite can reduce fast.The Yellow Emperor, a legendary ruler green pepper pickling out had both possessed the feature of traditional zymotic vegetables, had also well kept the original local flavor of capsicum and nutrition, and edible safety also significantly improves, and shelf-lives also obviously extends.
Biomaterial
The plant lactobacillus screening that the present invention uses is to the natural fermented micro-raw bacterium in the Yellow Emperor, a legendary ruler green pepper of Hainan, on December 12nd, 2011, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, depositary institution address is No. 3 Institute of Microorganism, Academia Sinica of No. 1, North Star West Road, Chaoyang District, BeiJing, China city institute, and Classification And Nomenclature is that plant lactobacillus (Lactobacillus plantarum) deposit number is CGMCC NO.5570.
specific embodiments
Embodiment 1: the acquisition of bacterial classification
1) get pickle ripe Yellow Emperor, a legendary ruler green pepper sample, pickle half ripe Yellow Emperor, a legendary ruler green pepper sample, fresh Yellow Emperor, a legendary ruler green pepper sample each join in right amount in aseptic physiological saline, shake up rear gradient dilution and be applied on MRS solid medium, cultivate 48 hours for 37 ℃.
2) bacterium colony growing is rule separated 3-5 time on fresh MRS solid medium, obtain the bacterial strain of purifying, the bacterial strain being separated to is done to Physiology and biochemistry and 16S rDNA evaluation, and do respectively glycerine pipe and inclined-plane preservation.
3) activation to bacterial strain and access in the liquid MRS substratum of 25mL, in 37 ℃ of standing cultivations 24-30h, centrifugal collection thalline, washs thalline with aseptic physiological saline, is diluted to concentration 10 9-10 10cFU/mL.
4) fresh Yellow Emperor, a legendary ruler green pepper is cleaned, chopping, the cold water of quality such as add, by 3) in the seed liquor for preparing by inoculum size 3% access of plant lactobacillus seed liquor.
5) sealing and fermenting: by the material mixing up 30 ℃ of fermentation 20-24h in the container of sealing.Stop fermentation, and add 12%(mass ratio) high salt sterilization.
6) according to the spectrophotometry in the mensuration > > of GB5009.33-2010 < < nitrite in food and nitrate, measure respectively the nitrite content in Yellow Emperor, a legendary ruler green pepper.It is maximum that nitrite content ability falls in one lactobacillus plantarum, and the rear remaining nitrite content of fermentation is minimum is 3.2 μ g/g.This bacterial strain is preserved in to Chinese microorganism strain preservation center, and deposit number is CGMCCNO.5570.
Embodiment 2 Yellow Emperor, a legendary ruler green pepper fermentative production
1) preparation of substratum
Strain activation and culture base: MRS solid medium
Lactobacillus solution substratum: MRS liquid nutrient medium
2) activation of bacterial strain: by plant lactobacillus separated, that the preserve activation of ruling on solid MRS substratum.
3) in the liquid MRS substratum of the plant lactobacillus access 25mL having activated the preparation of seed liquor: by 2), in 37 ℃ of standing cultivation 24-30h, centrifugal collection thalline, the physiological saline washing thalline with aseptic, is diluted to concentration 10 9-10 10cFU/mL.
4) fresh Yellow Emperor, a legendary ruler green pepper is cleaned, chopping, the cold water of quality such as add, by 3) in the seed liquor for preparing by inoculum size 3% access of plant lactobacillus seed liquor.
5) sealing and fermenting: by the material mixing up 30 ℃ of fermentation 20-24h in the container of sealing.Stop fermentation, and add 12%(mass ratio) high salt sterilization.
6) fermentation stop after according to the nitrite content in the spectrophotometry Yellow Emperor, a legendary ruler green pepper in the mensuration > > of GB5009.33-2010 < < nitrite in food and nitrate, the nitrite content in Yellow Emperor, a legendary ruler green pepper is 5.2 μ g/g.
Embodiment 3: Yellow Emperor, a legendary ruler green pepper fermentative production
1) preparation of substratum
Strain activation and culture base: MRS solid medium
Lactobacillus solution substratum: MRS liquid nutrient medium
2) activation of bacterial strain: by plant lactobacillus separated, that the preserve activation of ruling on solid MRS substratum.
3) in the liquid MRS substratum of the plant lactobacillus access 25mL having activated the preparation of seed liquor: by 2), in 37 ℃ of standing cultivation 24-30h, centrifugal collection thalline, the physiological saline washing thalline with aseptic, is diluted to concentration 10 9-10 10cFU/mL.
4) fresh Yellow Emperor, a legendary ruler green pepper is cleaned, chopping, the cold water of quality such as add, by 3) in the seed liquor for preparing by inoculum size 4% access of plant lactobacillus seed liquor.
5) sealing and fermenting: by the material mixing up 30 ℃ of fermentation 20-24h in the container of sealing.Stop fermentation, and add the high salt sterilization of 12% (mass ratio).
6) fermentation stop after according to the nitrite content in the spectrophotometry Yellow Emperor, a legendary ruler green pepper in the mensuration > > of GB5009.33-2010 < < nitrite in food and nitrate, the nitrite content in Yellow Emperor, a legendary ruler green pepper is 4.2 μ g/g.
Embodiment 4: Yellow Emperor, a legendary ruler green pepper fermentative production
1) preparation of substratum
Strain activation and culture base: MRS solid medium
Lactobacillus solution substratum: MRS liquid nutrient medium
2) activation of bacterial strain: by plant lactobacillus separated, that the preserve activation of ruling on solid MRS substratum.
3) in the liquid MRS substratum of the plant lactobacillus access 25mL having activated the preparation of seed liquor: by 2), in 37 ℃ of standing cultivation 24-30h, centrifugal collection thalline, the physiological saline washing thalline with aseptic, is diluted to concentration 10 9-10 10cFU/mL.
4) fresh Yellow Emperor, a legendary ruler green pepper is cleaned, chopping, the cold water of quality such as add, by 3) in the seed liquor for preparing by inoculum size 5% access of plant lactobacillus seed liquor.
5) sealing and fermenting: by the material mixing up 30 ℃ of fermentation 20-24h in the container of sealing.Stop fermentation, and add 12%(mass ratio) high salt sterilization.
6) fermentation stop after according to the nitrite content in the spectrophotometry Yellow Emperor, a legendary ruler green pepper in the mensuration > > of GB5009.33-2010 < < nitrite in food and nitrate, the nitrite content in Yellow Emperor, a legendary ruler green pepper is 4.8 μ g/g.
Embodiment 5 bone mud fermentative production
1) preparation of substratum
Strain activation and culture base: MRS solid medium
Lactobacillus solution substratum: MRS liquid nutrient medium
2) activation of bacterial strain: by plant lactobacillus separated, that the preserve activation of ruling on solid MRS substratum.
3) in the liquid MRS substratum of the plant lactobacillus access 25mL having activated the preparation of seed liquor: by 2), in 37 ℃ of standing cultivation 24-30h, centrifugal collection thalline, the physiological saline washing thalline with aseptic, is diluted to concentration 10 9-10 10cFU/mL.
4) bone mud is smashed to pieces, in the ratio of 1:5, adding cold water to regulate initial pH is 6.8, by 3) in the seed liquor for preparing by inoculum size 2.5% access of plant lactobacillus seed liquor.
5) fermentation: by the material mixing up 37 ℃ of fermentation 20-24h in the container of sealing.Stop fermentation.
6) fermentation stop after according to the nitrite content in the spectrophotometry bone mud in the mensuration > > of GB5009.33-2010 < < nitrite in food and nitrate, the nitrite content in bone mud is 6.7 μ g/g.

Claims (1)

1. the method for a plant lactobacillus fast degradation fermenting process nitrite, the plant lactobacillus seed liquor that it is characterized in that accessing during the fermentation after activated, enlarged culturing is fermented, the inoculum size volume percent of seed liquor is 3%-5%, leavening temperature is 30 ℃, fermentation time is 20-24h, adds 12% high salt seasoning and kill remaining microorganism after fermentation ends; Described plant lactobacillus is preserved in Chinese microorganism strain preservation center on December 12nd, 2011, and deposit number is CGMCC NO.5570; Described fermentation is with Yellow Emperor, a legendary ruler green pepper fermentative production Hainan special product high peppery property Yellow Emperor, a legendary ruler green pepper;
Concrete steps are:
The first step, the preparation of substratum:
Bacterial strain activation medium, i.e. MRS substratum: extractum carnis 10g/L, peptone 10g/L, yeast powder 5g/L, K 2hPO 42g/L, ammonium citrate 2g/L, sodium acetate 5g/L, glucose 20g/L, tween 80 1ml/L, MgSO 47H 2o0.58g/L, MnSO 44H 2o0.25g/L, pH=6.2-6.4,20g/L agar;
Liquid seed culture medium: MRS substratum does not add agar and is liquid seed culture medium;
Second step, the activation of bacterial strain: by the plant lactobacillus of the preserving activation of ruling on solid MRS substratum;
The 3rd step, the preparation of seed liquor: in liquid seed culture medium, in 37 ℃ of standing cultivation 24-30h, centrifugal collection thalline, washs thalline with aseptic physiological saline, is diluted to concentration 10 by the inoculation having activated in second step 9-10 10cFU/mL;
The 4th step, by fresh Yellow Emperor, a legendary ruler green pepper clean, chopping, the cold water of quality the seed liquor preparing by the inoculum size access of mass volume ratio 3%-5% such as add;
The 5th step, sealing and fermenting: by the material mixing up 30 ℃ of fermentation 20-24h in the container of sealing, stop fermentation;
The 6th step, high salt is antibacterial: after fermentation ends, 12% increase salt and suppress remaining microorganism in mass ratio.
CN201210180129.6A 2012-05-31 2012-05-31 Lactobacillus plantarum and method for fast degrading nitrite produced in fermentation by the lactobacillus plantarum CN102899262B (en)

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