CN1940078B - Production of biological antiseptic agent phenyllactic acid - Google Patents
Production of biological antiseptic agent phenyllactic acid Download PDFInfo
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- CN1940078B CN1940078B CN2006100884309A CN200610088430A CN1940078B CN 1940078 B CN1940078 B CN 1940078B CN 2006100884309 A CN2006100884309 A CN 2006100884309A CN 200610088430 A CN200610088430 A CN 200610088430A CN 1940078 B CN1940078 B CN 1940078B
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Abstract
Production of biological antiseptic benzyl lactic acid is carried out by taking lactic acid as bacteria, taking phenylalanine, phenyl-pyruvic acid or sodium phenyl-pyruvic acid as conversion substrate, adding into fermentation culture medium with carbon source, nitrogen source and inorganic salt, preparing benzyl lactic acid by biological conversion, culturing lactic acid in MRS culture medium at 25-40degree and inspecting. The fermentation culture medium carbon source is selected from glucose, maltose, lactose or cane sugar; nitrogen source is selected from corn liquor, yeast cream, protein peptone and soy mill cake. The concentration of benzyl lactic acid reaches to 50-150mg/100ml. It's safe, reliable and has excellent sterilizing function.
Description
Technical field
The present invention relates to a kind of preparation method of biological antiseptic agent phenyllactic acid, be specifically related to a kind of method that the microbial fermentation bio-transformation prepares phenyllactic acid, belong to technical field of food biotechnology.
Background technology
Phenyllactic acid (phenyllactic acid) is a kind of new type natural sanitas of discovered in recent years, and 1998 confirm that first phenyllactic acid has very strong restraining effect to Listeria monocytogenes (Listeria monocytogenes).Prove that now phenyllactic acid can suppress multiple food-borne pathogens such as streptococcus aureus (Staphylococcusaureus), pathogenic colon bacillus (Escherichia coli O157:H7); Particularly the fungi that causes food spoilage is comprised that toxigenic fungi has the restraining effect of wide spectrum, its bacteriostatic activity is better than common sanitas.Phenyllactic acid is safe, is a kind of natural bacteriostatic material.It is present in the natural honey, and is nontoxic to the humans and animals cell, is continue first-generation antibacterial substance that milk-acid bacteria produces such as lactic acid, acetic acid, third generation natural bacteriostatic material in addition such as s-generation antibacterial substance such as Nisin (lacticin) bacteriocin of etc.ing.At present Nisin and tennecetin have been used for foodstuffs industry, but have problem such as, physico-chemical property difference narrow such as antimicrobial spectrum, can only suppress G except that milk-acid bacteria as Nisin
+Bacterium, and to most G
-Bacterium, yeast and mould be effect not; Tennecetin only has restraining effect to mould.Compare with tennecetin with Nisin, phenyllactic acid is a kind of new type natural sanitas, and wider antimicrobial spectrum is arranged, and can suppress food-borne pathogens, spoilage organism, and particularly its pollution that can suppress fungi can prolong the shelf-lives of food; Solvability is good, be easy to spread in various food systems; Stability is high, have broad pH scope and thermostability; These advantages make it hold out broad prospects in Application in Food Industry.
The preparation of phenyllactic acid at present has the synthetic and bioconversion method of chemical catalysis.Chemical synthesis has many adverse factors, for example produces multiple by product and chemical pollutant, and the numerous by products that form make separation, purification step become complicated.Recent study personnel prepare phenyllactic acid in the method for seeking bio-transformation, because milk-acid bacteria is the microorganism of GRAS (being known as safety), become the focus of current research with the synthetic phenyllactic acid of safe bacterial strain.Recently some milk-acid bacteria of report can utilize the MRS substratum to produce phenyllactic acid, but yield poorly, the individual difference of bacterial classification is very big, production peak is 9.9mg/100ml.Produce phenyllactic acid about milk-acid bacteria at present and still do not have relevant report.
Summary of the invention
The preparation method who the purpose of this invention is to provide a kind of biological antiseptic agent phenyllactic acid, particular content relates to a kind of method of utilizing the lactobacillus-fermented bio-transformation to prepare phenyllactic acid.The milk-acid bacteria that the present invention adopts can be converted into phenyllactic acid with phenyl-pyruvic acid or Sodium.beta.-phenylpyruvate, and transformation efficiency height, by product are few, have obtained unusual effect by the fermentation conversion, for the bio-transformation of phenyllactic acid provides a new way.
Technical scheme of the present invention: with milk-acid bacteria (Lactobacillus plantarum, Lactobacilluspentose) as bacterial classification, with phenyl-pyruvic acid or Sodium.beta.-phenylpyruvate is conversion of substrate, adopt carbon source, nitrogenous source and inorganic salt to form fermention medium, the fermentation method bio-transformation prepares phenyllactic acid.
(1) bacterial classification: adopt milk-acid bacteria
Lactobacillus?plantarum?AS1.550；Lactobacillus?plantarum?AS1.557；
Lactobacillus?plantarum?CICC6003；Lactobacillus?plantarum?CICC6014；
Lactobacillus?pentose?BCRC?11053；Lactobacillus?pentose?BCRC15317。
When transforming as strain bio, select for use Lactobacillus plantarum AS1.550 or Lactobacillus pentose BCRC15317 effect good especially with milk-acid bacteria.
(2) substratum, in g/100ml:
Storage medium: glucose 1.5, yeast extract paste 1, lime carbonate 1.5, agar 1.5, pH 6.5-7.0.
Seed culture medium: glucose 1, peptone 1, yeast extract paste 1, anhydrous sodium acetate 1, potassiumphosphate 0.02, sal epsom 0.02, manganous sulfate 0.005, pH 6.5-7.0.
(3) preparation technology:
Culture presevation: used culture presevation on storage medium, 4 ℃ of preservations, two week switchings once.
Seed culture: change seed culture medium over to from storage medium and need activate 2 times, cultivate 12h for each 30 ℃.
Fermentation: substrate phenyl-pyruvic acid or Sodium.beta.-phenylpyruvate concentration in substratum is 0.1%-0.4% (W/V), seed culture fluid is with the inoculum size of 1%-3%, insert and add different carbon sources, in the fermention medium that nitrogenous source and inorganic salt are formed, 250ml triangular flask liquid amount is 200ml, carbon source interpolation level is the 0.5%-3% (W/V) of fermention medium, nitrogenous source interpolation level is the 0.5%-3.5% (W/V) of fermention medium, the fermention medium composition is counted with g/100ml: phenyl-pyruvic acid or Sodium.beta.-phenylpyruvate 0.1-0.4, carbon source 0.5-3.0, nitrogenous source 0.5-3.5, anhydrous sodium acetate 1, potassiumphosphate 0.02, sal epsom 0.02, manganous sulfate 0.005, pH 6.2-6.8,25-40 ℃ leaves standstill the fermented liquid that cultivation 12-24h obtains containing phenyllactic acid.Used carbon source is glucose, maltose, lactose or sucrose, and used nitrogenous source is that in corn steep liquor, yeast extract paste, peptone, the soybean meal one or more are used with.
Preferred fermention medium is in g/100ml: phenyl-pyruvic acid or Sodium.beta.-phenylpyruvate 0.3, and glucose 2, corn steep liquor 1, yeast extract paste 1, anhydrous sodium acetate 1, potassiumphosphate 0.02, sal epsom 0.02, manganous sulfate 0.005, pH 6.2-6.4 are better.
The method that detects phenyllactic acid is as follows:
It is centrifugal to get fermented liquid, and supernatant liquor is through filtering with microporous membrane (0.22 μ m), and HPLC analyzes on the filtrate.HPLC condition: Agilentl100 chromatographic column: ZORBAX 300SB C
18(5 μ m, 4.6 * 250mm), moving phase is (A) 0.05% trifluoroacetic acid/water and (B) 0.05% trifluoroacetic acid/methyl alcohol, and 0,20,23,25,30 minutes elutriant A/B volume ratio of different time sections consists of carried out gradient elution in 10: 90,0: 100,0: 100,10: 90,10: 90.Detector: UV-detector, 210nm, column temperature: 30 ℃, flow velocity: 1ml/min, sample size: 10 μ l, phenyllactic acid standard specimen concentration: 0.5%.
Used bacterial classification is Lactobacillus plantarum AS1.550; Lactobacillus plantarumAS1.557; Lactobacillus plantarum CICC6003; Lactobacillus plantarum CICC6014; Lactobacillus pentose BCRC11053; Lactobacillus pentose BCRC15317, above bacterial classification is all open, the expression of AS numbering (China General Microbiological Cuture Collection Center.CGMCC.Institute of Micribiology, Chinese Academy of Science: Chinese microorganism strain council common micro-organisms center).CICC numbering expression (China Center of Industrial Culture: Chinese industrial microbial strains preservation administrative center).BCRC numbering expression (Bioresource Collection andResearch Center: Biological resources preservation research centre, Taiwan).Thereafter numbering is represented deposit number, specifically checks each bacterial classification database webpage.
Beneficial effect of the present invention: the invention provides a kind of method of utilizing the lactobacillus-fermented bio-transformation to prepare phenyllactic acid, show that the output of 12h phenyllactic acid can reach 50-150mg/100ml by the method for fermentation.Adopt described milk-acid bacteria phenyl-pyruvic acid or Sodium.beta.-phenylpyruvate can be converted into phenyllactic acid, transformation efficiency height, by product are few, have obtained unusual effect by the fermentation conversion, for the bio-transformation of phenyllactic acid provides a new way.
Embodiment
By experiment, obtained the principal element of influence fermentation conversion phenyllactic acid productive rate:
Embodiment 1 different strain is to the influence of phenyllactic acid output
In fermention medium, add 0.3% phenyl-pyruvic acid (analytical pure), the above-mentioned preferred substratum of by specification carries out fermenting experiment to some milk-acid bacterias of primary election, behind the fermentation 12h fermented liquid is detected, the result is as shown in table 1, Lactobacillus plantarum AS1.550, the phenyllactic acid productive rate of Lactobacillus pentose BCRC15317 is higher, and different strain has difference comparatively significantly to the generation of phenyllactic acid.
Table 1 different strain is to producing the influence of phenyllactic acid
Embodiment 2 fermentation times are to producing the influence of phenyllactic acid
The described fermentation condition of by specification, add 0.3% phenyl-pyruvic acid,, find that fermentation 12h phenyllactic acid content is the highest by the detection that Lactobacillusplantarum AS1.550 is produced phenyllactic acid in the different fermentations time, along with fermentation time prolongs, phenyllactic acid content has decline slightly.So determine that fermentation time is 12h, fermentation time is as shown in table 2 to the influence of producing phenyllactic acid.
Table 2 fermentation time is to producing the influence of phenyllactic acid
Embodiment 3 different substrate phenyl-pyruvic acids or Sodium.beta.-phenylpyruvate are to producing the influence of phenyllactic acid
The described fermentation condition of by specification, add 0.3% different substrate phenyl-pyruvic acids or Sodium.beta.-phenylpyruvate respectively, by Lactobacillus plantarum AS1.550 is produced phenyllactic acid behind fermentation 12h detection, phenyllactic acid content was the highest when Sodium.beta.-phenylpyruvate was substrate.Different substrate phenyl-pyruvic acids or Sodium.beta.-phenylpyruvate are as shown in table 3 to the influence of producing phenyllactic acid.
Different substrate phenyl-pyruvic acids of table 3 or Sodium.beta.-phenylpyruvate are to producing the influence of phenyllactic acid
Claims (2)
1. the preparation method of a phenyllactic acid is characterized in that with milk-acid bacteria being conversion of substrate as bacterial classification with phenyl-pyruvic acid or Sodium.beta.-phenylpyruvate, adopts carbon source, nitrogenous source and inorganic salt to form fermention medium, and the fermentation method bio-transformation prepares phenyllactic acid;
(1) bacterial classification: adopt milk-acid bacteria Lactobacillus plantarum AS1.550;
(2) substratum, in g/100ml:
Storage medium: glucose 1.5, yeast extract paste 1, lime carbonate 1.5, agar 1.5, pH 6.5-7.0;
Seed culture medium: glucose 1, peptone 1, yeast extract paste 1, anhydrous sodium acetate 1, potassiumphosphate 0.02, sal epsom 0.02, manganous sulfate 0.005, pH 6.5-7.0;
(3) preparation technology:
Culture presevation: used culture presevation on storage medium, 4 ℃ of preservations, two week switchings once;
Seed culture: change seed culture medium over to from storage medium and need activate 2 times, cultivate 12h for each 30 ℃;
Fermentation: substrate phenyl-pyruvic acid or the Sodium.beta.-phenylpyruvate concentration in substratum is 0.1%-0.4% (W/V), seed culture fluid is with the inoculum size of 1%-3%, insert and add different carbon sources, in the fermention medium that nitrogenous source and inorganic salt are formed, 250ml triangular flask liquid amount is 200ml, carbon source interpolation level is the 0.5%-3% (W/V) of fermention medium, nitrogenous source interpolation level is the 0.5%-3.5% (W/V) of fermention medium, the fermention medium composition is counted with g/100ml: phenyl-pyruvic acid or Sodium.beta.-phenylpyruvate 0.1-0.4, carbon source 0.5-3.0, nitrogenous source 0.5-3.5, anhydrous sodium acetate 1, potassiumphosphate 0.02, sal epsom 0.02, manganous sulfate 0.005, pH 6.2-6.8,25-40 ℃ leaves standstill cultivation 12-24h, obtain containing the fermented liquid of phenyllactic acid, used carbon source is a glucose, maltose, lactose or sucrose, used nitrogenous source are corn steep liquor, yeast extract paste, peptone, in the soybean meal one or more are used with.
2. the preparation method of phenyllactic acid according to claim 1 is characterized in that fermention medium is in g/100ml: phenyl-pyruvic acid or Sodium.beta.-phenylpyruvate 0.3, glucose 2, corn steep liquor 1, yeast extract paste 1, anhydrous sodium acetate 1, potassiumphosphate 0.02, sal epsom 0.02, manganous sulfate 0.005, pH 6.2-6.4.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106497824A (en) * | 2016-09-27 | 2017-03-15 | 陈福生 | One plant height produces the preparation method of the Acetobacter gluconicum and its phenyllactic acid of phenyllactic acid |
| CN104560800B (en) * | 2014-12-24 | 2018-04-06 | 江南大学 | A kind of phenylpyruvic acid reductase and its application in asymmetric syntheses (R) phenyllactic acid |
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|---|---|---|---|---|
| CN101508960B (en) * | 2009-03-04 | 2011-01-19 | 常熟理工学院 | Rhizopus and uses thereof |
| CN101906392B (en) * | 2010-06-23 | 2012-02-01 | 常熟理工学院 | Lactobacillussp.W2 strain and application thereof |
| CN106399195B (en) * | 2016-11-04 | 2019-09-03 | 浙江工业大学 | A kind of Lactobacillus casei and its application |
| CN108384730B (en) * | 2018-01-15 | 2021-02-02 | 浙江工业大学 | Lactobacillus paracasei and application thereof in conversion synthesis of phenyllactic acid |
| CN109170486B (en) * | 2018-09-07 | 2022-01-11 | 中国海洋大学 | Phenyllactic acid composite biological preservative and preparation method thereof |
| CN110200899B (en) * | 2019-07-05 | 2022-03-25 | 欧诗漫生物股份有限公司 | Preservative and preparation method and application thereof |
| CN111996146B (en) * | 2020-08-31 | 2021-06-15 | 山东宝来利来生物工程股份有限公司 | Lactobacillus plantarum for high yield of phenyllactic acid and application thereof |
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2006
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Non-Patent Citations (3)
| Title |
|---|
| Francesca Valerio等.Production of phenyllactic acid by lactic acid bacteria: anapproach to the selection of strains contributing to foodquality and preservation.FEMS microbiology Letters233.2004,233289-295. * |
| Paola Lavermicocca等.Purification and Characterization of Novel AntifungalCompounds from the Sourdough Lactobacillus plantarumStrain 21B.Applied and Environmental Microbiology66 9.2000,66(9),4084-4090. |
| Paola Lavermicocca等.Purification and Characterization of Novel AntifungalCompounds from the Sourdough Lactobacillus plantarumStrain 21B.Applied and Environmental Microbiology66 9.2000,66(9),4084-4090. * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104560800B (en) * | 2014-12-24 | 2018-04-06 | 江南大学 | A kind of phenylpyruvic acid reductase and its application in asymmetric syntheses (R) phenyllactic acid |
| CN106497824A (en) * | 2016-09-27 | 2017-03-15 | 陈福生 | One plant height produces the preparation method of the Acetobacter gluconicum and its phenyllactic acid of phenyllactic acid |
| CN106497824B (en) * | 2016-09-27 | 2019-10-01 | 陈福生 | One plant height produces the gluconacetobacter of phenyllactic acid and its preparation method of phenyllactic acid |
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