CN104059868B - Pediococcus pentosaceus R1 strain with in-vitro antioxidation activity and application thereof as antioxidant in fermented meat products - Google Patents
Pediococcus pentosaceus R1 strain with in-vitro antioxidation activity and application thereof as antioxidant in fermented meat products Download PDFInfo
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Abstract
The invention relates to a Pediococcus pentosaceus R1 strain with in-vitro antioxidation activity and application thereof as an antioxidant in fermented meat products, belonging to the technical field of microbes. The Pediococcus pentosaceus R1 is collected at China General Microbiological Culture Collection Center (CGMCC), wherein the culture collection number is CGMCC No.9171, the collection date is May 16th, 2014, and the classification designation is Pediococcus pentosaceus. The Pediococcus pentosaceus R1 has favorable antioxidation activity in in-vitro antioxidation tests; the complete cell and intracellular extract have high capacity for removing hydroxy free radicals, and can inhibit the lipids from peroxidation; and the cell surface substances have antioxidation activity. Therefore, the Pediococcus pentosaceus R1 can be used as an antioxidant in fermented meat products to postpone the product deterioration caused by oxidation in the fermentation and storage process, thereby reducing the consumption of the antioxidant.
Description
Technical field
The invention belongs to microbial technology field, is related to one plant of with antioxidant activity penta filtered out from air-dry sausage
Sugar-tablet coccus and its fermentation meat in as antioxidant application.
Background technology
Lactic acid bacteria is one of modal microorganism in fermentation meat product, and it can reduce meat pH value, improve organizational structure,
Promote the formation of local flavor, while the growth of putrefaction bacteria can be suppressed, it is ensured that the edible safety of fermentation meat product.It is excellent in order to obtain
Used as leaven, Chinese scholars have carried out in these areas more research to good strain.But, fermentation meat product is in storage
During fat oxidative rancidity can occur, be the one of the main reasons of its quality deterioration.At present, used in fermentation meat product
Antioxidant, such as Butylated hydroxyanisole (BHA), 2,6- di-tert-butyl-4-methy phenols (BHT), tert-butyl hydroquinone
(TBHQ), delaying meat productss to aoxidize deterioration, but there is use cost in some antioxidants for propylgallate (PG) and VE etc.
Height, oxidation resistance is weak, and in addition toxicologic study shows that some antioxidants have potential safety hazard.Thus it is possible to be had
The starter for fermentative meat of antioxidant activity is another effective way for solving meat productss oxidation deterioration, but at present with regard to fermented meat
The research of lactic acid bacteria antioxidant activity is less in product.
The content of the invention
It is an object of the invention to provide one plant of Pediococcus pentosaceus with antioxidation activity in vitro, may be used on fermented meat system
In product, delay the product quality deterioration that the over oxidation in meat productss storage is caused.
Lactic acid bacteria --- the Pediococcus pentosaceuss R1 (Pediococcus that the present invention is isolated from fermenting air-dry sausage
Pentosaceus R1), it is deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), address:
Yard 1, BeiChen xi Road, Chaoyang District, Beijing City institute of microbiology of the Chinese Academy of Sciences, its culture presevation is numbered and is:CGMCC No.9171, protect
The date is hidden on 05 16th, 2014, the Classification And Nomenclature of strain is Pediococcus pentosaceuss (Pediococcus pentosaceus).
Pediococcus pentosaceuss R1 in the present invention has following biological characteristicses:
(1) morphological characteristic:Pediococcus pentosaceuss R1 Gram’s staining result is the positive, and cell is rounded, paired or tetrad
Arrangement, without brood cell, without pod membrane, atrichia.
(2) colony characteristicses:Pediococcus pentosaceuss R1 is good in MRS cultured on solid medium, and colonial morphology is circular, milky white
Color, smooth, projection, neat in edge is opaque.
(3) physiological and biochemical property:Negative catalase, does not produce indole, hydrogen sulfide and ammonia, does not reduce nitrate, not water
Solution arginine, can glucose fermentation, azymic sorbose, 6-(.alpha.-D-galactosido)-D-glucose., xylitol, sucrose, Lactose, melezitose, arabinose, wood
Sugar, rhamnose, Raffinose, Sorbitol, Mannitol;Can be from cellobiose, Esculin, Tan sugar, ribose, glucose, galactose, fruit
Sugar, mannose fermentation and acid.
The Pediococcus pentosaceuss R1 of the present invention is presented preferable antioxidant activity in Oxidation Resistance Test in vitro, its full cell and
Intracellular extract has stronger Scavenging activity to hydroxyl radical free radical, can anti-lipid peroxidation, and cell surface material has
Antioxidant activity.Therefore, Pediococcus pentosaceuss R1 can delay fermenting and preserving as the antioxidant in fermentation meat
Because of the product deterioration that oxidation is caused in journey, and then reduce the usage amount of antioxidant.
Preservation information:
Depositary institution's full name:China Committee for Culture Collection of Microorganisms's common micro-organisms center;
Depositary institution is referred to as:CGMCC;
Depositary institution address:Yard 1, BeiChen xi Road, Chaoyang District, Beijing City institute of microbiology of the Chinese Academy of Sciences;
Deposit number:CGMCC No.9171;
Preservation date:On 05 16th, 2014.
Description of the drawings
Fig. 1 is survival conditions of the Pediococcus pentosaceuss R1 in variable concentrations hydrogenperoxide steam generator;
Fig. 2 is Pediococcus pentosaceuss R1 somatic cells and intracellular extract to hydroxyl radical free radical Scavenging activity;
Fig. 3 is Pediococcus pentosaceuss R1 somatic cells and intracellular extract reducing power;
Fig. 4 is Pediococcus pentosaceuss R1 somatic cells and intracellular extract inhibition of peroxidation;
Fig. 5 is that removing energy of the somatic cells to hydroxyl radical free radical after different disposal is carried out to Pediococcus pentosaceuss R1 cell surfaces
Power.
Fig. 6 is the air-dried intestinal fermentation TBARS value of the 9th day of natural fermentation air-dry sausage and inoculation Pediococcus pentosaceuss
(TBARS)。
Specific embodiment
Technical scheme is further described below in conjunction with the accompanying drawings, but is not limited thereto, it is every to this
Inventive technique scheme is modified or equivalent, without deviating from the spirit and scope of technical solution of the present invention, all should be covered
In protection scope of the present invention.
First, EXPERIMENTAL DESIGN
Pediococcus pentosaceuss R1 (inoculum concentration 3%) is inoculated in MRS (Man Rogosa and Sharp) fluid medium, and 37
DEG C culture 12h.In 4 DEG C, 10,000 × g centrifugation 5min obtain somatic cells to fermentation liquid.Somatic cells are divided into into two parts, one
Somatic cells distilled water is divided to suspend again after washing twice, as somatic cells suspension;Another part, after ultrasonication from
Supernatant, as intracellular extract are obtained after heart 5min (4 DEG C, 10,000 × g).By investigating tolerance of the somatic cells to hydrogen peroxide
Property, full cell and intracellular extract are to hydroxyl radical free radical Scavenging activity, reducing power, lipid peroxidation capacity, and thalline is thin
Cellular surface polyphenoils Quality Research is investigating its anti-oxidant vigor.
2nd, hydrogen peroxide toleration
By somatic cells be added to containing variable concentrations (0,0.2,0.4,0.6,0.8,1.0,1.2mM) hydrogen peroxide life
In reason saline, somatic cells initial concentration is about 107Cfu/mL, 37 DEG C of culture 8h, coats on MRS agar culture mediums, 37 DEG C
Culture 48h is counted.
3rd, hydroxyl radical free radical Scavenging activity
0.1mL prepare liquids mix (FeCl containing 0.1mmol/L with 1mL reaction buffers3, 0.104mmol/L EDTA,
1.5mmol/L H2O2, 2.5mmol/L deoxyriboses, 0.1 institute of microbiology of the Chinese Academy of Sciences, its culture presevation numbering is:CGMCC
No.9171, preservation date is on 05 16th, 2014, and the Classification And Nomenclature of strain is Pediococcus pentosaceuss (Pediococcus
pentosaceus)。
Pediococcus pentosaceuss R1 in the present invention has following biological characteristicses:
(1) morphological characteristic:Pediococcus pentosaceuss R1 Gram’s staining result is the positive, and cell is rounded, paired or tetrad
Arrangement, without brood cell, without pod membrane, atrichia.
(2) colony characteristicses:Pediococcus pentosaceuss R1 is good in MRS cultured on solid medium, and colonial morphology is circular, milky white
Color, smooth, projection, neat in edge is opaque.
(3) physiological and biochemical property:Negative catalase, does not produce indole, hydrogen sulfide and ammonia, does not reduce nitrate, not water
Solution arginine, can glucose fermentation, azymic sorbose, 6-(.alpha.-D-galactosido)-D-glucose., xylitol, sucrose, Lactose, melezitose, arabinose, wood
Sugar, rhamnose, Raffinose, Sorbitol, Mannitol;Can be from cellobiose, Esculin, Tan sugar, ribose, glucose, galactose, fruit
Sugar, mannose fermentation and acid.
The Pediococcus pentosaceuss R1 of the present invention is presented preferable antioxidant activity in Oxidation Resistance Test in vitro, its full cell and
Intracellular extract has stronger Scavenging activity to hydroxyl radical free radical, can anti-lipid peroxidation, and cell surface material has
Antioxidant activity.Therefore, Pediococcus pentosaceuss R1 can delay fermenting and preserving as the antioxidant in fermentation meat
Because of the product deterioration that oxidation is caused in journey, and then reduce the usage amount of antioxidant.
Preservation information:
Depositary institution's full name:China Committee for Culture Collection of Microorganisms's common micro-organisms center;
Depositary institution is referred to as:CGMCC;
Depositary institution address:Yard 1, BeiChen xi Road, Chaoyang District, Beijing City institute of microbiology of the Chinese Academy of Sciences;
Deposit number:CGMCC No.9171;
Preservation date:On 05 16th, 2014.
Description of the drawings
Fig. 1 is survival conditions of the Pediococcus pentosaceuss R1 in variable concentrations hydrogenperoxide steam generator;Removal S-layer albumen,
With sodium metaperiodate (sodium periodate) (20mg/mL) oxidative cell surface polysaccharide, process these albumen of cell surface and
After polysaccharide material, its hydroxyl radical free radical Scavenging activity is investigated.
7th, the making of air-dry sausage
(1) supplementary material:
Lean meat (pig buttockss meat, pig tenterloin) 4.5kg, fat meat (back pork) 0.5kg , Qu wine (Yuquan great Qu) 0.15kg, nitrous
Sour sodium 0.0045kg, glucose 0.25kg, gourmet powder 0.075kg, salt 0.125kg, (mixed spices include mixed spices 0.025kg:
Pericarpium Zanthoxyli, Fructus Anisi Stellati, Cortex cinnamomi japonici (Ramulus Cinnamomi), Flos Caryophylli etc.).
(2) prepared by leaven
The Pediococcus pentosaceuss for having activated are inoculated in MRS culture medium carries out fermentation culture, 37 DEG C of culture 12h.Fermentation liquid
It is standby after 10,000g centrifugation 5min under the conditions of 4 DEG C.During addition leaven, lactic acid bacteria final concentration of 107Cfu/g meat stuffings.
(3) air-dry sausage fabrication processing
Raw meat (rejecting the connective tissues such as lymph, tendon, blood vessel) → dice (meat cubelets of 1cm square) → pickle (salt adding
With flavouring agent etc.) → with leaven (matched group without) → mix filling → record → fermented air-dried.
8th, TBARS value (TBARS) is determined
Take 2g samples to be put in test tube, addition 1.5mL thiobarbituricacidα- solution, 7.5mL trichloroacetic acids-hydrochloric acid solution,
30min is reacted after mixing in boiling water bath, is cooled down, taken 5mL samples and add isopyknic chloroform, 10min, 532nm are centrifuged under 1000g
Read light absorption value in place.TBARS values represent that computing formula is as follows with the milligram number of malonaldehyde in every kilogram of lipid oxidation sample:
In formula:A- absorbances, V is sample volume (mL);
The molecular weight 72.063 of M- malonaldehyde;
ε-molar absorption coefficient 156000 (L/mol.cm);
1- light paths 1 (cm);
The quality (g) of m- meat samples.
9th, result and analysis
Hydrogen peroxide is a kind of strong oxidizer, and it not only can cause directly destruction to cell tissue, be also used as hydroxyl
The precursor of free radical participates in oxidizing process indirectly.Therefore, the tolerance for studying bacterial strain hydrogen peroxide can be used as its antioxygen of measurement
Change an index of activity.As shown in figure 1, concentration of hydrogen peroxide is in below 1.0mM, bacterium number change not significantly because with low
During the hydrogen peroxide treatment lactic acid bacteria somatic cells of concentration, the oxidant defense system of its own can protected from oxidant
Harm.As the 1.4mM that increases to of concentration of hydrogen peroxide, bacterium number only declines 0.5 logarithm period, illustrates Pediococcus pentosaceuss to peroxide
The toleration for changing hydrogen is stronger.
Knowable to Fig. 2,3 and 4, Pediococcus pentosaceuss whole-cell component is to radical scavenging activity, reducing power and suppression
Lipid peroxy rate has respectively reached 54.01%, 0.045 and 29.58%;Intracellular extract group is respectively 40.33%, 0.37 He
57.07%
Fig. 5 is that Jing difference protease and LiCl remove somatic cells surface protein, and uses sodium periodate oxidation cell surface
Result of the somatic cells to hydroxyl radical free radical Scavenging activity after polysaccharide.Test shows, radical scavenging activity after trypsin treatment
It is most notable that power declines, and the basic protein composition for illustrating cell surface plays main antioxidation.
Fig. 6 is that Pediococcus pentosaceuss are inoculated in into the air-dry sausage after fermentation TBARS values of the 9th day, is natural fermentation with matched group
Air-dry sausage is compared, and its TBARS is substantially low, illustrates that the air-dry sausage degree of oxidation for being inoculated with Pediococcus pentosaceuss in the process is relatively low,
The bacterial strain serves good antioxidation.
Test by more than, it is determined that the Pediococcus pentosaceuss R1 filtered out from the fermenting air-dry sausage of Harbin has stronger
Antioxidant activity, can as fermentation meat product in potential antioxidant.
Claims (2)
1. one plant of Pediococcus pentosaceuss R1 with antioxidation activity in vitro, is preserved in China Committee for Culture Collection of Microorganisms
Common micro-organisms center, culture presevation is numbered and is:CGMCCNo.9171.
2. the Pediococcus pentosaceuss R1 with antioxidation activity in vitro as claimed in claim 1 in fermentation meat as antioxidation
The application of agent.
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CN110373353A (en) * | 2019-07-22 | 2019-10-25 | 东北农业大学 | A kind of mixed culture fermentation agent being able to ascend less salt air-dry sausage flavor |
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CN110964673B (en) * | 2019-12-30 | 2022-03-04 | 上海应用技术大学 | Pediococcus pentosaceus with high oxidation resistance and whitening effect and application thereof |
CN118126908B (en) * | 2024-05-10 | 2024-06-25 | 善恩康生物科技(苏州)有限公司 | Pediococcus pentosaceus capable of improving fatigue and motor function and product thereof |
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CN1566322A (en) * | 2003-06-26 | 2005-01-19 | 河南双汇投资发展股份有限公司 | Lactobacillus pentosus strain, ferment produced thereby and the use of ferment in meat ware |
JP2006291146A (en) * | 2005-04-14 | 2006-10-26 | Taiyo Corp | Antioxidant |
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CN1566322A (en) * | 2003-06-26 | 2005-01-19 | 河南双汇投资发展股份有限公司 | Lactobacillus pentosus strain, ferment produced thereby and the use of ferment in meat ware |
JP2006291146A (en) * | 2005-04-14 | 2006-10-26 | Taiyo Corp | Antioxidant |
JP2008104375A (en) * | 2006-10-24 | 2008-05-08 | Taiyo Corp | Peroxide degrading enzyme |
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Cited By (1)
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CN110373353A (en) * | 2019-07-22 | 2019-10-25 | 东北农业大学 | A kind of mixed culture fermentation agent being able to ascend less salt air-dry sausage flavor |
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