CN106497825A

CN106497825A - The gluconobacter sp of one plant of product brown pigment and its application in liquid fermentation edible vinegar

Info

Publication number: CN106497825A
Application number: CN201610853676.4A
Authority: CN
Inventors: 陈福生; 陈亨业; 王斌
Original assignee: Individual
Current assignee: Individual
Priority date: 2016-09-27
Filing date: 2016-09-27
Publication date: 2017-03-15

Abstract

The invention discloses the acetic acid bacteria and its and Acetobacter pasteurianus of one plant of product brown pigment（Acetobacter pasteurianusCGMCC 1.41）Application in mixed fermentation production brown and excellent in flavor liquid fermentation edible vinegar.The gluconobacter sp that the present invention is provided（Gluconobactersp.）Bacterial strain FBFS 82, was preserved in China typical culture collection center on July 11st, 2016, and preserving number is CCTCC M2016387.At 30 °C, under the conditions of 170 r/min, 6 d of mixes liquid fermentation, can produce acidity 4.3% to the gluconobacter sp FBFS 82 and Acetobacter pasteurianus CGMCC 1.41 of present invention offer, and tart flavour is soft, the brown brown liquid fermentation edible vinegar of better flavor.Correlational study has no report.

Description

The gluconobacter sp of one plant of product brown pigment and its application in liquid fermentation edible vinegar

Technical field

The invention belongs to technical field of food biotechnology, and it is related to the gluconobacter sp of one plant of product brown pigment（Gluconobacter sp.）FBFS 82 and its and Acetobacter pasteurianus（Acetobacter pasteurianusCGMCC 1.41, current China's food The conventional bacterial strain of vinegar production）Application in mixes liquid fermentation vinegar.

Background technology

Vinegar is one kind acid with materials such as grain, fruit, drinks etc. be starch-containing, saccharide, ethanol as fermenting raw materials Property flavoring agent.By the mode of production point, the vinegar of China can be divided into Solid-state fermentation vinegar and liquid fermentation edible vinegar.Solid-state fermentation vinegar It is the traditional vinegar of China, soft have the advantages that color and luster sauce red, with rich flavor, mellow in taste, tart flavour, but there is also production week Phase length, floor space are big, high labor intensive the shortcomings of.Liquid deep layer vinegar is as production floor space is little, automatization in recent years Degree is high, labor intensity is little, rate of producing acid is fast, low production cost the advantages of, therefore increasingly by domestic institute of vinegar manufacturer Using.According to incompletely statistics, the yield of current China's liquid fermentation edible vinegar has accounted for the 50% or so of vinegar total output.But liquid Fermentation vinegar local flavor is single, tart flavour zest strong, and color and luster is boring, does not meet the consumption habit of consumer of China, so mesh The liquid fermentation edible vinegar of front China is mainly used as raw material, by with conventional solid-state ferment vinegar mixing preparation or add food flavouring The mode such as agent and coloring agent is producing the edible vinegar of low side.Therefore the local flavor and color and luster for how improving liquid fermentation edible vinegar is China Liquid fermentation edible vinegar problem in the urgent need to address.

Present invention separation screening from soil obtains one plant of acetic acid bacteria gluconobacter sp FBFS 82 that can produce brown pigment, And there is provided itself and method of 1.41 mixed fermentations of Acetobacter pasteurianus CGMCC to increase liquid fermentation edible vinegar color and local flavor.

Content of the invention

The present invention provides one plant of gluconobacter sp（Gluconobactersp.）Bacterial strain FBFS 82, its deposit number is： CCTCC M 2016387.

Above-mentioned bacterial strains are isolated from soil by this laboratory.

Second object of the present invention is to provide a kind of by gluconobacter sp FBFS 82 and Acetobacter pasteurianus CGMCC 1.41 Method of the mixed fermentation to improve liquid fermentation edible vinegar color and local flavor.

The method that the present invention is provided, comprises the steps：Using above-mentioned gluconobacter sp FBFS 82 and Acetobacter pasteurianus 1.41 mixes liquid fermenting and producing vinegars of CGMCC.

In said method, the condition of the fermentation is 30 °C, and 170 r/min cultivate 6 d.

In said method, liquid culture medium formula is as follows：100 mg/mL of glucose, 10 mg/mL of yeast extract, anhydrous second 60 mg/mL of alcohol, is prepared with distilled water.

In said method, respectively by 5%（v/v）82 seed liquor of gluconobacter sp FBFS and Acetobacter pasteurianus CGMCC 1.41 Seed liquor is accessed in culture medium.

In said method, 82 seed liquor of gluconobacter sp FBFS is that the inclined-plane thalline for taking the ring bacterial strain accesses seed liquor training In foster base, at 30 °C, 170 r/min cultivate 24 h；1.41 seed liquor of Acetobacter pasteurianus CGMCC is take the ring bacterial strain oblique Face thalline is accessed in seed liquid culture medium, and at 30 °C, 170 r/min cultivate 24 h.

In said method, the seed liquid culture medium of gluconobacter sp FBFS 82 is as follows：100 mg/mL of glucose, yeast extract 10 mg/mL, are prepared with distilled water.The slant medium of gluconobacter sp FBFS 82 is as follows：100 mg/mL of glucose, yeast soak 10 mg/mL of cream, 15 mg/mL of agar, 30 mg/mL of Calcium Carbonate, are prepared with distilled water.The kind of Acetobacter pasteurianus CGMCC 1.41 Sub- liquid culture medium is as follows：100 mg/mL of glucose, 10 mg/mL of yeast extract, 60 mg/mL of dehydrated alcohol, are matched somebody with somebody with distilled water System.The slant medium of Acetobacter pasteurianus CGMCC 1.41 is as follows：100 mg/mL of glucose, 10 mg/mL of yeast extract, anhydrous 60 mg/mL of ethanol, 15 mg/mL of agar, 30 mg/mL of Calcium Carbonate, are prepared with distilled water.

Above-mentioned bacterial strains gluconobacter sp FBFS 82 is preserved in China typical culture collection center on July 11st, 2016 （Referred to as CCTCC, address is：Wuhan City, Hubei Province Hongshan District Bayi Road）, deposit number is：CCTCC M 2016387, classification It is named as gluconobacter sp（Gluconobactersp.）.

The experiment proves that, gluconobacter sp FBFS 82 can produce brown pigment, itself and Acetobacter pasteurianus CGMCC 1.41 in 30 °C, 6 d of mixed fermentation under the conditions of 170 r/min, it is 4.3% that can produce acidity, the liquid fermentation edible vinegar of local flavor and excellent color.

Description of the drawings

The colonial morphology of Fig. 1 gluconobacter sps FBFS 82.

The change of total acid content during Fig. 2 Vinegar Fermentations.

The composition of organic acid in Fig. 3 vinegars.

The color of Fig. 4 vinegars.

Specific embodiment

Below example facilitates a better understanding of the present invention, but does not limit the present invention.Experiment in following embodiments Method, if no special instructions, is conventional method.In following embodiments, test material used, if no special instructions, is all from Routine biochemistry reagent shop.Quantitative test in following examples, is respectively provided with three times and repeats to test, results averaged.Below The present invention is further described in conjunction with accompanying drawing.

The detection of total acid content during 1 Vinegar Fermentation of embodiment：By specification methods described, by acid-base titrations In measure sweat, the different fermentations time issues the total acid content of zymotic fluid, finds acidity all the entering with the time in fermentation liquid OK, first rise and maintain an equal level afterwards.During Acetobacter pasteurianus 1.41 independent fermentations of CGMCC, total acid is up to 4%；Gluconobacter sp FBFS 82 individually sends out During ferment, total acid is up to 3%；During two plants of bacterium mixed fermentation, total acid is up to 4.3%（Fig. 2）.

In examples detailed above, in detection fermentation liquid, the method for total acid content comprises the steps：Take appropriate fermentation liquid（1-5 mL） 20 mL are diluted to distilled water, with 1% phenolphthalein as indicator, are titrated to the standard solution of sodium hydroxide of 0.1 mol/L micro- red 30 s is colour-fast for color, calculates fermentation liquid acidity by the volume of the sodium hydroxide solution for consuming.

Flavor components in 1 vinegar of table

Note：-：Do not detect.

The composition of organic acid in 2 vinegar of embodiment：By specification methods described is fermented to obtain vinegar, by high performance liquid chromatography The content of organic acid in detection vinegar, it is found that Acetobacter pasteurianus CGMCC 1.41 can produce acetic acid and a small amount of citric acid does not produce winestone Acid, and gluconobacter sp FBFS 82 can produce citric acid and a small amount of tartaric acid and hardly produce acetic acid, thus both have certain mutual Benefit property, makes the vinegar after mixed fermentation while containing acetic acid, tartaric acid and citric acid, and wherein acetic acid and tartaric content be all Content when individually fermenting higher than two plants of bacterium, makes that the content of organic acid in mixed fermentation vinegar is more and species is more enriched（Figure 3）.

In examples detailed above, in detection vinegar, the high performance liquid chromatography of organic acid content comprises the steps：Vinegar is by height Speed centrifugation（10000 r/min, 10min）Thalline is removed, after crossing 0.22 μm of film, efficient liquid phase chromatographic analysis is carried out.

Above-mentioned efficient liquid phase chromatographic analysis condition is：Liquid chromatograph Shimadzu LC-20AT, chromatographic column inertsil 4.6 × 250 mm of ODS-3, mobile phase is：0.01 mol/L disodium hydrogen phosphates, are adjusted to pH 2.7 with 1 mol/L phosphoric acid, stream 1 mL/min of speed, 10 μ L of sample size, 40 °C of column temperature, 210 nm of Detection wavelength.

The identification of flavor components in 3 vinegar of embodiment：By specification methods described is fermented to obtain vinegar, by gas chromatogram- In GC-MS detection vinegar, the species of flavor components, it is found that the vinegar after mixed fermentation contains esters, aldehydes, alcohols, acid Multiple flavor components such as class, and generate four kinds of opportunistic pathogen strains（FBFS82 and CGMCC 1.41）The flavor components not having, such as have The ethyl phenylacetate of Mel fragrance, and the aldehyde C-9 with fragrance such as Flos Rosae Rugosae, Citrus（Table 1）.

In examples detailed above, in detection vinegar, the GC-MS of flavor components species comprises the steps：Food Flavor components in vinegar first pass through solid-phase microextraction extraction, detect by gas chromatograph-mass spectrometer (GC-MS).

The method of above-mentioned solid phase micro-extraction is：Accurately measure 8 mL vinegars to be placed in the sample bottle of 15 mL, add 25 g chlorine Change sodium（Reduce the dissolubility of flavor substance）, it is placed in 40 °C of waters bath with thermostatic control, 100 r/min magnetic agitation.By SPME extracting heads Be inserted into the head space part of sample bottle, to sample in 30 min of flavor components adsorption and enrichment, after draw back fiber head.Above-mentioned gas phase Chromatograph-mas spectrometer condition is：SH-RXI-5SIL MS capillary columns；Carrier gas is He；Shunting 20:1,24 mL/ of overall flow rate Min, 1 mL/min of column flow；Column temperature is 35 °C；Injector temperature is 230 °C；Column temperature is arranged：Initial temperature is 35 °C, with 10 °C/min is warming up to 250 °C.Mass spectrometric interface temperature is 250 °C, and ion source temperature is 200 °C, and ionization mode is EI, 70 eV of electron energy, quality of scanning scope 33-450 amu.

Claims

1. gluconobacter sp（Gluconobactersp.）FBFS 82, its deposit number is：CCTCC M 2016387.

2. gluconobacter sp FBFS 82 described in claim 1 and 1.41 mixed fermentations of Acetobacter pasteurianus CGMCC production brown and The method of excellent in flavor vinegar.

3. a kind of method for producing brown vinegar, gluconobacter sp FBFS 82 and Acetobacter pasteurianus described in fermentation claim 1 1.41 mixes liquids of CGMCC are fermented.

4. method as claimed in claim 3, it is characterised in that：Gluconobacter sp FBFS 82 and Acetobacter pasteurianus CGMCC 1.41 For fermentation strain, in 30 °C, 6 d of liquid culture under 170 r/min；The nutrient media components is：100 mg/mL of glucose, ferment 10 mg/mL of female extractum, 60 mg/mL of dehydrated alcohol, are prepared with distilled water.

5. method as claimed in claim 4, it is characterised in that：The fermentation is by gluconobacter sp FBFS described in claim 1 82 and Acetobacter pasteurianus CGMCC 1.41 seed liquor is seeded in fermentation medium described in claim 4 respectively and cultivates, inoculation Measure as 5%（v/v）.

6. the described two seed liquor of claim 5 are respectively by Acetobacter gluconicum FBFS 82 described in claim 1 and Pasteur's vinegar bar The slant culture of bacterium CGMCC 1.41 is taken in the 500 mL triangular flasks that a ring thalline is accessed equipped with 50 mL seed culture mediums, and 30 °C, 160 r/min cultivate 24h；82 seed liquid culture mediums of the gluconobacter sp FBFS are：100 mg/mL of glucose, yeast soak 10 mg/mL of cream, is prepared with distilled water；82 slant mediums of gluconobacter sp FBFS are：100 mg/mL of glucose, yeast extract 10 mg/mL, 15 mg/mL of agar, 30 mg/mL of Calcium Carbonate are prepared with distilled water；1.41 seed liquor of Acetobacter pasteurianus CGMCC is trained Foster base is：100 mg/mL of glucose, 10 mg/mL of yeast extract, 60 mg/mL of ethanol, are prepared with distilled water；Acetobacter pasteurianus 1.41 slant mediums of CGMCC are：100 mg/mL of glucose, 10 mg/mL of yeast extract, 60 mg/mL of ethanol, agar 15 Mg/mL, 30 mg/mL of Calcium Carbonate are prepared with distilled water.

7. the vinegar for being prepared by arbitrary methods described in claim 3-6.