CN113755399B - Method for screening strains with fragrance producing and mutton smell inhibiting functional characteristics and application of method - Google Patents

Method for screening strains with fragrance producing and mutton smell inhibiting functional characteristics and application of method Download PDF

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CN113755399B
CN113755399B CN202111180784.7A CN202111180784A CN113755399B CN 113755399 B CN113755399 B CN 113755399B CN 202111180784 A CN202111180784 A CN 202111180784A CN 113755399 B CN113755399 B CN 113755399B
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preservation
smell
aroma
mutton
milk
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CN113755399A (en
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田怀香
杨睿
陈臣
于海燕
黄娟
袁海彬
娄新曼
胡阳
郑国茂
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Shanghai Institute of Technology
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • A23V2400/00—Lactic or propionic acid bacteria
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    • A23V2400/169—Plantarum
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    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/41—Pediococcus
    • A23V2400/413—Acidilactici

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Abstract

The invention discloses a strain screening method with the function of inhibiting the smell of mutton and application thereof. The strain screening method comprises the following steps: activating a bacterial sample, enriching and culturing, separating pure seeds, preparing fermented milk and measuring the aroma production performance; the screening basis comprises the following aspects: taking the identification results of artificial sensory and key aroma compounds as indexes, and analyzing the compounds with fruity, nutty, milk aroma and other aroma notes by GC-MS to be 2-heptanone, benzaldehyde, ethyl butyrate, ethyl caproate, isoamylol and the like; fatty acid compounds such as caprylic acid and caproic acid have characteristic smell of mutton smell; the invention screens out strains with the characteristic of producing fragrance and inhibiting the smell of mutton by utilizing the strain screening method, wherein the strains comprise lactococcus lactis SD-3, pediococcus acidilactici DN-1 and lactobacillus plantarum GM-6.

Description

Method for screening strains with fragrance producing and mutton smell inhibiting functional characteristics and application of method
Technical Field
The invention relates to a strain screening method with the characteristic of producing fragrance and inhibiting the smell of mutton and application thereof, belonging to the technical field of microorganisms.
Background
The goat milk has high nutritive value, is rich in vitamins, mineral elements and active substances, and has higher protein and amino acid contents than cow milk. In addition, the milk sugar has low lactose content and is easy to absorb, is a good nutrition product for modern human health, and is called as king in milk. However, the goat milk and the dairy products thereof have unique flavor, and the corresponding products often have the problem of over-strong goat smell, thereby further reducing the acceptance of consumers and restricting the further development of the goat milk industry.
In order to improve and promote the flavor quality of the goat dairy product, at present, an exogenous addition way and an endogenous aroma-enhancing way are mainly adopted for regulation and control. But compared with the conventional approaches of external flavoring such as chemical flavoring, flavoring agent and the like, the method achieves the purpose of internal flavoring through microbial fermentation, is safer and healthier, and is not limited by raw materials.
The aroma-producing microorganisms are microorganisms capable of fermenting and metabolizing to produce aroma substances in the growth process, most of the aroma-producing microorganisms are lactic acid bacteria, bifidobacteria, saccharomycetes and moulds, and the aroma substances produced by different aroma-producing microorganisms are different. Through the identification of the characteristic flavor and the fragrance quality research of the traditional acid-coagulated cheese goat cheese milk cake, the fact that the attributes of milk fragrance and fruit fragrance are easily accepted by consumers and the attributes of mutton smell and over-heavy sour taste are not easily accepted is found. Meanwhile, methyl ketone substances such as 2-heptanone and 2-nonanone with milk flavor, benzaldehyde with nut and almond flavors, ethyl butyrate with fruit flavor, etc. have effect of inhibiting rank odor.
Therefore, in the case of fermentation strains, screening of aroma-enhancing (increasing of flavor substances such as milk flavor and fruit flavor) and mutton smell-inhibiting (reducing of the content of fatty acids such as caproic acid and caprylic acid) strains can be used for improving and enhancing the flavor quality of goat dairy products.
Disclosure of Invention
The technical problem to be solved by the invention is as follows: aiming at the problem that the traditional process of goat milk products has a serious mutton smell, a method for screening strains with the function of producing fragrance and inhibiting the mutton smell is provided.
In order to solve the problems, the invention provides a strain screening method with the function of producing fragrance and inhibiting the smell of mutton, which comprises the following steps:
step 1: respectively taking samples from a plurality of fresh milk cakes into a sterile test tube in a sterile environment, adding sterile water, fully oscillating and dissolving uniformly to obtain bacterial suspension, diluting the bacterial suspension into bacterial solutions with different concentrations by using the sterile water, sequentially coating the bacterial suspensions on an MRS agar culture medium for constant-temperature culture, and after bacterial growth, selecting a single bacterial colony, streaking, purifying and culturing to obtain a single bacterial strain;
step 2: respectively inoculating the obtained single strains to MRS liquid culture media, and fermenting at constant temperature to respectively obtain fermentation liquors of the single strains;
and 3, step 3: respectively centrifuging the obtained fermentation liquor, respectively washing precipitates obtained by centrifuging for multiple times by using sterile water, then respectively adding sterile water with the same volume as the bacterial suspension in the step 1 into the washed precipitates, and performing vortex oscillation to obtain a fermentation culture solution;
and 4, step 4: inoculating the fermentation culture solution obtained in the step 3 into sterile goat milk, performing fermentation culture at 37 ℃ until the pH =4.6 is completely fermented, and naturally cooling to room temperature to obtain fermented milk to be detected;
and 5: and (3) taking a fermented milk sample to be detected, respectively carrying out sensory evaluation primary screening and GC-MS aroma determination, and identifying strains to obtain the strains with the function of producing aroma and inhibiting the mutton smell.
Preferably, the composition of the MRS agar medium in step 1 comprises: 10.0g/L of peptone, 8.0g/L of beef extract powder, 4.0g/L of yeast extract powder, 20.0g/L of glucose, 2.0g/L of dipotassium phosphate, 2.0g/L of diammonium hydrogen citrate, 5.0g/L of sodium acetate, 0.2g/L of magnesium sulfate, 0.04g/L of manganese sulfate, 14.0g/L of agar and 801.0g/L of tween; the MRS liquid culture medium in the step 2 comprises the following components: 10.0g/L of peptone, 10.0g/L of beef extract powder, 5.0g/L of yeast extract powder, 20.0g/L of glucose, 0.1g/L of magnesium sulfate, 5.0g/L of sodium acetate, 2.0g/L of dipotassium hydrogen phosphate, 2.0g/L of ammonium citrate, 0.05g/L of manganese sulfate and 801.0g/L of Tween.
Preferably, the concentration of the bacterial suspension in the step 1 is 0.1g/mL; the bacterial liquids with different concentrations respectively have the concentration of 10 -2 、10 -3 、10 -4 、10 -5 And 10 -6 The bacterial liquid of (4).
Preferably, the conditions for the incubation at constant temperature in step 1 are: the temperature is 37 ℃, and the time is 12h; the conditions of constant-temperature fermentation in the step 2 are as follows: the temperature is 37 ℃ and the time is 12h.
Preferably, the conditions for centrifugation in step 3 are: the temperature is 4 ℃, the rotating speed is 6000rpm, and the time is 15min; the oscillation conditions are as follows: the rotation speed is 6000rpm, and the time is 10min.
Preferably, the volume ratio of the fermentation culture solution to the sterile goat milk in the step 4 is 5%.
Preferably, the sensory evaluation in the step 5 comprises evaluation indexes of aroma receptivity and aroma intensity; the GC-MS aroma determination comprises the following steps: extracting flavor substances in the fermented milk to be detected by adopting headspace solid-phase microextraction, analyzing and identifying the flavor substances by adopting GC-MS, and analyzing the generation and content change of key aroma components.
Preferably, the key aroma components include 2-heptanone, benzaldehyde, hexanoic acid, and octanoic acid.
The invention also provides a strain with the function of producing fragrance and inhibiting the smell of mutton, which is obtained by screening the strain with the function of producing fragrance and inhibiting the smell of mutton by adopting the screening method and comprises lactococcus lactis SD-3, pediococcus acidilactici DN-1 and lactobacillus plantarum GM-6;
the preservation number of the Lactococcus lactis SD-3 is CCTCC M2021743, the Latin name is Lactococcus lactis SD-3, the preservation unit is the China center for type culture preservation, the preservation time is 2021 years, 6 months and 23 days, and the preservation address is the preservation center of Wuhan university at Lophania mountain of Wuchang, hubei province;
the preservation number of the Pediococcus acidilactici DN-1 is CCTCC M2021794, the Latin name is Pediococcus lactis DN-1, the preservation unit is the China center for type culture preservation, the preservation time is 2021 years, 6 months and 28 days, and the preservation address is the preservation center of Wuhan university at Lophania Lophanthogama mountain of Wuhan city, hubei province;
the preservation number of the Lactobacillus plantarum GM-6 is CCTCC M2021744, the latin name is Lactobacillus plantarum GM-6, the preservation unit is the China center for type culture preservation, the preservation time is 2021 years, 6 months and 23 days, and the preservation address is the preservation center of Wuhan university at Lophania Lophanthogona mountain of Wuchang, hubei province.
The invention also provides application of the strain with the function of producing fragrance and inhibiting the smell of mutton in fermented goat milk.
GC-MS aroma determination shows that the fermented milk sample prepared from lactococcus lactis SD-3 has higher contents of 2-heptanone, benzaldehyde and ester compounds, the sensory evaluation result is optimal, the overall acceptance and the aroma acceptance sensory scores are highest, the milk aroma is most abundant, and the types and the total amount of aroma compounds are more abundant;
GC-MS aroma determination shows that a fermented milk sample prepared from pediococcus acidilactici DN-1 contains a large amount of key aroma compounds such as 2-heptanone, isoamylol and the like, and the contents of fatty acid compounds such as butyric acid, caproic acid, caprylic acid and the like are low;
GC-MS aroma measurement shows that the aroma acceptance degree of a fermented milk sample prepared by the lactobacillus plantarum GM-6 is high, the content of aldehyde compounds is high, and the flavor of nuts is obvious.
Compared with the prior art, the invention has the beneficial effects that:
1. according to the strain screening method with the function characteristics of producing fragrance and inhibiting the smell of mutton, the screened strain can ensure that a fermentation sample can endogenously produce more substances with milk fragrance and fruit fragrance and having the function of inhibiting the smell of mutton, the generation of the smell of mutton fatty acid is reduced to a certain extent, the flavor quality of the fermented dairy product is optimized on the basis of no need of exogenous addition of fragrance substances, and the production cost for preparing the fermented dairy product is reduced;
2. three strains of Lactococcus lactis SD-3, pediococcus acidilactici DN-1 and Lactobacillus plantarum GM-6 fermented goat milk with the fragrance producing and mutton smell inhibiting functional characteristics screened by the screening method disclosed by the invention generate 3-hydroxy-2-butanone, 2-heptanone, 2-octanone and 2-nonanone with higher contents, and the ketone fragrance substances have the fragrance characteristics of cream, cheese, coconut, fruity fragrance, sweet fragrance and the like; meanwhile, the sour congealed cheese goat cheese contains high-content almond-flavored benzaldehyde and fruit-flavored isoamylol, and is used as an odor component with a goat smell inhibiting effect and a positive influence in the characteristic flavor identification process of the sour congealed cheese goat cheese; meanwhile, by analyzing the aroma components of the fermented samples of different strains, it can be seen that the samples obtained by SD-3, DN-1 and GM-6 fermentation produced fatty acids with cowy odor, and the content of the fatty acids was lower than that of the fermented milk sample prepared by the commercial fungus powder YF-L811 in the comparative example. Namely, from the generation condition of the aroma substances, when the three lactic acid bacteria are applied to the fermented goat milk, the three lactic acid bacteria have more obvious characteristics of producing aroma and inhibiting the smell of the goat milk;
3. the goat milk fermented by the lactococcus lactis SD-3, the pediococcus acidilactici DN-1 and the lactobacillus plantarum GM-6 obtained by screening is higher than a fermented milk sample prepared by the commercial fungus YF-L811 in the aspects of sensory acceptability evaluation indexes such as color, texture, taste acceptability, aroma acceptability, overall acceptability and the like.
Drawings
FIG. 1 is a radar chart of sensory evaluation of fermented goat milk in examples 2, 3, and 4 and a control example.
Deposit description
The preservation number of Lactococcus lactis SD-3 is CCTCC M2021743, the latin name is Lactococcus lactis SD-3, the preservation unit is China center for type culture preservation, the preservation time is 2021, 6 and 23 days, and the preservation address is Wuhan university preservation center in Loganian mountain of Wuchang, wuhan, hubei;
the preservation number of the Pediococcus acidilactici DN-1 is CCTCC M2021794, the Latin name is Pediococcus lactis DN-1, the preservation unit is the China center for type culture preservation, the preservation time is 2021 years, 6 months and 28 days, and the preservation address is the preservation center of Wuhan university at Lophania Lophanthogama mountain of Wuchang city, hubei province;
the preservation number of the Lactobacillus plantarum GM-6 is CCTCC M2021744, the latin name is Lactobacillus plantarum GM-6, the preservation unit is the China center for type culture preservation, the preservation time is 2021 years, 6 months and 23 days, and the preservation address is the preservation center of Wuhan university at Lophania Lophanthogona mountain of Wuchang, hubei province.
Detailed Description
In order to make the invention more comprehensible, preferred embodiments are described in detail below with reference to the accompanying drawings.
The MRS agar medium used in the examples of the present invention comprises the following components: 10.0g/L of peptone, 8.0g/L of beef extract powder, 4.0g/L of yeast extract powder, 20.0g/L of glucose, 2.0g/L of dipotassium phosphate, 2.0g/L of diammonium hydrogen citrate, 5.0g/L of sodium acetate, 0.2g/L of magnesium sulfate, 0.04g/L of manganese sulfate, 14.0g/L of agar and 801.0g/L of tween.
The MRS liquid culture medium used in the embodiments of the present invention comprises the following components: 10.0g/L of peptone, 10.0g/L of beef extract powder, 5.0g/L of yeast extract powder, 20.0g/L of glucose, 0.1g/L of magnesium sulfate, 5.0g/L of sodium acetate, 2.0g/L of dipotassium hydrogen phosphate, 2.0g/L of ammonium citrate, 0.05g/L of manganese sulfate and 801.0g/L of tween.
The aroma compounds in the fermented milk obtained in the embodiments of the invention are extracted by adopting headspace solid-phase microextraction (HS-SPME), and analyzed and measured by adopting a gas chromatography-mass spectrometry (GC-MS) technology, and the steps are as follows:
accurately weighing 4.0g of fermented milk in a 15mL extraction bottle, adding 100 μ L of 20 mg/L2-octanol serving as an internal standard substance, placing the internal standard substance into a rotor, balancing the internal standard substance in a constant-temperature water bath kettle at 60 ℃ for 5min, inserting an extraction head device (75 μm CAR/PDMS) aged for 20min into the 15mL extraction bottle, and extracting the mixture at 60 ℃ for 40min. Then, after the extraction is finished, the extraction head is pushed into a gas phase injection port at 250 ℃ for resolving for 5 minutes, and the extracted volatile compounds are analyzed and identified by combining GC-MS.
The gas chromatography conditions were as follows: a chromatographic column: HP-INNOWAX (60 m.times.0.25 mm.times.0.25 μm); sample inlet temperature: 250 ℃; temperature programming: keeping the temperature at 40 ℃ for 4min, heating the temperature to 100 ℃ at 4 ℃/min, keeping the temperature for 2min, heating the temperature to 150 ℃ at 3 ℃/min, heating the temperature to 230 ℃ at 10 ℃/min, and keeping the temperature for 5min; carrier gas: helium (99.99% pure); flow rate: 1mL/min; and (3) sample introduction mode: no split-flow sample introduction. Mass spectrum conditions: electron bombardment energy: 70eV; ion source temperature: 230 ℃; temperature of the quadrupole rods: 150 ℃; emission current: 35 muA; scanning speed: 1.9scans/s; mass scan range: 30-450amu. After each analysis, the extraction head was aged in an aging instrument at 250 ℃ for 20min. Each sample was analyzed in duplicate 3 times.
The characterization of all volatile compounds was first searched using the NIST11 and wiley7n.1 mass spectral libraries and the compounds were initially determined based on information such as degree of match, ion fragmentation etc. And calculating the Retention Index (RI) by using the retention time of the C6-C30 normal paraffin standard substance. The volatile compounds are finally qualified by combining the two methods.
Sensory evaluation descriptors and reference standards in fermented milk obtained by screening strains in each example of the present invention are as follows:
(1) And (3) checking the sensory acceptability: the acceptance test was carried out in separate sensory compartments on three-digit randomly coded fermented milk samples, in turn, by 10 evaluators having a habit of eating and consuming goat milk products, the room temperature being kept at 20 ℃. The 5 indexes of color, texture, aroma, taste and overall acceptance of the sample are respectively evaluated. The samples were scored using a 9-point preference scale (numbers 1-9 represent extremely annoying, very annoying, comparatively unpleasant, somewhat unpleasant, neither unpleasant nor liked, somewhat liked, comparatively liked, very liked, and extremely liked), respectively.
(2) Sensory evaluation process: 15 screened and trained sensory evaluators evaluated the 10 aroma attributes of the fermented milk, and the aroma characteristics and references were as follows: "milk flavor", "sour flavor", "fruit flavor", "wine flavor", "sulfur flavor", "sour and rancid flavor", "animal (goat flavor)", "fat flavor", "nut flavor", and "cheese flavor". The definitions of these 10 aroma descriptors and references are as follows: milk flavor, 0.002% diacetyl water solution; sour, 0.06% lactic acid aqueous solution; a "fruity flavor" of 0.002% ethyl hexanoate aqueous solution; "wine-flavored", 30% ethanol aqueous solution; "Sulfur smell", mashed egg shell; "sour rot smell", 0.1% aqueous butyric acid solution; "animal (goat flavor)", fresh goat milk; "fatty flavor", salad oil; nut flavor, raw nuts; cheese flavor, fresh coconut meat or butter. The samples to be tested were coded numerically randomly, and 5g of each sample was cut into uniformly sized particles and weighed out and stored in a lidded, odorless glass container (50 mL total volume). And arranging according to a random sequence, and sequentially sending to an evaluator for sensory evaluation testing. Each fragrance attribute is rated on an intensity scale of 0-10 points (intensity specification: 0= no intensity or no fragrance perceived, 5= medium intensity, 10= very strong).
The identification of the strains in the embodiments of the invention adopts the following method:
16S rDNA sequence identification is carried out on the strain. Before identification, PCR amplification is carried out on the DNA sequence, and the reaction system is as follows: mu.L of 27F primer (5 'GAGAGTTTGATCCTGGCTCCAG-3') (SEQ ID NO: 1); 2 μ L of 1492r primer (5 'TACGGCTACCTTGTTACGAGAC-3') (SEQ ID NO: 2); 2 mu L of bacterial liquid; 19 μ L of ddH2O;25 μ L of 2 Xgold Star Best Mixed. The PCR reaction program is: (1) 95 ℃ for 10min, (2) 94 ℃ for 30s, (3) 55 ℃ for 30s, (4) 72 ℃ for 1.5min, repeating the steps (2) - (4) for 30 cycles, (5) 72 ℃ for 5min, and (6) storing at 4 ℃. And then carrying out gel electrophoresis detection on the PCR product by using 1.0% agarose, carrying out electrophoresis in 0.5 × TAE electrophoresis solution for 20min under the voltage of 100V, and then detecting the extraction effect on a gel imager, wherein if a clear band is visible and no obvious nonspecific amplification exists, the success of PCR can be judged. Then the 16S rDNA sequence of the strain is compared with the known strain sequence in GenBank.
Example 1
The strain was isolated from a fermented milk sample and fermented milk was prepared as follows:
(1) Taking 1g of sample (the sample is a plurality of fresh milk cakes (the ingredients of the milk cakes are goat milk and fermented whey acid water) collected from Yunnan Dali, taking the sample back to a laboratory after sampling, placing the sample in a refrigerator for sealing and storing, and treating as soon as possible) to a sterile test tube, adding 9mL of normal saline into the test tube, fully oscillating and dissolving uniformly to obtain the dilution of 10 -1 The bacterial liquid of (a);
(2) Taking the dilution obtained in the step (1) as 10 -1 Adding 1mL of the bacterial solution into another sterile test tube, adding 9mL of physiological saline into the test tube, fully oscillating and dissolving uniformly to obtain a dilution gradient of 10 -2 The bacterial liquid of (a);
repeating the above steps 4 times to obtain dilution of 10 -3 Bacterial liquid 10 -4 Bacterial liquid 10 -5 Bacterial liquid and 10 -6 Bacterial liquid;
(3) Respectively taking 0.2mL of bacterial liquid from the five test tubes obtained in the step (2), coating the bacterial liquid on an MRS agar medium plate, and placing the plate at the constant temperature of 37 ℃ for culturing for 2 to 3 days;
(4) Selecting 60 isolated plant single colonies obtained after the plate culture in the step (3) by using an inoculating needle, respectively inoculating the single colonies to an MRS liquid culture medium, and fermenting at a constant temperature of 37 ℃ for 12 hours to respectively obtain fermentation liquor of 60 isolated plants;
(5) Centrifuging 60 isolate fermentation liquor corresponding to the step (4) at 6000rpm for 15min at 4 ℃, respectively, washing precipitates obtained by centrifuging with sterile water for 2-3 times, respectively adding sterile water with the same volume as the original bacterial suspension into the washed precipitates, and performing vortex oscillation at 6000rpm for 10min for later use;
(6) Stirring about 1L goat milk for 30min at 50 deg.C; then heating to 60 ℃, and carrying out homogenization under the constant volume of 1L and 180bar; then water bath is carried out at 95 ℃, and the time is counted for 10min when the temperature of the sample is 85 ℃; then, it was cooled to 44 ℃ with ice.
(7) And (3) taking 60 parts of the sterile goat milk obtained in the step (6), inoculating the strain culture solution obtained in the step (5) according to the volume ratio of 5%, adding commercial fungus powder YF-L811 (5 mg/L for assisting curd), controlling the temperature to be 37 ℃, performing fermentation culture until the curd fermentation is finished (pH = 4.6), and naturally cooling to 25 ℃ to obtain the fermented milk.
Example 2
Screening strains from a fermented milk sample, comprising the steps of:
(1) Isolation of strains and preparation of fermented milks: the same procedure as in example 1;
(2) And (2) carrying out sensory evaluation primary screening and GC-MS aroma determination on the fermented milk sample obtained in the step (1), screening to obtain lactobacillus SD-3 with the characteristic of producing aroma and inhibiting the smell of mutton, carrying out 16S rDNA sequence identification on the strain SD-3, comparing the sequence with a known strain sequence in GenBank, wherein the SD-3 is Lactococcus lactis (Lactococcus lactis) with the preservation number of CCTCC M2021743.
Example 3
Screening strains from a fermented milk sample, comprising the steps of:
(1) Isolation of strains and preparation of fermented milks: the same procedure as in example 1;
(2) And (2) carrying out sensory evaluation preliminary screening and GC-MS aroma determination on the fermented milk sample obtained in the step (1), screening to obtain a lactic acid bacteria DN-1 with the characteristic of producing aroma and inhibiting the smell of mutton, identifying a 16S rDNA sequence of the strain DN-1, comparing the sequence with a known strain sequence in GenBank, wherein DN-1 is Pediococcus lactis (Pediococcus lactis) with the preservation number of CCTCC M2021794, and the sequence is shown as SEQ ID NO 4.
Example 4
Screening strains from a fermented milk sample, comprising the steps of:
(1) Isolation of strains and preparation of fermented milks: the operation steps are the same as those of the example 1;
(2) And (2) carrying out sensory evaluation primary screening and GC-MS aroma determination on the fermented milk sample obtained in the step (1), and screening to obtain the lactic acid bacteria GM-6 with the function of producing aroma and inhibiting the smell of mutton. 16S rDNA sequence identification is carried out on the strain GM-6, the sequence is shown as SEQ ID NO:5, and the sequence is compared with the known strain sequence in GenBank, the GM-6 is Lactobacillus plantarum (Lactobacillus plantarum), and the preservation number is CCTCC M2021744.
Comparative example
Commercial powder YF-L811 was inoculated into sterile goat milk, fermented at 37 ℃ until curd fermentation was completed (pH = 4.6), and naturally cooled to 25 ℃ to obtain control fermented milk. And (3) carrying out sensory evaluation primary screening and GC-MS aroma determination on a control fermented milk sample, and analyzing and identifying the sensory evaluation results of key aroma compounds such as 2-heptanone and isoamylol, content changes of fatty acid compounds such as butyric acid, caproic acid and caprylic acid, and the characteristic milk aroma, fruit aroma, nut aroma, mutton smell, sour taste and sour rot taste of the fermented milk.
Sensory indexes and aroma components of the fermented goat milks of the above examples 2, 3 and 4 and comparative example were measured, and the results of GC-MS identification are shown in Table 1, and sensory acceptability of the fermented goat milks of different bacterial species is evaluated, and the results are shown in Table 2.
TABLE 1 GC-MS measurement of fermented goat milks of different bacterial species
Figure BDA0003297180120000091
Figure BDA0003297180120000101
TABLE 2 evaluation results of sensory acceptability of goat milk fermented by different strains
Color Texture of Taste acceptance Acceptance of fragrance Overall acceptance
SD-3 5.4±0.52 5.6±0.70 5.4±0.52 7.4±0.84 7.5±0.97
DN-1 5.5±0.53 6.0±0.82 4.7±0.95 6.9±0.74 6.7±0.67
GM-6 5.6±0.70 4.1±0.74 6.6±0.84 6.8±0.92 7.0±0.67
Comparative example 5.3±0.67 4.3±0.67 5.0±0.94 5.4±0.70 5.2±0.63
As can be seen from Table 1, the goat milk fermented by Lactococcus lactis (Lactococcus lactis) SD-3, pediococcus acidilactici (Pediococcus lactis) DN-1 and Lactobacillus plantarum (Lactobacillus plantarum) GM-6 produces high contents of 3-hydroxy-2-butanone, 2-heptanone, 2-octanone and 2-nonanone, and the ketone fragrance substances have fragrance characteristics of cream, cheese, coconut, fruity and sweet fragrance and the like; meanwhile, the sour congeal cheese goat cheese contains high-content almond-flavored benzaldehyde and fruit-flavored isoamylol, and is used as an odor component with a goat odor inhibiting effect and a positive influence in the characteristic flavor identification process of the sour congeal cheese goat cheese. Meanwhile, by analyzing the aroma components of the fermentation samples of different strains, the content of the fatty acid with the cowy smell generated by the samples obtained by SD-3, DN-1 and GM-6 fermentation is also lower compared with the fermented milk sample prepared by the commercial fungus powder YF-L811 in the comparative example. Namely, from the generation of the aroma substances, the three strains of lactic acid bacteria have more obvious aroma producing and mutton odor inhibiting properties when applied to the fermented goat milk.
As can be seen from Table 2, the goat milk fermented by Lactococcus lactis subsp lactis SD-3, pediococcus acidilactici DN-1 and Lactobacillus plantarum GM-6 is higher than the fermented milk sample prepared from the commercial powder YF-L811 in the sensory acceptability evaluation indexes such as color, texture, taste acceptability, aroma acceptability and overall acceptability.
The sensory evaluation radar profiles of examples 2, 3, 4 and comparative example are shown in FIG. 1 according to the determined descriptive sensory analysis indexes, and it can be seen from FIG. 1 that the yoghurts obtained by SD-3, DN-1 and GM-6 fermentations are superior in milk flavor, fruit flavor and nut flavor. According to the aroma quality evaluation of the yogurt goat cheese cake, the milk aroma, the fruit aroma and the nut aroma are three types of aroma attributes accepted and favored by consumers. In addition, the strength of the cowy smell, the sour taste and the sour taste of the fermented milk obtained by the three strains is lower than that of the fermented milk obtained by the commercial powder fermentation. Therefore, the fragrance producing and smell inhibiting performance of the lactic acid bacteria is good.
In conclusion, the strain obtained by screening the method for screening the strain with the function of producing fragrance and inhibiting the smell of mutton can ensure that a fermentation sample can endogenously produce more substances with milk fragrance and fruit fragrance and having the function of inhibiting the smell of mutton, reduce the generation of smell fatty acid components to a certain extent, optimize the flavor quality of the fermented dairy product on the basis of not needing exogenous addition of fragrance substances and reduce the production cost for preparing the fermented dairy product.
While the present invention has been described in detail in connection with the above-described embodiments, it is to be understood that the present invention is not limited to those precise embodiments, and that various changes and modifications may be effected therein by one of ordinary skill in the pertinent art without departing from the scope or spirit of the present invention.
Sequence listing
<110> Shanghai applied technology university
<120> strain screening method with function of producing fragrance and inhibiting mutton smell and application thereof
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gagagtttga tcctggctca g 21
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aaacgaatgc taataccgca taaaaacttt aaacacaagt tttaagtttg aaagatgcaa 180
ttgcatcact caaagatgat cccgcgttgt attagctagt tggtgaggta aaggctcacc 240
aaggcgatga tacatagccg acctgagagg gtgatcggcc acattgggac tgagacacgg 300
cccaaactcc tacgggaggc agcagtaggg aatcttcggc aatggacgaa agtctgaccg 360
agcaacgccg cgtgagtgaa gaaggttttc ggatcgtaaa actctgttgg tagagaagaa 420
cgttggtgag agtggaaagc tcatcaagtg acggtaacta cccagaaagg gacggctaac 480
tacgtgccag cagccgcggt aatacgtagg tcccgagcgt tgtccggatt tattgggcgt 540
aaagcgagcg caggtggttt attaagtctg gtgtaaaagg cagtggctca accattgtat 600
gcattggaaa ctggtagact tgagtgcagg agaggagagt ggaattccat gtgtagcggt 660
gaaatgcgta gatatatgga ggaacaccgg tggcgaaagc ggctctctgg cctgtaactg 720
acactgaggc tcgaaagcgt ggggagcaaa caggattaga taccctggta gtccacgccg 780
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agcactccgc ctggggagta cgaccgcaag gttgaaactc aaaggaattg acgggggccc 900
gcacaagcgg tggagcatgt ggtttaattc gaagcaacgc gaagaacctt accaggtctt 960
gacatactcg tgctattcct agagatagga agttccttcg ggacacggga tacaggtggt 1020
gcatggttgt cgtcagctcg tgtcgtgaga tgttgggtta agtcccgcaa cgagcgcaac 1080
ccctattgtt agttgccatc attaagttgg gcactctaac gagactgccg gtgataaacc 1140
ggaggaaggt ggggatgacg tcaaatcatc atgcccctta tgacctgggc tacacacgtg 1200
ctacaatgga tggtacaacg agtcgcgaga cagtgatgtt tagctaatct cttaaaacca 1260
ttctcagttc ggattgtagg ctgcaactcg cctacatgaa gtcggaatcg ctagtaatcg 1320
cggatcagca cgccgcggtg aatacgttcc cgggccttgt acacaccgcc cgtcacacca 1380
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aagtct 1446
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tattgaatca ttagacggct agctcctaaa ggttacccca ccggctttgg gtgttacaaa 60
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gatccgcgat tactagcgat tccgacttcg tgtaggcgag ttgcagccta cagtccgaac 180
tgagaatggt tttaagagat tagctaaacc tcgcggtttc gcaactcgtt gtaccatcca 240
ttgtagcacg tgtgtagccc aggtcataag gggcatgatg atttgacgtc gtccccacct 300
tcctccggtt tgtcaccggc agtctcacta gagtgcccaa ctgaatgctg gcaactagta 360
ataagggttg cgctcgttgc gggacttaac ccaacatctc acgacacgag ctgacgacaa 420
ccatgcacca cctgtcattc tgtccccgaa gggaacgcct aatctcttag gttggcagaa 480
gatgtcaaga cctggtaagg ttcttcgcgt agcttcgaat taaaccacat gctccaccgc 540
ttgtgcgggc ccccgtcaat tcttttgagt ttcaaccttg cggtcgtact ccccaggcgg 600
attacttaat gcgttagctg cagcactgaa gggcggaaac cctccaacac ttagtaatca 660
tcgtttacgg catggactac cagggtatct aatcctgttc gctacccatg ctttcgagcc 720
tcagcgtcag ttacagacca gacagccgcc ttcgccactg gtgttcttcc atatatctac 780
gcatttcacc gctacacatg gagttccact gtcctcttct gcactcaagt ctcccagttt 840
ccaatgcact tcttcggttg agccgaaggc tttcacatta gacttaaaag accgcctgcg 900
ctcgctttac gcccaataaa tccggataac gcttgccacc tacgtattac cgcggctgct 960
ggcacgtagt tagccgtggc tttctggtta aataccgtca ctgggtgaac agttactctc 1020
acccacgttc ttctttaaca acagagcttt acgagccgaa acccttcttc actcacgcgg 1080
cgttgctcca tcagacttgc gtccattgtg gaagattccc tactgctgcc tcccgtagga 1140
gtctgggccg tgtctcagtc ccaatgtggc cgattaccct ctcaggtcgg ctacgcatca 1200
tcgccttggt gagccgttac ctcaccaact agctaatgcg ccgcgggtcc atccagaagt 1260
gatagcagag ccatctttta aaagaaaacc aggcggtttt ctctgttata cggtattagc 1320
atctgtttcc aggtgttatc ccctgcttct gggcaggtta cccacgtgtt actcacccgt 1380
ccgccactca cttcgtgtta aaatctcatt cagtgcaagc acgtcctgat caattaacgg 1440
aagttcgtcg actgcattat agcgccccat tgag 1474
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<213> Artificial Sequence (Artificial Sequence)
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ttaggcggct ggttcctaaa aggttacccc accgactttg ggtgttacaa actctcatgg 60
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ttactagcga ttccgacttc atgtaggcga gttgcagcct acaatccgaa ctgagaatgg 180
ctttaagaga ttagcttact ctcgcgagtt cgcaactcgt tgtaccatcc attgtagcac 240
gtgtgtagcc caggtcataa ggggcatgat gatttgacgt catccccacc ttcctccggt 300
ttgtcaccgg cagtctcacc agagtgccca acttaatgct ggcaactgat aataagggtt 360
gcgctcgttg cgggacttaa cccaacatct cacgacacga gctgacgaca accatgcacc 420
acctgtatcc atgtccccga agggaacgtc taatctctta gatttgcata gtatgtcaag 480
acctggtaag gttcttcgcg tagcttcgaa ttaaaccaca tgctccaccg cttgtgcggg 540
cccccgtcaa ttcctttgag tttcagcctt gcggccgtac tccccaggcg gaatgcttaa 600
tgcgttagct gcagcactga agggcggaaa ccctccaaca cttagcattc atcgtttacg 660
gtatggacta ccagggtatc taatcctgtt tgctacccat actttcgagc ctcagcgtca 720
gttacagacc agacagccgc cttcgccact ggtgttcttc catatatcta cgcatttcac 780
cgctacacat ggagttccac tgtcctcttc tgcactcaag tttcccagtt tccgatgcac 840
ttcttcggtt gagccgaagg ctttcacatc agacttaaaa aaccgcctgc gctcgcttta 900
cgcccaataa atccggacaa cgcttgccac ctacgtatta ccgcggctgc tggcacgtag 960
ttagccgtgg ctttctggtt aaataccgtc aatacctgaa cagttactct cagatatgtt 1020
cttctttaac aacagagttt tacgagccga aacccttctt cactcacgcg gcgttgctcc 1080
atcagacttt cgtccattgt ggaagattcc ctactgctgc ctcccgtagg agtttgggcc 1140
gtgtctcagt cccaatgtgg ccgattaccc tctcaggtcg gctacgtatc attgccatgg 1200
tgagccgtta ccccaccatc tagctaatac gccgcgggac catccaaaag tgatagccga 1260
agccatcttt caagctcgga ccatgcggtc caagttgtta tgcggtatta gcatctgttt 1320
ccaggtgtta tcccccgctt ctgggcaggt ttcccacgtg ttactcacca gttcgccact 1380
cactcaaatg taaatcatga tgcaagcacc aatcaatacc agagttcgtt cgacttgcat 1440
gtat 1444

Claims (4)

1. The strain with the characteristic of producing fragrance and inhibiting the smell of mutton is Lactococcus lactis SD-3, the preservation number of the Lactococcus lactis SD-3 is CCTCC M2021743, the latin name is Lactococcus lactis SD-3, the preservation unit is the China center for type culture preservation, the preservation time is 2021, 6 and 23 days, and the preservation address is the preservation center of Wuhan university at Lophania martensii mountain of Wuhan city, hubei province.
2. The strain with the fragrance producing and mutton smell inhibiting functional characteristics is Pediococcus acidilactici DN-1, the preservation number of the Pediococcus acidilactici DN-1 is CCTCC M2021794, the latin name is Pediococcus lactis DN-1, the preservation unit is China center for type culture preservation, the preservation time is 2021 year, 6 months and 28 days, and the preservation address is the preservation center of Wuhan university in Lojia mountain of Wuchang, hubei province.
3. The strain with the characteristic of producing fragrance and inhibiting the smell of mutton is Lactobacillus plantarum GM-6, the preservation number of the Lactobacillus plantarum GM-6 is CCTCC M2021744, the latin name is Lactobacillus plantarum GM-6, the preservation unit is the China type culture preservation center, the preservation time is 2021 year, 6 months and 23 days, and the preservation address is the preservation center of Wuchang Lojia mountain university of Wuhan city, hubei province.
4. The use of a strain having the functional property of producing fragrance and suppressing mutton smell as claimed in any one of claims 1 to 3 in fermented goat milk.
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