Lactobacillus saké H30-5, fermentation agent and its application in improving the quality of fermented surimi

Abstract

The present invention relates to the field of microbial fermentation technology, and in particular to sake widely swayed lactobacillus H30-5, fermentation agent and its application in improving the quality of fermented fish paste. The present invention screens sake widely swayed lactobacillus H30-5 from tilapia naturally fermented fish paste, and effectively improves the texture and flavor of tilapia fermented fish paste by adding bacteria and fermenting, providing important technical support for the high-value utilization of tilapia. The experimental data of the present invention show that the tilapia fermented fish paste produced by using sake widely swayed lactobacillus H30-5 has a unique flavor and taste. Compared with the tilapia naturally fermented fish paste without adding bacteria, the gel strength, hardness, elasticity, chewiness and pleasant volatile flavor substance content of the tilapia fermented fish paste obtained by adding bacteria and fermenting are significantly increased, and the content of volatile flavor substances with fishy smell is significantly reduced. The obtained fermented fish paste products have broad market application prospects.

Description

Technical Field

[0001] The invention relates to the technical field of microbial fermentation, in particular to sake lactobacillus H30-5, a fermentation agent and application thereof in improving the quality of fermented fish paste. Background Art

[0002] Due to the low gel strength of freshwater fish paste, the fish paste products produced by the traditional hot gel method have poor texture and strong fishy smell, which greatly limits the high-value utilization of freshwater fish. The use of microbial fermentation can effectively improve the gel properties of freshwater fish paste and the content of nutrients such as free amino acids and free fatty acids in the product. In addition, the production process does not require rinsing, which will not cause the loss of nutrients in the fish paste, which is conducive to energy conservation and emission reduction. However, due to the complex microbial flora in the fish paste raw materials, the quality and flavor of the fermented fish paste are difficult to control, and the metabolism of corrupt microorganisms is prone to produce harmful substances such as biogenic amines, which increases the difficulty of the application of fish paste fermentation.

[0003] Lactic acid bacteria fermentation can effectively inhibit the growth of spoilage microorganisms and the production of biogenic amines, but compared with naturally fermented surimi, its gel strength, texture and flavor are not significantly improved. Therefore, it is urgent to find a microorganism that can effectively improve the texture (gel strength, hardness, elasticity and chewiness, etc.) and flavor of fermented surimi. Summary of the invention

[0004] The purpose of the present invention is to provide sake lactobacillus H30-5, fermentation agent and its application in improving the quality of fermented fish paste, so as to solve the problems existing in the above-mentioned prior art. The strain or fermentation agent provided by the present invention is conducive to improving the gel strength, hardness, elasticity and chewiness of fermented fish paste, increasing the content of pleasant volatile flavor substances, and reducing the content of volatile flavor substances with fishy smell.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] The invention provides a strain of Latilactobacillus sakei H30-5, and the preservation number of the Latilactobacillus sakei H30-5 is CGMCC No.29840.

[0007] The present invention provides the use of the above-mentioned Lactobacillus sakei H30-5 in preparing a fermentation agent for improving the quality of fermented fish paste.

[0008] Preferably, the fermented surimi comprises fermented tilapia surimi.

[0009] The invention provides a fermentation bacterial agent for improving the quality of fermented fish paste. The effective components of the fermentation bacterial agent include the above-mentioned Lactobacillus sakei H30-5.

[0010] Preferably, the effective viable count of Lactobacillus sakei H30-5 in the fermentation agent is 10 8 -10 9 CFU / mL.

[0011] Preferably, the fermented surimi comprises fermented tilapia surimi.

[0012] The present invention provides the use of the above-mentioned Lactobacillus sakei H30-5 or the above-mentioned starter in improving the quality of fermented fish paste.

[0013] Preferably, the fermented surimi comprises fermented tilapia surimi.

[0014] The present invention provides a method for improving the quality of fermented surimi, comprising the following steps:

[0015] The above-mentioned fermentation bacteria agent is used to carry out bacterial fermentation on the fish paste.

[0016] Preferably, the temperature of the bacterial fermentation is 10-35°C and the time is 12-36h;

[0017] The fermentation agent is based on the effective viable count of 10 5 -10 8 The final concentration of CFU / g was added to the surimi for fermentation.

[0018] Further preferably, the method for preparing the surimi is: taking the back muscles of fresh tilapia, rinsing them with cold water, draining the water, and beating them into surimi with a meat grinder, adding water, glucose, sucrose and salt to the surimi, and stirring them at 10° C. for 1-5 minutes to obtain the surimi.

[0019] Further preferably, the amount of water added is 0-10% of the mass of the surimi; the amount of glucose added is 0.5-2% of the mass of the surimi; the amount of sucrose added is 0.5-2% of the mass of the surimi; and the amount of salt added is 0.5-3% of the mass of the surimi.

[0020] The present invention discloses the following technical effects:

[0021] The present invention uses a screening culture medium to screen out sake-wide Lactobacillus H30-5 from naturally fermented tilapia surimi, and effectively improves the texture and flavor of tilapia fermented surimi by adding bacteria and fermenting, providing important technical support for the high-value utilization of tilapia. The experimental data of the present invention show that the tilapia fermented surimi produced by using sake-wide Lactobacillus H30-5 has a unique flavor and taste. Compared with the naturally fermented tilapia surimi without bacteria, the gel strength, hardness, elasticity, chewiness and pleasant volatile flavor substance content of the tilapia fermented surimi obtained by adding bacteria and fermenting are significantly increased, and the content of volatile flavor substances with fishy smell is significantly reduced. The obtained fermented surimi products have broad market application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0023] Figure 1 This is the molecular evolutionary tree of sake-sake Lactobacillus H30-5;

[0024] Figure 2 The growth and acid production characteristics of Lactobacillus sakei H30-5 in MRS medium;

[0025] Figure 3 The salt tolerance of sake lactobacillus H30-5.

[0026] Biological deposit information

[0027] Latilactobacillus sakei H30-5 has been deposited in the General Microbiology Center of China Culture Collection Administration (CGMCC), with the deposit number CGMCC No.29840. The deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the deposit time is February 1, 2024. DETAILED DESCRIPTION

[0028] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0029] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0030] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0031] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0032] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0033] Example 1

[0034] (1) Isolation and screening of strains

[0035] The sake-dispersed Lactobacillus H30-5 was screened and isolated from the naturally fermented tilapia surimi. The screening method was as follows: the tilapia surimi that had been naturally fermented for 30 hours was added to sterile physiological saline for homogenization. The homogenate was gradiently diluted and spread on an MRS plate (10.0 g peptone, 5.0 g beef powder, 20.0 g glucose, 4.0 g yeast powder, 5.0 g sodium acetate, 2.0 g disodium hydrogen phosphate, 0.20 g magnesium sulfate, 2.0 g triammonium citrate, 0.05 g manganese sulfate, 1 mL Tween 80, 15.0 g agar and 1000 mL water), and cultured anaerobically at 30°C for 2 days, and single colonies were selected to obtain the target strain H30-5.

[0036] (2) Molecular biological identification of strains

[0037] Figure 1 As shown, it was identified as Latilactobacillus sakei.

[0038] The growth and acid production of strain H30-5 in MRS liquid medium (10.0 g peptone, 5.0 g beef powder, 20.0 g glucose, 4.0 g yeast powder, 5.0 g sodium acetate, 2.0 g disodium hydrogen phosphate, 0.20 g magnesium sulfate, 2.0 g triammonium citrate, 0.05 g manganese sulfate, 1 mL Tween 80 and 1000 mL water) are shown in Figure 2. Figure 2 As shown in the figure, the bacteria grows fast, enters the logarithmic growth phase in 9 hours, and enters the stable phase in 24 hours. The acid production of the strain is consistent with the growth. At the same time, different concentrations of sodium chloride were added to the MRS liquid medium to determine the salt tolerance of the strain. The results are shown in the figure. Figure 3 As shown, the bacteria has strong salt tolerance and can grow and produce acid well at a salt concentration of 0-4%.

[0039] The strain was named Latilactobacillus sakei H30-5 and was deposited in the General Microbiology Center of China Culture Collection Administration (CGMCC) on February 1, 2024. The deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC No. 29840.

[0040] Example 2

[0041] (1) Preparation of fermentation agent: The sake-sake Lactobacillus H30-5 was anaerobically cultured at 30°C for 12 h, centrifuged at 4°C and 12000 g for 10 min, and resuspended in sterile saline to obtain a fermentation agent. The effective viable count of sake-sake Lactobacillus H30-5 in the fermentation agent was 10 9 CFU / mL;

[0042] (2) Preparation of surimi: The back muscles of fresh tilapia were taken, rinsed with cold water, drained, and minced with a meat grinder to make surimi. Based on the weight of the surimi, 1 wt.% glucose, 1 wt.% sucrose and 0.5 wt.% salt were added to the surimi. The fermentation agent prepared in step (1) was added at the same time, so that the final concentration of Lactobacillus sakei H30-5 in the resulting mixture was 10 8 CFU / g, chopped with a chopper for 5 min at 10°C;

[0043] (3) Surimi fermentation: The surimi was stuffed into sausage casings using a sausage stuffer and fermented at 10°C for 12 h to obtain fermented surimi.

[0044] Naturally fermented tilapia surimi was used as control.

[0045] The texture results of tilapia fermented surimi are shown in Table 1.

[0046] Table 1 Survey results of texture of fermented tilapia surimi

[0047] texture Fermentation with bacteria Natural fermentation Gel strength (g×cm) 392.1 36.1 Hardness(g) 183.0 62.0 Elasticity(mm) 12.4 11.5 Chewing properties (mJ) 13.6 4.2

[0048] As shown in Table 1, fermentation with Lactobacillus sakei H30-5 can significantly improve the texture of fermented tilapia surimi, with the gel strength, hardness, elasticity and chewiness reaching 392.1 g×cm, 183.0 g, 12.4 mm and 13.6 mJ, respectively, which are 9.9 times, 1.0 times, 7.8% and 2.2 times higher than those of naturally fermented surimi.

[0049] The content of volatile flavor substances was then analyzed using HS-SPME-GC-MS, and the results are shown in Table 2.

[0050] Table 2 Detection results of volatile flavor substances

[0051]

[0052]

[0053] The results are shown in Table 2. After fermentation with bacteria, the contents of pleasant volatile flavor substances such as alcohols, ketones, aldehydes, esters, and furans in tilapia surimi were significantly increased, while the contents of fishy volatile flavor substances such as trimethylamine and methyl mercaptan were significantly reduced.

[0054] At the same time, 16S rRNA gene high-throughput sequencing technology was used to analyze the microbial diversity in surimi. The results showed that after fermentation with added bacteria, the proportion of the genus Latilactobacillus reached 92.4%, and the proportion of spoilage microorganisms was significantly reduced.

[0055] Example 3

[0056] (1) Preparation of fermentation agent: The sake-sake Lactobacillus H30-5 was anaerobically cultured at 30°C for 24 h, centrifuged at 4°C and 12000 g for 10 min, and resuspended in sterile saline to obtain a fermentation agent. The effective viable cell count of sake-sake Lactobacillus H30-5 in the fermentation agent was 10 8 CFU / mL;

[0057] (2) Preparation of surimi: The back muscles of fresh tilapia were taken, rinsed with cold water, drained, and minced with a meat grinder to make surimi. Based on the weight of the surimi, 5 wt.% of water, 0.5 wt.% of glucose, 0.5 wt.% of sucrose, and 2 wt.% of salt were added to the surimi. The fermentation agent prepared in step (1) was added at the same time, so that the final concentration of Lactobacillus saké H30-5 in the resulting mixture was 105 CFU / g of fermentation bacteria, chopped with a chopper for 2 minutes at 10°C;

[0058] (3) Surimi fermentation: The surimi was stuffed into sausage casings using a sausage stuffer and fermented at 25°C for 30 h to obtain fermented surimi.

[0059] Naturally fermented tilapia surimi was used as control.

[0060] The texture results of tilapia fermented surimi are shown in Table 3.

[0061] Table 3 Survey results of texture of fermented tilapia surimi

[0062] texture Fermentation with bacteria Natural fermentation Gel strength (g×cm) 560.4 118.4 Hardness(g) 687.5 206.0 Elasticity(mm) 14.1 13.2 Chewing properties (mJ) 48.7 20.7

[0063] As shown in Table 3, fermentation with Lactobacillus sakei H30-5 can significantly improve the texture of fermented tilapia surimi, with gel strength, hardness, elasticity and chewiness reaching 560.4 g×cm, 687.5 g, 14.1 mm and 48.7 mJ, respectively, which are 3.7 times, 2.3 times, 6.8% and 1.4 times higher than those of naturally fermented surimi.

[0064] The content of volatile flavor substances was then analyzed using HS-SPME-GC-MS, and the results are shown in Table 4.

[0065] Table 4 Detection results of the content of volatile flavor substances

[0066] Volatile flavor substances (mg / kg) Odor description Fermentation with bacteria Natural fermentation 1-Octen-3-ol Mushroom aroma 1.02 0.17 1-Octanol Strong oily smell with citrus and rose notes 0.23 0.09 1-Hexanol Green, fruity, mellow, sweet 2.61 0.63 1-Nonanol Strong rose and orange aroma with oily smell 0.27 0.08 trans-2-octen-1-ol Greasey smell 0.11 0.03 2,3-Butanedione Sweet cream, buttery aroma 0.88 0.27 2,3-Pentanedione Butter, popcorn flavor 0.05 0.01 3-Hydroxy-2-butanone Sweet, dairy, and fatty aroma 5.48 5.20 Nonanal Waxy, sweet orange, fatty, floral 0.43 0.24 Hexanal Strong green, grassy, ​​vegetable, and fruity aromas 1.95 0.32 Octanal Sweet orange, oil, honey aroma 0.18 0.04 Heptanal Sweet apricot, nutty aroma 0.36 0.09 Valeraldehyde Fruity, bready aroma 0.18 0.01 Benzaldehyde Sweet, almond, fruity 0.29 0.06 Ethyl hexanoate Fruity 0.03 0.01 2-n-Pentylfuran Bean aroma, meaty aroma 0.19 0.01 Trimethylamine Fishy smell 0.52 1.16 Methyl mercaptan Bad smell 0.04 0.06

[0067] As shown in Table 4, after fermentation with bacteria, the contents of pleasant volatile flavor substances such as alcohols, ketones, aldehydes, esters, furans, etc. in tilapia surimi were significantly increased, while the contents of fishy volatile flavor substances such as trimethylamine and methyl mercaptan were significantly reduced.

[0068] At the same time, 16S rRNA gene high-throughput sequencing technology was used to analyze the microbial diversity in surimi. The results showed that after fermentation with added bacteria, the proportion of the genus Latilactobacillus reached 59.2%, and the proportion of spoilage microorganisms was significantly reduced.

[0069] Example 4

[0070] (1) Preparation of fermentation agent: Sake Lactobacillus H30-5 was anaerobically cultured at 30°C for 36 h, centrifuged at 4°C and 12000 g for 10 min, and resuspended in sterile saline to obtain a fermentation agent. The effective viable count of Sake Lactobacillus H30-5 in the fermentation agent was 10 8 CFU / mL;

[0071] (2) Preparation of surimi: The back muscles of fresh tilapia were taken, rinsed with cold water, drained, and minced with a meat grinder to make surimi. Based on the weight of the surimi, 10 wt.% of water, 2 wt.% of glucose, 2 wt.% of sucrose, and 3 wt.% of salt were added to the surimi. The fermentation agent prepared in step (1) was added at the same time, so that the final concentration of Lactobacillus sakei H30-5 in the resulting mixture was 10 6 CFU / g of fermentation bacteria, chopped with a chopper at 10°C for 1 min;

[0072] (3) Surimi fermentation: The surimi was stuffed into sausage casings using a sausage stuffer and fermented at 35°C for 36 h to obtain fermented surimi.

[0073] Naturally fermented tilapia surimi was used as control.

[0074] The texture results of tilapia fermented surimi are shown in Table 5.

[0075] Table 5 Survey results of texture of fermented tilapia surimi

[0076] texture Fermentation with bacteria Natural fermentation Gel strength (g×cm) 821.1 461.5 Hardness(g) 686.0 316.5 Elasticity(mm) 14.4 13.4 Chewing properties (mJ) 47.5 30.8

[0077] As shown in Table 5, fermentation with Lactobacillus sakei H30-5 can significantly improve the texture of fermented tilapia surimi, with the gel strength, hardness, elasticity and chewiness reaching 821.1 g×cm, 686.0 g, 14.4 mm and 47.5 mJ, respectively, which are improved by 77.9%, 1.2 times, 7.5% and 54.2% compared with naturally fermented surimi.

[0078] The content of volatile flavor substances was then analyzed using HS-SPME-GC-MS, and the results are shown in Table 6.

[0079] Table 6 Detection results of volatile flavor substances

[0080]

[0081]

[0082] As shown in Table 6, after fermentation with bacteria, the contents of pleasant volatile flavor substances such as alcohols, ketones, aldehydes, esters, furans, etc. in tilapia surimi were significantly increased, while the contents of fishy volatile flavor substances such as trimethylamine and methyl mercaptan were significantly reduced.

[0083] At the same time, 16S rRNA gene high-throughput sequencing technology was used to analyze the microbial diversity in surimi. The results showed that after fermentation with added bacteria, the proportion of the genus Latilactobacillus reached 68.1%, and the proportion of spoilage microorganisms was significantly reduced.

[0084] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A strain of Lactobacillus sakei H30-5, characterized in that: The preservation number of the sake lactobacillus H30-5 is CGMCC No.29840.

2. The use of the Lactobacillus sakei H30-5 according to claim 1 in preparing a fermentation agent for improving the quality of fermented fish paste, characterized in that: The fermented surimi includes fermented tilapia surimi.

3. A fermentation agent for improving the quality of fermented surimi, characterized in that: The active ingredients of the fermentation bacteria agent include the sake lactobacillus H30-5 described in claim 1.

4. The fermentation agent according to claim 3, characterized in that: The effective viable count of Lactobacillus saké H30-5 in the fermentation agent is 10 8 -10 9 CFU / mL.

5. The fermentation agent according to claim 3, characterized in that: The fermented surimi includes fermented tilapia surimi.

6. Use of the sake-sake Lactobacillus H30-5 according to claim 1 or the fermentation agent according to any one of claims 3 to 5 in improving the quality of fermented surimi, characterized in that: The fermented surimi includes fermented tilapia surimi.

7. A method for improving the quality of fermented surimi, characterized in that: The following steps are involved: The fermentation agent according to any one of claims 3 to 5 is used to ferment the fish paste; the fermented fish paste includes tilapia fermented fish paste.

8. The method according to claim 7, characterized in that The temperature of the bacterial fermentation is 10-35°C and the time is 12-36h; The fermentation agent is based on the effective viable count of 10 5 -10 8 The final concentration of CFU / g was added to the surimi for fermentation.

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